Identification of Inflammatory Mediators in Saliva Samples From Hospitalized Newborns: Potential Biomarkers?

Saliva measurements serve as a noninvasive tool for clinically monitoring newborns (NB) and children, a vulnerable population with promising potential for both research and clinical practice. Saliva acts as a repository for various inflammatory biomarkers involved in diverse biological functions. Particularly for children, it offers numerous advantages when compared to plasma and urine sampling. Nevertheless, there is a significant knowledge gap regarding detectable levels of cytokines in the saliva of newborns and children, as well as studies aiming to assess the relationship of this content with physiological and pathological processes. OBJECTIVES: To characterize the levels of 11 inflammatory mediators (IFNg, IL1b, IL2, IL4, IL6, IL8, IL10, IL12, IL17, TNF, and VEGF) in saliva samples from NB on the first and second day of hospitalization in the Neonatal Intensive Care Unit (NICU). METHOD: Exploratory study, descriptive, nested within a primary clinical, observational, and prospective study, conducted in the NICU of a public hospital in São Paulo, Brazil. Demographic data and vital signs were recorded in the clinical records of 90 NB, and five saliva samples from 5 NB were collected between the first and second day of life (D1-D2) at approximately 8-hr intervals (8-9 am, 4-5 pm, and 11-12 pm). Saliva samples were used for the measurement of 11 cytokines (IFNg, IL1b, IL2, IL4, IL6, IL8, IL10, IL12, IL17, TNF, and VEGF). RESULTS: Five NBs participated in this exploratory study, and the vital signs showed variability from the first (D1) to the second day (D2) of hospitalization, variability similar to that of the total population of the primary study. The presence and levels of the 11 cytokines were detected in the saliva samples, as well as a statistical correlation between 10 cytokines (IFNg, IL1b, IL2, IL4, IL6, IL10, IL12, IL17, TNF, and VEGF) and vital signs. CONCLUSIONS: The novelty of measuring inflammatory mediators in saliva samples from hospitalized NBs in the NICU is highlighted, providing support and new perspectives for the development of clinical and experimental research and an opportunity for developing and implementing new salivary biomarkers in different population segments.

Alexis retractor efficacy in transthoracic thoracoscopically assisted discectomy for thoracic disc herniations.

PURPOSE: A transthoracic anterior or lateral approach for giant thoracic disc herniations is a complex operation which requires optimal exposure and maximal visualisation. Traditional metal rigid retractors may inflict significant skin trauma especially with prolonged operative use and limit the working angles of endoscopic instrumentation at depth. We pioneer the use of the Alexis retractor in transthoracic thoracoscopically assisted discectomy for the first time. METHODS: The authors describe and demonstrate the technical use of the Alexis retractor during operative cases. Patient positioning, clinical rationale and operative nuances are elucidated for readers to gain an appreciation of the transthoracic approach to thoracic disc herniations. RESULTS: The advantages of the Alexis retractor include minimally invasive circumferential flexible retraction, facilitation of bimanual instrument use, diminished risk of surgical site infections and reduced rib retraction leading to less postoperative pain. CONCLUSION: Use of the flexible and intuitive Alexis retractor maximises operative exposure and is an effective adjunct when performing complex transthoracic approaches for thoracic disc herniations.

Neutrophil Extracellular Trap-related Biomarkers Are Increased in the Synovial Fluid of Patients With Periprosthetic Joint Infections.

BACKGROUND: The diagnosis of periprosthetic joint infection (PJI) is a major challenge in clinical practice. The role of neutrophils in fighting infection has been increasingly understood, and one mechanism of action of these cells is neutrophil extracellular traps. However, little is known about this process in PJI. QUESTIONS/PURPOSES: (1) Are the biomarkers of neutrophil extracellular trap formation (citrullinated histone H3 [H3Cit], cell-free DNA [cf-DNA], and myeloperoxidase [MPO]) increased in the synovial fluid of patients with PJI? (2) What is the diagnostic accuracy of biomarkers of neutrophil extracellular trap formation for PJI? METHODS: Between May 2020 and March 2021, 43 patients who underwent revision THA or TKA were enrolled in this study. Eleven patients were excluded and 32 patients were categorized into the PJI group (n = 16) or non-PJI group (n = 16) according to the 2018 Second International Consensus Meeting on Musculoskeletal Infection criteria. There were 15 men and 17 women in this study, with a median (range) age of 70 years (60 to 80 years). Twenty-seven patients had TKA and five had THA. We measured cf-DNA, MPO, and H3Cit in synovial fluid. The sensitivity, specificity, and receiver operating characteristic curve were calculated for each biomarker using the Musculoskeletal Infection Society criteria as the gold standard for diagnosis and considering a clinical surveillance of 2 years for patients in the non-PJI group. RESULTS: Patients with PJI had higher levels of synovial fluid cf-DNA (median [range] 130 ng/µL [18 to 179] versus 2 ng/µL [0 to 6]; p < 0.001), MPO (1436 ng/µL [55 to 3996] versus 0 ng/µL [0 to 393]; p < 0.001), and H3Cit (2115 ng/µL [5 to 2885] versus 3 ng/µL [0 to 87]; p < 0.001) than those in the non-PJI group. In receiver operating characteristic curve analyses, we observed near-perfect performance for all biomarkers evaluated, with an area under the curve of 1 (95% CI 0.9 to 1), 0.98 (95% CI 0.9 to 1), and 0.94 (95% CI 0.8 to 0.99) for cf-DNA, MPO, and H3Cit, respectively. The sensitivity for detecting PJI using synovial fluid was 100% for cf-DNA, 94% for MPO, and 88% for H3Cit. The specificity was 100% for cf-DNA and MPO, and 88% for H3Cit. CONCLUSION: Our results show that neutrophils in the periprosthetic microenvironment release neutrophil extracellular traps as part of the bactericidal arsenal to fight infection. These results allow a better understanding of the cellular and molecular processes that occur in this microenvironment, enabling the design of more assertive strategies for identifying new biomarkers and improving the available ones. Novel studies are needed to define whether and how neutrophil extracellular trap-related biomarkers can be useful for diagnosing PJI. LEVEL OF EVIDENCE: Level II, diagnostic study.

