hACE2 upregulation and participation of macrophages and clear cells in the immune response of epididymis to SARS-CoV-2 in K18-hACE2 mice.

BACKGROUND: The severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) virus caused the coronavirus disease 2019 pandemic, and the prevalence of deaths among men is higher than among women. The epididymis, divided into caput, corpus, and cauda, shows a region-specific immunity. The K18-hACE2 mouse expresses human angiotensin-converting enzyme 2 (hACE2), the receptor that allows SARS-CoV-2 infection. However, studies using this transgenic mouse to evaluate the impact of this viral infection in epididymis have not yet been performed. OBJECTIVES: We evaluated the expression of hACE2 in the epididymis of SARS-CoV-2-infected K18-hACE2 mice, and assessed the epididymal immune response, focusing on F4/80+ mononuclear phagocytes and tumor necrosis factor-alpha expression. MATERIALS AND METHODS: The following analyses were performed in the epididymal sections of infected mice: epithelial height and duct diameter, birefringent collagen, Terminal deoxynucleotidyl Transferase-mediated dUTP Nick End Labelling, immunoreactions for detection of hACE2, spike, FGF, V-ATPase, F4/80, tumor necrosis factor-alpha, and iNOS. Viral particles were identified under electron microscopy. hACE2, Rigi, Tgfb1 and Tnfa expression were also evaluated by real-time quantitative polymerase chain reaction. RESULTS: All epididymal regions expressed hACE2, which increased in all epididymal regions in the infected mice. However, the caput appeared to be the most infected region. Despite this, the caput region showed minimal changes while the cauda showed significant epithelial changes associated with increased iNOS immunoexpression. The F4/80+ mononuclear phagocyte area increased significantly in both stroma and epithelium. In addition to the epithelial and stromal mononuclear phagocytes, tumor necrosis factor-alpha was also detected in clear cells, whose cytoplasm showed a significant increase of this cytokine in the infected animals. DISCUSSION AND CONCLUSION: The K18-hACE2 mouse is a useful model for evaluating the impact of SARS-CoV-2 infection in the epididymis. The infection induced hACE2 upregulation, favoring the virulence in the epididymis. The epididymal regions responded differentially to infection, and the activation of F4/80+ mononuclear phagocytes associated with the increased tumor necrosis factor-alpha immunolabeling in clear cells indicates a role of clear cells/mononuclear phagocytes immunoregulatory mechanisms in the epididymal immune response to SARS-CoV-2 infection.

Global landscape of nicotine and tobacco products additives policies.

INTRODUCTION: Product design is a key tobacco industry strategy to recruit consumers and hinder cessation. An important technology in tobacco product design is the use of additives including those that create characterising flavours, that is, flavours that are perceived by taste and or smell by the user. To curb youth use, many countries are implementing policies to limit the use of flavours and other additives in tobacco products. This paper reviews the global landscape of tobacco additives policies including those focusing only on flavouring additives, updating a previous review. METHODS: We gathered additives policy data from available online sources of tobacco control policies including parties' reports to the WHO Framework Convention on Tobacco Control (where WHO refers to World Health Organization), the website Tobacco Control Laws, government websites, searches with the Google search engine and consultations with the WHO to ensure comprehensiveness of the assessment. The policies were classified according to their objective and characteristics: Labelling regulations, sales bans and tobacco product design regulations. We classified and organised the regulations according to their stated intent. RESULTS: We found that 62 countries had tobacco products' additives-related regulations at the regional, national or subnational levels. Most of these regulations focused on flavouring additives especially bans on the selling, importing and manufacturing tobacco products with characterising flavours. CONCLUSIONS: The data demonstrate various global policies addressing additives and/or flavours and a range of definitions of additives and flavours. Establishing common definitions and standards could facilitate cooperation between countries and regions.

Demographic characteristics and clinical-radiological correlation in patients with indications for Total Knee Arthroplasty: A cross-sectional study.

