Lack of evidence for longitudinal dissociation of the atrioventricular conduction axis.

Longitudinal dissociation of the aggregated specialized cardiomyocytes within the non-branching portion of atrioventricular conduction axis has proved a controversial topic for both morphologists and electrophysiologists. We have now used morphological methods, including three-dimensional assessment, to revisit, in human, canine, and bovine hearts, the presence or absence of interconnections between the aggregated cardiomyocytes making up the non-branching bundle. We analyzed three datasets from human and canine hearts, and two from bovine hearts, using longitudinal and orthogonal serial histological sections. In addition, we assessed three hearts using translucent India ink injected specimens, permitting assessment of the three-dimensional arrangement of the cardiomyocytes. Using the longitudinal sections, we found numerous oblique interconnections between the groups of specialized cardiomyocytes. When assessing orthogonal sections, we noted marked variation in the grouping of the cardiomyocytes. We interpreted this finding as evidence of bifurcation and convergence of the groups seen in the longitudinal sections. The three-dimensional assessment of the bovine material confirmed the presence of the numerous interconnections. The presence of multiple connections between the cardiomyocytes in the non-branching bundle rules out the potential for longitudinal dissociation.

Maternofetal outcomes in early-onset gestational diabetes: does weight gain matter?

AIM: Women with early-onset gestational diabetes mellitus (GDM) have overall lower gestational weight gain (GWG) compared to those with later-onset GDM, albeit with usually worse maternofetal outcomes. We intent to investigate the association between inadequate GWG and maternofetal outcomes in pregnant women with early-onset GDM. METHODS: We performed a retrospective study of women with early-onset GDM based on the National Registry of GDM. Three study groups were defined according to the recommendations of the Institute of Medicine for GWG: excessive GWG (eGWG), adequate (aGWG) or insufficient (iGWG). RESULTS: A total of 8040 pregnant women were included: 27% (n = 2170) eGWG, 31% (n = 2492) aGWG and 42% (n = 3378) iGWG. Preeclampsia (4.3 vs 3 vs 1.6%, p < 0.001), polyhydramnios (3.1 vs 2.3 vs 1.8%, p = 0.008) and cesarean section (37.4 vs 34.1 vs 29.5%, p < 0.001) were significantly more frequent among women with eGWG. Additionally, there was a higher frequency of macrosomia (8.1 vs 3.6 vs 2.4%, p < 0.001), large-for-gestational-age (8.2 vs 3.7 vs 2.6%, p < 0.001) and birth trauma (2.6 vs 1.5 vs 1.1%, p < 0.001) in this group. On the other hand, fetal death (0.2 vs 0.2 vs 0.5%, p = 0.04), small-for-gestational-age (9 vs 10.3 vs 14.9, p < 0.001) and preterm delivery (5.6 vs 7.1 vs 7.5%, p = 0.03) were more frequent in iGWG group. CONCLUSIONS: Over two-thirds of pregnant women with early-onset GDM had inappropriate GWG, which was significantly associated with adverse maternofetal outcomes. Weight management must be a focus of special attention in women with early-onset GDM, beyond glycemic control, to achieve healthy pregnancy outcomes.

Assessing intermittent saline inflows in urban water systems.

Urban water drainage systems' primary function is to transport sanitary or stormwater. The intrusion of saline waters has recognized detrimental effects. Especially in coastal areas, saline inflows can compromise performance by increasing the risk of untreated discharges, weakening the structural condition of concrete or metallic components, reducing the effectiveness of wastewater treatment processes and limiting the potential reuse for irrigation. Performance deterioration can be prevented by an early assessment of exposure to saline water, followed by timely actions to control its causes and consequences. The paper describes a procedure for diagnosing undue saline inflows. The procedure is based on the determination of saline inflow's magnitude, acceptance levels, and contribution to the system's performance. Contextual factors and performance indicators, and their reference values, are selected for the assessment. Options to address the problem are proposed, depending on the results. These options can relate to organizational, operational, and structural actions. Application to a case study allowed to validate the method and discuss the results. Here, saline volumes entering the system are quite relevant (almost 30%), posing problems regarding corrosion, treatment plant operation and significant concrete exposure to intermittent saline waters.