Survival outcomes of intracranial extraventricular neurocytomas: a systematic review and individual patient data meta-analysis.

OBJECTIVE: The rarity of intracranial extraventricular neurocytomas (EVNs) has precluded accurate definition of its surgical characteristics to date. The authors present the first survival analysis of this unique entity that aims to clarify tumor characteristics, surgical outcomes, and efficacy of postoperative adjuvant therapy. METHODS: A systematic review was conducted in accordance with the Preferred Reporting Items for Systematic Reviews and Meta-Analyses guidelines. Searches of the MEDLINE, Embase, Scopus, Google Scholar, and Cochrane Database of Systematic Reviews databases were performed from inception to date. Primary outcomes were progression-free survival (PFS) and overall survival (OS). Prognostic variables were age, sex, tumor consistency, extent of resection, and receipt of postoperative adjuvant therapy. Survival data were analyzed using Kaplan-Meier survival curves and the log-rank test to compare dichotomized cohorts. Multivariate Cox regression models were constructed, interrogated with Schoenfeld residuals, and subsequently utilized to identify independent prognostic factors. Risk of bias was assessed with the Mayo Clinic instrument. RESULTS: Five hundred fourteen articles were initially retrieved, which was distilled to 10 included articles consisting of 101 cases of intracranial EVNs. The 5-year OS rate was 90.4% (95% CI 81.8%-99.8%) and the PFS rate was 48.6% (95% CI 34.46%-68.8%). The median PFS was 60 months. Patients younger than 50 years of age experienced superior OS (p = 0.03) and PFS (p < 0.01). Gross-total resection (GTR) was superior to subtotal resection (STR) in reducing mortality (p < 0.01). Adjuvant therapy following either STR or GTR did not significantly improve survival. CONCLUSIONS: Intracranial EVNs are rare tumors that portend a poorer prognosis than central neurocytomas, despite both being WHO grade 2 tumors. Complete surgical extirpation is the cornerstone of management. There is no clearly established role for adjuvant postoperative therapy, but each case should be managed on an individual basis.

Impaired macrophage and memory T-cell responses to Bacillus Calmette-Guerin nonpolar lipid extract.

Introduction: The attenuation of BCG has led to the loss of not only immunogenic proteins but also lipid antigens. Methods: Thus, we compared the macrophage and T-cell responses to nonpolar lipid extracts harvested from BCG and Mycobacterium tuberculosis (Mtb) to better understand the role of BCG lipids in the already known diminished responses of the vaccine strain. Results: Relative to Mtb, nonpolar lipid extract from BCG presented a reduced capacity to trigger the expression of the genes encoding TNF, IL-1b, IL-6 and IL-10 in RAW 264.7 macrophages. Immunophenotyping of PBMCs isolated from healthy individuals revealed that lipids from both BCG and Mtb were able to induce an increased frequency of CD4+ and CD8+ T cells, but only the lipid extract from Mtb enhanced the frequency of CD4-CD8-double-negative, γσ+, CD4+HLA-DR+, and γσ+HLA-DR+ T cells relative to the nonstimulated control. Interestingly, only the Mtb lipid extract was able to increase the frequency of CD4+ memory (CD45RO+) T cells, whereas the BCG lipid extract induced a diminished frequency of CD4+ central memory (CD45RO+CCR7-) T cells after 48 h of culture compared to Mtb. Discussion: These findings show that the nonpolar lipids of the BCG bacilli presented diminished ability to trigger both proinflammatory and memory responses and suggest a potential use of Mtb lipids as adjuvants to increase the BCG vaccine efficacy.

First report of fibropapillomatosis (FP) and Chelonid alphaherpesvirus 5 (ChHV5) in a green sea turtle (Chelonia mydas) from the historically fibropapillomatosis-free Fernando de Noronha Archipelago, Northeastern Brazil / Primeiro caso de fibropapilomatose (FP) e de Chelonid alphaherpesvirus 5 (ChHV5) em tartaruga-verde (Chelonia mydas) no Arquipélago de Fernando de Noronha, Nordeste do Brasil