OBJECTIVE: The objective of this study is to correlate clinical symptoms and functionality, using the KOOS (Knee Injury and Osteoarthritis Outcome Score) and KSS (Knee Society Score System) scores, with the radiographic changes, using the Kellgren-Lawrence classification, in patients with knee osteoarthritis and indications for Total Knee Arthroplasty (TKA). METHODS: 120 patients (189 knees) with gonarthrosis and indications for TKA were included in the study. Demographic questionnaires were applied, and clinical and functional assessment was carried out using the KOOS and KSS scores. Knee radiographs were taken and graded according to Kellgren-Lawrence. The clinical scores were compared with the radiographic classification to establish a correlation between these two measurements. Statistical analysis was performed using the τ-Kendall correlation test. RESULTS: Weak and inversely proportional correlations were found between the clinical scores and the radiographic classification. Among clinical scores evaluated, KSS showed the highest correlation with Kellgren-Lawrence classification (τ = -0.356; p < 0.001), followed by KOOS-quality of life (τ = -0.176; p = 0.004), KOOS-total score (τ = -0.166; p = 0.004), KOOS-function in daily living (τ = -0.160; p = 0.005) and KOOS-symptoms (τ = -0.159; p = 0.006). KOOS-pain (τ = -0.149; p = 0.01) and KOOS-sport and recreation function (τ = -0.142; p = 0.025) scores had the weakest correlations. CONCLUSION: There is a weak correlation between the clinical-functional scores of TKA candidates and their radiographic classification by Kellgren-Lawrence. Among clinical scores evaluated, KSS had the strongest negative correlation with the radiographic classification.

Aortic aneurysm: Correlations with phenotypes associated with connective tissue dysplasia.

An aortic aneurysm is a localized enlargement that exceeds the normal diameter of the vessel by 50 %, posing a risk due to the likelihood of rupture. The cause of aortic aneurysm, especially in young people, is connective tissue dysplasia, a condition characterized by defects in the assembly of collagen and elastin proteins, leading to changes in elastic properties and disruption of the formation of organs and their systems. The article presents data confirming the relationship between many morphological manifestations of connective tissue dysplasia (e.g., funnel-shaped deformation of the sternum, scoliosis of the thoracic spine, abdominal hernias, arterial tortuosity, striae of atypical localization) and the risk of aortic aneurysm formation. The literature suggests that the identified combinations of some external manifestations of connective tissue dysplasia deserve special attention and may be constitutional markers for the possible development of aortic aneurysm, which is a promising direction for further research in this area.

Analysis of anticipatory and compensatory postural adjustment in women of different age groups using surface electromyography.

Postural balance is crucial for daily activities, relying on the coordination of sensory systems. Balance impairment, common in the elderly, is a leading cause of mortality in this population. To analyze balance, methods like postural adjustment analysis using electromyography (EMG) have been developed. With age, women tend to experience reduced mobility and greater muscle loss compared to men. However, few studies have focused on postural adjustments in women of different ages using EMG of the lower limbs during laterolateral and anteroposterior movements. This gap could reveal a decrease in muscle activation time with aging, as activation time is vital for postural adjustments. This study aimed to analyze muscle activation times in women of different ages during postural adjustments. Thirty women were divided into two groups: young and older women. A controlled biaxial force platform was used for static and dynamic balance tests while recording lower limb muscle activity using EMG. Data analysis focused on identifying muscle activation points and analyzing postural adjustment times. Results showed significant differences in muscle activation times between the two groups across various muscles and platform tilt conditions. Younger women had longer muscle activation times than older women, particularly during laterolateral platform inclinations. In anteroposterior movements, older women exhibited longer activation times compared to their laterolateral performance, with fewer differences between the groups. Overall, older women had shorter muscle activation times than younger women, suggesting a potential indicator of imbalance and increased fall risk.

Effect of Periodontal Instrumentation on Tooth Structure Exposure to Ionizing Radiation: In Vitro Study.

