Smell loss associated with SARS-CoV-2 is not clinically different from other viruses: a multicenter cohort study.

BACKGROUND: Although the COVID-19 pandemic has increased the prevalence of cases with olfactory loss, other respiratory viruses can also cause this condition. We aimed to compare the prevalence of acute SARS-CoV-2 infection and other respiratory viruses in patients with sudden smell loss, and to assess the impact of SARS-CoV-2 viral load and co-infection on olfactory symptoms. METHODS: Patients with sudden smell loss were recruited in a multicenter prospective cohort study in 15 hospitals in Brazil. Clinical questionnaire, Connecticut Chemosensory Clinical Research Center (CCCRC) olfactory test and nasopharyngeal swab to perform a PCR-based respiratory viral panel were collected at first visit (day 0) and 30 and 60 days after recruitment. RESULTS: 188 of 213 patients presented positive test result for SARS-CoV-2, among which 65 were co-infected with other respiratory viruses (e.g., rhinovirus, enterovirus, and parainfluenza). 25 had negative test results for SARS-CoV-2. Patients in both SARSCoV-2 and non-SARS-CoV-2 groups had objective anosmia (less than 2 points according to the psychophysical olfactory CCCRC) at day 0, with no significant difference between them. Both groups had significant smell scores improvement after 30 and 60 days, with no difference between them. Co-infection with other respiratory viruses, and SARS-CoV-2 viral load did not impact olfactory scores. CONCLUSION: Patients with sudden smell loss associated with SARS-CoV-2 and other respiratory viruses had similar presentation, with most participants initiating with anosmia, and total or near total recovery after 60 days. SARS-CoV-2 viral load and co-infections with other respiratory viruses were not associated with poorer olfactory outcomes.

New host and geographical records for Parafilaroides normani (Nematoda: Filaroididae) Dailey, 2009 in South American fur seal, Arctocephalus australis, from southern Brazil.

Lungworms are a common finding in seals and fur seals around the world. However, from existing records, the biogeographical distribution of filaroid helminths appears to be restricted, and these parasites are endemic in only certain areas and species, mainly in the Northern Hemisphere. The occurrence of infection in pinniped species in the Southern Hemisphere is scarce. The objective of this work is to verify the prevalence of lungworms in Arctocephalus australis in waters off the southern coast of Brazil. Twenty subadult specimens of A. australis found recently dead on the southern coast of Brazil were necropsied and their lungs were examined. Parasitic cysts were found in only one specimen (prevalence of 5%). The helminths were morphologically identified as Parafilaroides normani (Metastrongyloidea: Filaroididae). This helminth species has been reported in pinnipeds from Australia, New Zealand and South Africa. This is the first record of P. normani in A. australis and for the western South Atlantic, providing additional data regarding the biogeographic distribution of the parasite.

Molecular Dynamics of Cyclodextrins in Water Solutions from NMR Deuterium Relaxation: Implications for Cyclodextrin Aggregation.

The aggregation of the most common natural cyclodextrins (α-, ß-, and γ-) in aqueous solutions is addressed by studying the CD-CD interactions using deuterium relaxation rates for deuterium labeled CDs. Relaxation times (T1) and their corresponding relaxation rates (R1 = 1/T1) provide information about the rotational correlation times of CDs and serve as a proxy for solute-solute interactions. Measured T1's for α-, ß-, and γ-CD at the lowest CD concentrations were in agreement with predictions of a hydrodynamic model for toroids, in particular with regard to the dependence of T1 on CD size. On the other hand, the dependence of T1's with respect to the increase in CD concentration could not be explained by hydrodynamic or direct interaction between CD molecules, and it is suggested that there is an equilibrium between monomeric and dimeric CD to account for the observed concentration dependence. No evidence in favor of large aggregates of CDs involving a non-negligible fraction was found for the investigated CDs.

Recombinant expression of Ixolaris, a Kunitz-type inhibitor from the tick salivary gland, for NMR studies.