Fibropapillomatosis (FP) is an infectious disease caused by Chelonid alphaherpesvirus 5 (ChHV5). Nevertheless, its clinical manifestations are considered multifactorial. Due to its relevance, FP is currently monitored in sea turtle populations in the United States, Australia, Caribbean, and Brazil. Between 2000 and 2020, the TAMAR Project/ TAMAR Project Foundation analyzed the prevalence of FP in nine states and oceanic islands along the Brazilian coast, including Fernando de Noronha Archipelago (FNA), a historically FP-free area. A total of 4,435 green sea turtles (Chelonia mydas) were monitored from 2010 to 2016. Additionally, in 2012 and 2014, 43 FP-free skin samples were analyzed for ChHV5 using a qualitative PCR for the UL30 polymerase (pol) sequence. In 2015, a bilateral ocular nodule characterized as an FP tumor was reported in one of the monitored individuals undergoing rehabilitation. Tissue samples were collected following surgical removal of the tumor. Characterization of a 454 bp UL30 polymerase gene revealed a ChHV5 sequence previously reported in other areas of the Atlantic Brazilian coast. In the years following this finding from January 2017 to March 2020, a total of 360 C. mydas were monitored in the same area and no FP tumors were detected. This is the first report of FP and the first detection of ChHV5 in FNA, a finding of great concern considering this site's historical absence of FP occurrence. This study highlights the importance of monitoring this disease in historically FP-free areas of the Brazilian Atlantic coast.(AU)

A fibropapilomatose (FP) é uma doença infecciosa causada pelo Chelonid alphaherpesvirus 5 (ChHV5). No entanto, as manifestações clínicas da doença são consideradas multifatoriais. Esta doença é monitorada atualmente em populações de tartarugas marinhas nos EUA, Austrália, Caribe e Brasil. Desde 2000, o Projeto TAMAR/Fundação Projeto TAMAR analisa a presença de FP em nove estados da costa brasileira e ilhas oceânicas, incluindo o arquipélago de Fernando de Noronha, uma área historicamente livre de FP. Um total de 4.435 indivíduos de Chelonia mydas foram monitorados de 2010 a 2016 e 43 amostras de pele foram analisadas para detectar ChHV5 em 2012 e 2014 com o objetivo de avaliar a presença do vírus em tecidos sem FP, usando uma PCR qualitativa para detecção de sequências do gene da UL30 polimerase. Em 2015, uma tartaruga verde (C. mydas) foi relatada com um nódulo ocular bilateral caracterizado como FP. Amostras de tecido foram coletadas durante sua reabilitação e procedimento cirúrgico para remover o tumor. A caracterização parcial de uma sequência de 454 bp do gene UL30 polimerase detectou ChHV5 anteriormente relatado em outras áreas da costa atlântica brasileira. Após estes achados, de janeiro de 2017 a março de 2020, um total de 360 indivíduos de C. mydas foram monitorados e nenhum caso de FP foi registrado. Este é o primeiro relato de FP e a primeira caracterização de ChHV5 no arquipélago de Fernando de Noronha, uma questão preocupante e que ressalta a importância do monitoramento desta doença em áreas historicamente livres de FP na costa atlântica brasileira.(AU)

Pneumonia tuberculosa: relato de caso / Tuberculous pneumonia: case report / Neumonía tuberculosa: reporte de caso

A tuberculose é uma doença infecciosa causada pelo Mycobacterium tuberculosis muito disseminada no Brasil. Tem preferência pelo acometimento pulmonar, embora vários outros órgãos possam ser afetados, como vasos linfáticos, trato geniturinário, ossos, articulações, sistema nervoso central, trato gastrointestinal e pericárdio. A pneumonia tuberculosa é uma forma rara, que se apresenta com consolidação alveolar, principalmente em lobo superior, podendo mimetizar um quadro de pneumonia pneumocócica. O objetivo deste ensaio é relatar o caso clínico de um indivíduo do sexo masculino com diagnóstico de pneumonia tuberculosa no Hospital Geral Roberto Santos (HGRS) e fazer uma breve revisão da literatura sobre o tema. O método utilizado para isso será o relato de caso clínico, com informações obtidas por meio de revisão de dados contidos em prontuário médico. Os resultados obtidos foram: paciente do sexo masculino, 19 anos, previamente hígido, não tabagista, natural de Recife, procedente de Salvador, admitido em agosto de 2019 no serviço de emergência do HGRS, com história de tosse produtiva de secreção amarelada, febre não mensurada e calafrios. O relato de caso descrito mostra uma apresentação atípica, rara e pouco comum de pneumonia tuberculosa/tuberculose pulmonar. A procura por diagnósticos diferenciais em quadros com resposta ruim à terapêutica inicialmente instituída é fundamental para evitar atrasos entre a realização do diagnóstico correto e a introdução das devidas medidas terapêuticas.

Tuberculosis is a highly prevalent infectious disease in Brazil, caused by Mycobacterium tuberculosis. Despite mostly affecting the lungs, it may also affect other organs, such as lymphatic vessels, genitourinary tract, bones, joints, central nervous system, gastrointestinal tract, and pericardium. A rare form of the disease, tuberculous pneumonia presents with alveolar consolidation, especially in the upper lobe, possibly mimicking pneumococcal pneumonia. This study sought to report the clinical case of a male individual diagnosed with Tuberculous Pneumonia at the Hospital Geral Roberto Santos, as well as to perform a brief literature review of subject. The study was conducted with data collected from the medical records of a male patient aged 19 years old, previously healthy, non-smoker, born in Recife and living in Salvador, admitted in August 2019 to the emergency service of Hospital Geral Roberto Santos with a history of productive cough yellowish sputum, unmeasured fever, and chills. The case report here described shows an atypical, rare, and uncommon presentation of Tuberculous Pneumonia/Pulmonary Tuberculosis. To avoid delays between the correct diagnosis and the introduction of the respective therapeutic measures, health professionals should search for differential diagnoses in conditions with poor response to the initially instituted therapy.

