Evaluation of reference genes for gene expression analysis by real-time quantitative PCR (qPCR) in different tissues from mice infected by Ascaris suum.

Human ascariasis is the most prevalent helminth infection, affecting 445 million people worldwide. To better understand the impact of the immune system on the pathophysiology of individuals infected with Ascaris suum, mice have been used as experimental models. The RT-qPCR technique is a critical auxiliary tool of investigation used to quantify mRNA levels. However, proper normalization using reference genes is essential to ensure reliable outcomes to avoid analytical errors and false results. Despite the importance of reference genes for experimental A. suum infection studies, no specific reference genes have been identified yet. Therefore, we conducted a study to assess five potential reference genes (GAPDH, 18s, ACTB, B2M, and HPRT1) in different tissues (liver, lungs, small and large intestines) affected by A. suum larval migration in C57BL/6j mice. Tissue collection was carried out to analyze parasite burden and confirm the presence of larvae during the peak of migration in each tissue. Upon confirmation, we analyzed different genes in the tissues and found no common gene with stable expression. Our results highlight the importance of analyzing different genes and using different software programs to ensure reliable relative expression results. Based on our findings, B2M was ranked as the ideal reference gene for the liver, while 18S was the most stable gene in the lung and small intestine. ACTB, or a combination of ACTB with GAPDH, was deemed suitable as reference genes for the large intestine due to their stable expression and less variation between the control and infected groups. To further demonstrate the impact of using different reference genes, we normalized the expression of a chemokine gene (CXCL9) in all tissues. Significant differences in CXCL9 expression levels were observed between different groups in all tissues except for the large intestine. This underscores the importance of selecting appropriate reference genes to avoid overestimating target gene expression levels and encountering normalization-related issues that can lead to false results. In conclusion, our study highlights the significance of using reliable reference genes for accurate RT-qPCR analysis, especially in the context of A. suum infection studies in different tissues. Proper normalization is crucial to ensure the validity of gene expression data and avoid potential pitfalls in interpreting results.

The long-lasting Ascaris suum antigens in the lungs shapes the tissue adaptation modifying the pulmonary architecture and immune response after infection in mice.

Ascariasis is the most prevalent helminth affecting approximately 819 million people worldwide. The acute phase of Ascariasis is characterized by larval migration of Ascaris spp., through the intestinal wall, carried to the liver and lungs of the host by the circulatory system. Most of the larvae subsequently transverse the lung parenchyma leading to tissue injury, reaching the airways and pharynx, where they can be expectorated and swallowed back to the gastrointestinal tract, where they develop into adult worms. However, some larvae are trapped in the lung parenchyma inciting an inflammatory response that causes persistent pulmonary tissue damage long after the resolution of infection, which returns to tissue homeostasis. However, the mechanism by which chronic lung disease develops and resolves remains unknown. Here, using immunohistochemistry, we demonstrate that small fragments and larval antigens of Ascaris suum are deposited and retained chronically in the lung parenchyma of mice following a single Ascaris infection. Our results reveal that the prolonged presence of Ascaris larval antigens in the lung parenchyma contributes to the persistent immune stimulation inducing histopathological changes observed chronically following infection, and clearly demonstrate that larval antigens are related to all phases of tissue adaptation after infection: lung injury, chronic inflammation, resolution, and tissue remodeling, in parallel to increased specific humoral immunity and the recovery of lung function in mice. Additional insight is needed into the mechanisms of Ascaris antigen to induce chronic immune responses and resolution in the host lungs following larval migration.

Enteropathogenic Escherichia coli modulates the virulence and pathogenicity of Entamoeba dispar.