Improving body image at scale among Brazilian adolescents: study protocol for the co-creation and randomised trial evaluation of a chatbot intervention.

BACKGROUND: Body image concerns are prevalent among Brazilian adolescents and can lead to poor psychological and physical health. Yet, there is a scarcity of culturally-appropriate, evidence-based interventions that have been evaluated and made widely available. Chatbot technology (i.e., software that mimics written or spoken human speech) offers an innovative method to increase the scalability of mental health interventions for adolescents. The present protocol outlines the co-creation and evaluation of a body image chatbot for Brazilian adolescents via a partnership between academics, industry organisations and the United Nations Children's Fund (UNICEF). METHODS: A two-armed fully remote randomised controlled trial will evaluate the chatbot's effectiveness at improving body image and well-being. Adolescent girls and boys (N = 2800) aged 13-18 years recruited online will be randomly allocated (1:1) into either: 1) a body image chatbot or 2) an assessment-only control condition. Adolescents will engage with the chatbot over a 72-hour period on Facebook Messenger. Primary outcomes will assess the immediate and short-term impact of the chatbot on state- and trait-based body image, respectively. Secondary outcomes will include state- and trait-based affect, trait self-efficacy and treatment adherence. DISCUSSION: This research is the first to develop an evidence-informed body image chatbot for Brazilian adolescents, with the proposed efficacy trial aiming to provide support for accessible, scalable and cost-effective interventions that address disparities in body image prevalence and readily available resources. TRIAL REGISTRATION NUMBER: NCT04825184 , registered 30th March 2021.

Early Interaction of Alternaria infectoria Conidia with Macrophages.

Fungi of the genus Alternaria are ubiquitous indoor and outdoor airborne agents, and individuals are daily exposed to their spores. Although its importance in human infections and, particularly in respiratory allergies, there are no studies of how Alternaria spp. spores interact with host cells. Our aim was to study the early interaction of Alternaria infectoria spores with macrophages, the first line of immune defense. RAW 264.7 macrophages were infected with A. infectoria conidia, and the internalization and viability of conidia once inside the macrophages were quantified during the first 6 h of interaction. Live cell imaging was used to study the dynamics of this interaction. TNF-α production was quantified by relative gene expression, and the concentration of other cytokines (IL-1α, IL-1ß, IL-6, IL-4, IL-10, IL-17, GM-CSF and INF-γ) and a chemokine, MIP-1α, was quantified by ELISA. Conidia were rapidly internalized by macrophages, with approximately half internalized after 30 min of interaction. During the first 6 h of interaction, macrophages retained the ability to mitotically divide while containing internalized conidia. The classical macrophage-activated morphology was absent in macrophages infected with conidia, and TNF-α and other cytokines and chemokines failed to be produced. Thus, macrophages are able to efficiently phagocyte A. infectoria conidia, but, during the first 6 h, no effective antifungal response is triggered, therefore promoting the residence of these fungal conidia inside the macrophages.

Early maternal separation promotes alterations in the thermoregulatory profile of adult Wistar rats.

Stressful lifelong events may influence psychiatric diseases, like depression and anxiety. Interestingly, depressed patients have dysfunction of thermoregulatory cooling mechanisms. Thus, understanding the mechanisms related to the thermoregulatory changes in stress-related pathologies is important to better understand the symptoms and treatments for those diseases. However, the influence of early-life stress on the thermoregulatory profile of adults is unknown. In this study, we aimed to evaluate the thermoregulatory profile of adult male Wistar rats submitted to early-life stress by maternal separation (MS). On postnatal days 2-14, rats were submitted daily to MS for 3 h per day. At 3-4 months of age, anxiety-like behavior was evaluated using the open field test and elevated plus maze, depression-like behavior was evaluated using the forced swim test and thermoregulatory profile were also evaluated. In the behavioral tests, MS animals exhibited anxiety- and depression-like behaviors, and had higher core body temperatures during dark period of the circadian cycle, when compared to controls. In addition, MS animals presented higher hyperthermic and vasoconstriction responses than control animals when exposed to the warmth environment, and engaged in cold-seeking behavior whenever possible to select their preferred ambient temperature. The results suggest that, besides emotional alterations, MS induces a change in the thermoregulatory profile of rats that persists into adulthood.