This study aimed to evaluate the effects of different scaling protocols on the morphology and roughness of the root dentin substrate exposure to ionizing radiation. One hundred and thirty extracted bovine incisors were randomly divided into two groups (n=65): non-irradiated (NIR) and irradiated (IR). Each group was initially subdivided into three subgroups according to the type of non-surgical periodontal protocol: NIT: no instrumentation; HS: hand scaling with 15 apical-coronal instrument movements; US: ultrasonic scaling with 15 apical-coronal cycles. Subsequently, all samples were subjected to the prophylaxis protocol, being subdivided into the following groups: NIT/PP: prophylaxis with a fine prophylactic paste using a rubber cup for 15 seconds; HS/PP: Hand scaling followed by the prophylaxis protocol; US/PP: Ultrasonic scaling followed by the prophylaxis protocol. The roughness of the root dentin surface was measured with a profilometer (Ra/Rz - µm), and the morphology of the dentin surfaces was analyzed using scanning electron microscopy (SEM). The analyses were conducted before and after the prophylaxis protocol. In the absence of prophylaxis, the roughest surfaces were observed after ultrasonic instrumentation followed by hand instrumentation for both IR and NIR groups. No difference in Ra and RZ values between HS/PP and US/PP was observed for both substrates. For the IR group, the prophylaxis resulted in similar Ra and RZ values for both instrumentation groups in comparison to no instrumentation. Ordinal logistic regression showed that both HS and US resulted in higher scores than NIT, irrespective of IR presence. In conclusion, the IR showed a rougher root surface for both HS and US in comparison to NIR. However, the prophylaxis procedure significantly reduced the roughness of root surfaces after both instrumentation procedures.

Health care for patients with long COVID: a scoping review.

OBJECTIVE: To map the scientific evidence on the care offered to health service users with Long Covid-19. METHOD: This is a scoping review based on the methods of the Joanna Briggs Institute. Primary studies were included, in Portuguese, English and Spanish, published between December 2019 and June 2023, in the Virtual Health Library, Web of Science, Scopus, PUBMED, SciELO and LITCovid LongCovid databases. RESULTS: Of the ١٣ articles analyzed, it stands out that the care provided to patients with Long Covid is associated with drug prescription, indication of physical exercises, telerehabilitation and physiotherapy. CONCLUSION: A fragmentation was identified in the care provided to users of health services with Long Covid, with care directed only at isolated symptoms, without addressing the biopsychosocial care that people with this condition need.

Microfragmented Adipose Tissue Associated With Collagen Membrane in the Treatment of Focal Knee Cartilage Defect.

Focal articular cartilage defects are an important factor that leads to dysfunction of the knee joint. Several different surgical approaches have been tried, most of them showing poor results in the long term. The use of orthobiologics in the context of focal chondral lesion has emerged as a potential tool in the treatment of this condition. In this article, we present a surgical technique for the treatment of focal chondral lesions using a collagen membrane associated with microfragmented adipose tissue graft.

Application Techniques and Concentrations of Ascorbic Acid to Reduce Saline Stress in Passion Fruit.

Salinity restricts the growth of irrigated fruit crops in semi-arid areas, making it crucial to find ways to reduce salt stress. One effective strategy is using eliciting substances like ascorbic acid. In this context, the objective of this study was to evaluate the effects of application methods and concentrations of ascorbic acid on the morphophysiology and production of sour passion fruit irrigated with saline water. The experiment was organized using a factorial randomized block design (3 × 3 × 2) with three application methods (soaking, spraying, and soaking and spraying), three concentrations of ascorbic acid (0, 0.8, and 1.6 mM) and two levels of electrical conductivity of irrigation water-ECw (0.8 and 3.8 dS m-1). Foliar spraying of ascorbic acid at a concentration of 0.8 mM mitigated the effects of salt stress on the relative water content of leaves, the synthesis of photosynthetic pigments, gas exchange, and total production of sour passion fruit when irrigated with ECw of 3.8 dS m-1. Plants grown with water of 0.8 dS m-1 and under foliar application of 0.8 mM of ascorbic acid achieved the maximum growth in stem diameter and the greatest volume of pulp in the fruits.