Amoebiasis is a disease caused by Entamoeba histolytica, affecting the large intestine of humans and occasionally leading to extra-intestinal lesions. Entamoeba dispar is another amoeba species considered commensal, although it has been identified in patients presenting with dysenteric and nondysenteric colitis, as well as amoebic liver abscess. Amoebic virulence factors are essential for the invasion and development of lesions. There is evidence showing that the association of enterobacteria with trophozoites contributes to increased gene expression of amoebic virulence factors. Enteropathogenic Escherichia coli is an important bacterium causing diarrhea, with high incidence rates in the world population, allowing it to interact with Entamoeba sp. in the same host. In this context, this study aims to evaluate the influence of enteropathogenic Escherichia coli on ACFN and ADO Entamoeba dispar strains by quantifying the gene expression of virulence factors, including galactose/N-acetyl-D-galactosamine-binding lectin, cysteine proteinase 2, and amoebapores A and C. Additionally, the study assesses the progression and morphological aspect of amoebic liver abscess and the profile of inflammatory cells. Our results demonstrated that the interaction between EPEC and ACFN Entamoeba dispar strains was able to increase the gene expression of virulence factors, as well as the lesion area and the activity of the inflammatory infiltrate. However, the association with the ADO strain did not influence the gene expression of virulence factors. Together, our findings indicate that the interaction between EPEC, ACFN, and ADO Entamoeba dispar strains resulted in differences in vitro and in vivo gene expression of Gal/GalNAc-binding lectin and CP2, in enzymatic activities of MPO, NAG, and EPO, and consequently, in the ability to cause lesions.

Bioactive potential of Bio-C Temp demonstrated by systemic mineralization markers and immunoexpression of bone proteins in the rat connective tissue.

Intracanal medications are used in endodontic treatment due to their antibacterial activity and ability to induce the periapical repair. Among the intracanal medications, the Calen (CAL; SS. White, Brazil) is a calcium hydroxide-based medication that provides an alkaline pH and releases calcium, exerting an antimicrobial activity. Bio-C Temp (BIO; Angelus, Brazil), a ready-to-use bioceramic intracanal medication, was designed to stimulate the mineralized tissues formation. Here, we investigated the bioactive potential of BIO in comparison to the CAL in the rat subcutaneous. Polyethylene tubes filled with medications, and empty tubes (control group, CG) were implanted in the subcutaneous tissue of rats. After 7, 15, 30 and 60 days, the blood was collected for calcium (Ca+2) and alkaline phosphatase (ALP) measurement, and the capsules around the implants were processed for morphological analyses. The data were submitted to two-way ANOVA and Tukey test (p < 0.05). At 7, 15 and 30 days, the ALP level was grater in BIO and CAL than in CG (p < 0.0001). At 7 and 15 days, greater Ca+2 level was seen in the serum of CAL samples. From 7 to 60 days, an increase in the number of fibroblasts, osteocalcin- and osteopontin-immunolabelled cells was observed in BIO and CAL groups (p < 0.0001). In all periods, BIO and CAL specimens showed von Kossa-positive structures. Moreover, ultrastructural analysis revealed globules of mineralization in the capsules around the BIO and CAL specimens. Thus Bio-C Temp caused an increase in the ALP, osteocalcin and osteopontin, which may have allowed the formation of calcite, suggesting bioactive potential.

Participation of fibroblast growth factor-1 and interleukin-10 in connective tissue repair following subcutaneous implantation of bioceramic materials in rats.