La tuberculosis es una enfermedad infecciosa causada por Mycobacterium tuberculosis muy prevalente en Brasil. Tiene preferencia por la afectación pulmonar, aunque pueden estar implicados muchos otros órganos, como vasos linfáticos, tracto genitourinario, huesos, articulaciones, sistema nervioso central, tracto gastrointestinal y pericardio. La neumonía tuberculosa es una forma rara, que se presenta con consolidación alveolar, especialmente en el lóbulo superior, que puede simular un cuadro de neumonía neumocócica. El objetivo de este ensayo es informar el caso clínico de un varón diagnosticado de Neumonía Tuberculosa en el Hospital Geral Roberto Santos y realizar una breve revisión de la literatura sobre el tema. El método utilizado para ello será elreporte de caso clínico con información obtenida mediante revisión de datos contenidos en historias clínicas. Los resultados obtenidos fueron: paciente varón de 19 años, previamente sano, no fumador, nacido en Recife, residente en Salvador, ingresado en agosto de 2019 en el servicio de urgencias del Hospital Geral Roberto Santos, con antecedente de tos productiva amarillenta, fiebre no medida y escalofríos. El reporte de caso aquí descrito muestra una presentación atípica, rara y poco común de neumonía tuberculosa/tuberculosis pulmonar. La búsqueda de diagnósticos diferenciales en condiciones con mala respuesta a la terapia inicialmente instituida es fundamental para evitar retrasos entre el diagnóstico correcto y la introducción de las respectivas medidas terapéuticas.

ANALYSIS OF TISSUE BIOPSY AND JOINT ASPIRATION IN THE DIAGNOSIS OF PERIPROSTHETIC HIP INFECTIONS: CROSS-SECTIONAL STUDY.

OBJECTIVE: To evaluate sensitivity, specificity, accuracy, positive predictive value, and negative predictive value of preoperative joint aspiration (PJA) and periarticular tissue percutaneous biopsy (PTPB), as well as their combination, in the diagnosis of infection after total hip arthroplasty. METHODS: This cross-sectional study (Level of Evidence II) was conducted with prospective data on 29 patients submitted to PJA with PTPB at the National Institute of Orthopedics and Traumatology from September 2015 to January 2016. Specimens obtained during the procedures underwent microbiological analyses, and the results were compared with those obtained in subsequent revision arthroplasty surgeries. RESULTS: PJA, PTPB, and their combination reached values of 78%, 73%, 89% for sensitivity, respectively; 72%, 90%, 94% for specificity; and 76%, 80%, 90% for accuracy. CONCLUSIONS: PJA combined with PTPB was sensitive, specific, and effective in diagnosing periprosthetic hip infection. Level of Evidence II, Prospective Cross-Sectional Study.

OBJETIVO: Avaliar a sensibilidade, especificidade, acurácia, valor preditivo positivo e valor preditivo negativo dos métodos diagnósticos aspirado articular pré-operatório (AAPO), biópsia percutânea de tecidos periarticulares (BPTP) e ambos associados na infecção pós-artroplastia total de quadril (IPATQ). MÉTODOS: Trata-se de um estudo transversal (Nível de Evidência II) com coleta prospectiva de dados obtidos de 29 pacientes submetidos a AAPO com BPTP no Instituto Nacional de Ortopedia e Traumatologia durante o período de setembro de 2015 à janeiro de 2016. Foram comparados os resultados das análises microbiológicas dos espécimes obtidos por meio da BPTP e do AAPO com os obtidos intraoperatoriamente nas cirurgias subsequentes de revisão das artroplastias. RESULTADOS: Encontramos uma sensibilidade da AAPO, BPTP e ambos, respectivamente de 78%, 73%, 89%, uma especificidade de 72%, 90%, 94% e uma acurácia de 76%, 80%, 90%. CONCLUSÕES: O procedimento de AAPO com BPTP para diagnóstico de infecção periprotética de quadril é sensível, específico e eficaz. Nível de Evidência II, Estudo Transversal Prospectivo.

Analysis of tissue biopsy and joint aspiration in the diagnosis of periprosthetic hip infections: cross-sectional study / Análise da biópsia de tecidos periarticulares e do aspirado articular no diagnóstico de infecções periprotéticas do quadril: estudo transversal

ABSTRACT Objective: To evaluate sensitivity, specificity, accuracy, positive predictive value, and negative predictive value of preoperative joint aspiration (PJA) and periarticular tissue percutaneous biopsy (PTPB), as well as their combination, in the diagnosis of infection after total hip arthroplasty. Methods: This cross-sectional study (Level of Evidence II) was conducted with prospective data on 29 patients submitted to PJA with PTPB at the National Institute of Orthopedics and Traumatology from September 2015 to January 2016. Specimens obtained during the procedures underwent microbiological analyses, and the results were compared with those obtained in subsequent revision arthroplasty surgeries. Results: PJA, PTPB, and their combination reached values of 78%, 73%, 89% for sensitivity, respectively; 72%, 90%, 94% for specificity; and 76%, 80%, 90% for accuracy. Conclusions: PJA combined with PTPB was sensitive, specific, and effective in diagnosing periprosthetic hip infection. Level of Evidence II, Prospective Cross-Sectional Study

