Toll-like receptors 2, 4, and 9 modulate promoting effect of COPD-like airway inflammation on K-ras-driven lung cancer through activation of the MyD88/NF-ĸB pathway in the airway epithelium.

Introduction: Toll-like receptors (TLRs) are an extensive group of proteins involved in host defense processes that express themselves upon the increased production of endogenous damage-associated molecular patterns (DAMPs) and pathogen-associated molecular patterns (PAMPs) due to the constant contact that airway epithelium may have with pathogenic foreign antigens. We have previously shown that COPD-like airway inflammation induced by exposure to an aerosolized lysate of nontypeable Haemophilus influenzae (NTHi) promotes tumorigenesis in a K-ras mutant mouse model of lung cancer, CCSPCre/LSL-K-rasG12D (CC-LR) mouse. Methods: In the present study, we have dissected the role of TLRs in this process by knocking out TLR2, 4, and 9 and analyzing how these deletions affect the promoting effect of COPD-like airway inflammation on K-ras-driven lung adenocarcinoma. Results: We found that knockout of TLR 2, 4, or 9 results in a lower tumor burden, reduced angiogenesis, and tumor cell proliferation, accompanied by increased tumor cell apoptosis and reprogramming of the tumor microenvironment to one that is antitumorigenic. Additionally, knocking out of downstream signaling pathways, MyD88/NF-κB in the airway epithelial cells further recapitulated this initial finding. Discussion: Our study expands the current knowledge of the roles that TLR signaling plays in lung cancer, which we hope, can pave the way for more reliable and efficacious prevention and treatment modalities for lung cancer.

An FBN1 deep intronic variant is associated with pseudoexon formation and a variable Marfan phenotype in a five generation family.

Exome sequencing of genes associated with heritable thoracic aortic disease (HTAD) failed to identify a pathogenic variant in a large family with Marfan syndrome (MFS). A genome-wide linkage analysis for thoracic aortic disease identified a peak at 15q21.1, and genome sequencing identified a novel deep intronic FBN1 variant that segregated with thoracic aortic disease in the family (LOD score 2.7) and was predicted to alter splicing. RT-PCR and bulk RNA sequencing of RNA harvested from fibroblasts explanted from the affected proband revealed an insertion of a pseudoexon between exons 13 and 14 of the FBN1 transcript, predicted to lead to nonsense mediated decay (NMD). Treating the fibroblasts with an NMD inhibitor, cycloheximide, greatly improved the detection of the pseudoexon-containing transcript. Family members with the FBN1 variant had later onset aortic events and fewer MFS systemic features than typical for individuals with haploinsufficiency of FBN1. Variable penetrance of the phenotype and negative genetic testing in MFS families should raise the possibility of deep intronic FBN1 variants and the need for additional molecular studies.

Prevalence and Factors Associated with Anxiety and Depression in Peruvian Medical Students.

During medical training, students are exposed to stressors that deteriorate mental health. This study determined depression and anxiety prevalence and their associated factors in students from four medical schools in northern Peru. A cross-sectional study was conducted among medical students from Lambayeque, Peru. The Goldberg Anxiety and Zung Depression tests were applied. The dependent variables were depression and anxiety, and their association with covariates (age, sex, type of university, socioeconomic level, experience, family problem, and physical activity) was evaluated. Prevalence ratios were calculated using generalized linear models. Of 482 students, the prevalence of anxiety was 61.8% and depression was 22.0%. A severe level of anxiety was observed in 6.2% in the group between 16 and 20 years old. It was observed that private university students had higher frequencies of having depression (PR = 2.01) and anxiety (PR = 1.35); males had a lower risk of anxiety (PR = 0.82), but higher risk of depression compared to females (PR = 1.45). Performing physical activity decreased the prevalence of depression (PR = 0.53); however, it increased the frequency of anxiety (PR = 1.26). Having family problems increased the prevalence of anxiety (PR = 1.26). Medical students from private universities experienced higher rates of depression and anxiety. Depression and anxiety were associated with gender and physical activity. These findings highlight the importance of mental health promotion due to its link with quality of life and academic performance.