AIM: To evaluate whether the bioceramic materials Bio-C Pulpo (Bio-C, Angelus) and mineral trioxide aggregate (MTA) Repair HP (MTA-HP, Angelus) induce fibroblast proliferation and release of interleukin-10 (IL-10), an anti-inflammatory cytokine, stimulating connective tissue remodelling. The tissue response of Bio-C and MTA-HP was compared with the White MTA (WMTA; Angelus) since studies have demonstrated that WMTA induces tissue repair. METHODOLOGY: Bio-C, MTA-HP and WMTA were inserted into polyethylene tubes and implanted in the subcutaneous tissue of Holtzman rats for 7, 15, 30 and 60 days. As a control group (CG), empty tubes were implanted subcutaneously. The number of fibroblasts (FB), Ki-67-, fibroblast growth factor-1- (FGF-1) and IL-10-immunolabelled cells and collagen content in the capsules was obtained. The data were subjected to two-way anova followed by Tukey's test (p ≤ .05). RESULTS: At 7 days, significant differences in the number of FB were not detected amongst Bio-C, MTA-HP and WMTA groups (p Ëƒ .05). The capsules of all groups exhibited a significant increase in the number of FB and content of collagen over time. From 7 to 60 days, a significant reduction in the number of FGF-1- and Ki-67-immunolabelled cells was seen in the capsules of all specimens. In all periods, no significant difference in the number of FGF-1-immunolabelled cells was detected between Bio-C and CG specimens. At 60 days, significant differences in the immunoexpression of FGF-1 were not observed amongst the groups. At 7 and 15 days, the highest immunoexpression for Ki-67 was present in Bio-C specimens whilst, after 30 and 60 days, no significant difference was observed amongst the bioceramic materials. At 7 days, few IL-10 immunolabelled cells were present in the capsules of all specimens whereas, at 60 days, a significant increase in the IL-10-immunostaining was present in all groups. At 60 days, the Bio-C, MTA-HP and WMTA groups showed a greater number of IL-10-immunolabelled cells than in the CG specimens (p < .0001). CONCLUSIONS: Bio-C, MTA-HP and WMTA stimulate fibroblast proliferation, leading to the formation of collagen-rich capsules. FGF-1 and IL-10 may mediate the remodelling of capsules around Bio-C, MTA-HP and WMTA bioceramic materials.

Control Attitudes Scale-Revised-Brazilian Version.

BACKGROUND: Heart failure demands self-care skills and behaviors that can be negatively impacted by a low level of perceived control (PC), a belief about having the necessary resources to deal with negative events. Having valid and reliable instruments to measure PC is important to support interventions that improve self-care and related outcomes. The Control Attitudes Scale-Revised (CAS-R) was developed in the United States to measure PC in cardiac conditions. In Brazil, there are no instruments available to measure this construct. OBJECTIVE: The aims of this study were to translate and adapt the CAS-R to the Brazilian population and to assess the content validity of the adapted version. METHODS: The CAS-R was translated, back-translated, and assessed by an expert committee for linguistic equivalences. An agreement > 80% was considered adequate. Content validity (clarity, theoretical relevance, and practical pertinence) was assessed by both an expert professional panel (n = 6-8) and a panel of patients with heart failure (n = 40). A content validity coefficient > 0.70 was considered acceptable. RESULTS: The translations to Brazilian Portuguese were considered consistent with the original CAS-R. In the third round of linguistic equivalence assessment, all items achieved acceptable agreement, except for 2 items. After modifications to the instrument to achieve adequate equivalences, the adapted version had a final content validity coefficient of 0.93. Most patients were able to understand the instrument. CONCLUSIONS: The CAS-R-Brazilian version is equivalent to the original CAS-R and has satisfactory evidence of content validity. Additional psychometric testing will be performed to allow for the assessment of PC in individuals with heart failure in Brazil.

Biocompatibility, bioactive potential, porosity, and interface analysis calcium silicate repair cements in a dentin tube model.

