Radiation dose in barium swallow examinations: an Oxford tertiary care centre experience.

AIM: To assess the radiation dose in standard barium swallow studies (BaSS) performed at a large tertiary referral centre, benchmarked against the national diagnostic reference level (NDRL), and to provide evidence to support reducing radiation dose through improving operator's training. MATERIALS AND METHODS: In this retrospective observational study, 1,004 adult BaSS (M:F = 411:593) were assessed. The radiation dose was first compared with NDRL, and then stratified by the operator's training level and compared using analysis of variance (ANOVA). The operators were categorised as radiology residents, advanced radiography practitioners, GI radiology fellows, and GI radiology consultants. Further comparative analysis is performed by comparing the test radiation dose when stratified based on the referrer's specialty, and whether they were performed for female patients of childbearing age. RESULTS: The findings show 68.2% of BaSS performed are within the NDRL and demonstrated statistically significant correlation (p<0.0001) between radiation dose and operator's training level, and for female patients of childbearing age (p<0.002) and the rest. The median dose was lower with more senior operators. There was no significant correlation between radiation dose and the referrer's specialty. CONCLUSION: The study presents evidence to support revisiting consultant-supervised BaSS training, in the interests of lowering the radiation exposure to patients and improving compliance with the NDRL.

A three-decade survey of Brazilian Fusarium oxysporum f. sp. lycopersici races assessed by pathogenicity tests on differential tomato accessions and by molecular markers.

AIM: Physiological race determination of 143 Fusarium oxysporum f. sp. lycopersici (FOL) isolates collected along 30 years in major tomato-producing regions of Brazil. MATERIALS AND RESULTS: Physiological races were determined via root-dipping inoculation of differential tomato accessions and by the PCR-based marker system of Hirano and Arie (2006). According to pathogenicity/virulence assays, five race 1, 23 race 2 and 115 race 3 isolates were identified. FOL race 1 and 2 isolates prevailed up to early 2000s. Afterwards, the large majority of the isolates was classified as the invasive race 3. Novel reports of race 3 were done in five states, thus expanding its geographical distribution. Using this PCR-based marker system, a precise discrimination was observed for all race 3 isolates. However, all race 1 and 2 isolates displayed only the cosmopolitan race 1-specific amplicon pattern. CONCLUSION: The development and/or validation of novel race-specific marker systems are necessary to allow a precise discrimination of the potentially endemic Brazilian FOL race 2. SIGNIFICANCE AND IMPACT OF THE STUDY: The present characterization of isolates indicates that distinct evolutionary mechanisms are acting to select new FOL races and/or genetic variants across agroecosystems around the globe.

Performance and Meat Quality of Broiler Chickens Reared on two Different Litter Materials and at two Stocking Densities.

1. This study evaluated the performance and meat quality of broiler chickens reared on two litter materials and at two stocking densities.2. The chicks were allotted in a completely randomised design in a 2 × 2 factorial arrangement with two litter materials (wood shavings or rice straw) and two stocking densities (24 or 30 kg/m2), with six replicates per treatment. Broiler performance, carcase yield, prime cuts, blood plasma proteins, the enzymes alanine aminotransferase and aspartate aminotransferase, edible viscera, immune organs, intestine weight and length, abdominal fat, breast meat colour, pH, weight loss by defrosting, weight loss by cooking and shear force were evaluated.3. The density of 24 kg/m2 positively influenced feed consumption and weight gain. The type of litter had a positive influence on feed conversion, with a lower value for birds raised on wood shavings.4. Breast production was improved in birds reared at a density of 24 kg/m2 when compared to birds reared at a density of 30 kg/m2. The rice straw bedding negatively affected abdominal fat (resulting in higher percentages) when compared to birds reared on wood shavings.5. The types of litter affected the enzyme aspartate aminotransferase in birds raised on wood shavings, although these values were not increased to the point of influence on the physiological functions of broilers.6. The colour of the breast meat, pH, temperature, weight loss after thawing or cooking, shear force, cholesterol, triglycerides, albumin, total proteins, glucose or alanine aminotransferase content were not influenced by the stocking densities or bedding.7. The use of wood shavings as poultry bedding at a stocking density of 24 kg/m2resulted in the best performance for broiler chickens at 42 days of age.

Fcγ-RI, Fcγ-RII and IL-10 as predictive biomarkers for post-therapeutic cicatrization time in monocytes from cutaneous leishmaniasis patients.