Pseudoplagiostoma humilis sp. nov., a New Fungal Species Causing Shoot Blight and Dieback in Anacardium humile in Brazil.

The cajuzinho do cerrado (Anacardium humile-Anacardiaceae), a shrub species native to Brazil, is harvested for multiple uses in food and medicine. Members of a harvesting community, near the municipality of Bonito de Minas, Minas Gerais state, Brazil reported characteristic symptoms of shoot blight and dieback reducing pseudofruit and seed production by this plant. This study aimed to identify the etiological agent of this disease. Two fungal isolates were obtained from symptomatic leaf samples and morphologically and molecularly characterized. The fungus was identified, based on morphological analyses, as a probable new species of Pseudoplagiostoma. Phylogenetic analyses based on a combination of DNA sequence data (nuc rDNA ITS1-5.8S-ITS2 region, tef1-α and tub2), confirmed this hypothesis. The isolates obtained were allocated to a distinct, well-supported clade (IB = 0.99, ML = 100%), placed as a unique lineage here proposed as a new species named Pseudoplagiostoma humilis. The pathogenicity test confirmed that this new species was the causal agent of shoot blight and dieback on A. humile. This is the fourteenth Pseudoplagiostoma species reported in the world and the third in Brazil.

Optimized protocol for collecting root canal biofilms for in vitro studies.

Endodontic retreatment is often necessitated by several factors, including the persistence of microorganisms in the root canal system (RCS). Their complex organization in biofilms increases their pathogenic potential, necessitating new disinfection strategies. This study aimed to standardize a new in vitro protocol for collecting biofilm from the RCS. Thirty-four bovine incisors were used in the study, divided into two experimental groups with two collection steps each: (a) biofilm collection protocol and (b) absorbent paper points protocol. Twelve specimens from each group were selected for counting colony-forming units (CFUs), while eight specimens were prepared for scanning electron microscopy (SEM). Two additional specimens served as sterilization controls to ensure that experiments were free of contamination. The coronal region was removed and standardized at 15 mm. After preparation with ProTaper up to F5, the apical foramen was sealed with composite resin, and the roots were stabilized with acrylic resin in 1.5-mL Eppendorf tubes. The specimens were sterilized and inoculated with Enterococcus faecalis NTCT 775 every 24 h for 21 days. After this period, each group underwent biofilm collection protocols, and CFU and scanning electron microscopy (SEM) data were analyzed. The Shapiro-Wilk test was performed to assess the normality of log-transformed data, and the results indicated a normal distribution for all groups, allowing parametric testing. The Levene test was used to evaluate the equality of variances. The proposed biofilm collection method yielded significantly higher CFU counts compared with the absorbent paper points method, particularly when analyzed on a log10 scale. An independent samples t-test confirmed a statistically significant difference between the two methods (p < 0.0001). The proposed protocol achieved an efficiency rate of 95.85 % ± 1.15 %, whereas the absorbent paper points protocol yielded a lower efficiency of 5.46 % ± 1.37 %. Therefore, the biofilm collection protocol proposed in this study proved to be more effective for biofilm removal from the RCS.

Pediatric Organ Donation: 16-Year Experience of PICU/ICU of a Third Level Hospital in Portugal, 2006-2021.

Although pediatric organ donation represents a small proportion of overall organ donation, children and adolescents make a significant contribution to the pool of donated organs. In this study 252 solid organs were collected from children and adolescent. Two hundred and two recipients benefited from 62 pediatric organ donors, with a recipient/donor ratio of 3.3.