OBJECTIVES: This study is to evaluate biocompatibility, bioactive potential, porosity, and dentin/material interface of Bio-C Repair (BIOC-R), MTA Repair HP (MTAHP), and Intermediate Restorative Material (IRM). MATERIALS AND METHODS: Dentin tubes were implanted into subcutaneous of rats for 7, 15, 30, and 60 days. Thickness of capsules, number of inflammatory cells (ICs), interleukin-6 (IL-6), osteocalcin (OCN), and von Kossa were evaluated. Porosity and material/dentin interface voids were also analyzed. Data were submitted to ANOVA and Tukey's tests (p < 0.05). RESULTS: IRM capsules were thicker and contained greater ICs and IL-6-immunopositive cells at 7 and 15 days. BIOC-R capsules exhibited higher thickness and ICs at 7 days and greater IL-6 at 7 and 15 days than MTAHP (p < 0.05). At 30 and 60 days, no significant difference was observed among the groups. OCN-immunopositive cells, von Kossa-positive, and birefringent structures were observed in BIOC-R and MTAHP. MTAHP exhibited higher porosity and interface voids (p < 0.05). CONCLUSIONS: BIOC-R, MTAHP, and IRM are biocompatible. Bioceramics materials demonstrate bioactive potential. MTAHP presented the highest porosity and presence of voids. CLINICAL RELEVANCE: BIOC-R and MTAHP have adequate biological properties. BIOC-R demonstrated lower porosity and presence of voids, which may represent better sealing for its clinical applications.

Comparison of Bio-C Pulpo and MTA Repair HP with White MTA: effect on liver parameters and evaluation of biocompatibility and bioactivity in rats.

AIM: To evaluate the tissue response promoted by Bio-C Pulpo (Bio), MTA Repair HP (MTA-HP) and White MTA (WMTA) and whether these materials cause liver changes in a rat experimental model. METHODOLOGY: Polyethylene tubes filled with Bio, MTA-HP and WMTA, and empty tubes (control group, CG) were implanted into the subcutaneous tissues of rats for 7, 15, 30 and 60 days. Inflammatory reaction score (IRS), capsule thickness, number of inflammatory cells (IC), von Kossa reaction, interleukin-6 (IL-6) and alkaline phosphatase (ALP) immunohistochemistry reactions were performed. Combined methods, von Kossa followed by immunohistochemistry for detection of ALP, were performed. At 60 days, the serum glutamic-oxaloacetic transaminase (GOT) and glutamic-pyruvic transaminase (GPT) levels were measured and liver fragments were collected for histological analysis; the data were assessed by one-way ANOVA analysis followed by Sidak's post-test. The biocompatibility and bioactivity data were subjected to the two-way ANOVA analysis followed by Tukey post hoc test, except the IRS. The IRS data were subjected to the Kruskal-Wallis ANOVA non-parametric test followed by Dunn's test (p ≤ .05). RESULTS: No significant difference was detected in serum GOT and GPT concentrations and in the number of hepatocytes among the experimental and CG samples. Although Bio-C Pulpo had the highest IC and IL-6-immunolabelled cells (p < 0.0001) at all periods, no significant difference was observed in the IRS among the materials, except at 60 days. In this period, the WMTA had lower IRS. All groups had a significant reduction in the capsule thickness and in the number of IC and IL-6-immunolabelled cells over time. Bio-C Pulpo, MTA-HP and WMTA specimens had greater immunoexpression of ALP than CG (p < .0001). At all periods, von Kossa-positive and birefringent structures were observed in the capsules around the materials. ALP-immunolabelled cells were also seen near von Kossa-positive structures. CONCLUSIONS: Bio-C Pulpo, MTA-HP and WMTA materials did not cause morphological changes in the liver and no significant alteration in the serum GOT and GPT levels. Moreover, these bioceramic materials were biocompatible and exhibited bioactive potential. However, Bio-C Pulpo induced greater inflammatory infiltrate than MTA-HP and WMTA at all periods.

Effect of ionizing radiation and cariogenic biofilm challenge on root-dentin caries.