Cutaneous leishmaniasis (CL) treatment is based on therapy with Glucantime® , yet, there are few laboratory methods to monitor its success. In this study, ex vivo and in vitro evaluations of peripheral blood monocytes were performed in a longitudinal study to characterize the impact of Glucantime® on overall phenotypic/functional features of these cells from CL patients to identify predictive biomarkers for post-therapeutic monitoring by flow cytometry. The ex vivo evaluation from CL patients demonstrated a modulatory profile before treatment, with a decrease in TLR-2, FcγRII, HLA-DR, CD86, IFN-γR, TNF, IL-12, NO, and an increase in FcγRIII and IL-10R. Conversely, treatment changes some of these biomarker expressions by decreasing FcγRIII and IL-10R and increasing IFN-γR, IL-12 and NO. Moreover, an in vitro analysis of these patients showed a reduced phagocytic capacity of Leishmania braziliensis and higher levels of IL-10 and TGF-ß modulating functional profile. Regardless of the compromised L. braziliensis phagocytic capacity, treatment re-established the production of IL-12, IL-10, TGF-ß and NO at the basal level. Notably, monocytes from patients with early cicatrization showed enhanced FcγRI and FcγRII expressions and reduced IL-10, which was further corroborated by a baseline fold change analysis. Finally, the logistic regression model emphasized the performance of FcγRI, FcγRII and IL-10 as robust predictive biomarkers for post-therapeutic cicatrization during cutaneous leishmaniasis.

Low performance on mathematical tasks in preschoolers: the importance of domain-general and domain-specific abilities.

BACKGROUND: Different domain-specific and domain-general cognitive precursors play a key role in the development of mathematical abilities. The contribution of these domains to mathematical ability changes during development. Primary school-aged children who show mathematical difficulties form a heterogeneous group, but it is not clear whether this also holds for preschool low achievers (LAs) and how domain-specific and domain-general abilities contribute to mathematical difficulties at a young age. The aim of this study was to explore the cognitive characteristics of a sample of preschool LAs and identify sub-types of LAs. METHODS: 81 children were identified as LAs from 283 preschoolers aged 3 to 5 years old and were assessed on a number of domain-general and domain-specific tasks. RESULTS: Cluster analysis revealed four subgroups of LAs in mathematics: (1) a weak processing sub-type; (2) a general mathematical LAs sub-type; (3) a mixed abilities sub-type; and (4) a visuo-spatial deficit sub-type. Whilst two of the groups showed specific domain-general difficulties, none showed only domain-specific difficulties. CONCLUSIONS: Current findings suggest that preschool LAs constitute a heterogeneous group and stress the importance of domain-general factors for the development of mathematical abilities during the preschool years.

Lipoxin A4 selectively programs the profile of M2 tumor-associated macrophages which favour control of tumor progression.

In tumor microenvironments, the macrophage population is heterogeneous, but some macrophages can acquire tumor-promoting characteristics. These tumor-associated macrophages (TAM) exhibit an M2-like profile, with deficient production of NO and ROS, characteristics of pro-inflammatory M1 cytotoxic macrophages. Lipoxins (LX) and 15-epi-lipoxins are lipid mediators which can induce certain features of M2 macrophages in mononuclear cells, but their effects on TAM remain to be elucidated. This study tested the hypothesis that ATL-1, a synthetic analogue of 15-epi-lipoxin A4 , could modulate TAM activity profile. We show that human macrophages (MΦ) differentiated into TAM-like cells after incubation with conditioned medium from MV3, a human melanoma lineage cell. Contrasting with the effects observed in other M2 subsets and M1 profile macrophages, ATL-1 selectively decreased M2 surface markers in TAM, suggesting unique behavior of this particular M2 subset. Importantly, these results were replicated by the natural lipoxins LXA4 and the aspirin induced 15-epi-LXA4 (ATL). In parallel, ATL-1 stimulated TAM to produce NO by increasing the iNOS/arginase ratio and activated NADPH oxidase, triggering ROS production. These alterations in TAM profile induced by ATL-1 led to loss of the anti-apoptotic effects of TAM on melanoma cells and increased their cytotoxic properties. Finally, ATL-1 was found to inhibit tumor progression in a murine model in vivo, which was accompanied by alterations in TAM profile and diminished angiogenesis. Together, the results show an unexpected effect of lipoxin, which induces in TAM a change from an M2- to an M1-like profile, thereby triggering tumor cell apoptosis and down-modulating the tumor progression.