Ixolaris is an anticoagulant protein identified in the tick saliva of Ixodes scapularis. Ixolaris contains 2 Kunitz like domains and binds to Factor Xa or Factor X as a scaffold for inhibition of the Tissue Factor (TF)/Factor VIIa (FVIIa). In contrast to tissue factor pathway inhibitor (TFPI), however, Ixolaris does not bind to the active site cleft of FXa. Instead, complex formation is mediated by the FXa heparin-binding exosite. Due to its potent and long-lasting antithrombotic activity, Ixolaris is a promising agent for anticoagulant therapy. Although numerous functional studies of Ixolaris exist, three-dimensional structure of Ixolaris has not been obtained at atomic resolution. Using the pET32 vector, we successfully expressed a TRX-His6-Ixolaris fusion protein. By combining Ni-NTA chromatography, enterokinase protease cleavage, and reverse phase HPLC (RP-HPLC), we purified isotopically labeled Ixolaris for NMR studies. 1D 1H and 2D 15N-1H NMR analysis yielded high quality 2D 15N-1H HSQC spectra revealing that the recombinant protein is folded. These studies represent the first steps in obtaining high-resolution structural information by NMR for Ixolaris enabling the investigation of the molecular basis for Ixolaris-coagulation factors interactions.

Speech and language therapy service delivery: overcoming limited provision for children.

OBJECTIVES: To test an alternative Speech and Language Therapy (SLT) service delivery model based on partnerships between a University and local schools and charities, and to report on the impact and feasibility of intervention based on long-term outcome measures and three case studies with individual analysis of Reliable Change. STUDY DESIGN: The following six-step model was tested: 1-establishing partnerships; 2-flagging children; 3-pre-treatment SLT assessment; 4-reporting and discussion with parents and teachers; 5-treatment; 6-post-treatment assessment. Case studies are presented. METHODS: A partnership was established with one kindergarten in a pre-test and a total of 25 kindergartens during the second phase of the process. A group of 139 children were then flagged and assessed. The following long-term outcomes (18 months post-therapy) were investigated: phonetic-phonological standardised test percentiles and raw scores; receptive and expressive language percentiles and raw scores according to a standardised language test; percentage of syllables stuttered; duration of stuttering moments; academic achievement in norm-tests' core areas (mathematics, Portuguese language and social studies). Case studies and a 95% credible interval analysis to assess Reliable Change are presented. RESULTS: Seventy five (54%) children needed SLT support. Fifty (67%) of those children returned to the clinic for long-term assessments and the analysis of all outcome measures showed significant improvements in their performance, 18 months post-therapy. Case Studies Reliable Change analysis revealed a statistically significant improvement, which also clearly shows the feasibility and the positive impact of the intervention. CONCLUSIONS: This specialised and differentiated care network constitutes an alternative delivery system of SLT services that addresses the lack of support currently experienced by children and their families. The long-term outcome measures and the 95% credible interval analysis are reliable methods to determine the impact of interventions.

Acute and chronic malnutrition and their predictors in children aged 0-5 years in São Tomé: a cross-sectional, population-based study.

OBJECTIVE: Undernutrition is an important cause of morbidity and mortality in infants and children worldwide. The aim of this study was to evaluate the nutritional status and their predictors in children from 0 to 5 years of age in São Tomé. STUDY DESIGN: A cross-sectional study was conducted in São Tomé Island. METHODS: A total of 1285 individuals were enrolled between January and May 2011. Children were measured, and height for age (HAZ), weight for height (WHZ) and body mass index (BAZ) Z-score were computed. Global acute undernutrition is defined as weight for height <-1 Z-score (wasting < -2 Z-scores) and global chronic undernutrition as length/height for age <-1 Z-score (stunting < -2 Z-scores). Relevant information was collected from individual health bulletins, namely gestational age and birth weight, as well as weight at 6, 12, 18 and 24 months for all individual above these ages. Mothers were invited to answer a specific questionnaire. RESULTS: A high percentage of global acute undernutrition (30.9% in <24 months and 21.9% in ≥24 months) and global chronic undernutrition (32.5% in <24 months and 41.1% in ≥24 months) was observed. Appropriate birth weight for gestational age (AGA) is significantly associated with lower odds for both acute (OR 0.485 [95% CI 0.299-0.785]) and chronic undernutrition (OR 0.427 [95% CI 0.270-0.675]) in children >12 months. Weight gain above 0.67 Z-score in the first semester of life was strongly related to lower odds for both acute (OR 0.109 [95% CI 0.040-0.291]) and chronic undernutrition (OR 0.379 [95% CI 0.187-0.770]) in children >12 months of age. Similarly, mother's education seems to protect against acute (>12 months: OR 0.448 [95% CI 0.244-0.825]; >24 months: OR 0.186 [95% CI 0.064-0.540]) and chronic undernutrition (>12 months: OR 0.389 [95% CI 0.232-0.653]; >24 months: OR 0.324 [95% CI 0.171-0.625]). All logistic regressions were adjusted for all children (gender, age, gestational age, birth weight, breastfeeding, begin consumption alcohol), mothers (age, height, body mass index, educational level, number of previous pregnancies, smoking during pregnancy, alcohol consumption during pregnancy) and household (number of siblings and persons at home) variables included in the protocol. CONCLUSIONS: Birth weight, nutritional status, and the mother's education and weight gain particularly in the first year of life were important factors protecting against undernutrition during infancy and childhood. These results emphasize the importance of women's nutrition and of adequate birth weight and particularly weight gain during the first year of life in order to prevent wasting and stunting in childhood.