RESUMO Objetivo: Avaliar a sensibilidade, especificidade, acurácia, valor preditivo positivo e valor preditivo negativo dos métodos diagnósticos aspirado articular pré-operatório (AAPO), biópsia percutânea de tecidos periarticulares (BPTP) e ambos associados na infecção pós-artroplastia total de quadril (IPATQ). Métodos: Trata-se de um estudo transversal (Nível de Evidência II) com coleta prospectiva de dados obtidos de 29 pacientes submetidos a AAPO com BPTP no Instituto Nacional de Ortopedia e Traumatologia durante o período de setembro de 2015 à janeiro de 2016. Foram comparados os resultados das análises microbiológicas dos espécimes obtidos por meio da BPTP e do AAPO com os obtidos intraoperatoriamente nas cirurgias subsequentes de revisão das artroplastias. Resultados: Encontramos uma sensibilidade da AAPO, BPTP e ambos, respectivamente de 78%, 73%, 89%, uma especificidade de 72%, 90%, 94% e uma acurácia de 76%, 80%, 90%. Conclusões: O procedimento de AAPO com BPTP para diagnóstico de infecção periprotética de quadril é sensível, específico e eficaz. Nível de Evidência II, Estudo Transversal Prospectivo.

LPS-induced immunomodulation and hormonal variation over time in toads.

The inflammatory response is a complex process that relies on interactions among multiple endocrine and immune modulators. Studies incorporating time-related and integrative endocrine and immune responses to an immune challenge might shed light on the characterization of the phases of the inflammatory response in anurans. The present study investigated time-related changes (1, 3, 6, and 18 h post-challenge) in plasma corticosterone (CORT), melatonin (MEL) and testosterone (T) levels, phagocytosis percentage (PP), plasma bacterial killing ability (BKA), and neutrophil to lymphocyte ratio (NLR) following a lipopolysaccharide (LPS) immune challenge in Rhinella diptycha toads. Our results showed the response to LPS injection was characterized by increased CORT, PP, BKA, and NLR, with a concomitant decrease in plasma MEL and T. Increased CORT was more pronounced at 6 and 18 h, while increased NLR was observed only 18 h post-LPS injection. Meanwhile, plasma MEL and T decreased independently of the time post-LPS injection. Additionally, toads in better body condition showed higher BKA and PP in the LPS-treated group, regardless of the time postinjection. Our results show that toads (R. diptycha) were sensitive to the LPS challenge, mounting an inflammatory response, which started quickly (after 1 h) and developed over time and was influenced by body condition. These results demonstrate a time-related hormonal and immune variation as a consistent pattern of activation of the immune system, as well as of hypothalamic-pituitary-adrenal/interrenal and immune-pineal axes following an immune challenge more deeply studied in mammals, suggesting the evolutionary conservation of the regulatory mechanisms for tetrapod vertebrates.

Angioarchitecture of collateral arteries of the aortic arch of Antillean manatee (Trichechus manatus manatus Linnaeus, 1758).

The aortic arrangement is an important structure associated with the maintenance of homeostasis. Based on this information, this study was conducted to describe the collateral arteries of the aortic arch of Antillean manatee and define the standard model for the species. Three specimens, an adult male, adult female and a male neonate, all strandings on the coast of the state of Rio Grande do Norte, Brazil, were used. The study was performed in the Laboratory of Morphophysiology of Vertebrates of the Federal University of Rio Grande do Norte, where in situ photographs were taken to demonstrate their topography. Subsequently, their hearts were removed and fixed in 10% formaldehyde and after 72 hr were dissected and analysed, obtaining schematic drawings and photographs of the vascular arrangement. The aortic arch was represented by three collateral arteries identified as the brachiocephalic trunk, left common carotid artery and left subclavian artery. This arrangement was similar to that found for other sirenians, and yet, for other mammals like hooded seal, murine, margarita island capuchin, black-handed tamarin, Mongolian gerbil and human. The morphological similarity presented in this study with different species of mammals, including humans, may contribute valuable information from an evolutionary point of view.

STAT1-NFκB crosstalk triggered by interferon gamma regulates noradrenaline-induced pineal hormonal production.

Melatonin production by pineal glands is modulated by several immune signals. The nuclear translocation of nuclear factor kappa-B (NFκB) homodimers, lacking transactivation domains, once induced by lipopolysaccharide (LPS) or tumor necrosis factor (TNF), inhibits the expression of Aanat gene and the synthesis of noradrenaline (NA)-induced melatonin. Interferon gamma (IFN-γ), on the other hand, increases melatonin synthesis. Furthermore, this cytokine activates the signal transducer as well as the activator of transcription 1 (STAT1) pathway, which was never evaluated as a melatonin synthesis modulator before. Reports demonstrated that IFN-γ might also activate NFκB. The present study evaluated the role of STAT1-NFκB crosstalk triggered by IFN-γ regarding the regulation of NA-induced pineal glands' hormonal production. Moreover, IFN-γ treatment increased NA-induced Aanat transcription, in addition to the synthesis of N-acetylserotonin (NAS) and melatonin. These effects were associated with STAT1 nuclear translocation, confirmed by the co-immunoprecipitation of STAT1 and Aanat promoter. Pharmacological STAT1 enhancement augmented NA-induced Aanat transcription as well as NAS and melatonin production. Additionally, IFN-γ induced the nuclear translocation of RelA-NFκB subunits. The blockade of this pathway prevented IFN-γ effects on the pineal function. The present data show that STAT1 and NFκB crosstalk controls melatonin production through a synergistic mechanism, disclosing a new integrative mechanism regarding pineal hormonal activity control.