NK cells control HIV-1 infection of macrophages through soluble factors and cellular contacts in the human decidua.

BACKGROUND: During the first trimester of pregnancy, HIV-1 in utero transmission is rare despite the permissivity of the placenta and the decidua (the uterine mucosa during pregnancy) to infection. In the decidua from the first trimester of pregnancy, macrophages (dMs) are the HIV-1 main target cells. Decidual natural killer (dNK) cells account for 70 % of decidual leukocytes. They display distinct phenotype and functions compared to peripheral NK cells. At the periphery, NK cells are involved in the control of HIV-1 infection. In this study, we investigate whether human decidual natural killer (dNK) cells control dM HIV-1 infection. RESULTS: Autologous cocultures of infected dMs with dNK cells reveal that dNK cells strongly inhibit dM HIV-1 infection. The addition of dNK cells to dMs at different times after infection suggests that the control occurs before the complete establishment of the infection. Double chamber cocultures show that cellular contacts are necessary for an optimal control of infection. Nevertheless, soluble factors secreted by dMs and dNK cells in double chamber cocultures partially inhibit dM HIV-1 infection, indicating that soluble factors have also a role in the control of infection. IFN-γ secretion is increased in infected and uninfected cocultures. We show that IFN-γ is involved in the control of dM HIV-1 infection by dNK cells. CONCLUSIONS: These results demonstrate that human dNK cells inhibit efficiently HIV-1 infection in dMs in vitro, and highlight the role of innate immune determinants in the control of HIV-1 transmission.

Laryngeal papillomatosis in children: The impact of late recognition over evolution.

To assess the impact of the delay in recognition of the initial symptoms of laryngeal papillomatosis in children over the evolution of the disease. Retrospective study of patients with respiratory papillomatosis referred from general pediatric practices to a tertiary hospital with pediatric laryngology specialization. Gender, age at time of diagnosis, symptom duration, sites affected at the time of diagnosis, treatment, and evolution of the disease over time were evaluated. From January 2003 to December 2013, 21 patients (15 females and 6 males) were identified and followed for at least 3 years. The average age at which symptoms first appeared was 40.2 months, and the average age at the time of initial treatment was 76 months. The most frequent clinical manifestation was hoarseness. The most common site of involvement was the glottis followed by the supraglottis and subglottis, respectively. Three of the 21 patients required tracheostomy. The average time from symptom onset to diagnosis was 52.3 months. There were no fatalities. On average 3.7 procedures were performed per patient. Patients presenting more than 1 year from the symptom onset to the time of first treatment required a greater number of procedures to control disease. Delay in diagnosis can have negative clinical consequences due to disease progression. The later the diagnosis the more surgeries are needed to control the disease. Involvement of the subglottic larynx is a risk factor for emergent tracheostomy.

A protein extract and a cysteine protease inhibitor enriched fraction from Jatropha curcas seed cake have in vitro anti-Toxoplasma gondii activity.

Toxoplasma gondii is a parasite of great medical and veterinary importance that has worldwide distribution and causes toxoplasmosis. There are few treatments available for toxoplasmosis and the search for plant extracts and compounds with anti-Toxoplasma activity is of utmost importance for the discovery of new active drugs. The objective of this study was to investigate the action of a protein extract and a protease inhibitor enriched fraction from J. curcas seed cake on developing tachyzoites of T. gondii-infected Vero cells. The protein extract (JcCE) was obtained after solubilization of the J. curcas seed cake with 100 mM sodium borate buffer, pH 10, centrifugation and dialysis of the resulting supernatant with the extracting buffer. JcCE was used for the in vitro assays of anti-Toxoplasma activity at 0.01, 0.1, 0.5, 1.5, 3.0 and 5.0 mg/ml concentration for 24 h. The results showed that JcCE reduced the percentage of infection and the number of intracellular parasites, but had no effect on the morphology of Vero cells up to 3.0 mg/mL. The cysteine protease inhibitor enriched fraction, which was obtained after chromatography of JcCE on Sephadex G-75 and presented a unique protein band following SDS-PAGE, reduced both the number of T. gondii infected cells and intracellular parasites. These results suggest that both JcCE and the cysteine protease inhibitor enriched fraction interfere with the intracellular growth of T. gondii.

First Report of Leaf Spot Caused by Myrothecium roridum on Coffea canephora in Brazil.