BACKGROUND: Pediatric organ donors represent a small but important portion of the deceased donor pool, helping both children and adults in the transplant waitlist. Despite this, pediatric donation remains an overlooked subject of research. METHODS: Retrospective, single­center, descriptive study. All brain death patients under 18 years old who were admitted to the Intensive Care Unit (ICU) between January 1st, 2006, and December 31st, 2021, and who were eligible for organ donation were included. RESULTS: Between January 2006 and December 2021, 200 children/adolescent died in the ICU. From those, 62 patients (31%) were considered eligible for organ donation. The mean age of the donors at the time of death was 8.8 years. Sixty­three per cent were male. The most frequent cause of death was traumatic brain injury (n = 36). Two hundred and fifty organs were collected benefitting 202 persons with a recipient/donor ratio of 3.3. Kidneys were the most frequent organ donated (n = 116), followed by liver (n = 56) and heart (n = 34). The median number of organs donated per child was four, with a minimum of 1 organ and maximum of 8. CONCLUSIONS: Pediatric organ donation represents a small proportion of overall organ donation, but children and adolescents have important impact on the lives they save. The field of pediatric organ donation needs more research to better understand the contribution of the pediatric population to both adults and children who wait for an organ.

Catastrophic thrombotic events with partial bilateral amputation of legs and fingers in a 12-year-old girl with COVID-19 in Brazil: case report.

The first case of COVID-19 was detected in Dec 2019, in China. The disease shortly evolved into a pandemic and imposed an unparalleled health and social burden on mankind. Severe forms of COVID-19 mainly affect adults, especially the elderly and those with comorbidities. We report a severe case of COVID-19 in a previously healthy 12-year-old female who was admitted to the emergency room on May 26, 2020, with fever, abdominal pain, vomiting, and diarrhea. During the hospital stay, she tested positive for SARS-CoV-2 and developed multiple organ failure and catastrophic thrombotic events resulting in bilateral amputation of legs and fingers. She was discharged from the hospital for outpatient follow-up after 107 days. By the time this report was written, the patient was undergoing prosthesis prescription and training and regaining her independence to walk.

O primeiro caso de covid-19 foi detectado em dezembro de 2019, na China. A doença rapidamente evoluiu para uma pandemia e trouxe um fardo social e de saúde sem paralelo para a humanidade. As formas graves de covid-19 afetam principalmente adultos, especialmente idosos com comorbidades. Relatamos um caso grave de covid-19 em uma paciente de 12 anos de idade, previamente saudável, que deu entrada no pronto-socorro em 26/05/2020 com febre, dores abdominais, vômitos e diarreia. Durante a investigação, apresentou teste positivo para SARS-CoV-2 e evoluiu com falência múltipla de órgãos e eventos trombóticos catastróficos, resultando em amputação bilateral de pernas e quirodáctilos. Ela recebeu alta hospitalar para acompanhamento ambulatorial após 107 dias. No momento da redação deste relato, a paciente estava sendo submetida a protetização e recuperando marcha independente.

Generation and characterization of cortical organoids from iPSC-derived dental pulp stem cells using traditional and innovative approaches.

Cortical organoids derived from human induced pluripotent stem cells (hiPSCs) represent a powerful in vitro experimental system to investigate human brain development and disease, often inaccessible to direct experimentation. However, despite steady progress in organoid technology, several limitations remain, including high cost and variability, use of hiPSCs derived from tissues harvested invasively, unexplored three-dimensional (3D) structural features and neuronal connectivity. Here, using a cost-effective and reproducible protocol as well as conventional two-dimensional (2D) immunostaining, we show that cortical organoids generated from hiPSCs obtained by reprogramming stem cells from human exfoliated deciduous teeth (SHED) recapitulate key aspects of human corticogenesis, such as polarized organization of neural progenitor zones with the presence of outer radial glial stem cells, and differentiation of superficial- and deep-layer cortical neurons and glial cells. We also show that 3D bioprinting and magnetic resonance imaging of intact cortical organoids are alternative and complementary approaches to unravel critical features of the 3D architecture of organoids. Finally, extracellular electrical recordings in whole organoids showed functional neuronal networks. Together, our findings suggest that SHED-derived cortical organoids constitute an attractive model of human neurodevelopment, and support the notion that a combination of 2D and 3D techniques to analyze organoid structure and function may help improve this promising technology.

