Neuronal nitric oxide synthase and its interaction with soluble guanylate cyclase is a key factor for the vascular dysfunction of experimental sepsis.

OBJECTIVE: Vascular dysfunction plays a central role in sepsis, and it is characterized by hypotension and hyporesponsiveness to vasoconstrictors. Nitric oxide is regarded as a central element of sepsis vascular dysfunction. The high amounts of nitric oxide produced during sepsis are mainly derived from the inducible isoform of nitric oxide synthase 2. We have previously shown that nitric oxide synthase 2 levels decrease in later stages of sepsis, whereas levels and activity of soluble guanylate cyclase increase. Therefore, we studied the putative role of other relevant nitric oxide sources, namely, the neuronal (nitric oxide synthase 1) isoform, in sepsis and its relationship with soluble guanylate cyclase. We also studied the consequences of nitric oxide synthase 1 blockade in the hyporesponsiveness to vasoconstrictors. DESIGN: Randomized controlled prospective experimental study. SETTING: Academic research laboratory. SUBJECTS: Female Wistar rats submitted to cecal ligation and puncture method. INTERVENTIONS: 1) Six, 12, and 24 hours after cecal ligation and puncture, vascular reactivity to phenylephrine (3 and 30 nmol/kg) before and after 7-nitroindazole (45 µmol/kg, s.c.) or aminoguanidine (30 µmol/kg, s.c.) administration was evaluated. 2) Protein levels and interaction between nitric oxide synthase 1 and soluble guanylate cyclase were determined. 3) Six, 12, and 24 hours after cecal ligation and puncture, thoracic aorta segments were stimulated with phenylephrine in the presence or absence of 7-nitroindazole and cyclic guanosine monophosphate accumulation was determined. 4) After 24 hours of cecal ligation and puncture, norepinephrine was infused (10 µg/kg/min) in the presence or absence of 7-nitroindazole or S-methyl-L-thiocitrulline (1 µmol/kg, IV) and mean arterial pressure was registered. MEASUREMENTS AND MAIN RESULTS: 1) Both nitric oxide synthase 1 and soluble guanylate cyclase are expressed in higher levels in vascular tissues during sepsis; 2) both proteins physically interact and nitric oxide synthase 1 blockade inhibits cyclic guanosine monophosphate production; 3) pharmacological blockade of nitric oxide synthase 1 using 7-nitroindazole or S-methyl-L-thiocitrulline reverts the hyporesponsiveness to phenylephrine and increases the vasoconstrictor effect of norepinephrine and phenylephrine. CONCLUSIONS: Sepsis induces increased expression and physical association of nitric oxide synthase 1/soluble guanylate cyclase and a higher production of cyclic guanosine monophosphate that together may help explain sepsis-induced vascular dysfunction. In addition, selective inhibition of nitric oxide synthase 1 restores the responsiveness to vasoconstrictors. Therefore, inhibition of nitric oxide synthase 1 (and possibly soluble guanylate cyclase) may represent a valuable alternative to restore the effectiveness of vasopressor agents during late sepsis.

Exploring Social Impairment in Those with Opioid Use Disorder: Linking Impulsivity, Childhood Trauma, and the Prefrontal Cortex.

