Dinophysis acuminata or Dinophysis acuta: What Makes the Difference in Highly Stratified Fjords?

Dinophysis acuminata and D. acuta, which follows it seasonally, are the main producers of lipophilic toxins in temperate coastal waters, including Southern Chile. Strains of the two species differ in their toxin profiles and impacts on shellfish resources. D. acuta is considered the major cause of diarrhetic shellfish poisoning (DSP) outbreaks in Southern Chile, but there is uncertainty about the toxicity of D. acuminata, and little information on microscale oceanographic conditions promoting their blooms. During the austral summer of 2020, intensive sampling was carried out in two northern Patagonian fjords, Puyuhuapi (PUY) and Pitipalena (PIT), sharing D. acuminata dominance and D. acuta near detection levels. Dinophysistoxin 1 (DTX 1) and pectenotoxin 2 (PTX 2) were present in all net tow samples but OA was not detected. Although differing in hydrodynamics and sampling dates, D. acuminata shared behavioural traits in the two fjords: cell maxima (>103 cells L-1) in the interface (S ~ 21) between the estuarine freshwater (EFW)) and saline water (ESW) layers; and phased-cell division (µ = 0.3-0.4 d-1) peaking after dawn, and abundance of ciliate prey. Niche analysis (Outlying Mean Index, OMI) of D. acuta with a high marginality and much lower tolerance than D. acuminata indicated an unfavourable physical environment for D. acuta (bloom failure). Comparison of toxin profiles and Dinophysis niches in three contrasting years in PUY-2020 (D. acuminata bloom), 2018 (exceptional bloom of D. acuta), and 2019 (bloom co-occurrence of the two species)-shed light on the vertical gradients which promote each species. The presence of FW (S < 11) and thermal inversion may be used to provide short-term forecasts of no risk of D. acuta blooms and OA occurrence, but D. acuminata associated with DTX 1 pose a risk of DSP events in North Patagonian fjords.

Association of polymorphisms in long pentraxin 3 and its plasma levels with COVID-19 severity.

COVID-19 is an infectious respiratory disease caused by SARS-CoV-2. Pentraxin 3 (PTX3) is involved in the activation and regulation of the complement system, demonstrating an important role in the pathogenesis of COVID-19. The aim was to evaluate the association of single nucleotide polymorphisms in PTX3 and its plasma levels with the severity of COVID-19. This is a retrospective cohort study, carried out between August 2020 and July 2021, including patients with confirmed COVID-19 hospitalized in 2 hospitals in the Northeast Region of Brazil. Polymorphisms in PTX3 (rs1840680 and rs2305619) were determined by real-time PCR. PTX3 plasma levels were measured by ELISA. Serum levels of interleukin (IL)-6, IL-8, and IL-10 were determined by flow cytometry. A multivariate logistic regression model was used to identify parameters independently associated with COVID-19 severity. P values < 0.05 were considered significant. The study included 496 patients, classified as moderate (n = 267) and severe (n = 229) cases. The PTX3 AA genotype (rs1840680) was independently associated with protection against severe COVID-19 (P = 0.037; odds ratio = 0.555). PTX3 plasma levels were significantly associated with COVID-19 severity and mortality (P < 0.05). PTX3 levels were significantly correlated with IL-6, IL-8, IL-10, C-reactive protein, total leukocytes, neutrophil-to-lymphocyte ratio, urea, creatinine, ferritin, length of hospital stay, and higher respiratory rate (P < 0.05). Our results revealed a protective effect of the PTX3 AA genotype (rs1840680) on the development of severe forms of COVID-19. Additionally, PTX3 plasma levels were associated with the severity of COVID-19. The results of this study provide evidence of an important role of PTX3 in the immunopathology of COVID-19.

Current Medical Treatment for Alcohol-Associated Liver Disease.