Optimization and evaluation of a chitosan-coated PLGA nanocarrier for mucosal delivery of Porphyromonas gingivalis antigens.

Recent advances in understanding Alzheimer's disease (AD) suggest the possibility of an infectious etiology, with Porphyromonas gingivalis emerging as a prime suspect in contributing to AD. P. gingivalis may invade systemic circulation via weakened oral/intestinal barriers and then cross the blood-brain barrier (BBB), reaching the brain and precipitating AD pathology. Based on the proposed links between P. gingivalis and AD, a prospective approach is the development of an oral nanovaccine containing P. gingivalis antigens for mucosal delivery. Targeting the gut-associated lymphoid tissue (GALT), the nanovaccine may elicit both mucosal and systemic immunity, thereby hampering P. gingivalis ability to breach the oral/intestinal barriers and the BBB, respectively. The present study describes the optimization, characterization, and in vitro evaluation of a candidate chitosan-coated poly(lactic-co-glycolic acid) (PLGA-CS) nanovaccine containing a P. gingivalis antigen extract. The nanocarrier was prepared using the double emulsion solvent evaporation method and optimized for selected experimental factors, e.g. PLGA amount, surfactant concentration, w1/o phase ratio, applying a d-optimal statistical design to target the desired physicochemical criteria for its intended application. After nanocarrier optimization, the nanovaccine was characterized in terms of particle size, polydispersity index (PdI), ζ-potential, encapsulation efficiency (EE), drug loading (DL), morphology, and in vitro release profile, as well as for mucoadhesivity, stability under simulated gastrointestinal conditions, antigen integrity, in vitro cytotoxicity and uptake using THP-1 macrophages. The candidate PLGA-CS nanovaccine demonstrated appropriate physicochemical, mucoadhesive, and antigen release properties for oral delivery, along with acceptable levels of EE (55.3 ± 3.5 %) and DL (1.84 ± 0.12 %). The integrity of the encapsulated antigens remained uncompromised throughout NPs production and simulated gastrointestinal exposure, as confirmed by SDS-PAGE and Western blotting analyses. Furthermore, the nanovaccine showed effective in vitro uptake, while exhibiting low cytotoxicity. Taken together, these findings underscore the potential of PLGA-CS NPs as carriers for adequate antigen mucosal delivery, paving the way for further investigations into their applicability as vaccine candidates against P. gingivalis.

Zinc pathogenic importance in correcting immunity and restoring public health in the post-COVID period: An overview.

CONTEXT: The problem of correcting immune system function and compensating for co-morbidities becomes particularly clinically significant in the post-COVID period. There is evidence that certain trace elements in the human body, particularly zinc ions, play a critical role in restoring the function of the immune system and internal organs. OBJECTIVE: To analyze the mechanisms of zinc action maintaining the body homeostasis in order to justify pathogenetically the inclusion of zinc drugs in the therapy of patients in the post-COVID period. METHODS: Data from Elsevier, Global Health, PubMed-NCBI, Embase, MEDLINE, Scopus, Research gate, RSCI Scopus, Cochrane Library, Google Academy, e-LIBRARY.RU and CyberLeninka were used. RESULTS: This review showed that the importance of zinc in maintaining body homeostasis in the post-COVID period is determined by its multifaceted effect on all parts of the immune system, its anti-inflammatory activity, antimicrobial properties and participation in the restoration of internal organ function. Elimination of zinc deficiency in the post-COVID period is essential to support immunity, compensate for comorbidities and reduce the risk of complications. The impossibility of synthesizing zinc in the body requires its constant intake in sufficient quantities. Zinc levels are significantly reduced after infectious diseases, as this element is specifically distributed to organs and tissues to maintain immunological and metabolic functions. The degree of zinc deficiency is associated with the severity of COVID-19 and the post-COVID period. It is pathogenetically justified to prescribe zinc drugs in the post-COVID period, the choice of which should take into account comorbidities and severity of hypozincemia. CONCLUSION: Regularly administered therapy with zinc drugs in the post-COVID period will help correct the population immunity and restore public health.