OBJECTIVES: To evaluate the effect of ionizing radiation and cariogenic biofilm challenge using two continuous flow models, normal and reduced salivary flow, on the development of initial root-dentin caries lesions. MATERIALS AND METHODS: Microcosm biofilms were grown under two salivary flow rates (0.06 and 0.03 mL min-1) and exposed to 5% sucrose (3 × daily, 0.25 mL min-1, 6 min) dripped over non-irradiated and irradiated root-dentin blocks for up to 7 days. The vibration modes of root dentin, matrix/mineral (M/M), and carbonate/mineral (C/M) ratios were evaluated by FTIR. The mineral density was assessed by micro-CT. RESULTS: With normal salivary flow, FTIR revealed an increase in the organic matrix (amide III) and a decrease in the mineral phase (ν4, ν2 PO43-, AII + ν2 CO32-, C/M) in caries lesions. Irradiated dentin exhibited a reduction in the mineral phase (ν1, ν3 PO43-, ν2 CO32-, C/M). Differences in mineral densities were not significant. With reduced salivary flow, FTIR also revealed increased organic matrix (amide III) for irradiated caries lesions and decrease in mineral phase (v4, v2 PO43-, v2 CO32-, and C/M) in caries lesions. ν1, ν3 PO43- precipitated on the surface of irradiated dentin and a lower mineral density was observed. CONCLUSIONS: Initial caries lesions differed between non-irradiated and irradiated dentin and between normal and reduced salivary flow rates. Significant mineral loss with exposure of the organic matrix and low mineral density were observed for irradiated dentin with a reduced salivary flow rate. CLINICAL RELEVANCE: Ionizing radiation associated with a reduced salivary flow rate enhanced the progression of root-dentin caries.

Use of polyclonal IgY antibodies to detect serum immune complexes in patients with active hookworm infection.

Definitive diagnosis of hookworm infection is usually based on the microscopic detection of eggs in a stool sample; however, several cases display a low or irregular egg output. Serodiagnosis can be a useful tool to identify these cases, but conventional tests do not differentiate past from active infections. The aim of this study was to obtain and apply egg yolk polyclonal immunoglobulin (IgY) antibodies to detect immune complexes (ICs) in serum samples from patients infected with hookworm. Hens were immunized with Ancylostoma ceylanicum saline extract, their eggs were collected and then IgY antibodies were extracted and purified. Antibody purity was tested by 12% sodium dodecyl sulphate polyacrylamide gel electrophoresis and specificity was assessed by immunoblotting and immunofluorescence. IgY production was evaluated by kinetics enzyme-linked immunosorbent assay (ELISA). Sandwich ELISA tested the ability of IgY to detect ICs in serum samples, from which diagnostic parameters were calculated. Antibody responses increased steadily from day 7 to 42. In the immunoblotting assay, IgY recognized two protein complexes. The immunofluorescence assay showed no staining in control samples. The sandwich ELISA presented a very high diagnostic value, with a sensitivity of 90% and a specificity of 86.7%. Our pioneer strategy highlights the potential use of egg yolk IgY as a diagnostic test to detect active hookworm infection.

IgY antibody and human neurocysticercosis: a novel approach on immunodiagnosis using Taenia crassiceps hydrophobic antigens.

Human neurocysticercosis (NCC) is a worldwide neglected disease caused by Taenia solium metacestode and responsible for various complications and neurological disorders. This study aimed to evaluate the use of specific immunoglobulin Y (IgY) produced by laying hens immunized with a hydrophobic fraction of Taenia crassiceps metacestodes (hFTc) in NCC diagnosis. Egg yolk IgY antibodies were fractionated, purified and characterized. Enzyme-linked immunosorbent assay (ELISA) was carried out to evaluate the production kinetics and avidity maturation of anti-hFTc IgY antibodies throughout the IgY obtention process. Antigen recognition tests were carried out by Western blotting and immunofluorescence antibody test using purified and specific anti-hFTc IgY antibodies for detection of parasitic antigens of T. crassiceps and T. solium metacestodes. Sandwich ELISA was performed to detect circulating immune complexes formed by IgG and parasitic antigens in human sera. The results showed high diagnostic values (93.2% sensitivity and 94.3% specificity) for immune complexes detection in human sera with confirmed NCC. In conclusion, specific IgY antibodies produced from immunized hens with hFTc antigens were efficient to detect T. solium immune complexes in human sera, being an innovative and potential tool for NCC immunodiagnosis.

Educational Intervention Using WhatsApp on Medication Adherence in Hypertension and Diabetes Patients: A Randomized Clinical Trial.