Epitope mapping by solution NMR spectroscopy.

Antibodies play an ever more prominent role in basic research as well as in the biotechnology and pharmaceutical sectors. Characterizing their epitopes, that is, the region that they recognize on their target molecule, is useful for purposes ranging from molecular biology research to vaccine design and intellectual property protection. Solution NMR spectroscopy is ideally suited to the atomic level characterization of intermolecular interfaces and, as a consequence, to epitope discovery. Here, we illustrate how NMR epitope mapping can be used to rapidly and accurately determine protein antigen epitopes. The basic concept is that differences in the NMR signal of an antigen free or bound by an antibody will identify epitope residues. NMR epitope mapping provides more detailed information than mutagenesis or peptide mapping and can be much more rapid than X-ray crystallography. Advantages and drawbacks of this technique are discussed together with practical considerations.

Genetic diversity and population structure of Synthesium pontoporiae (Digenea, Brachycladiidae) linked to its definitive host stocks, the endangered Franciscana dolphin, Pontoporia blainvillei (Pontoporiidae) off the coast of Brazil and Argentina.

Pontoporia blainvillei (Gervais and d'Orbigny, 1844) is an endangered small cetacean endemic to South America with four Franciscana Management Areas (FMA) recognized as different population stocks. The role of the intestinal parasite Synthesium pontoporiae (Digenea: Brachycladiidae) as a possible biological marker to differentiate P. blainvillei stocks was evaluated using nuclear and mitochondrial DNA markers. Internal transcribed sequence 1 and 2 (ITS1 and ITS2) regions of S. pontoporiae did not show intraspecific variability. The mitochondrial NADH dehydrogenase subunit 3 (ND3) and cytochrome oxidase subunit I (COI) gene sequences suggested lack of population structure in S. pontoporiae and population expansion. The apparent panmixia of S. pontoporiae may be due to the high mobility of one or more of its intermediary hosts. Alternatively, it may be due to the small sample size. This result is incongruent with the previously proposed FMA.

Bet v 1--a Trojan horse for small ligands boosting allergic sensitization?

BACKGROUND: Birch pollen allergy represents the main cause of winter and spring pollinosis in the temperate climate zone of the northern hemisphere and sensitization towards Bet v 1, the major birch pollen allergen, affects over 100 million allergic patients. The major birch pollen allergen Bet v 1 has been described as promiscuous acceptor for a wide variety of hydrophobic ligands. OBJECTIVE: In search of intrinsic properties of Bet v 1, which account responsible for the high allergenic potential of the protein, we thought to investigate the effects of ligand-binding on immunogenic as well as allergenic properties. METHODS: As surrogate ligand of Bet v 1 sodium deoxycholate (DOC) was selected. Recombinant and natural Bet v 1 were characterised physico-chemically as well as immunologically in the presence or absence of DOC, and an animal model of allergic sensitization was established. Moreover, human IgE binding to Bet v 1 was analysed by nuclear magnetic resonance (NMR) spectroscopy. RESULTS: Ligand-binding had an overall stabilizing effect on Bet v 1. This translated in a Th2 skewing of the immune response in a mouse model. Analyses of human IgE binding on Bet v 1 in mediator release assays revealed that ligand-bound allergen-induced degranulation at lower concentrations; however, in basophil activation tests with human basophils ligand-binding did not show this effect. For the first time, human IgE epitopes on Bet v 1 were determined using antibodies isolated from patients' sera. The IgE epitope mapping of Bet v 1 demonstrated the presence of multiple binding regions. CONCLUSIONS AND CLINICAL RELEVANCE: Deoxycholate binding stabilizes conformational IgE epitopes on Bet v 1; however, the epitopes themselves remain unaltered. Therefore, we speculate that humans are exposed to both ligand-bound and free Bet v 1 during sensitization, disclosing the ligand-binding cavity of the allergen as key structural element.