Coffea canephora (conilon coffee) represents approximately 30% of the coffee marketed worldwide. The state of Espírito Santo is the largest conilon coffee-producing state in Brazil. In 2013 and 2014, leaves with a leaf spot were observed on most of the conilon coffee seedlings in a commercial nursery in Laranja da Terra, Espírito Santo, Brazil. The infected leaves were deposited in the VIC Herbarium (VIC 42482) and a pure single-spore culture of the pathogen was deposited in the culture collection of the Universidade Federal de Viçosa (Accession No. COAD 1729). The initial symptoms were circular, brown to dark brown lesions with yellow margins occurring on both leaf surfaces. In high humidity, concentric rings formed and the lesions expanded rapidly to reach up to 30 mm in diameter, and later became dark brown with a grayish center. Black sporodochia with white, and marginal mycelial tuffs bearing black spore masses were observed in the older lesions. These symptoms were consistent with those of Myrothecium leaf spot reported on Coffea spp. (3). Microscopic observation revealed aseptate, hyaline, and cylindrical conidia, rounded at both ends, greenish to black in mass, and 5 to 6 µm long and 1 to 2 µm wide. The symptoms and morphological characteristics described above matched the description of Myrothecium roridum Tode (4). To confirm this identification, DNA was extracted using a Wizard Genomic DNA Purification Kit and the sequence of an internal transcribed spacer (ITS) region was obtained and deposited in GenBank (Accession No. KJ815095). The sequence of the ITS region exhibited 100% identity over 561 bp with another M. roridum sequence in GenBank (JF343832). To verify the pathogenicity of the fungus, healthy leaves of the C. canephora clones 12v and 14 (four seedlings each) were wounded superficially with a sterilized needle and inoculated by spraying them with a suspension of M. roridum conidia (106 conidia ml-1). The seedlings were covered with plastic bags and incubated in a growth chamber at 25°C under a photoperiod of 12 h light/12 h dark for 5 days. The control seedlings were sprayed with distilled water and incubated similarly. Fifteen days after inoculation, symptoms in all inoculated seedlings were consistent with those initially observed on the naturally infected seedlings, whereas the controls remained healthy. Re-isolation and identification confirmed Koch's postulates. M. roridum has a wide host range, and symptoms were similar to those reported in other hosts of the pathogen in Brazil (2,3). There is only one report of M. roridum on C. canephora in Colombia (1); however, this pathogen was previously reported on C. arabica in Brazil, Colombia, Costa Rica, Guatemala, India, Indonesia, Puerto Rico, and the Virgin Islands (1,3). To our knowledge, this is the first report of a leaf spot caused by M. roridum on conilon coffee in Brazil. The cultivation of conilon coffee is increasing and the reported leaf spot disease affects the quality of the seedlings in nurseries. It is therefore important to conduct a thorough study of management strategies for this disease. References: (1) D. F. Farr and A. Y. Rossman. Fungal Databases. Syst. Mycol. Microbiol. Lab. ARS, USDA. Retrieved from http://nt.ars-grin.gov/fungaldatabases , 27 May 2014. (2) A. M. Quezado Duval et al. Braz. J. Microbiol. 41:246, 2010. (3) S. F. Silveira et al. Fitopatol. Bras. 32:440, 2007. (4) M. Tulloch. Mycol. Pap. No. 130. CMI, Wallingford, UK, 1972.

Tubed myocutaneous pectoralis major flap: A rescue option in esophagus reconstruction.

This article presents a complex case of total pharyngoesophageal defect reconstruction in a patient with recurrent supraglottic squamous cell carcinoma. After failed attempts with free microsurgical techniques due to complications, a tubed myocutaneous pectoralis major flap was successfully employed. The procedure aimed to achieve alimentary continuity, speech, and swallowing functionality. Despite initial challenges and concerns about stenosis, the reconstruction of a total pharyngolarygectomy defect resulted in a good functional outcome, although minor deficits in neck mobility and aesthetic donor zone deformities were noted. The case highlights the broad armamentarium of reconstruction techniques that plastic and aesthetic surgery teams must be trained, namely the myocutaneous pectoralis major flap which in selected cases can be the end solution.

Glycemic variability after mechanical thrombectomy for anterior circulation acute ischemic stroke. / Variabilidad glucémica tras trombectomía mecánica en el ictus isquémico agudo de la circulación anterior.