Background: Low adherence to pharmacological therapy by patients with diabetes mellitus and hypertension is frequent and represents a worldwide challenge. Introduction: This study examined the effect of an educational intervention using WhatsApp® messaging on medication adherence of patients with hypertension and diabetes. Materials and Methods: A randomized clinical trial was performed with 403 patients who had diabetes and/or hypertension and who had enrolled in the 33 Basic Health Units of Maringá-PR, Brazil. The patients were randomly assigned to either the intervention group (n = 203), which received usual care (multi-professional educational appointments according to each Unit schedule) plus 55 audio, image or text WhatsApp messages about health care promotion, with an emphasis on medication adherence, or the control group. The control group (n = 200) only received usual care. Medication adherence, as measured by the Morisky-Green Test, was compared through the chi-square test after 16 weeks. Relative risk (RR) was used as a measure of effect size. Results: After the follow-up period (4 months), 67.5% of the patients in the intervention group were adherent versus 58.5% in the control group (RR: 1.15, 95% confidence interval = 0.99-1.34, p = 0.077). Discussion: Although the effect of the intervention was not statistically significant, there was a clinically significant impact associated with a 15% increase in medication adherence. Conclusion: Given the complexity of adherence to the use of antidiabetic and antihypertensive medications, educational interventions using WhatsApp could be useful as a reinforcement to increase adherence to medication.

Improving the cost effectiveness of enhanced green fluorescent protein production using recombinant Escherichia coli BL21 (DE3): Decreasing the expression inducer concentration.

Green fluorescent protein (GFP) is a globular protein used as biosensor and biomarker in medical and industrial fields. However, due to the expensive production costs of expressing proteins using high-cost inducers like isopropyl-ß-d-1-thiogalactopyranoside (IPTG), the number of GFP applications are still scarce. This work studied the production of enhanced GFP (EGFP) using Escherichia coli BL21 (DE3) [pLysS; pET28(a)], aiming to increase its yield and reduce costs. First, the influence of agitation rate, induction time, and concentration of IPTG in the production of EGFP was evaluated, but only the first two parameters were significant. Subsequently, aiming to reduce costs related to the use of inducer, the IPTG concentration (0.005, 0.010, and 0.025 mM) was decreased and, interestingly, the production levels were maintained or increased. These results show that a proper choice of production conditions, particularly through the decrease of inducer concentration, is effective to reduce the upstream production costs and guarantee high EGFP expression.

A preliminary comparison between the effects of red and infrared laser irradiation on viability and proliferation of SHED.

The aim of this preliminary study was to compare the effects of different energy densities from red and infrared low-level laser (LLL) on viability and proliferation of stem cells from human exfoliated deciduous teeth (SHED). SHED were irradiated with red laser (R) or infrared laser (IR) set with the following dosimetry: 1.2 J/cm2 (0.05 J), 2.5 J/cm2 (0.1 J), 5.0 J/cm2 (0.2 J), and 7.5 J/cm2 (0.3 J). Positive (C+) and negative (C-) control groups comprised non-irradiated cells. Data were analyzed by two-way ANOVA followed by Tukey's test (P < 0.05). At 24- and 48-h period, group R5.0 showed significantly higher cell viability rates than R1.2 and R2.5. At 48 h, R2.5 also revealed lower proliferation than R5.0. Comparing to the C+ group, R2.5 exhibited lower viability at 72 h, and proliferation at 24 and 48 h. Groups R1.2, IR1.2, and IR5.0 were less viable at 24 h, while R1.2, IR2.5, and IR5.0 revealed lower proliferative capacity at 48 h. Overall, our results showed that LLL can favor viability and proliferation of SHED, especially when cells receive red laser irradiation at 5.0 J/cm2. Therefore, according to this preliminary investigation, 5 J/cm2 applied by red LLL induced high rates of cell viability and proliferation, while the same irradiation dose using infrared laser led to negative effects. LLL irradiation with 1.2 and 2.5 J/cm2 was deleterious to metabolic activity and proliferation of SHED regardless of the type of laser. Further studies are necessary to gain in-depth knowledge about the effects of different wavelengths of LLL on SHED viability and proliferation.