Innovation results of IAM planning in urban water services.

The requirement to provide urban water services continuously while infrastructures are ageing, imposes the need for increasingly sustainable infrastructure asset management (IAM). To achieve and maintain adequate levels of service, the AWARE-P IAM methodology has been applied in collaborative projects launched by the National Civil Engineering Laboratory, in partnership with IST (Technical University of Lisbon), Addition (software company) and several water utilities. The objective of these projects is to support urban water utilities in the development, implementation and maintenance of IAM plans. To guarantee the success of IAM planning, following the AWARE-P IAM methodology, utilities are required to: consider that the infrastructure has system behaviour and lifespan is indefinite and guarantee the full-alignment of IAM planning with organisation objectives. By analysing the strategic and tactical plans of participating utilities, the proposed methodology principles are discussed and supported. The main innovation results from the implementation of IAM planning are also presented and discussed, including the challenges of setting up an IAM process, together with the major benefits and drawbacks that come up when developing IAM plans. The results were demonstrated by the effective implementation of 16 strategic and 14 tactical IAM plans by the participating utilities.

Factors associated with non-treatment of hypertension and gender differences at baseline in the ELSA-Brasil cohort.

The treatment of arterial hypertension (AH) contributes to the reduction of morbidity and mortality. Gender differences are likely to play a role, as non-treatment is associated with clinical and sociodemographic aspects. The aim of this study was to investigate the factors associated with non-treatment of AH and gender differences in hypertensive individuals from the ELSA-Brasil cohort. The study was conducted with 5,743 baseline hypertensive cohort participants. AH was considered if there was a previous diagnosis or if systolic blood pressure (SBP) was ≥140 and/or diastolic BP (DBP) was ≥90 mmHg. Sociodemographic and anthropometric data, lifestyle, comorbidities, and use of antihypertensive medications were evaluated through interviews and in-person measurements. Treatment with renin-angiotensin-aldosterone system inhibitors (RAASi) or other antihypertensive medications and non-treatment were evaluated with multivariate logistic regression. Non-treatment was observed in 32.8% of hypertensive individuals. Of the 67.7% treated individuals, 41.1% received RAASi. Non-treatment was associated with alcohol consumption in women (OR=1.41; 95%CI: 1.15-1.73; P=0.001), lowest schooling level in men (OR=1.70; 95%CI: 1.32-2.19; P<0.001), and younger age groups in men and women (strongest association in males aged 35-44 years: OR=4.58, 95%CI: 3.17-6.6, P<0.001). Among those using RAASi, a higher proportion of white, older individuals, and with more comorbidities was observed. The high percentage of non-treatment, even in this civil servant population, indicated the need to improve the treatment cascade for AH. Public health policies should consider giving special attention to gender roles in groups at higher risk of non-treatment to reduce inequities related to AH in Brazil.

Cytogenetic analysis of Lagria villosa (Coleoptera, Tenebrionidae): emphasis on the mechanism of association of the Xy(p) sex chromosomes.

The Xy(p) sex determination mechanism is the system most frequent and ancestral to Coleoptera. Moreover, the presence of argyrophilous material associated with the sex bivalent is described as being responsible for the maintenance and association of these chromosomes. There are no karyotype data available regarding the genus Lagria and no consensus in the literature regarding the argyrophilous material present in the lumen of sex bivalent. Therefore, the aim of this work was to investigate the mechanism of sex chromosome bivalent association in Lagria villosa by analyzing the argyrophilous nature of the material present in the Xy(p) lumen. It was also intended to characterize L. villosa cytogenetically. The analysis of meiotic cells showed 2n = 18 = 16+Xy(p) for males and 2n = 18 = 16+XX in females and the meiotic formula was 2n = 8(II)+Xy(p). The C-banding showed blocks of pericentromeric heterochromatin in all chromosomes except in the y(p) chromosome. In these regions, the use of fluorochromes revealed the presence of heterochromatin containing GC rich DNA sequences. The study of synaptonemal complex showed a gradual increase in the electron-density of the axial elements of the sex chromosomes and their association with strongly electron-dense material. The pepsin pretreatment revealed that the material impregnated by silver is protein.

Chromosomal organization of repetitive DNA in Sorubim lima (Teleostei; Pimelodidae).