Recognizing distant faces.

As an 'early alerting' sense, one of the primary tasks for the human visual system is to recognize distant objects. In the specific context of facial identification, this ecologically important task has received surprisingly little attention. Most studies have investigated facial recognition at short, fixed distances. Under these conditions, the photometric and configural information related to the eyes, nose and mouth are typically found to be primary determinants of facial identity. Here we characterize face recognition performance as a function of viewing distance and investigate whether the primacy of the internal features continues to hold across increasing viewing distances. We find that exploring the distance dimension reveals a qualitatively different salience distribution across a face. Observers' recognition performance significantly exceeds that obtained with the internal facial physiognomy, and also exceeds the computed union of performances with internal and external features alone, suggesting that in addition to the mutual configuration of the eyes, nose and mouth, it is the relationships between these features and external head contours that are crucial for recognition. We have also conducted computational studies with convolutional neural networks trained on the task of face recognition to examine whether this representational bias could emerge spontaneously through exposure to faces. The results provide partial support for this possibility while also highlighting important differences between the human and artificial system. These findings have implications for the nature of facial representations useful for a visual system, whether human or machine, for recognition over large and varying distances.

Convolutional Neural Network Applied to SARS-CoV-2 Sequence Classification.

COVID-19, the illness caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) virus belonging to the Coronaviridade family, a single-strand positive-sense RNA genome, has been spreading around the world and has been declared a pandemic by the World Health Organization. On 17 January 2022, there were more than 329 million cases, with more than 5.5 million deaths. Although COVID-19 has a low mortality rate, its high capacities for contamination, spread, and mutation worry the authorities, especially after the emergence of the Omicron variant, which has a high transmission capacity and can more easily contaminate even vaccinated people. Such outbreaks require elucidation of the taxonomic classification and origin of the virus (SARS-CoV-2) from the genomic sequence for strategic planning, containment, and treatment of the disease. Thus, this work proposes a high-accuracy technique to classify viruses and other organisms from a genome sequence using a deep learning convolutional neural network (CNN). Unlike the other literature, the proposed approach does not limit the length of the genome sequence. The results show that the novel proposal accurately distinguishes SARS-CoV-2 from the sequences of other viruses. The results were obtained from 1557 instances of SARS-CoV-2 from the National Center for Biotechnology Information (NCBI) and 14,684 different viruses from the Virus-Host DB. As a CNN has several changeable parameters, the tests were performed with forty-eight different architectures; the best of these had an accuracy of 91.94 ± 2.62% in classifying viruses into their realms correctly, in addition to 100% accuracy in classifying SARS-CoV-2 into its respective realm, Riboviria. For the subsequent classifications (family, genera, and subgenus), this accuracy increased, which shows that the proposed architecture may be viable in the classification of the virus that causes COVID-19.

Targeting IL-1ß as an immunopreventive and therapeutic modality for K-ras-mutant lung cancer.

K-ras-mutant lung adenocarcinoma (KM-LUAD) is associated with abysmal prognosis and is tightly linked to tumor-promoting inflammation. A human mAb, canakinumab, targeting the proinflammatory cytokine IL-1ß, significantly decreased the risk of lung cancer in the Canakinumab Anti-inflammatory Thrombosis Outcomes Study. Interestingly, we found high levels of IL-1ß in the lungs of mice with K-rasG12D-mutant tumors (CC-LR mice). Here, we blocked IL-1ß using an anti-IL-1ß mAb in cohorts of 6- or 14-week-old CC-LR mice to explore its preventive and therapeutic effect, respectively. IL-1ß blockade significantly reduced lung tumor burden, which was associated with reprogramming of the lung microenvironment toward an antitumor phenotype characterized by increased infiltration of cytotoxic CD8+ T cells (with high IFN-γ and granzyme B expression but low programmed cell death 1 [PD-1] expression) while suppressing neutrophils and polymorphonuclear (PMN) myeloid-derived suppressor cells. When querying the Cancer Genome Atlas data set, we found positive correlations between IL1B expression and infiltration of immunosuppressive PMNs and expression of their chemoattractant, CXCL1, and PDCD1 expressions in patients with KM-LUAD. Our data provide evidence that IL-1ß blockade may be a preventive strategy for high-risk individuals and an alternative therapeutic approach in combination with currently available treatments for KM-LUAD.