Decreasing length of limb immobilisation following nonelective transfemoral percutaneous coronary intervention: A randomised clinical trial.

AIMS AND OBJECTIVES: To assess the intensity and frequency of pain, use of analgesic drugs, and the incidence of paraesthesia, urinary retention and vascular complications upon decreasing affected limb immobilisation from 4-2 hrs after sheath removal in patients submitted to transfemoral percutaneous coronary intervention (PCI). BACKGROUND: After sheath removal from the femoral artery following urgent or emergency PCI, patients are maintained with limb immobilisation for a mean period of 4 hr. DESIGN: Randomised clinical trial (RCT) based on the CONSORT guidelines. METHOD: Randomised clinical trial was performed in patients with Acute Coronary Syndrome submitted to transfemoral PCI. The intervention group was submitted to a supine position with the head of the bed elevated (30-degree angle) with affected limb immobilisation for 2 hr after sheath removal and the control group for 4 hrs. The outcomes were pain complaints, need for analgesic drugs, incidence of paraesthesia, urinary retention and vascular complications. The outcomes were assessed immediately, 6, 12 and 24 hr after release from limb immobilisation before the patients were released from bed rest. RESULTS: A total of 150 patients (75 in each group) participated in the study. No significant differences in outcomes were observed between the groups, except in relation to the haematoma formation that was higher in the intervention group. CONCLUSION: A reduced length of limb immobilisation after sheath removal following PCI does not change the frequency and intensity of pain, need of analgesic drugs, urinary retention and paraesthesia. The incidence of haematoma was higher in the intervention group, without significant clinical manifestations. RELEVANCE TO CLINICAL PRACTICE: The results of this study can be considered for patients submitted to elective, urgent or emergency PCI, who have a lower risk of complications, thereby allowing for decreased periods of limb immobilisation.

By-Products of Camu-Camu [Myrciaria dubia (Kunth) McVaugh] as Promising Sources of Bioactive High Added-Value Food Ingredients: Functionalization of Yogurts.

Camu-camu (Myrciaria dubia (Kunth) McVaugh) is a fruit economically relevant to the Amazon region, mostly consumed in the form of processed pulp. Our aim was to perform an unprecedented comparative study on the chemical composition and bioactivities of the camu-camu pulp and industrial bio-residues (peel and seed), and then the most promising fruit part was further explored as a functionalized ingredient in yogurt. A total of twenty-three phenolic compounds were identified, with myricetin-O-pentoside and cyanindin-3-O-glucoside being the main compounds in peels, followed by p-coumaroyl hexoside in the pulp, and ellagic acid in the seeds. The peel displayed the richest phenolic profile among samples, as well as the most significant antibacterial (MICs = 0.625-10 mg/mL) and anti-proliferative (GI50 = 180 µg/mL against HeLa cells) activities. For this reason, it was selected to be introduced in a food system (yogurt). Taken together, our results suggest the possibility of using the camu-camu peel as a source of food additives.

Trypanosoma janseni n. sp. (Trypanosomatida: Trypanosomatidae) isolated from Didelphis aurita (Mammalia: Didelphidae) in the Atlantic Rainforest of Rio de Janeiro, Brazil: integrative taxonomy and phylogeography within the Trypanosoma cruzi clade.

