Procalcitonin Exerts a Mediator Role in Septic Shock Through the Calcitonin Gene-Related Peptide Receptor.

OBJECTIVES: Clinically, procalcitonin represents the most widely used biomarker of sepsis worldwide with unclear pathophysiologic significance to date. Pharmacologically, procalcitonin was shown to signal through both calcitonin receptor and calcitonin gene-related peptide receptor in vitro, yet the identity of its biologically relevant receptor remains unknown. DESIGN: Prospective randomized animal investigations and in vitro human blood studies. SETTING: Research laboratory of a university hospital. SUBJECTS: C57BL/6J mice and patients with post-traumatic sepsis. INTERVENTIONS: Procalcitonin-deficient mice were used to decipher a potential mediator role in experimental septic shock and identify the relevant receptor for procalcitonin. Cecal ligation and puncture and endotoxemia models were employed to investigate septic shock. Disease progression was evaluated through survival analysis, histology, proteome profiling, gene expression, and flow cytometry. Mechanistic studies were performed with cultured macrophages, dendritic cells, and gamma delta T cells. Main findings were confirmed in serum samples of patients with post-traumatic sepsis. MEASUREMENTS AND MAIN RESULTS: Procalcitonin-deficient mice are protected from septic shock and show decreased pulmonary inflammation. Mechanistically, procalcitonin potentiates proinflammatory cytokine expression in innate immune cells, required for interleukin-17A expression in gamma delta T cells. In patients with post-traumatic sepsis, procalcitonin positively correlates with systemic interleukin-17A levels. In mice with endotoxemia, immunoneutralization of interleukin-17A inhibits the deleterious effect of procalcitonin on disease outcome. Although calcitonin receptor expression is irrelevant for disease progression, the nonpeptide calcitonin gene-related peptide receptor antagonist olcegepant, a prototype of currently introduced antimigraine drugs, inhibits procalcitonin signaling and increases survival time in septic shock. CONCLUSIONS: Our experimental data suggest that procalcitonin exerts a moderate but harmful effect on disease progression in experimental septic shock. In addition, the study points towards the calcitonin gene-related peptide receptor as relevant for procalcitonin signaling and suggests a potential therapeutic application for calcitonin gene-related peptide receptor inhibitors in sepsis, which warrants further clinical investigation.

Falta de asociación entre inflamación subclínica evaluada por ultrasonido y niveles de citocinas en artritis reumatoide / Lack of association between ultrasound-defined subclinica inflammation and cytokines levels in rheumatoid arthritis

Antecedentes: La artritis reumatoide (AR) es una enfermedad autoinmune crónica que se caracteriza por proliferación sinovial, ruptura de cartílago y destrucción ósea. Los biomarcadores en AR no se utilizan en forma rutinaria para evaluar la inflamación y tampoco la remisión. El ultrasonido musculoesquelético (US) visualiza los cambios en las articulaciones y el daño morfoestructural, mejorando la evaluación de la sinovitis.Objetivo: Identificar y describir la inflamación subclínica en pacientes con AR en re-misión, utilizando US.Métodos: Se incluyeron pacientes con AR en remisión. Se realizó una evaluación clí-nica con DAS28; se tomó muestra de sangre para analizar citocinas. Un ecografista reumatólogo sin acceso a datos clínicos realizó un conteo ecográfico utilizando el sco-re-7. Se utilizaron parámetros de tendencia central, análisis de correlación bivariada y X cuadrado. Se estableció un nivel de confianza del 95% y, por tanto, cualquier valor p ≤0.05 se consideró significativo.Resultados: Se incluyeron 38 pacientes con AR. La edad media fue de 45,26±12,24 años. Los niveles de citocinas asociadas al tiempo de la AR desde la remisión, no fue-ron estadísticamente significativas. El ultrasonido en los pacientes evidenció al menos una de las lesiones elementales; en escala de grises, la sinovitis ocurrió en un 94,7%; sinovitis con señal Doppler de poder (DP) 52,6%; en cuanto a erosiones, se registra-ron, respectivamente, un 55,3% en escala de grises y un 15,8% con DP. DAS28 >2,04 fue positivo al asociarse con el recuento de articulaciones dolorosas y significativo (p=0,009). Conclusión: La asociación entre la sinovitis clínica y en ecografía no tiene correlación con los criterios de AR en remisión, independientemente de cuán estricta sea su aplicación.