Alcohol-associated liver disease is one of the main causes of chronic liver disease. It comprises a clinical-histologic spectrum of presentations, from steatosis, steatohepatitis, to different degrees of fibrosis, including cirrhosis and severe necroinflammatory disease, called alcohol-associated hepatitis. In this focused update, we aim to present specific therapeutic interventions and strategies for the management of alcohol-associated liver disease. Current evidence for management in all spectra of manifestations is derived from general chronic liver disease recommendations, but with a higher emphasis on abstinence and nutritional support. Abstinence should comprise the treatment of alcohol use disorder as well as withdrawal syndrome. Nutritional assessment should also consider the presence of sarcopenia and its clinical manifestation, frailty. The degree of compensation of the disease should be evaluated, and complications, actively sought. The most severe acute form of this disease is alcohol-associated hepatitis, which has high mortality and morbidity. Current treatment is based on corticosteroids that act by reducing immune activation and blocking cytotoxicity and inflammation pathways. Other aspects of treatment include preventing and treating hepatorenal syndrome as well as preventing infections although there is no clear evidence as to the benefit of probiotics and antibiotics in prophylaxis. Novel therapies for alcohol-associated hepatitis include metadoxine, interleukin-22 analogs, and interleukin-1-beta antagonists. Finally, granulocyte colony-stimulating factor, microbiota transplantation, and gut-liver axis modulation have shown promising results. We also discuss palliative care in advanced alcohol-associated liver disease.

Small interfering RNA for cancer treatment: overcoming hurdles in delivery.

In many ways, cancer cells are different from healthy cells. A lot of tactical nano-based drug delivery systems are based on the difference between cancer and healthy cells. Currently, nanotechnology-based delivery systems are the most promising tool to deliver DNA-based products to cancer cells. This review aims to highlight the latest development in the lipids and polymeric nanocarrier for siRNA delivery to the cancer cells. It also provides the necessary information about siRNA development and its mechanism of action. Overall, this review gives us a clear picture of lipid and polymer-based drug delivery systems, which in the future could form the base to translate the basic siRNA biology into siRNA-based cancer therapies.

Tissue Dependent Role of PTX3 During Ischemia-Reperfusion Injury.

Reperfusion of an ischemic tissue is the treatment of choice for several diseases, including myocardial infarction and stroke. However, reperfusion of an ischemic tissue causes injury, known as Ischemia and Reperfusion Injury (IRI), that limits the benefit of blood flow restoration. IRI also occurs during solid organ transplantation. During IRI, there is activation of the innate immune system, especially neutrophils, which contributes to the degree of injury. It has been shown that PTX3 can regulate multiple aspects of innate immunity and tissue inflammation during sterile injury, as observed during IRI. In humans, levels of PTX3 increase in blood and elevated levels associate with extent of IRI. In mice, there is also enhanced expression of PTX3 in tissues and plasma after IRI. In general, absence of PTX3, as seen in PTX3-deficient mice, results in worse outcome after IRI. On the contrary, increased expression of PTX3, as seen in PTX3 transgenic mice and after PTX3 administration, is associated with better outcome after IRI. The exception is the gut where PTX3 seems to have a clear deleterious role. Here, we discuss mechanisms by which PTX3 contributes to IRI and the potential of taming this system for the treatment of injuries associated with reperfusion of solid organs.

Elevated Pentraxin-3 Concentrations in Patients With Leprosy: Potential Biomarker of Erythema Nodosum Leprosum.

Background: Leprosy, the leading infectious cause of disability worldwide, remains a major public health challenge in the most severely affected countries despite the sharp decline in new cases in recent years. The search for biomarkers is essential to achieve a better understanding of the molecular and cellular mechanisms underlying the disease. Methods: Pentraxin-3 (PTX3) analyses of sera from 87 leprosy patients with or without reactions were conducted via enzyme-linked immunosorbent assay. In situ identification of PTX3 in skin lesion was confirmed by quantitative reverse-transcription polymerase chain reaction, immunohistochemistry, and immunofluorescence assays. Results: We found that PTX3 serum levels were higher in multibacillary patients when evaluated before the onset of acute erythema nodosum leprosum (ENL) and persistently elevated during reaction. Thalidomide treatment reduced PTX3 in the serum 7 days after starting treatment. In situ analyses have also demonstrated enhancement of PTX3 in ENL lesions and showed that treatment with thalidomide reduced its expression and the prominent neutrophilic infiltrate, a hallmark of the disease. Conclusions: In summary, our study provides in vivo evidence that PTX3 is enhanced during ENL but not in reversal reaction and provides a new molecular target in ENL pathogenesis.

Circulating levels of pentraxin-3 (PTX3) in patients with liver cirrhosis.