INTRODUCTION: Morbidity remains high among patients who undergo successful mechanical thrombectomy (MT) for anterior circulation large vessel occlusion (LVO). Stress hyperglycemia worsens the prognosis after acute ischemic stroke (AIS), but aggressively treating hyperglycemia does not improve the outcome. There is no consensus on how to best manage glycemia after AIS. Glycemic variability (GV) reflects glycemic fluctuations over time and could be the culprit. We aimed to elucidate how GV impacts outcome of AIS patients treated with MT. PATIENTS AND METHODS: This was a single-center retrospective study. We consecutively included AIS patients who received MT for anterior circulation LVO. We recorded discrete blood glucose measurements within the first 24 hours post thrombectomy, from which we calculated two measures of GV: standard deviation (SD) and coefficient of variation. Univariate and multivariate analyses were conducted to identify predictors of poor functional outcome (modified Ranking scale score 3-6) and mortality at 3-month follow-up. RESULTS: We included 657 patients. Patients with poor functional outcome (42.5%) and patients that died (14.8%) had significantly higher GV as measured by SD. In a multivariable model adjusted for confounders, higher SD was associated with mortality -adjusted odds ratio: 1.020 (95% CI 1.001-1.040)- but not with functional outcome -adjusted odds ratio for modified Ranking scale score 3-6: 1.007 (95% CI 0.990-1.025)-. CONCLUSIONS: Our results suggest that higher GV after MT for anterior circulation AIS is an independent risk factor for 3-month mortality. Future trials should evaluate the benefit of reducing GV in this setting.

TITLE: Variabilidad glucémica tras trombectomía mecánica en el ictus isquémico agudo de la circulación anterior.Introducción. La morbilidad de los pacientes con ictus isquémico agudo (IIA) sometidos a trombectomía mecánica (TM) exitosa permanece alta. La hiperglucemia empeora el pronóstico tras un IIA, pero tratarla agresivamente no mejora los resultados. No existe consenso sobre el tratamiento óptimo de la glucemia después de un IIA. La variabilidad glucémica (VG), que refleja las fluctuaciones glucémicas a lo largo del tiempo, puede ser un factor importante. Nuestro objetivo fue investigar cómo la VG afecta el resultado de pacientes con IIA tratados con TM. Pacientes y métodos. Realizamos un estudio retrospectivo unicéntrico que incluyó a pacientes con IIA que recibieron TM para la oclusión de un gran vaso de la circulación anterior. Se registraron mediciones discretas de glucemia en las primeras 24 horas postrombectomía, a partir de las cuales se calcularon dos medidas de VG: desviación estándar y coeficiente de variación. Se realizó un análisis univariado y multivariado para identificar predictores de resultado funcional desfavorable (escala de Rankin modificada: 3-6) y mortalidad a los tres meses. Resultados. Se incluyó a 657 pacientes. Los que tenían una puntuación en la escala de Rankin modificada = 3 (42,5%) y los fallecidos (14,8%) tuvieron una VG significativamente mayor medida por desviación estándar. En un modelo multivariado, una mayor desviación estándar se asoció de forma independiente con la mortalidad ­odds ratio ajustada: 1,02 (intervalo de confianza al 95%: 1,001-1,04)­ pero no con el resultado funcional ­odds ratio ajustada de la escala de Rankin modificada = 3: 1,007 (intervalo de confianza al 95%: 0,99-1,025)­. Conclusiones. Nuestros resultados sugieren que una mayor VG tras la TM para el IIA de la circulación anterior es un factor de riesgo independiente de mortalidad a los tres meses. Los futuros ensayos deben evaluar el beneficio de reducir la VG en este contexto.

Ultra-high pressure LC for astaxanthin determination in shrimp by-products and active food packaging.

Nowadays, there is increasing interest in natural antioxidants from food by-products. Astaxanthin is a potent antioxidant and one of the major carotenoids in crustaceans and salmonids. An ultra-high pressure liquid chromatographic method was developed and validated for the determination of astaxanthin in shrimp by-products, and its migration from new packaging materials to food simulants was also studied. The method uses an UPLC® BEH guard-column (2.1 × 5 mm, 1.7 µm particle size) and an UPLC® BEH analytical column (2.1 × 50 mm, 1.7 µm particle size). Chromatographic separation was achieved using a programmed gradient mobile phase consisting of (A) acetonitrile-methanol (containing 0.05 m ammonium acetate)-dichloromethane (75:20:5, v/v/v) and (B) ultrapure water. This method was evaluated with respect to validation parameters such as linearity, precision, limit of detection, limit of quantification and recovery. Low-density polyethylene films were prepared with different amounts of the lipid fraction of fermented shrimp waste by extrusion, and migration was evaluated into food simulants (isooctane and ethanol 95%, v/v). Migration was not detected under the tested conditions.