Background: Rheumatoid arthritis (RA) is a chronic autoimmune disease, character-ized by synovial proliferation, cartilage breakdown and bone destruction. Biomarkers are not routinely used to evaluate inflammation neither remission. Musculoskeletal ultrasound visualizes joint changes and morpho-structural damage improving the as-sessment of synovitis.Objective: To identify and describe subclinical inflammation in patients with RA in remission using US.Methods: RA patients in remission were included. A clinical evaluation and DAS28 score performed; a blood sample took to analyze cytokines. A rheumatologist ultraso-nographer blinded to clinical data performed a US 7-score joint count. Central tenden-cy parameters, bivariate correlation analysis, and X Square were used. A confidence level of 95% was set and, therefore, any p-value ≤0.05 was considered as significant.Results: 38 RA patients were included. Mean age was 45.26±12.24 years. Cytokines associated with the time since remission was not statistically significant. Patients dis-played at least one of US elementary lesions; gray-scale synovitis occurred in 94.7%; synovitis with PD signal 52.6%; gray-scale erosions 55.3% and erosions with PD 15.8% respectively. DAS28 >2.04 positive for tender joint count was significant (p=0.009).Conclusion: The association between the clinical and US synovitis does not correlate with RA remission criteria no matter how strict is its application.

Profile of glycated-hemoglobin, antioxidant vitamin and cytokine levels in pulmonary tuberculosis patients: A cross sectional study at Pulmonary Diseases Center Semarang City, Indonesia.

BACKGROUND: Uncontrolled blood glucose, which marked by high level of HbA1c, increases risk of pulmonary TB because of cellular immunity dysfunction. This study aimed to analyze profile of glycated hemoglobin, antioxidant vitamins status and cytokines levels in active pulmonary TB patients. METHODS: This was a cross sectional study, conducted at Pulmonary Diseases Center Semarang City, Indonesia. Study subject consisted of 62 pulmonary TB patients, diagnosed with positive acid fast bacilli and chest X-ray. ELISA was used to measure IFN-γ and IL-12. Status of antioxidant vitamins was determined by concentration of vitamin A and E using HPLC. Blood glucose control was determined by HbA1c concentration (HbA1c ≥7% is considered as uncontrolled). RESULTS: A significant difference of age between pulmonary tuberculosis patients with normal and uncontrolled blood glucose (p = 0.000) was showed, while all other characteristics (sex, education, occupation) did not differ with p = 0.050, 0.280, 0.380 respectively. Mean HbA1c was 7.25 ± 2.70%. Prevalence of uncontrolled glucose among pulmonary TB patients was 29%. Levels of IFN-γ and IL-12 did not differ according to HbA1c concentration (p = 0.159 and p = 0.965 respectively). Pulmonary tuberculosis patients with uncontrolled blood glucose has higher vitamin E (p = 0.006), while vitamin A did not differ significantly (p = 0.478). CONCLUSIONS: This study supports the importance of performing diabetes screening among pulmonary TB patients. Further study needs to be done to determine the feasibility of TB-DM co-management.

Immunopathogenesis of 2009 pandemic influenza

Three years after the pandemic, major advances have been made in our understanding of the innate and adaptive immune responses to the influenza A(H1N1)pdm09 virus and those responses’ contribution to the immunopathology associated with this infection. Severe disease is characterized by early secretion of proinflammatory and immunomodulatory cytokines. This cytokine secretion persisted in patients with severe viral pneumonia and was directly associated with the degree of viral replication in the respiratory tract. Cytokines play important roles in the antiviral defense, but persistent hypercytokinemia may cause inflammatory tissue damage and participate in the genesis of the respiratory failure observed in these patients. An absence of pre-existing protective antibodies was the rule for both mild and severe cases. A role for pathogenic immunocomplexes has been proposed for this disease. Defective T cell responses characterize severe cases of infection caused by the influenza A(H1N1)pdm09 virus. Immune alterations associated with accompanying conditions such as obesity, pregnancy or chronic obstructive pulmonary disease may interfere with the normal development of the specific response to the virus. The role of host immunogenetic factors associated with disease severity is also discussed in this review. In conclusion, currently available information suggests a complex immunological dysfunction/alteration that characterizes the severe cases of 2009 pandemic influenza. The potential benefits of prophylactic/therapeutic interventions aimed at preventing/correcting such dysfunction warrant investigation (AU)