BACKGROUND: Despite the circulating levels of PTX3 were related to the severity of various diseases, there are no studies investigating its role in patients with liver cirrhosis. We aimed to study PTX3 levels in patients with liver cirrhosis. MATERIAL AND METHODS: A prospective cohort study included 130 patients hospitalized for acute decompensation of liver cirrhosis, 29 stable cirrhotic outpatients and 32 healthy controls evaluated in a tertiary hospital in Southern Brasil. RESULTS: The median PTX3 level was significantly higher in stable cirrhotic patients compared to controls (2.6 vs. 1.1 ng/mL; p < 0.001), hospitalized cirrhotic patients compared to controls (3.8 vs. 1.1 ng/mL; p < 0.001), and hospitalized cirrhotic patients compared to stable cirrhotic patients (3.8 vs. 2.6 ng/mL; p = 0.001). A positive correlation was found between PTX3 and serum creatinine (r = 0.220; p = 0.012), Chronic Liver Failure - Sequential Organ Failure Assessment score (CLIF-SOFA) (r = 0.220; p = 0.010), MELD (r = 0.279; p = 0.001) and Child-Pugh score (r = 0.224; p = 0.010). Significantly higher levels of PTX3 were observed in patients on admission with ACLF (8.9 vs. 3.1 ng/mL; p < 0.001) and MELD score ≥ 20 (6.6 vs. 3.4 ng/mL; p = 0.002). Death within 90 days occurred in 30.8% of patients and was associated with higher levels of PTX3 (5.3 vs. 3.4 ng/mL; p = 0.009). The probability of Kaplan-Meier survival was 77.0% in patients with PTX-3 < 5.3 ng mL (upper tercile) and 53.5% in those with PTX3 ≥ 5.3 ng/mL (p = 0.002). CONCLUSION: These results indicate the potential for use of PTX3 as an inflammatory biomarker for the prognosis of patients with hepatic cirrhosis.

Kinetics of extracellular ATP in mastoparan 7-activated human erythrocytes.

BACKGROUND: The peptide mastoparan 7 (MST7) stimulated ATP release in human erythrocytes. We explored intra- and extracellular processes governing the time-dependent accumulation of extracellular ATP (i.e., ATPe kinetics). METHODS: Human erythrocytes were treated with MST7 in the presence or absence of two blockers of pannexin 1. ATPe concentration was monitored by luciferin-luciferase based real-time luminometry. RESULTS: Exposure of human erythrocytes to MST7 led to an acute increase in [ATPe], followed by a slower increase phase. ATPe kinetics reflected a strong activation of ATP efflux and a low rate of ATPe hydrolysis by ectoATPase activity. Enhancement of [ATPe] by MST7 required adhesion of erythrocytes to poly-D-lysin-coated coverslips, and correlated with a 31% increase of cAMP and 10% cell swelling. However, when MST7 was dissolved in a hyperosmotic medium to block cell swelling, ATPe accumulation was inhibited by 49%. Erythrocytes pre-exposure to 10µM of either carbenoxolone or probenecid, two blockers of pannexin 1, exhibited a partial reduction of ATP efflux. Erythrocytes from pannexin 1 knockout mice exhibited similar ATPe kinetics as those of wild type mice erythrocytes exposed to pannexin 1 blockers. CONCLUSIONS: MST7 induced release of ATP required either cell adhesion or strong activation of cAMP synthesis. Part of this release required cell swelling. Kinetic analysis and a data driven model suggested that ATP efflux is mediated by two ATP conduits displaying different kinetics, with one conduit being fully blocked by pannexin 1 blockers. GENERAL SIGNIFICANCE: Kinetic analysis of extracellular ATP accumulation from human erythrocytes and potential effects on microcirculation.

Correlação da cicloxigenase-2 e p53 com gradação histopatológica dos mastocitomas cutâneos caninos / Cyclooxygenase-2 and p53 correlation with histopathological grade in cutaneous mast cell tumors

O mastocitoma cutâneo canino é uma neoplasia observada na clínica de animais de companhia de grande importância devido a sua alta freqüência, comportamento clínico agressivo e dificuldade de excisão cirúrgica com margens seguras. De todos os métodos de classificação de masteocitomas e estabelecimento do prognóstico, a gradação histopatológica parece ser a de maior consistência preditiva. No entanto, o comportamento clínico de mastocitomas continua imprevisível e técnicas que auxiliem na determinação do prognóstico são necessárias. Foi realizado estudo retrospectivo baseado nos livros de registro do serviço de Patologia Veterinária da faculdade de Medicina Veterinária e Zootencia – UNESP, Botucatu, para seleção dos casos com diagnóstico de mastocitoma cutâneo canino, que foram graduados na histopatologia e submetidos a técnica imunoistoquímica para os anticorpos primários anti-p53 mutante e anti-COX2. Não foram observadas diferenças estatísticas entre os valores desses biomarcadores e a gradação histopatológicas, porém todos os graus de mastocitoma apresentaram elevados níveis de COX-2 e p-53 (maiores que 50%) o que corrobora com o comportamento biológico potencialmente maligno dessa neoplasia