Phonological and articulation treatment approaches in Portuguese children with speech and language impairments: a randomized controlled intervention study.

BACKGROUND: In Portugal, the routine clinical practice of speech and language therapists (SLTs) in treating children with all types of speech sound disorder (SSD) continues to be articulation therapy (AT). There is limited use of phonological therapy (PT) or phonological awareness training in Portugal. Additionally, at an international level there is a focus on collecting information on and differentiating between the effectiveness of PT and AT for children with different types of phonologically based SSD, as well as on the role of phonological awareness in remediating SSD. It is important to collect more evidence for the most effective and efficient type of intervention approach for different SSDs and for these data to be collected from diverse linguistic and cultural perspectives. AIMS: To evaluate the effectiveness of a PT and AT approach for treatment of 14 Portuguese children, aged 4.0-6.7 years, with a phonologically based SSD. METHODS & PROCEDURES: The children were randomly assigned to one of the two treatment approaches (seven children in each group). All children were treated by the same SLT, blind to the aims of the study, over three blocks of a total of 25 weekly sessions of intervention. Outcome measures of phonological ability (percentage of consonants correct (PCC), percentage occurrence of different phonological processes and phonetic inventory) were taken before and after intervention. A qualitative assessment of intervention effectiveness from the perspective of the parents of participants was included. OUTCOMES & RESULTS: Both treatments were effective in improving the participants' speech, with the children receiving PT showing a more significant improvement in PCC score than those receiving the AT. Children in the PT group also showed greater generalization to untreated words than those receiving AT. Parents reported both intervention approaches to be as effective in improving their children's speech. CONCLUSIONS & IMPLICATIONS: The PT (combination of expressive phonological tasks, phonological awareness, listening and discrimination activities) proved to be an effective integrated method of improving phonological SSD in children. These findings provide some evidence for Portuguese SLTs to employ PT with children with phonologically based SSD.

Broken anesthetic needle in the pterygomandibular space. Four case reports.

Needle breakage in the oral cavity during local anesthesia is a rare but possible complication. In the majority of cases this happens in the pterygomandibular space during an inferior alveolar nerve block and the causes are multifactorial. The treatment recommended is localization and removal of the extraneous body, as soon as possible, in a surgical setting so as to prevent possible migration of the metal fragment with consequent possible lesions to the surrounding anatomical structures. We present four case studies that have occurred since 2011.

Standardization of a phonetic-phonological test for European-Portuguese children.

OBJECTIVE: To develop and standardize a phonetic-phonological test (Teste Fonético-Fonológico-Avaliação da Linguagem Pré-Escolar, TFF-ALPE) for the assessment of European-Portuguese (EP) children's articulation and phonological abilities. PATIENTS AND METHODS: In order to standardize TFF-ALPE, 768 children aged 3;0-6;11 participated in this study. The standardization, validity and reliability of TFF-ALPE were analyzed. RESULTS: TFF-ALPE presents strong cohesion and has strong inter- and intrajudge reliability. There was also a strong correlation between the TFF-ALPE data and those obtained in other studies. The content validity was demonstrated by the description of the test domain and the items that comprise TFF-ALPE. CONCLUSION: TFF-ALPE is a valid and reliable phonetic-phonological assessment instrument that speech-language pathologists can use with EP-speaking children.

Macrophage LXRα gene therapy ameliorates atherosclerosis as well as hypertriglyceridemia in LDLR(-/-) mice.