Application of denaturing gradient gel electrophoresis for detection of bacterial and yeast communities along a salinity gradient in the estuary of the Cachoeira River in Brazil.

An estuary is a transition zone between freshwater and marine ecosystems, resulting in dilution of seawater. Estuaries are also considered environments of intense biological activity related to the processes of nutrient cycling. The aim of this study was to evaluate the microbial community composition along a salinity gradient in the estuary of the Cachoeira River, located in southern Bahia, Brazil. The analysis of bacterial and yeast communities was performed by determining the denaturing gradient gel electrophoresis band richness. Formation of zones with similar profiles of bands was observed, and the increasing richness at the intermediate zone demonstrated a clear spatial distinction of communities depending on salinity. In addition, the dissolved oxygen content, temperature, pH, salinity, and dissolved inorganic nutrient contents (NH3(+), NO2(-), NO3(-), PO4(-)) were determined. Nutrients were distributed in similar patterns, with decreasing concentrations as the salinity increases.

Endogenous sex hormones are not associated with subclinical atherosclerosis in menopausal women.

AIM: The aim of this paper was to compare the carotid intima-media thickness (IMT) in pre- and postmenopausal women and to evaluate the association between endogenous sex hormones, body fat distribution, and insulin resistance and the IMT. METHODS: This cross-sectional study included 145 women aged 45-65 yr, comprising 56 premenopausal (FSH<20IU/mL and regular menstrual cycles) and 89 postmenopausal (FSH>40IU/ml and amenorrheic). All patients were evaluated for lipid profile, estradiol and testosterone, insulin ratio (G/I), HOMA-IR, and ultrasound measurement of IMT. Each variable was assessed for correlation with IMT using the univariate model. RESULTS: No difference was observed in IMT between pre- and postmenopausal women. A positive and statistically significant correlation was found between IMT and FSH levels (rs=0.21, P<0.009) and HOMA (rs=0.16, P<0.04). A positive and statistically significant correlation was observed between testosterone and waist (rs=0.3, P<0.04). No correlation was found between IMT and time of menopause (r=0.02, P=0.19). CONCLUSION: Estradiol and testosterone are not associated with subclinical atherosclerosis in menopausal women. A positive correlation between IMT and FSH may reflect an association between low estrogen and IMT. Abdominal fat can be an important link between androgenic levels and cardiovascular risk.

Temperature estimation of a pair of trapped ions.

We apply estimation theory to a system formed by two interacting trapped ions. By using the Fisher matrix formalism, we introduce a simple scheme for estimation of the temperature of the longitudinal vibrational modes of the ions. We use the ions interaction to effectively infer the temperature of the individual ions, by optimising the interaction time evolution and by measuring only over one of the ions. We also investigate the effect of a non-thermal reservoir over the inference approach. The non-classicality of one of the ions vibrational modes, introduced due to a squeezed thermal reservoir, does not directly affect the inference of the individual temperatures, although allowing the modes to be entangled. To check actual experimental conditions, we analyze the temperature inference under heating due to surface-electrode noise.

MHV-68 Open Reading Frame 20 is a nonessential gene delaying lung viral clearance.

Recently, it has been demonstrated that the MHV-68 ORF20-encoded gene product induces cell-cycle arrest at the G2/M phase, followed by apoptosis. To study the role of this conserved gene in vivo, two independent ORF20-deficient MHV-68 viruses and their revertants were constructed. As the replication in vitro of both mutants followed similar kinetics to that of the wild-type and revertant viruses, ORF20 is therefore a nonessential virus gene. No cell cycle arrest could be observed upon infection of cells with wild type MHV-68 or mutant viruses. In addition, no major differences were detected between mock- and virus-infected cells when protein and inactivation levels of the mitotic promoter factor cdc2/cyclinB were analyzed. Following intranasal infection, the recovery of mutant, revertant and wild-type viruses in the lungs was similar. With the ORF20-deficient viruses, however, there was a significant delay of four days in clearance of virus from the lungs. Surprisingly, the magnitude and cell population distribution in the exudates of the lung was essentially similar to mice infected with wild-type, revertant or ORF20-deleted viruses. Subsequent establishment of latency was normal for both mutants, demonstrating that ORF20 does not play a critical role in establishment of a persistent infection. These results indicate that while expression of ORF20 may impact on the pathogenicity of the infection, the observed induction of G2/M arrest in ORF20-expressing cells may not be the primary function of ORF20 in the context of viral infection.