Interspaced repetitive DNA elements and segmental duplications have been extensively analyzed in fishes through physical chromosome mapping methods, providing a better comprehension of the structure and organization of the genome of this group. In order to contribute to this scenario, a sequence integration study of different classes of repetitive DNA with high resolution physical chromosome mapping was performed in Sorubim lima. Fluorescence in situ hybridization (FISH) and fiber-FISH with probes for 18S and 5S rRNA genes, TTAGGGn sequence and non-LTR retrotransposon family members Rex1, Rex3 and Rex6 showed that non-LTR elements may be dispersed in the chromosome set with relative concentration in heterochromatic regions, as shown by Rex1, or may even intercalate in 45S rDNA and the telomeric sequence, as found for Rex3 and Rex6. These results reinforce the presence of preferential regions of retroelement accumulation and contribute to a better understanding of the genomic organization of some repetitive DNA classes in fishes.

Dynamic calcium-mediated stress response and recovery signatures in the fungal pathogen, Candida albicans.

IMPORTANCE: Intracellular calcium signaling plays an important role in the resistance and adaptation to stresses encountered by fungal pathogens within the host. This study reports the optimization of the GCaMP fluorescent calcium reporter for live-cell imaging of dynamic calcium responses in single cells of the pathogen, Candida albicans, for the first time. Exposure to membrane, osmotic or oxidative stress generated both specific changes in single cell intracellular calcium spiking and longer calcium transients across the population. Repeated treatments showed that calcium dynamics become unaffected by some stresses but not others, consistent with known cell adaptation mechanisms. By expressing GCaMP in mutant strains and tracking the viability of individual cells over time, the relative contributions of key signaling pathways to calcium flux, stress adaptation, and cell death were demonstrated. This reporter, therefore, permits the study of calcium dynamics, homeostasis, and signaling in C. albicans at a previously unattainable level of detail.

Population structure and genetic diversity analysis in Gynaikothrips uzeli (Zimerman, 1909) (Thysanoptera: Phlaeothripidae) by RAPD markers.

Thrips are small insects (0.5-3.0 mm) with distinct habits and life histories characterized by haplodiploid sex determination. In general, low levels of genetic diversity have been reported in haplodiploid insects, although most reports focus on the order Hymenoptera. Therefore, we used RAPD markers to evaluate the structure and both inter- and intra-population genetic variability of Gynaikothrips uzeli (Thysanoptera: Phlaeothripidae). Six populations, three from Paraná state, southern Brazil, and three from Bahia, northeastern Brazil, were studied. Similarly to other haplodiploid insects, the genetic diversity of G. uzeli was reduced. This result is putatively related to the haplodiploid sex determination system, which yields little genetic variation, and to ecological traits of the studied species, such as the low dispersal abilities and life mode in leaf galls. All individuals were homogeneously clustered in their respective collection sites, forming two main groups in which populations from similar environments were more closely related. The analyzed populations were highly structured, and the genetic variation was higher among than within populations.

Urban water infrastructure asset management - a structured approach in four water utilities.

Water services are a strategic sector of large social and economic relevance. It is therefore essential that they are managed rationally and efficiently. Advanced water supply and wastewater infrastructure asset management (IAM) is key in achieving adequate levels of service in the future, particularly with regard to reliable and high quality drinking water supply, prevention of urban flooding, efficient use of natural resources and prevention of pollution. This paper presents a methodology for supporting the development of urban water IAM, developed during the AWARE-P project as well as an appraisal of its implementation in four water utilities. Both water supply and wastewater systems were considered. Due to the different contexts and features of the utilities, the main concerns vary from case to case; some problems essentially are related to performance, others to risk. Cost is a common deciding factor. The paper describes the procedure applied, focusing on the diversity of drivers, constraints, benefits and outcomes. It also points out the main challenges and the results obtained through the implementation of a structured procedure for supporting urban water IAM.

Physical mapping of 5S rDNA in two species of Knifefishes: Gymnotus pantanal and Gymnotus paraguensis (Gymnotiformes).