PIP4K2A and PIP4K2C transcript levels are associated with cytogenetic risk and survival outcomes in acute myeloid leukemia.

Phosphoinositide signaling pathway orchestrates primordial molecular and cellular functions in both healthy and pathologic conditions. Phosphatidylinositol-5-phosphate 4-kinase type 2 lipid kinase (PIP4K2) family, which compromises PIP4K2A, PIP4K2B and PIP4K2C, has drawn the attention in human cancers. Particularly in hematological malignancies, PIP4K2A was already described as an essential protein for a malignant phenotype, although the clinical and biological impact of PIP4K2B and PIP4K2C proteins have not being explored in the same extent. In the present study, we investigated the impact on clinical outcomes and gene network of PIP4K2A, PIP4K2B and PIP4K2C mRNA transcripts in acute myeloid leukemia (AML) patients included in The Cancer Genome Atlas (2013) study. Our results indicate that PIP4K2A and PIP4K2C, but not PIP4K2B, mRNA levels were significantly reduced in AML patients assigned to the favorable risk group (p < 0.05) and low levels of PIP4K2A and PIP4K2C positively affect clinical outcomes of AML patients (p < 0.05). Gene set enrichment analyses indicate that the expression of PIP4K2 genes is associated with biological process such as signal transduction, metabolism of RNA and genomic instability related-gene sets. In summary, our study provides additional evidence of the involvement of members of the PIP4K2 family, in particular PIP4K2A and PIP4K2C, in AML.

Night work effects on salivary cytokines TNF, IL-1ß and IL-6.

Shift work is unavoidable in modern societies, but at the same time disrupts biological rhythms and contributes to social distress and disturbance of sleep, health and well-being of shift workers. Shift work has been associated with some chronic diseases in which a chronic inflammatory condition may play a role. However, few studies investigating the association of cytokine and other inflammation markers with shift workers have been published in recent years. In this study we evaluated the effects of permanent night work on the production of tumor necrosis factor (TNF), interleukin-1ß (IL-1ß), IL-6 and melatonin in saliva. Another aim was to demonstrate the benefit of the use of salivary cytokines for studies in chronobiology, since it is an easy and non-invasive method that allows for sampling at several times. Thirty-eight healthy male workers, being 21 day workers and 17 night workers, agreed to participate in this study. Sleep was evaluated by actigraphy and activity protocols. Saliva was collected during three workdays approximately at the middle of the work shift and at bed and wake times of the main sleep episode. Saliva samples were then analyzed by enzyme-linked immunosorbent assay to measure TNF, IL-1ß, IL-6 and melatonin levels, and the results were submitted to non-parametric statistical analysis. The use of saliva instead of blood allowed for a greater number of samples from the same subjects, allowing identifying alterations in the daily production patterns of salivary cytokines TNF, IL-1ß and IL-6 that probably are linked to night work. Salivary TNF and IL-1ß levels were similar for day and night workers, with higher daily production after awakening, in the morning hours for day workers and in the afternoon for night workers. Both groups presented a significant daily variation pattern of these two cytokines. Day and night workers produced similar amounts of salivary IL-6. Nevertheless, the daily variation pattern observed among day workers, with a peak after awakening, was absent among night workers. Thus, in our study, night workers showed partially adjusted daily variation patterns for salivary TNF and IL-1ß, not seen for salivary IL-6. Results for salivary IL-6 could be better explained as a consequence of circadian disruption due to permanent night work. Our results suggest that the whole circadian system, including clocks and pineal gland, is involved in regulating cytokine profile in shift workers and that a coordinated production of these cytokines, important for an adequate inflammatory response, could be disturbed by shift work. The distinct effects that shift work may have on different cytokines could give some cues about the mechanisms involved in this association.

Desempenho da esterase leucocitária no diagnóstico rápido da infecção periprotética de quadril / Performance of leucocitary esterase in the rapid diagnosis of hip periprotetic infection