Association between myocardial mechanical dispersion and ventricular arrhythmogenicity in chagas cardiomyopathy.

Chagas disease is a major health concern in Latin America. Ventricular arrhythmia (VA) is a hallmark of Chagas cardiomyopathy (CCM), associated with worse prognosis. The present study aimed to verify the association between myocardial mechanical dispersion (MD) and ventricular arrhythmogenicity in CCM. In a cross-sectional study, 77 patients (55.8 ± 10.4 years) with CCM were evaluated. Global longitudinal strain (GLS) and MD were assessed by echocardiography, derived from the speckle tracking technique. Myocardial MD was measured from the onset of the Q/R wave on electrocardiogram to the peak longitudinal strain in 16 segments of the left ventricle. Frequency and complexity of ventricular extrasystoles (VES) were assessed by dynamic electrocardiography. The density and complexity of VES and the presence of non-sustained ventricular tachycardias (NSVTs) increase as MD increases. In logistic regression, MD was the only variable associated with the presence of paired VES and ventricular bigeminy. In addition, both MD and GLS were associated with the presence of NSVT (both, p < 0.01), and MD was independently associated with NSVT (OR 1.04, 95% CI 1.004-1.201, p = 0.031). In CCM, MD is associated with a higher density and complexity of VES, including NSVT.

Carangoides bartholomaei (Cuvier, 1833) stomach: a source of aspartic proteases for industrial and biotechnological applications.

The viscera and other residues from fish processing are commonly discarded by the fishing industry. These by-products can be a source of digestive enzymes with industrial and biotechnological potential. In this study, we aimed at the extraction, characterization, and application of acidic proteases from the stomach of Carangoides bartholomaei (Cuvier, 1833). A crude extract from the stomachs was obtained and submitted to a partial purification process by salting-out, which obtained a Purified Extract (PE) with a specific proteolytic activity of 54.0 U⋅mg-1. A purification of 1.9 fold and a yield of 41% were obtained. The PE presents two isoforms of acidic proteases and a maximum proteolytic activity at 45 °C and pH 2.0. The PE acidic proteolytic activity was stable in the pH range of 1.5 to 7.0 and temperature from 25 °C to 50 °C. Purified Extract kept 35% of its proteolytic activity at the presence of NaCl 15% (m/v) but was totally inhibited by pepstatin A. Purified Extract aspartic proteases presented high activity in the presence of heavy metals such as Cd2+, Hg2+, Pb2+, Al3+, and Cu2+. The utilization of PE as an enzymatic addictive in the collagen extraction from Nile tilapia scales has doubled the process yield. The results indicate the potential of these aspartic proteases for industrial and biotechnological applications.

Climate change vulnerability of cetaceans in Macaronesia: Insights from a trait-based assessment.

Over the last decades global warming has caused an increase in ocean temperature, acidification and oxygen loss which has led to changes in nutrient cycling and primary production affecting marine species at multiple trophic levels. While knowledge about the impacts of climate change in cetacean's species is still scarce, practitioners and policymakers need information about the species at risk to guide the implementation of conservation measures. To assess cetacean's vulnerability to climate change in the biogeographic region of Macaronesia, we adapted the Marine Mammal Climate Vulnerability Assessment (MMCVA) method and applied it to 21 species management units using an expert elicitation approach. Results showed that over half (62%) of the units assessed presented Very High (5 units) or High (8 units) vulnerability scores. Very High vulnerability scores were found in archipelago associated units of short-finned pilot whales (Globicephala macrorhynchus) and common bottlenose dolphins (Tursiops truncatus), namely in the Canary Islands and Madeira, as well as Risso's dolphins (Grampus griseus) in the Canary Islands. Overall, certainty scores ranged from Very High to Moderate for 67% of units. Over 50% of units showed a high potential for distribution, abundance and phenology changes as a response to climate change. With this study we target current and future information needs of conservation managers in the region, and guide research and monitoring efforts, while contributing to the improvement and validation of trait-based vulnerability approaches under a changing climate.