Mast cell tumor is a frequent neoplasia in small animal clinic, and it is very important due its high frequency, aggressive clinical behavior and difficult surgical excision with safe margins. Of all grading methods and prognostic establishment, the histopathological grading is the most confident for predicting the prognosis. Furthermore, the mast cell tumors present an unpredictable clinical behavior and new prognostic tools should be used to identify patients with higher risk of metastasis and death. A restropective study was made using the Pathology Service (from FMVZ, Unesp, Botucatu, SP) register book to select the mast cell tumors, that were graduated eccording to Patnaik et al. (2) and immunohistochemistry technique was applied for p-53 and COX-2 primary antibodies. No statistical differences were seen in both antibodies used and the hispothological grade, but all tumors had more than 50% of the neoplastic cells positive for each antibody, whinch in agreement with the potential malignant behavior of this neoplasia

Correlação da cicloxigenase-2 e p53 com gradação histopatológica dos mastocitomas cutâneos caninos / Cyclooxygenase-2 and p53 correlation with histopathological grade in cutaneous mast cell tumors

O mastocitoma cutâneo canino é uma neoplasia observada na clínica de animais de companhia de grande importância devido a sua alta freqüência, comportamento clínico agressivo e dificuldade de excisão cirúrgica com margens seguras. De todos os métodos de classificação de masteocitomas e estabelecimento do prognóstico, a gradação histopatológica parece ser a de maior consistência preditiva. No entanto, o comportamento clínico de mastocitomas continua imprevisível e técnicas que auxiliem na determinação do prognóstico são necessárias. Foi realizado estudo retrospectivo baseado nos livros de registro do serviço de Patologia Veterinária da faculdade de Medicina Veterinária e Zootencia UNESP, Botucatu, para seleção dos casos com diagnóstico de mastocitoma cutâneo canino, que foram graduados na histopatologia e submetidos a técnica imunoistoquímica para os anticorpos primários anti-p53 mutante e anti-COX2. Não foram observadas diferenças estatísticas entre os valores desses biomarcadores e a gradação histopatológicas, porém todos os graus de mastocitoma apresentaram elevados níveis de COX-2 e p-53 (maiores que 50%) o que corrobora com o comportamento biológico potencialmente maligno dessa neoplasia(AU)

Mast cell tumor is a frequent neoplasia in small animal clinic, and it is very important due its high frequency, aggressive clinical behavior and difficult surgical excision with safe margins. Of all grading methods and prognostic establishment, the histopathological grading is the most confident for predicting the prognosis. Furthermore, the mast cell tumors present an unpredictable clinical behavior and new prognostic tools should be used to identify patients with higher risk of metastasis and death. A restropective study was made using the Pathology Service (from FMVZ, Unesp, Botucatu, SP) register book to select the mast cell tumors, that were graduated eccording to Patnaik et al. (2) and immunohistochemistry technique was applied for p-53 and COX-2 primary antibodies. No statistical differences were seen in both antibodies used and the hispothological grade, but all tumors had more than 50% of the neoplastic cells positive for each antibody, whinch in agreement with the potential malignant behavior of this neoplasia(AU)

Modulação da expressão de genes pela pentraxina 3 / Modulation of gene expression by PTX3