Liver X receptors (LXRs) are implicated in the regulation of cholesterol homeostasis, inflammatory response and atherogenesis. Administration of LXR agonists inhibits the progress of atherosclerosis, and also increases plasma triglyceride levels, representing an obstacle to their use in treating this disease. The objective of this study was to develop an alternative approach that could overcome this obstacle. Eight-week-old low-density lipoprotein receptor-deficient (LDLR(-/-)) mice were transplanted with hematopoietic stem cell (HSC)-enriched bone marrow cells transduced with lentivectors expressing either green fluorescent protein (GFP) (Lenti-SP-GFP, control) or LXRα (Lenti-SP-LXRα) driven by a synthetic macrophage promoter. At 4 weeks post-transplant, the mice were fed with a Western diet for 8 weeks and then killed. Compared with Lenti-SP-GFP mice, the Lenti-SP-LXRα mice had a 30% reduction in atherosclerotic lesions, which was accompanied by increases in levels of macrophage expression of cholesterol efflux genes apolipoprotein E and ATP-binding cassette A1, as well as decreases in plasma inflammatory cytokines interleukin-6 and tumor necrosis factor-α. Intriguingly, a 50% reduction of plasma triglyceride level was also observed. We conclude that HSC-based macrophage LXRα gene therapy ameliorates the development of atherosclerosis along with an unexpected concomitant reduction of plasma triglyceride levels in LDLR(-/-) mice. These findings highlight the potential value of macrophage LXR expression as an avenue for therapeutic intervention against atherosclerosis.

Worldwide incidence and prevalence of pediatric onset systemic lupus erythematosus.

Compilation of worldwide data regarding the incidence and prevalence of pediatric-onset systemic lupus erythematosus (SLE) is needed in order to evaluate the scope of the disease in the pediatric population. A literature review was performed to unify the current data available on the global incidence and prevalence of pediatric-onset SLE. We examined 13 available epidemiological studies concentrated on the incidence and prevalence of pediatric-onset SLE. The available reports were predominantly from North America, Europe and Asia. The limited amount of studies available highlights the need for more epidemiological research in order to better comprehend the global scope of this disease.

Burden of compensated and decompensated cirrhosis: real world data from an Italian population-based cohort study.

OBJECTIVE: To quantify the annual healthcare resource utilization, costs and mortality rate for a large cohort of Italian patients with compensated (CC) and decompensated cirrhosis (DC). PATIENTS AND METHODS: A population-based cohort study was conducted through the data-linkage of mortality for all-cause, hospitalizations and outpatient drugs and service databases of the Campania Region. All adults hospitalized with cirrhosis diagnosis (2007-2015) were grouped in CC and DC (prevalent patients) on January 1, 2016 and followed for 1-year. Incident patients with DC (2015) were also retrieved and followed from discharge date up to 1-year. Negative binomial regression was used to estimate Incidence Rate Ratios (IRRs) for predictors of all-cause hospitalizations. Costs were evaluated from the Italian National Health Service perspective and expressed in euro patient/year. RESULTS: A total of 21,433 prevalent cirrhotic patients (57.1% CC and 42.9% DC) and 1,371 incident patients with DC were identified. During a 1-year, 21.5% of prevalent patients with CC were admitted for acute events, 26.8% of those with DC and 55.4% of incident patients with DC. Ascites (IRR=1.71;95% CI: 1.37-2.14) and hepatic encephalopathy (IRR=1.35; 95% CI: 1.04-1.77) at index admission were strong predictors of hospitalizations in incident DC patients. The 1-year mortality rate was respectively 5.8% and 10.1% for prevalent patients with CC and DC and 35.6% for incident patients with DC. Direct costs amounted to 3,194€ patient/year for the prevalent CC group and 4,001€ patient/year for the DC group and 13,806 € patient/year for incident individuals with DC. CONCLUSIONS: The burden of cirrhosis dramatically differs between CC and DC patients, especially after the first decompensation episode. Ascites and hepatic encephalopathy at index admission were strong predictors of hospitalizations in incident DC patients.

A novel biologic activity of thrombin: stimulation of monocyte chemotactic protein production.