Macroscopic Distribution, Histopathology and Viral Antigen Expression in Dogs with Canine Distemper Virus-induced Hyperkeratosis in Nasodigital and Other Regions.

Cutaneous hyperkeratosis is one of the many clinicopathological manifestations of canine distemper and is characterized by thickening and hardening of the skin, predominantly in nasodigital areas. Although this lesion may rarely affect other regions, this has been poorly characterized. Twelve dogs with canine distemper and cutaneous hyperkeratosis, necropsied at an anatomical pathology service, were investigated. Twenty-two cutaneous hyperkeratotic foci were observed on footpads (11/22), nasal planum (3/22), haired skin on the snout (2/22), periocular region (2/22), ventral abdomen (2/22), scrotum (1/22) and vulva (1/22). The dogs had one (5/12), two (4/12) or three (3/12) regions concomitantly affected. Orthokeratotic hyperkeratosis was a predominant histopathological feature in 17 dogs, occasionally accompanied by other lesions, including inclusion bodies (14/17), epidermal hyperplasia (9/17) and keratinocyte hydropic degeneration (6/17). Canine distemper virus antigen was expressed in at least one skin lesion in 10 dogs. Fourteen (14/17) hyperkeratotic foci were immunopositive while three (3/17) were immunonegative. Viral antigen expression was most common in the sweat glands (13/17), epidermis (11/17) and vascular endothelial cells or pericytes (8/17). Histological findings and antigen detection were similar among nasodigital and other regions. We emphasize the importance of clinicopathological recognition of these lesions for the initial suspicion of canine distemper, thereby facilitating early treatment.

Histopathological Features of Spinal Cord Lesions in Dogs with Distemper-Associated Demyelinating Leucoencephalomyelitis.

Demyelinating leucoencephalomyelitis (DLEM) is one of the pathological presentations of canine distemper, but its histological characteristics and topographical distribution of spinal cord injuries have been poorly explored. Seventeen dogs submitted for necropsy to a veterinary pathology service (2006-2008) and diagnosed with distemper-associated DLEM were investigated. Seventy-two of 231 spinal cord sections from these animals had histological lesions, which consistently affected the white matter. The lumbosacral region was mostly affected (13/17), followed by the thoracolumbar (11/17), cervical (9/17) and cervicothoracic (9/17) regions. Among the 72 affected cord sections, lesions in the lateral (42/72) and dorsal funiculi (31/72) were the most common. Demyelination (17/17), astrocytosis (17/17), microgliosis (17/17), gemistocytes (11/17) and non-suppurative inflammation (10/17) were frequently seen in the white matter. Grey matter changes were less common, and included gliosis (8/17), non-suppurative inflammation (7/17) and malacia (5/17). Acute lesions were most prevalent (13/17) but it was common (10/17) for the same dog to have lesions at different stages of evolution in distinct spinal cord regions. Pathological changes in several spinal cords did not correlate with the reported clinical signs and some dogs did not present with spinal cord-related clinical signs. Our results highlight important aspects of the distribution and morphology of spinal cord lesions in dogs with distemper-associated DLEM, and may assist clinicians and pathologists in the diagnosis of spontaneous cases of this important infectious disease and contribute to further studies concerning distemper pathogenesis.

Superconducting Long-Range Proximity Effect through the Atomically Flat Interface of a Bi2Te3 Topological Insulator.

We report on structural and electronic properties of superconducting nanohybrids made of Pb grown in the ultrahigh vacuum on the atomically clean surface of single crystals of topological Bi2Te3. In situ scanning tunneling microscopy and spectroscopy demonstrated that the resulting network is composed of Pb-nanoislands dispersed on the surface and linked together by an amorphous atomic layer of Pb, which wets Bi2Te3. As a result, the superconducting state of the system is characterized by a thickness-dependent superconducting gap of Pb-islands and by a very unusual position-independent proximity gap between them. Furthermore, the data analysis and DFT calculations demonstrate that the Pb-wetting layer leads to significant modifications of both topological and trivial electronic states of Bi2Te3, which are responsible for the observed long-range proximity effect.