A infecção articular periprotética (IAP) é uma das complicações mais frequentes e graves da artroplastia total de quadril (ATQ). A presença de IAP requer a adoção de medidas terapêuticas específicas, frequentemente havendo necessidade de cirurgia de revisão. O diagnóstico preciso e o tratamento adequado são imprescindíveis para evitar a perda funcional e a evolução para infecção sistêmica. Entretanto, o diagnóstico correto da IAP é um desafio, uma vez que ainda não há um método diagnóstico padrão ouro, e os critérios diagnósticos existentes se baseiam em achados cirúrgicos e testes sorológicos, histológicos e microbiológicos, que são imprecisos e demorados. Entre os exames preconizados, a pesquisa de esterase leucocitária (PEL) tem as vantagens de fornecer resultados rápidos, "à beira do leito" ("point of care"), e ser de baixo custo, além de apresentar boa acurácia, de acordo com estudos internacionais. Seu desempenho na população brasileira, no entanto, é pouco conhecido. Dessa forma, este estudo se propõe a avaliar o desempenho da PEL para identificação da IAP pós ATQ nos pacientes submetidos à cirurgia de revisão da ATQ no INTO. Foram incluídos os pacientes nos quais foi possível coletar líquido sinovial (LS) durante o procedimento cirúrgico. Para diagnóstico de IAP foram utilizados os critérios preconizados pela Sociedade de Infecção Musculoesquelética. O valor diagnóstico da PEL foi comparado aos exames de cultura microbiológico, avaliação histopatológica da membrana periprotética, velocidade de hemossedimentação, quantificação de proteína C reativa no plasma, contagem de células brancas no LS e percentual de polimorfonucleares no LS. Foram incluídos no estudo 53 pacientes, sendo 25 (54,3%) homens e 21 (45,6%) mulheres. Dezessete pacientes tiveram diagnóstico de IAP confirmado, dos quais quinze tiveram crescimento de microrganismos na cultura do tecido periprotético. Foi possível realizar a PEL em 42 pacientes, 15 do grupo infecção e 27 do grupo não infecção. A PEL do líquido sinovial apresentou valores de sensibilidade e especificidade de 53% e 100%, respectivamente. A sensibilidade da PEL foi inferior apenas à sensibilidade do teste microbiológico. A baixa sensibilidade encontrada sugere que a PEL não deve ser utilizada como exame de triagem de IAP, já que seu valor preditivo negativo baixo (79,4%) não permite que o resultado negativo seja utilizado para excluir a hipótese de infecção. A especificidade e o valor preditivo positivo alto, aliados à rapidez, baixo custo e possibilidade de ser utilizada no intra-operatório, faz da PEL um exame bastante útil dentro do arsenal diagnóstico da IAP. Quando positiva, a PEL confirma o diagnóstico de infecção, sendo, portanto, capaz de mudar a conduta do cirurgião durante uma cirurgia de revisão classificada inicialmente como asséptica, de acordo com a avaliação pré-operatória. Desta forma recomendamos o uso rotineiro da PEL em todas as revisões aparentemente assépticas de quadril

Periprosthetic Joint Infection (PJI) is one of the most frequent and severe complications of total hip arthroplasty (THA). The presence of PJI requires the adoption of specific therapeutic measures, often requiring revision surgery. Accurate diagnosis and adequate treatment are essential to restore patient function and inhibit evolution to systemic infection. However, the correct diagnosis of PJI is challenging, since there is not such a gold standard, highly accurate, diagnostic method for this disease. PJI diagnosis is currently based on the combination of surgical findings and serological, histological and microbiological tests that are imprecise and time consuming. Among the exams recommended, the test of leukocyte esterase (TLE) has the advantages of providing quick results, at bed side ("point of care"), with low cost, besides having good accuracy, according to international studies. Its performance in the Brazilian population, however, is unknown. Thus, this study proposes to evaluate the performance of the TLE for the identification of the PJI post THA in patients submitted to revision surgery of THA, treated at INTO. Patients in whom was possible to collect synovial fluid (SF) during the surgical procedure were included. The criteria recommended by the Musculoskeletal Infection Society were used for the diagnosis of PJI. The diagnostic value of TLE was compared to microbiological culture, histopathological evaluation of periprosthetic membrane, erythrocyte sedimentation rate, plasma C-reactive protein quantification, white cell count in SF and percentage of polymorphonuclear in SF. The study included 53 patients, of which 25 (54.3%) were men and 21 (45.6%) were women. Seventeen patients had the diagnosis of infection confirmed, of which fifteen had growth of microorganisms in the periprosthetic tissue culture. It was possible to perform TLE in 42 patients, 15 of the infection group and 27 of the non-infection group. The TLE presented values of sensitivity and specificity of 57% and 100%, respectively. The sensitivity of TLE was significantly reduced in comparison to sensitivity of microbiological test. The low sensitivity suggests that TLE should not be used as an IAP screening test, since its low negative predictive value (79.4%) does not allow that the negative result lead to the exclusion of the hypothesis of infection. However, specificity and high positive predictive value, combined with the rapidity, low cost and possibility of being used intraoperatively, make TLE a very useful exam within the diagnostic arsenal of PJI. When positive, TLE confirms the diagnosis of infection and is, therefore, able to change the surgeon's behavior during revision surgery initially classified as aseptic, according to the preoperative evaluation. In this way, we recommend the routine use of TLE in all apparently aseptic hip revisions

The occurrence of aflatoxigenic Aspergillus spp. in dairy cattle feed in Southern Brazil

ABSTRACT The presence of mycotoxins or related fungi in animal feed is a major problem for animal and human health. Silage and concentrated feed samples were collected from 21 dairy farms in the Western part of Paraná state in Southern Brazil. Water activity and pH of all samples were measured, and each sample was analyzed to check for the presence of aflatoxigenic Aspergillus. Water activity was observed to be lower in the concentrated feed samples. The pH was lower in the silage samples, indicating fermentation processes. Two silage samples and four concentrated feed samples were contaminated with Aspergillus spp. Seven isolates of Aspergillus spp. were obtained and their potential to produce aflatoxins was evaluated. Four of the isolates, two from the silage samples and two from the concentrated feed samples, produced the aflatoxins B1, B2, G1, and G2 in culture media. These isolates were identified as Aspergillus parasiticus and Aspergillus nomius. The presence of aflatoxigenic isolates of Aspergillus spp. in silage and concentrated feed samples is a matter of concern, because of the risk of aflatoxin production and contamination of the animal feed.