Physical mapping of 5S rDNA in 2 species of knifefishes, Gymnotuspantanal and G. paraguensis (Gymnotiformes), was performed using fluorescence in situ hybridization with a 5S rDNA probe. The 5S rDNA PCR product from the genomes of both species was also sequenced and aligned to determine non-transcribed spacer sequences (NTS). Both species under study had different patterns of 5S rDNA gene cluster distribution. While in the karyotype of G. pantanal two 5S rDNA-bearing pairs were observed, the karyotype of G. paraguensis possessed as many as 19 such pairs. Such multiplication of 5S rDNA gene clusters might be caused by the involvement of transposable elements because the NTS of G. paraguensis was 400 bp long with high identity (90%) with a mobile transposable element called Tc1-like transposon, described from the cyprinid fish Labeo rohita.

Repetitive DNA and meiotic behavior of sex chromosomes in Gymnotus pantanal (Gymnotiformes, Gymnotidae).

Neotropical fishes have a low rate of chromosome differentiation between sexes. The present study characterizes the first meiotic analysis of sex chromosomes in the order Gymnotiformes. Gymnotus pantanal - females had 40 chromosomes (14m/sm, 26st/a) and males had 39 chromosomes (15m/sm, 24st/a), with a fundamental number of 54 - showed a multiple sexual determination chromosome system of the type X(1)X(1)X(2)X(2)/X(1)X(2)Y. The heterochromatin is restricted to centromeres of all chromosomes of the karyotype. The meiotic behavior of sex chromosomes involved in this system in males is from a trivalent totally pared in the pachytene stage, with a high degree of similarity. The cells of metaphase II exhibit 19 and 20 chromosomes, normal disjunction of sex chromosomes and the formation of balanced gametes with 18 + Y and 18 + X(1)X(2) chromosomes, respectively. The small amount of heterochromatin and repetitive DNA involved in this system and the high degree of chromosome similarity indicated a recent origin of the X(1)X(1)X(2)X(2)/X(1)X(2)Y system in G. pantanal and suggests the existence of a simple ancestral system with morphologically undifferentiated chromosomes.

Two genetic stocks of Steindachneridion melanodermatum living in sympatry in nature and genetic variability of wild parents and F1 generation.

Steindachneridion melanodermatum is a large Brazilian catfish, highly prized for sport fishing and for its meat. Specimens of this species, both caught in nature from Iguacu River and F(1) fish born in captivity, were analyzed with regard to patterns of RAPD molecular markers. Genetic similarity ranged from 0.57 to 0.95; two groups were determined for the wild specimens. The results suggest different genetic lineages in sympatry in nature. Heterozygosity and percentage of polymorphic loci were 0.31 and 79% and 0.23 and 62%, respectively, for the two populations of wild specimens and 0.26 and 66%, respectively, for those born in captivity.

Mild periconceptional hyperglycemia: predictor of adverse fetomaternal outcomes in gestational diabetes?

AIM: To clarify whether mild first trimester hyperglycaemia (characteristic of early-onset GDM) is associated with higher incidence of congenital malformations and other adverse fetomaternal outcomes compared to women with second trimester hyperglycaemia (later-onset GDM). DESIGN AND METHODS: We analyzed the Portuguese National GDM database, containing data collected between 2011 and 2017. Two study groups were defined: Group 1-Women with GDM diagnosed during the first trimester (with fasting glycemia ≥ 92 and < 126 mg/dL); Group 2-Women with GDM diagnosed after the first 12 weeks of gestation, with either fasting glycemia or oral glucose tolerance test, according to the International Association of Pregnancy and Diabetes Study Group criteria. The fetomaternal characteristics of each group were compared. RESULTS: A total of 18.518 pregnant women diagnosed with GDM were included which 34.4% of them belonged to Group 1. Pregnant women from this group were significantly younger and had a higher median BMI than the women from the other group. Overall, there was no significant differences in maternal morbidity parameters between groups. Non-evolutive pregnancies were significantly more frequent along the present gestation in the group 1 (1.1% vs. 0.1%, p < 0.001), as was fetal death (0.6% vs. 0.2%, p < 0.001). Congenital malformations did not differ significantly between groups (3.2% vs. 2.8%, p = 0.155). CONCLUSIONS: The mild near conceptional hyperglycaemic state characteristic of an early-onset GDM seems to be associated with an increased prevalence of non-evolutive pregnancies and foetal deaths when compared to later-onset GDM.