Background: Challenges with social functioning, which is a hallmark of opioid use disorder (OUD), are a drawback in treatment adherence and maintenance. Yet, little research has explored the underlying mechanisms of this impairment. Impulsivity, a known risk factor for OUD, and corresponding neural alterations may be at the center of this issue. Childhood adversity, which has been linked to both impulsivity and poorer treatment outcomes, could also affect this relationship. This study aims to understand the relationship between impulsivity and social functioning in those recovering from OUD. Differences in the prefrontal cortex will be analyzed, as well as potential moderating effects of childhood trauma. Methods: Participants with (N = 16) and without (N = 19) social impairment completed a survey (e.g., social functioning, Barrat's Impulsivity Scale, Adverse Childhood Experiences (ACEs) and cognitive tasks while undergoing neuroimaging. Functional near infrared spectroscopy (fNIRS), a modern, portable, wearable and low-cost neuroimaging technology, was used to measure prefrontal cortex activity during a behavioral inhibition task (Go/No-Go task). Results: Those who social functioning survey scores indicated social impairment (n = 16) scored significantly higher on impulsivity scale (t(33)= -3.4, p < 0.01) and reported more depressive symptoms (t(33) = -2.8, p < 0.01) than those reporting no social impairment (n = 19). Social functioning was negatively correlated with impulsivity (r=-0.7, p < 0.001), such that increased impulsivity corresponded to decreased social functioning. Childhood trauma emerged as a moderator of this relationship, but only when controlling for the effects of depression, B=-0.11, p = 0.023. Although both groups had comparable Go/No-Go task performance, the socially impaired group displayed greater activation in the dorsolateral (F(1,100.8) = 7.89, p < 0.01), ventrolateral (F(1,88.8) = 7.33, p < 0.01), and ventromedial (F(1,95.6) = 7.56, p < 0.01) prefrontal cortex during impulse control. Conclusion: In addition to being more impulsive, individuals with social impairment exhibited differential activation in the prefrontal cortex when controlling responses. Furthermore, the impact of impulsivity on social functioning varies depending on ACEs demonstrating that it must be considered in treatment approaches. These findings have implications for addressing social needs and impulsivity of those in recovery, highlighting the importance of a more personalized, integrative, and trauma-informed approach to intervention.

Viremia and Inflammatory Cytokines in Dengue: Interleukin-2 as a Biomarker of Infection, and Interferon-α and -γ as Markers of Primary versus Secondary Infection.

The pathogenesis of Dengue virus (DENV) infection is complex and involves viral replication that may trigger an inflammatory response leading to severe disease. Here, we investigated the correlation between viremia and cytokine levels in the serum of DENV-infected patients. Between 2013 and 2014, 138 patients with a diagnosis of acute-phase DENV infection and 22 patients with a non-dengue acute febrile illness (AFI) were enrolled. Through a focus-forming assay (FFU), we determined the viremia levels in DENV-infected patients and observed a peak in the first two days after the onset of symptoms. A higher level of viremia was observed in primary versus secondary DENV-infected patients. Furthermore, no correlation was observed between viremia and inflammatory cytokine levels in DENV-infected patients. Receiver operating characteristic (ROC) curve analysis revealed that IL-2 has the potential to act as a marker to distinguish dengue from other febrile illnesses and is positively correlated with Th1 cytokines. IFN-α and IFN-γ appear to be potential markers of primary versus secondary infection in DENV-infected patients, respectively. The results also indicate that viremia levels are not the main driving force behind inflammation in dengue and that cytokines could be used as infection biomarkers and for differentiation between primary versus secondary infection.

Epidemiological study on dengue in southern Brazil under the perspective of climate and poverty.

Social and epidemiological aspects of dengue were evaluated in an important metropolitan area in southern Brazil, from August 2012 to September 2014. Demographic, clinical, serological data were collected from patients with acute dengue symptoms treated at public health system units (HSUs). A systematic approach to analyze the spatial and temporal distribution of cases was developed, considering the temporal cross-correlation between dengue and weather, and the spatial correlation between dengue and income over the city's census tracts. From the 878 patients with suggestive symptoms, 249 were diagnosed as positive dengue infection (28%). Considering the most statistically significant census tracts, a negative correlation was found between mean income and dengue (r = -0.65; p = 0.02; 95% CI: -0.03 to -0.91). The occurrence of dengue followed a seasonal distribution, and it was found to be three and four months delayed in relation to precipitation and temperature, respectively. Unexpectedly, the occurrence of symptomatic patients without dengue infection followed the same seasonal distribution, however its spatial distribution did not correlate with income. Through this methodology, we have found evidence that suggests a relation between dengue and poverty, which enriches the debate in the literature and sheds light on an extremely relevant socioeconomic and public health issue.

The citrus flavonoid naringenin impairs the in vitro infection of human cells by Zika virus.