Thrombin is a serine protease that is released at sites of vascular injury and exerts a variety of biologic effects on different cell types. Thrombin is postulated to play a role in the pathogenesis of a number of diseases including atherosclerosis, since it activates vascular smooth muscle and endothelial cells. Thrombin mediates these effects through a specific receptor that is upregulated in vascular cells in atherosclerosis. Atherosclerosis and glomerulosclerosis are characterized by the presence of monocyte-macrophages in the lesions. Monocyte chemotactic protein (MCP-1) is believed to be an important mediator of monocyte recruitment to the tissue and can be induced in a broad variety of cells including mesangial cells. We studied the effect of thrombin on MCP-1 production and gene expression in well-characterized human mesangial cells, vascular pericytes that play a central role in fibrosis of the glomerular microvascular bed. alpha thrombin stimulates MCP-1 production and gene expression in mesangial cells in a dose- and time-dependent manner. Experiments with diisopropylfluorophosphate thrombin and gamma thrombin demonstrate that this thrombin effect requires both receptor binding as well as catalytic activity, features consistent with the known properties of the recently characterized and cloned thrombin receptor. Moreover, a human thrombin receptor activating peptide (TRAP1-7) also stimulates MCP-1 production. Northern blot analysis demonstrated that mesangial cells express an mRNA transcript that hybridizes with labeled human thrombin receptor cDNA. These data describe a novel biologic activity of thrombin and suggest an additional mechanism by which this coagulation factor may participate in the progression of glomerulosclerosis, and by analogy, atherosclerosis.

Phylogeographical analysis reveals multiple conservation units in brook lampreys Lampetra planeri of Portuguese streams.

The populations of brook lamprey Lampetra planeri of Portuguese Rivers were analysed phylogeographically using a fragment of 644 bp of the mitochondrial control region of 158 individuals from six populations. Samples representing L. planeri and migratory lampreys Lampetra fluviatilis of rivers draining to the North Sea and the Baltic Sea were also included to assess the relationships of Portuguese samples. The data support a clear differentiation of all the populations studied. Several populations, which are isolated among themselves and also from the migratory lampreys, proved to be entirely composed of private haplotypes, a finding that supports some time of independent evolutionary history for these populations. This, combined with the geographic confinement to small water bodies, justifies the recognition of at least four conservation units in the Portuguese rivers Sado, São Pedro, Nabão and Inha.

Predictors and Trend of Ketoacidosis Hospitalization Rate in Type 2 Diabetes Mellitus Patients from 2006 to 2015 in Abruzzo Region, Italy.

AIMS: This study aimed to assess the trend of hospitalizations for DKA in adult patients with type 2 diabetes mellitus and its associated factors. DESIGN: A retrospective cross-sectional study was performed. Data were collected from hospital discharge records (HDRs) of patients (age ≥18) with either primary or secondary discharge diagnosis for DKA and type 2 diabetes from 2006 to 2015 in Abruzzo region. Age-adjusted hospitalization rates were computed by gender and standardized on the regional population in 2006. A logistic regression model was implemented using presence of DKA as dependent variable. RESULTS: We identified 160,366 HDRs with type 2 diabetes. Out of them, 1611 (1.00%) were due to DKA. The hospitalization rates for DKA increased both for male +115.9 and female +142.8%, from 2006 to 2015. The most significant predictors of DKA were age 18-44 (aOR=4.17), uncontrolled diabetes (aOR=1.79), trauma (aOR=1.38), any infection (aOR=1.68), liver disease (aOR=1.29), fluid and electrolyte disorders (aOR=2.09), psychosis (aOR=1.69). CONCLUSIONS: Trends of DKA in adult patients with type 2 diabetes has been increasing in both male and female. Multimorbidity is an open challenge for public health, therefore better coordination is needed among different specialist consultants to reduce the occurrence of this preventable complication.

Identification of monocyte chemotactic activity produced by malignant cells.

Human malignant cells secrete low molecular size proteins that attract peripheral blood monocytes and may be responsible for the accumulation of tumor-associated macrophages observed in vivo. Similar chemotactic proteins are secreted by cultured vascular smooth muscle cells. The predominant monocyte chemoattractants produced by tumor cells of differing origin were demonstrated to be related to smooth muscle cell-derived chemotactic factor. Thus, a single class of chemotactic proteins is produced by different cell types, which suggests a common mechanism for the recruitment of monocytes and macrophages. These results are significant in view of the potential of macrophages to affect tumor growth.