BACKGROUND: Didelphis spp. are a South American marsupial species that are among the most ancient hosts for the Trypanosoma spp. OBJECTIVES: We characterise a new species (Trypanosoma janseni n. sp.) isolated from the spleen and liver tissues of Didelphis aurita in the Atlantic Rainforest of Rio de Janeiro, Brazil. METHODS: The parasites were isolated and a growth curve was performed in NNN and Schneider's media containing 10% foetal bovine serum. Parasite morphology was evaluated via light microscopy on Giemsa-stained culture smears, as well as scanning and transmission electron microscopy. Molecular taxonomy was based on a partial region (737-bp) of the small subunit (18S) ribosomal RNA gene and 708 bp of the nuclear marker, glycosomal glyceraldehyde-3-phosphate dehydrogenase (gGAPDH) genes. Maximum likelihood and Bayesian inference methods were used to perform a species coalescent analysis and to generate individual and concatenated gene trees. Divergence times among species that belong to the T. cruzi clade were also inferred. FINDINGS: In vitro growth curves demonstrated a very short log phase, achieving a maximum growth rate at day 3 followed by a sharp decline. Only epimastigote forms were observed under light and scanning microscopy. Transmission electron microscopy analysis showed structures typical to Trypanosoma spp., except one structure that presented as single-membraned, usually grouped in stacks of three or four. Phylogeography analyses confirmed the distinct species status of T. janseni n. sp. within the T. cruzi clade. Trypanosoma janseni n. sp. clusters with T. wauwau in a well-supported clade, which is exclusive and monophyletic. The separation of the South American T. wauwau + T. janseni coincides with the separation of the Southern Super Continent. CONCLUSIONS: This clade is a sister group of the trypanosomes found in Australian marsupials and its discovery sheds light on the initial diversification process based on what we currently know about the T. cruzi clade.

Correlations between stress, anxiety and depression and sociodemographic and clinical characteristics among outpatients with heart failure.

Aim To describe and investigate correlations among anxiety, stress and depression and identify their relationship with sociodemographic and clinical characteristics of patients with heart failure. METHODS: This is an analytical cross-sectional study. Sociodemographic and clinical characteristics were collected, along with levels of anxiety, stress and depression from 309 outpatients. RESULTS: The mean levels of stress, anxiety and depression were correlated but low. Time since diagnosis, the disease's functional class, family income, and smoking influenced stress. Functional class and Chagas disease influenced anxiety and depression. Being unemployed and smoking influenced anxiety, while being a homeowner influenced depression. CONCLUSION: These findings should be considered when planning nursing interventions.

Excessive bleeding predictors after cardiac surgery in adults: integrative review.

AIMS AND OBJECTIVES: To integrate literature data on the predictors of excessive bleeding after cardiac surgery in adults. BACKGROUND: Perioperative nursing care requires awareness of the risk factors for excessive bleeding after cardiac surgery to assure vigilance prioritising and early correction of those that are modifiable. DESIGN: Integrative literature review. METHODS: Articles were searched in seven databases. Seventeen studies investigating predictive factors for excessive bleeding after open-heart surgery from 2004-2014 were included. RESULTS: Predictors of excessive bleeding after cardiac surgery were: Patient-related: male gender, higher preoperative haemoglobin levels, lower body mass index, diabetes mellitus, impaired left ventricular function, lower amount of prebypass thrombin generation, lower preoperative platelet counts, decreased preoperative platelet aggregation, preoperative platelet inhibition level >20%, preoperative thrombocytopenia and lower preoperative fibrinogen concentration. Procedure-related: the operating surgeon, coronary artery bypass surgery with three or more bypasses, use of the internal mammary artery, duration of surgery, increased cross-clamp time, increased cardiopulmonary bypass time, lower intraoperative core body temperature and bypass-induced haemostatic disorders. Postoperative: fibrinogen levels and metabolic acidosis. CONCLUSIONS: Patient-related, procedure-related and postoperative predictors of excessive bleeding after cardiac surgery were identified. RELEVANCE TO CLINICAL PRACTICE: The predictors summarised in this review can be used for risk stratification of excessive bleeding after cardiac surgery. Assessment, documentation and case reporting can be guided by awareness of these factors, so that postoperative vigilance can be prioritised. Timely identification and correction of the modifiable factors can be facilitated.