Interplay between estrogen and Stat3/NF-κB-driven immunomodulation in lung cancer.

K-ras mutant lung adenocarcinoma (LUAD) is the most common type of lung cancer, displays abysmal prognosis and is tightly linked to tumor-promoting inflammation, which is increasingly recognized as a target for therapeutic intervention. We have recently shown a gender-specific role for epithelial Stat3 signaling in the pathogenesis of K-ras mutant LUAD. The absence of epithelial Stat3 in male K-ras mutant mice (LR/Stat3Δ/Δ mice) promoted tumorigenesis and induced a nuclear factor-kappaB (NF-κB)-driven pro-tumor immune response while reducing tumorigenesis and enhancing anti-tumor immunity in female counterparts. In the present study, we manipulated estrogen and NF-κB signaling to study the mechanisms underlying this intriguing gender-disparity. In LR/Stat3Δ/Δ females, estrogen deprivation by bilateral oophorectomy resulted in higher tumor burden, an induction of NF-κB-driven immunosuppressive response, and reduced anti-tumor cytotoxicity, whereas estrogen replacement reversed these changes. On the other hand, exogenous estrogen in males successfully inhibited tumorigenesis, attenuated NF-κB-driven immunosuppression and boosted anti-tumor immunity. Mechanistically, genetic targeting of epithelial NF-κB activity resulted in reduced tumorigenesis and enhanced the anti-tumor immune response in LR/Stat3Δ/Δ males, but not females. Our data suggest that estrogen exerts a context-specific anti-tumor effect through inhibiting NF-κB-driven tumor-promoting inflammation and provide insights into developing novel personalized therapeutic strategies for K-ras mutant LUAD.

The future of bioenergy.

Energy from biomass plays a large and growing role in the global energy system. Energy from biomass can make significant contributions to reducing carbon emissions, especially from difficult-to-decarbonize sectors like aviation, heavy transport, and manufacturing. But land-intensive bioenergy often entails substantial carbon emissions from land-use change as well as production, harvesting, and transportation. In addition, land-intensive bioenergy scales only with the utilization of vast amounts of land, a resource that is fundamentally limited in supply. Because of the land constraint, the intrinsically low yields of energy per unit of land area, and rapid technological progress in competing technologies, land intensive bioenergy makes the most sense as a transitional element of the global energy mix, playing an important role over the next few decades and then fading, probably after mid-century. Managing an effective trajectory for land-intensive bioenergy will require an unusual mix of policies and incentives that encourage appropriate utilization in the short term but minimize lock-in in the longer term.

Opioid Analgesics Do Not Improve Abdominal Pain or Quality of Life in Crohn's Disease.

BACKGROUND: Abdominal pain and opioid analgesic use are common in Crohn's disease (CD). AIMS: We sought to identify factors associated with abdominal pain in CD and evaluate the impact of opioid analgesics on pain and quality-of-life scores in this setting. METHODS: We performed a longitudinal cohort study using a prospective, consented IBD natural history registry from a single academic center between 2009 and 2013. Consecutive CD patients were followed for at least 1 year after an index visit. Data were abstracted regarding pain experience (from validated surveys), inflammatory activity (using endoscopic/histologic findings), laboratory studies, coexistent psychiatric disorders, medical therapy, opioid analgesic, and tobacco use. RESULTS: Of 542 CD patients (56.6% women), 232 (42.8%) described abdominal pain. Individuals with pain were more likely to undergo surgery and were more frequently prescribed analgesics and/or antidepressants/anxiolytics. Elevated ESR (OR 1.79; 95%CI 1.11-2.87), coexistent anxiety/depression (OR 1.87; 95%CI 1.13-3.09), smoking (OR 2.08; 95%CI 1.27-3.40), and opioid use (OR 2.46; 95%CI 1.33-4.57) were independently associated with abdominal pain. Eighty patients (14.8%) were prescribed opioids, while 31 began taking them at or after the index visit. Patients started on opioids demonstrated no improvement in abdominal pain or quality-of-life scores on follow-up compared to patients not taking opioids. CONCLUSIONS: Abdominal pain is common in CD and is associated with significant opioid analgesic utilization and increased incidence of anxiety/depression, smoking, and elevated inflammatory markers. Importantly, opioid use in CD was not associated with improvement in pain or quality-of-life scores. These findings reinforce the limitations of currently available analgesics in IBD and support exploration of alternative therapies.