Interplay among steroids, body condition and immunity in response to long-term captivity in toads.

Stressful experiences can promote harmful effects on physiology and fitness. However, stress-mediated hormonal and immune changes are complex and may be highly dependent on body condition. Here, we investigated captivity-associated stress effects, over 7, 30, 60, and 90 days on plasma corticosterone (CORT) and testosterone (T) levels, body index, and innate immunity (bacterial killing ability and phagocytosis of peritoneal cells) in toads (Rhinella icterica). Toads in captivity exhibited elevated CORT and decreased T and immunity, without changes in body index. The inter-relationships between these variables were additionally contrasted with those obtained previously for R. schneideri, a related species that exhibited extreme loss of body mass under the same captive conditions. While T and phagocytosis were positively associated in both species, the relationship between CORT and bacterial killing ability was dependent on body index alterations. While CORT and bacterial killing ability were positively associated for toads that maintained body index, CORT was negatively associated with body index in toads that lost body mass over time in captivity. In these same toads, body index was positively associated with bacterial killing ability. These results demonstrate that steroids-immunity inter-relationships arising from prolonged exposure to a stressor in toads are highly dependent on body condition.

The occurrence of aflatoxigenic Aspergillus spp. in dairy cattle feed in Southern Brazil.

The presence of mycotoxins or related fungi in animal feed is a major problem for animal and human health. Silage and concentrated feed samples were collected from 21 dairy farms in the Western part of Paraná state in Southern Brazil. Water activity and pH of all samples were measured, and each sample was analyzed to check for the presence of aflatoxigenic Aspergillus. Water activity was observed to be lower in the concentrated feed samples. The pH was lower in the silage samples, indicating fermentation processes. Two silage samples and four concentrated feed samples were contaminated with Aspergillus spp. Seven isolates of Aspergillus spp. were obtained and their potential to produce aflatoxins was evaluated. Four of the isolates, two from the silage samples and two from the concentrated feed samples, produced the aflatoxins B1, B2, G1, and G2 in culture media. These isolates were identified as Aspergillus parasiticus and Aspergillus nomius. The presence of aflatoxigenic isolates of Aspergillus spp. in silage and concentrated feed samples is a matter of concern, because of the risk of aflatoxin production and contamination of the animal feed.

Captivity effects on immune response and steroid plasma levels of a Brazilian toad (Rhinella schneideri).

Stressors can increase plasma glucocorticoid (GC) levels and decrease plasma androgen levels in different species of vertebrates. GCs can have immune-enhancing or immunosuppressive effects, which are dependent upon stress duration and intensity. The worldwide decline in amphibian populations is strongly linked to an array of different stressors. The impacts of stress on GCs, androgens, and the immune response are important to clarify and should lead to the better development of conservation strategies. The present study in adult male toads of Rhinella schneideri investigated the effects of captivity (7, 30, and 60 days) on plasma corticosterone (CORT) and plasma testosterone (T), as well as innate immune responses, specifically humoral and cell mediated responses, as indicated by bacterial killing ability (BKA) and phagocytosis by peritoneal cells, respectively. Captivity increased CORT threefold and decreased T versus controls. CORT maintained a threefold elevation throughout the captivity period, while body mass and T gradually decreased with time in captivity. BKA was lower at day 30, versus days 7 and 60, while peritoneal cell phagocytic efficiency decreased after day 30, remaining low at day 60. Moreover, phagocytosis efficiency was positively associated with T and body condition, suggesting that the effects of chronic stress on reproductive potential and immune response might be associated with the state of energetic reserves.

Melatoninergic System in Parkinson's Disease: From Neuroprotection to the Management of Motor and Nonmotor Symptoms.

Melatonin is synthesized by several tissues besides the pineal gland, and beyond its regulatory effects in light-dark cycle, melatonin is a hormone with neuroprotective, anti-inflammatory, and antioxidant properties. Melatonin acts as a free-radical scavenger, reducing reactive species and improving mitochondrial homeostasis. Melatonin also regulates the expression of neurotrophins that are involved in the survival of dopaminergic neurons and reduces α-synuclein aggregation, thus protecting the dopaminergic system against damage. The unbalance of pineal melatonin synthesis can predispose the organism to inflammatory and neurodegenerative diseases such as Parkinson's disease (PD). The aim of this review is to summarize the knowledge about the potential role of the melatoninergic system in the pathogenesis and treatment of PD. The literature reviewed here indicates that PD is associated with impaired brain expression of melatonin and its receptors MT1 and MT2. Exogenous melatonin treatment presented an outstanding neuroprotective effect in animal models of PD induced by different toxins, such as 6-hydroxydopamine (6-OHDA), 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP), rotenone, paraquat, and maneb. Despite the neuroprotective effects and the improvement of motor impairments, melatonin also presents the potential to improve nonmotor symptoms commonly experienced by PD patients such as sleep and anxiety disorders, depression, and memory dysfunction.