The Zika virus (ZIKV) is an arthropod-borne virus that belongs to the Flaviviridae family. The ZIKV infection is usually asymptomatic or is associated with mild clinical manifestations; however, increased numbers of cases of microcephaly and birth defects have been recently reported. To date, neither a vaccine nor an antiviral treatment has become available to control ZIKV replication. Among the natural compounds recognized for their medical properties, flavonoids, which can be found in fruits and vegetables, have been found to possess biological activity against a variety of viruses. Here, we demonstrate that the citrus flavanone naringenin (NAR) prevented ZIKV infection in human A549 cells in a concentration-dependent and ZIKV-lineage independent manner. NAR antiviral activity was also observed when primary human monocyte-derived dendritic cells were infected by ZIKV. NAR displayed its antiviral activity when the cells were treated after infection, suggesting that NAR acts on the viral replication or assembly of viral particles. Moreover, a molecular docking analysis suggests a potential interaction between NAR and the protease domain of the NS2B-NS3 protein of ZIKV which could explain the anti-ZIKV activity of NAR. Finally, the results support the potential of NAR as a suitable candidate molecule for developing anti-ZIKV treatments.

Análise do perfil clínico e imunológico de pacientes com dengue em fase aguda na região norte do Paraná

A dengue é uma doença multifatorial causada pela infecção por um dos quatro sorotipos do vírus da dengue (DENV 1-4). Neste trabalho avaliou-se diferentes características (viremia, citocinas e sorotipo viral) da infecção por DENV em pacientes do município de Cambé, região Norte do Paraná. Para tanto, foram obtidas 888 amostras de soros de pacientes com suspeita clínica de infecção, provenientes de unidades de pronto atendimento de Cambé, durante os anos de 2013 e 2014. As análises sorológicas (ELISA anti-IgM, anti-IgG e NS1) demonstraram que 264 representavam casos de infecção aguda (NS1+ e/ou IgM+). Observou-se uma correlação positiva entre condições climáticas (pluviosidade e temperatura) e a densidade populacional do mosquito vetor com a incidência da doença. A média de idade dos pacientes com dengue foi de 33 anos, e não houve prevalência de infecção de acordo com a cor ou gênero. Adicionalmente, os sintomas mais frequentes foram febre (87%), cefaléia (78%) e mialgia (70%), e observou-se uma baixa prevalência de sinais hemorrágicos (6%). O isolamento viral em células C6/36 confirmou a presença da partícula viral em 145 das 264 (55%) amostras. Além disso, houve uma alta correspondência entre as técnicas de ELISA NS1 e a detecção da partícula viral. Ademais, o sorotipo viral foi determinado em 138 das 145 amostras positivas para o isolamento viral pela técnica de RT-PCR One Step. Deste modo, observou-se a soroprevalência do sorotipo-1 do DENV (137 amostras) no período do estudo, com apenas uma amostra de DENV-4. Ainda, 7 amostras não foram amplificadas pelo protocolo de análise e estão sendo testadas com outros primers e para outros vírus. Adicionalmente, a viremia foi determinada pela técnica de titulação em células C6/36 nas amostras com sorotipo viral definido (138). Nestes pacientes, a viremia média foi de 1,0 x 105 FFUC6/36/mL, sendo esta mais alta nos primeiros três dias da doença. Adicionalmente, determinou-se os níveis de citocinas inflamatórias no soro destes pacientes e pode-se observar um perfil misto Th1/Th2, marcado pela produção aumentada de IFN-γ, IL-6 e IL-4 nos pacientes com dengue comparado aos saudáveis. Finalmente, com o sequenciamento da região codificadora da proteína E seguida de análise filogenética demonstrou a co-circulação de duas linhagens (1b e 2) de DENV-1 de genótipo V em Cambé no período de estudo. Assim, conclui-se que diversos fatores podem influenciar na incidência e na patologia da dengue. Deste modo, a análise combinada de fatores epidemiológicos, bem como a caracterização de cepas virais e da resposta imune dos pacientes, é capaz de contribuir para o esclarecimento dos diferentes aspectos envolvidos na imunopatologia da dengue, e no surgimento de novos surtos da doença.