Reliability and validity of body weight and body image perception in children and adolescents from the South American Youth/Child Cardiovascular and Environmental (SAYCARE) Study.

OBJECTIVE: To assess the reliability and validity of body weight (BW) and body image (BI) perception reported by parents (in children) and by adolescents in a South American population. DESIGN: Cross-sectional study. BW perception was evaluated by the question, 'Do you think you/your child are/is: severely wasted, wasted, normal weight, overweight, obese?' BI perception was evaluated using the Gardner scale. To evaluate reliability, BW and BI perceptions were reported twice, two weeks apart. To evaluate validity, the BW and BI perceptions were compared with WHO BMI Z-scores. Kappa and Kendall's tau-c coefficients were obtained. SETTING: Public and private schools and high schools from six countries of South America (Argentina, Peru, Colombia, Uruguay, Chile, Brazil).ParticipantsChildren aged 3-10 years (n 635) and adolescents aged 11-17 years (n 400). RESULTS: Reliability of BW perception was fair in children's parents (κ=0·337) and substantial in adolescents (κ=0·709). Validity of BW perception was slight in children's parents (κ=0·176) and fair in adolescents (κ=0·268). When evaluating BI, most children were perceived by parents as having lower weight. Reliability of BI perception was slight in children's parents (κ=0·124) and moderate in adolescents (κ=0·599). Validity of BI perception was poor in children's parents (κ=-0·018) and slight in adolescents (κ=0·023). CONCLUSIONS: Reliability of BW and BI perceptions was higher in adolescents than in children's parents. Validity of BW perception was good among the parents of the children and adolescents with underweight and normal weight.

Different Munc18 proteins mediate baseline and stimulated airway mucin secretion.

Airway mucin secretion is necessary for ciliary clearance of inhaled particles and pathogens but can be detrimental in pathologies such as asthma and cystic fibrosis. Exocytosis in mammals requires a Munc18 scaffolding protein, and airway secretory cells express all 3 Munc18 isoforms. Using conditional airway epithelial cell-deletant mice, we found that Munc18a has the major role in baseline mucin secretion, Munc18b has the major role in stimulated mucin secretion, and Munc18c does not function in mucin secretion. In an allergic asthma model, Munc18b deletion reduced airway mucus occlusion and airflow resistance. In a cystic fibrosis model, Munc18b deletion reduced airway mucus occlusion and emphysema. Munc18b deficiency in the airway epithelium did not result in any abnormalities of lung structure, particle clearance, inflammation, or bacterial infection. Our results show that regulated secretion in a polarized epithelial cell may involve more than one exocytic machine at the apical plasma membrane and that the protective roles of mucin secretion can be preserved while therapeutically targeting its pathologic roles.

Understanding the Complexity of the Tumor Microenvironment in K-ras Mutant Lung Cancer: Finding an Alternative Path to Prevention and Treatment.

Kirsten rat sarcoma viral oncogene (K-ras) is a well-documented, frequently mutated gene in lung cancer. Since K-ras regulates numerous signaling pathways related to cell survival and proliferation, mutations in this gene are powerful drivers of tumorigenesis and confer prodigious survival advantages to developing tumors. These malignant cells dramatically alter their local tissue environment and in the process recruit a powerful ally: inflammation. Inflammation in the context of the tumor microenvironment can be described as either antitumor or protumor (i.e., aiding or restricting tumor progression, respectively). Many current treatments, like immune checkpoint blockade, seek to augment antitumor inflammation by alleviating inhibitory signaling in cytotoxic T cells; however, a burgeoning area of research is now focusing on ways to modulate and mitigate protumor inflammation. Here, we summarize the interplay of tumor-promoting inflammation and K-ras mutant lung cancer pathogenesis by exploring the cytokines, signaling pathways, and immune cells that mediate this process.