Satellite DNA and chromosomes in Neotropical fishes: methods, applications and perspectives.

Constitutive heterochromatin represents a substantial portion of the eukaryote genome, and it is mainly composed of tandemly repeated DNA sequences, such as satellite DNAs, which are also enriched by other dispersed repeated elements, including transposons. Studies on the organization, structure, composition and in situ localization of satellite DNAs have led to consistent advances in the understanding of the genome evolution of species, with a particular focus on heterochromatic domains, the diversification of heteromorphic sex chromosomes and the origin and maintenance of B chromosomes. Satellite DNAs can be chromosome specific or species specific, or they can characterize different species from a genus, family or even representatives of a given order. In some cases, the presence of these repeated elements in members of a single clade has enabled inferences of a phylogenetic nature. Genomic DNA restriction, using specific enzymes, is the most frequently used method for isolating satellite DNAs. Recent methods such as C(0)t-1 DNA and chromosome microdissection, however, have proven to be efficient alternatives for the study of this class of DNA. Neotropical ichthyofauna is extremely rich and diverse enabling multiple approaches with regard to the differentiation and evolution of the genome. Genome components of some species and genera have been isolated, mapped and correlated with possible functions and structures of the chromosomes. The 5SHindIII-DNA satellite DNA, which is specific to Hoplias malabaricus of the Erythrinidae family, has an exclusively centromeric location. The As51 satellite DNA, which is closely correlated with the genome diversification of some species from the genus Astyanax, has also been used to infer relationships between species. In the Prochilodontidae family, two repetitive DNA sequences were mapped on the chromosomes, and the SATH 1 satellite DNA is associated with the origin of heterochromatic B chromosomes in Prochilodus lineatus. Among species of the genus Characidium and the Parodontidae family, amplifications of satellite DNAs have demonstrated that these sequences are related to the differentiation of heteromorphic sex chromosomes. The possible elimination of satellite DNA units could explain the genome compaction that occurs among some species of Neotropical Tetraodontiformes. These topics are discussed in the present review, showing the importance of satellite DNA analysis in the differentiation and karyotype evolution of Actinopterygii.

Measurement of the absolute gamma emission intensities from the decay of Th-229 in equilibrium with progeny.

There are few long-lived radionuclides yielding high intensity gamma-rays emission with energies ranging from 100 keV to 500 keV that can be applied as radioactive gamma standard to calibrate HPGe detectors. Furthermore, this energy range represents the main emitted energies of the majority of radionuclides used in nuclear medicine. The Brazilian National Laboratory for Ionizing Radiation Metrology (LNMRI/IRD/CNEN) has attempted to identify radionuclides that have the potential to be used as a calibration source due to their long half-life as well as their emission spectrum. Hence, LNMRI promotes standardization studies of gamma-emitting radionuclides that meet these criteria on order to disseminate them. Thorium-229, with its well-defined energies and relatively high intensities, is one such candidate radionuclide for the energy and full-energy peak efficiency calibration of high-purity gamma spectrometers. Thorium-229 was standardized by the method of 4παß(LS)-γ(NaI(Tl)) live timed anticoincidence counting. The emission intensities of gamma-rays associated with the decay of 229Th have been determined by HPGe gamma ray spectrometry with accuracy and precision. The results are in agreement with current literature data.

Aerobic bacterial microflora of Broad-snouted caiman (Caiman latirostris) oral cavity and cloaca, originating from parque Zoológico Arruda Câmara, Paraíba, Brazil.

The objective of this study was to isolate and identify the aerobic bacterial microflora from the oral cavity mucosa and cloaca's samples, collected from Broad-snouted caiman (Caiman latirostris), born and bred in captivity at Parque Zoológico Arruda Câmara, João Pessoa, Paraíba, Brazil. The most common bacteria were Staphylococcus sp. (14.74%), Corynebacterium sp. (13.68%), Escherichia coli (13.68%) and Shigella sp.(11.58%), and the less common were Citrobacter sp. (1.05%), Klebsiella pneumoniae (1.05%) and Salmonella sp. (1.05%).This emphasizes the importance of these microorganisms' participation in infectious processes (sepsis) and injuries caused by crocodilians.