Introdução: PTX3 é uma proteína de fase aguda produzida por monócitos/macrófagos, células dendríticas, endoteliais, de músculo liso, neutrófilos e fibroblastos em reposta à sinais pró-inflamatórios primários e agonistas de receptores do tipo Toll. PTX3 tem um papel preponderante na modulação da resposta inflamatória in vivo, entretanto seus mecanismos de ação ainda não foram elucidados. O objetivo deste estudo foi a identificação de alvos moleculares da ação de PTX3. Métodos: Utilizamos a tecnologia de microarrays de cDNAs (plataforma do ILPC/SP contendo 4.608 cDNAs humanos) e de oligonucleotídeos (array do FCCC com 16.128 oligos murinos) para estudar o perfil de genes modulados por PTX3(10&*61549;g/ml por 3h) em fibroblastos humanos (GM637) e macrófagos murinos (residentes (RES) e recrutados por tioglicolato (TIO)), respectivamente. Resultados: Nos fibroblastos GM637 foram identificados 37 genes modulados por PTX3 com p<0,05 (t-Student) e razão de expressão (RE) >1,3. A expressão de alguns desses genes foi validada por QPCR, entretanto todos os valores de RE encontrados pelas duas metodologias foram <1,55. Da análise empregando macrófagos RES, 239 genes regulados por PTX3 com p<0,05 (t-Student) foram detectados dos quais 36 com RE > 1,8 (31 induzidos e 5 inibidos). Dentre esses, genes relacionados ao transporte de íons (Kcnn2, Cacna1c), endocitose (Amph) e regulação da pressão sanguínea (Edn1). Nos macrófagos TIO foram identificados 687 genes modulados por PTX3 com p<0,05 (t-Student), dos quais 28 apresentaram RE>1,8 e incluem elementos envolvidos na atividade hormonal (Prl), peptidase (Corin), transporte mediado por vesículas (Vamp3) e circulação sanguínea (Pon1). Da comparação do perfil de expressão entre TIO e RES 792 genes com p<0,05 (t-Student) foram identificados dos quais 34 com RE>3,0. Foram encontrados elementos envolvidos com endocitose (Ly75), resposta à hipóxia (Hif1a), metabolismo de lipídeos (Lpl, Lip, Aadacl1), adesão celular (Icam2, Alcam), citocina (Il1b). Conclusões: O caldo tioglicolato se mostrou um potente modulador da expressão gênica em macrófagos, regulando a expressão de inúmeros genes relacionados à resposta imune. PTX3 é uma proteína pleiotrópica envolvida na imunidade inata e inflamação não apenas como um receptor de reconhecimento de padrões (PRR), mas também como importante modulador da expressão gênica. Suporte financeiro: FAPESP.

Introduction: PTX3 is a secreted glycoprotein produced by monocytes\macrophages, dendritic, endothelial, smooth muscle cells and fibroblasts in response to pro-inflammatory signals and microbial moieties. Our previous studies showed that PTX3 has a key role in modulating inflammatory response in vivo and the identification of the PTX3 molecular targets is essential for the understanding of its action. The aim of this study is to identify genes modulated by PTX3. Methods: We used cDNA (a ILPC-SP plataform containing 4.608 human sequences) and oligo microarray (a FCCC biochip containing 16.128 murines oligos) to identify genes modulated by PTX3 (10 g/ml for 3h) in human fibroblasts (GM637) and murine macrophages (resident (RES) and thioglycollate elicited (TIO)), respectively. Results: We identified 37 genes modulated by PTX3 with p<0,05 (t-Student) and fold change >1,3, in GM637 human fibroblasts. From the comparison between resident macrophages (RESC) and resident macrophages treated with PTX3 239 differently expressed genes were identified with p<0,05 (t-Student) and 36 genes with fold change >1,8 (31 up and 5 down modulated by PTX3). Some of the identified genes are elements related to endocytosis (Amph), regulation of blood pressure (Edn1) and ion transport (Kcnn2, Cacna1c). A total of 687 genes with p<0,05 (t-Student) were modulated by PTX3 in TIO elicited macrophages, among them 28 genes with fold change >1,8. We found elements involved in peptidase activity (Corin), hormone activity (Prl), vesicle-mediated transport (Vamp3) and blood circulation (Pon1) between the PTX3 modulated genes in TIO macrophages. After the comparison between TIO X RES macrophages 792 genes with p<0,05 (tStudent), and 34 genes with fold change >3,0 were detected. Some of these genes are related to endocytosis (Ly75), response to hypoxia (Hif1a), lipid metabolic process (Lpl, Lip, Aadacl1), cell adhesion (Icam2, Alcam) and cytokine (Il1b). Conclusion: Thioglycollate broth is a potent modulator of gene expression in peritoneal macrophages, regulating genes with pivotal role in innate immune response. Both human fibroblasts and murine peritoneal macrophages are responsive to PTX3. PTX3 is a pleiotropic protein involved in the innate immune response and inflammation not only as a soluble pattern recognition receptor (PRR) but also as a modulator of the gene expression. Supported by: FAPESP