Graduação histológica e aspectos clínico-patológicos relacionados em 22 meningiomas de cães / Histological grading and related clinical and pathological aspects of 22 canine meningioma

Vinte e dois casos de meningiomas em cães, diagnosticados num período de aproximadamente 18 anos, foram revisados. Os neoplasmas foram graduados e classificados histologicamente de acordo com os critérios da Organização Mundial da Saúde (OMS de 2007) para meningiomas em humanos adaptados para cães, em Grau I (G-I; benignos), Grau II (G-II; atípicos) e Grau III (G-III; anaplásico ou maligno). Dos protocolos de necropsias foram retiradas adicionalmente informações referentes ao sexo, idade, raça, evolução clínica, sinais clínicos, localização anatômica e achados macroscópicos. Os meningiomas intracranianos supratentoriais foram os mais frequentes em relação às demais localizações intracranianas ou intraespinhais. Os intracranianos caracterizaram-se principalmente por sinais clínicos de alteração tálamo-cortical. Os intraespinhais caracterizaram-se principalmente por causarem ataxia. Meningiomas G-I foram os mais frequentes (63,6%) nos 22 cães, seguidos pelos G-III (22,7%) e G-II (13,6%). Os G-I caracterizaram-se por ter o subtipo psammomatoso como o mais frequente, mais de um padrão morfológico em um mesmo tumor, e um terço deles apresentando áreas de invasão do tecido nervoso, 71,4% dos casos acometendo fêmeas, uma média de idade de 11 anos, cães com raça definida como os mais acometidos e por ter o maior tempo de sobrevivência após a manifestação dos sinais clínicos. Os meningiomas G-II caracterizaram-se por ter o subtipo cordoide como o mais frequente, invasão do tecido nervoso em um terço dos casos, somente fêmeas acometidas, uma média de idade de 12 anos, dois terços dos cães acometidos sem raça definida e o tempo máximo de sobrevivência de 20 dias. Os meningiomas G-III caracterizaram-se por ter o subtipo papilar como o mais frequente, invasão do tecido nervoso em 80% dos casos, 60% dos casos acometendo fêmeas, uma média de idade de 8 anos, 80% dos cães acometidos da raça Boxer e o tempo máximo de sobrevivência de 90 dias. Este estudo permitiu estabelecer uma relação entre os três graus histológicos observados em 22 casos de meningiomas em cães com vários parâmetros clínico-epidemiológicos e patológicos, fornecendo informações úteis para um melhor conhecimento da correlação entre a graduação histológica e a evolução clínica desses neoplasmas.(AU)

Twenty two cases of meningiomas in dogs, diagnosed in about 18 years, were analyzed. The neoplasms were histologicaly classified and graded according to the World and Health Organization (WHO of 2007) for human meningiomas, adapted for dogs, in Grade I (G-I; benign), Grade II (G-II; atypical), and Grade III (G-III; anaplastic or malignant). Additional data about gender, age, breed, skull conformation, clinical course and signs, anatomic localization, gross and histological findings were obtained from the necropsy reports. Intracranial and supratentorial meningiomas were the most frequent in relation to the other intracranial or intraspinal sites. The intracranial ones were characterized mainly by clinical signs of thalamic-cortical alteration. Intraspinal ones were mainly characterized by ataxia. G-I meningiomas were the most frequent (63.6%) in dogs, followed by G-III (22.7%) and G-II (13.6%). GI were characterized by having the psammomatous subtype as the most frequent, more than one morphological pattern in the same tumor, one third presenting areas of invasion of nervous tissue, 71.4% of cases involving females, a mean age of 11 years, pure breed dogs as the most affected ones and for having the longest survival time after the manifestation of clinical signs. G-II meningiomas were characterized by having the chordoid subtype as the most frequent, invasion of nervous tissue in one third of cases, only females affected, a mean age of 12 years, two-thirds of the dogs affected were mongrels and the maximum survival time of 20 days. The G-III meningiomas were characterized by having the papillary subtype as the most frequent, invasion of the nervous tissue in 80% of the cases, 60% of the cases involving females, a mean age of 8 years, 80% of dogs affected were Boxers and the maximum survival time of 90 days. In conclusion, this study allowed to establish a relationship between the three histological grades observed in 22 cases of meningiomas in dogs with various clinical-epidemiological and pathological parameters, providing useful information for a better understanding of the correlation between the histological grading and the clinical evolution of these neoplasms.(AU)

Caracterização histoquímica no diagnóstico da babesiose bovina por Babesia bovis / Histochemical characterization in the diagnosis of bovine babesiosis by Babesia bovis

A babesiose é uma doença hemolítica transmitida por carrapatos e causada por protozoários intraeritrocitários do gênero Babesia. Esta é uma doença de incidência elevada na Região Sul do Brasil e responsável por perdas econômicas consideráveis. O diagnóstico clínico-patológico pode ser feito através da demonstração do parasito pelo exame de esfregaços sanguíneos ou de tecidos frescos. Entretanto, com frequência, somente órgãos fixados em formol são remetidos para laboratórios de patologia, o que impossibilita a realização do esfregaço. O principal objetivo deste estudo foi encontrar técnicas histoquímicas alternativas e capazes de aprimorar a evidenciação de Babesia bovis intraeritrocitária em tecidos fixados em formol. Para este estudo, foram analisadas retrospectivamente amostras de tecidos de 50 casos de necropsias de bovinos, as quais haviam sido fixadas em formol e processadas rotineiramente para histopatologia. Os casos foram divididos em um grupo controle, constituído por 12 casos de babesiose cerebral com substância cinzenta encefálica róseo-cereja característica (grupo A), e 38 casos sugestivos de tristeza parasitária bovina (grupo B), conforme os protocolos de necropsias. Foram testadas as técnicas histoquímicas de Azul Alciano, Azul de Metileno, Azul de Toluidina, Giemsa, Gram (método de McCallum-Goodpasture), Grocott, Ácido Periódico de Schiff e Ziehl-Neelsen. Dentre estas, observou-se que as técnicas de Azul de Metileno e Azul de Toluidina permitiram observar características morfológicas e tintoriais de maneira mais nítida, auxiliando na identificação de B. bovis. Adicionalmente, foram estabelecidos vários parâmetros clínico-epidemiológicos e anatomopatológicos da babesiose por B. bovis.(AU)

Babesiosis is a hemolytic tick-borne disease caused by intraerythrocytic protozoal parasites of the genus Babesia. This is a disease of high incidence in the southern Brazil and responsible for considerable economic losses. Clinical-pathological diagnosis can be made by demonstrating the parasite by examining blood smears or fresh tissues. However, frequently, only formalin-fixed organs are sent to pathology laboratories, which makes it impossible to perform the smears. The main objective of this study was to find alternative histochemical techniques capable to improve the identification of intraerythrocytic Babesia bovis in histological sections. For this study, tissue samples from 50 bovine necropsy cases were retrospectively analyzed, which had been fixed in formalin and routinely processed for histopathology. The cases were divided into a control group, consisting of 12 cases of cerebral babesiosis with characteristic pink-cherry gray matter (group A), and 38 cases suggestive of cattle tick fever (group B), according to necropsy protocols. Histochemical techniques of Alcian Blue, Methylene Blue, Toluidine Blue, Giemsa, Gram (McCallum-Goodpasture method), Grocott, Periodic Acid of Schiff and Ziehl-Neelsen were tested. Among these, it was observed that the techniques of Methylene Blue and Toluidine Blue allowed to observe morphological and dye characteristics in a clearer way, aiding in the identification of B. bovis. In addition, several clinical-epidemiological and anatomopathological parameters of babesiosis caused by B. bovis were established.(AU)

Impact of prediagnostic smoking and smoking cessation on colorectal cancer prognosis: a meta-analysis of individual patient data from cohorts within the CHANCES consortium.

Background: Smoking has been associated with colorectal cancer (CRC) incidence and mortality in previous studies and might also be associated with prognosis after CRC diagnosis. However, current evidence on smoking in association with CRC prognosis is limited. Patients and methods: For this individual patient data meta-analysis, sociodemographic and smoking behavior information of 12 414 incident CRC patients (median age at diagnosis: 64.3 years), recruited within 14 prospective cohort studies among previously cancer-free adults, was collected at baseline and harmonized across studies. Vital status and causes of death were collected for a mean follow-up time of 5.1 years following cancer diagnosis. Associations of smoking behavior with overall and CRC-specific survival were evaluated using Cox regression and standard meta-analysis methodology. Results: A total of 5229 participants died, 3194 from CRC. Cox regression revealed significant associations between former [hazard ratio (HR) = 1.12; 95 % confidence interval (CI) = 1.04-1.20] and current smoking (HR = 1.29; 95% CI = 1.04-1.60) and poorer overall survival compared with never smoking. Compared with current smoking, smoking cessation was associated with improved overall (HR<10 years = 0.78; 95% CI = 0.69-0.88; HR≥10 years = 0.78; 95% CI = 0.63-0.97) and CRC-specific survival (HR≥10 years = 0.76; 95% CI = 0.67-0.85). Conclusion: In this large meta-analysis including primary data of incident CRC patients from 14 prospective cohort studies on the association between smoking and CRC prognosis, former and current smoking were associated with poorer CRC prognosis compared with never smoking. Smoking cessation was associated with improved survival when compared with current smokers. Future studies should further quantify the benefits of nonsmoking, both for cancer prevention and for improving survival among CRC patients, in particular also in terms of treatment response.

Increased Atrial ß-Adrenergic Receptors and GRK-2 Gene Expression Can Play a Fundamental Role in Heart Failure After Repair of Congenital Heart Disease with Cardiopulmonary Bypass.

Surgeries to correct congenital heart diseases are increasing in Brazil and worldwide. However, even with the advances in surgical techniques and perfusion, some cases, especially the more complex ones, can develop heart failure and death. A retrospective study of patients who underwent surgery for correction of congenital heart diseases with cardiopulmonary bypass (CPB) in a university tertiary-care hospital that died, showed infarction in different stages of evolution and scattered microcalcifications in the myocardium, even without coronary obstruction. CPB is a process routinely used during cardiac surgery for congenital heart disease. However, CPB has been related to increased endogenous catecholamines that can lead to major injuries in cardiomyocytes. The mechanisms involved are not completely understood. The aim of this study was to evaluate the alterations induced in the ß-adrenergic receptors and GRK-2 present in atrial cardiomyocytes of infants with congenital heart disease undergoing surgical repair with CPB and correlate the alterations with functional and biochemical markers of ischemia/myocardial injury. The study consisted of right atrial biopsies of infants undergoing surgical correction in HC-FMRPUSP. Thirty-three cases were selected. Atrial biopsies were obtained at the beginning of CPB (group G1) and at the end of CPB (group G2). Real-time PCR, Western blotting, and immunofluorescence analysis were conducted to evaluate the expression of ß1, ß2-adrenergic receptors, and GRK-2 in atrial myocardium. Cardiac function was evaluated by echocardiography and biochemical analysis (N-terminal pro-brain natriuretic peptide (NT-ProBNP), lactate, and cardiac troponin I). We observed an increase in serum lactate, NT-proBNP, and troponin I at the end of CPB indicating tissue hypoxia/ischemia. Even without major clinical consequences in cardiac function, these alterations were followed by a significant increase in gene expression of ß1 and ß2 receptors and GRK-2, suggesting that this is one of the mechanisms responsible for the exacerbated response of cardiomyocytes to circulating catecholamines. These alterations could explain the irreversible myocardial damage and lipid peroxidation of membranes classically attributed to catecholamine excess, observed in some infants who develop heart failure and postoperative death. Although other factors may be involved, this study confirms that CPB acts as a potent inducer of increased gene expression of ß- adrenergic receptors and GRK-2, making the myocardium of these infants more susceptible to the effects of circulating endogenous catecholamines, which may contribute to the development of irreversible myocardial damage and death.