El posterior estudio de la gripe pándemica de 2009 hasta la actualidad ha dado como fruto un mayor conocimiento de las respuestas inmunológicas innatas y adaptativas al virus de la gripe A(H1N1)pdm09 y de la contribución de las respuestas a la inmunopatología asociada con esta infección. Se ha comprobado que la enfermedad grave se caracteriza por la secreción de citocinas proinflamatorias e inmunomoduladoras desde el principio de la enfermedad. En los pacientes con neumonía viral grave la secreción de citocinas se mantuvo y estaba relacionada directamente con el grado de replicación viral en el tracto respiratorio. Si bien las citocinas tienen un papel importante en la defensa antiviral, sin embargo la persistencia de hipercitoquinemia puede dañar el tejido inflamatorio y ser uno de los motivos que provocan el fracaso respiratorio observado en estos pacientes. En casi todos los casos leves y graves de enfermedad se observó ausencia de anticuerpos protectores preexistentes. Se ha propuesto que los inmunocomplejos patogénicos tienen un papel en esta enfermedad. Los casos graves de infección causados por el virus de la gripe A(H1N1)pdm09 se caracterizan por una respuesta defectuosa de las células T. En el desarrollo normal de la respuesta específica al virus pueden interferir trastornos inmunológicos asociados con situaciones concomitantes como obesidad, embarazo o enfermedad pulmonar obstructiva crónica. Los autores de esta revisión plantean también el papel que tienen los factores inmunogenéticos del huésped en la gravedad de la enfermedad. La gripe pandémica de 2009, de acuerdo con la información disponible 3 años más tarde, se caracteriza por una disfunción/alteración inmunológica compleja, por lo que los potenciales beneficios de las intervenciones terapéuticas deben ir dirigidos a corregir dicha disfunción y siempre garantizados mediante la investigación (AU)

Erythropoietin in cardiorenal anemia syndrome.

Incidents of heart and renal failure (HF, RF) together, are increasing in our country and all over the world, so a great attention has been dedicated to this problem recently. These diseases together have shown bad results because of the process of accelerated arteriosclerosis, structural changes of myocardium, oxidative stress, inflammation, increased activities of sympathetic nervous system (SNS), increased activities of a renin-angiotensin-aldosterone system (RAAS) (1). These factors are crucial in the development of patho-physiological process and consequential development of anemia, that together with heart and renal failure through interaction, cause serious disorder that we call the cardio-renal anemia syndrome (2). We examined effects of erythropoietin (Epoetin beta) at 90 (60 men and 30 women) pre-dialysed and dialysed patients with HF signs during a period of three years in individual dozes of 2000-6000 units subcutaneous (sc) weekly. Using computer S PLUS and SAS multiple variant analysis we have got correlations by Pearson. Epoetin beta significantly develops anemia parameters: number of erythrocytes (r=0.51779; p<0.0001), hemoglobin (r=0.38811; p<0.0002), MCV (r=0.59876; p<0.0001) at patients with HF. Positive effects are seen at NYHA class (r=0.59906; p<0.0001), on quality of life before and after prescribing medicine. Parameters of renal functions are improving: more urea (r =0.45557; p<0.0001) than creatinine (r=0.26397; p<0.00119) and potassium values K(+)) are not changed significantly (r=0.02060; p<0.8471). Epoetin beta has been useful in treatment of pre-dialysed and dialysed patients with HF and anemia by improving functional ability of myocardium and quality of life.

Batch immunoextraction method for efficient purification of aromatic cytokinins.

A range of benzylaminopurines naturally occur in plants and exhibit high biological activity. Others have been synthesized, such as 6-(2-hydroxy-3-methoxybenzylamino)purine riboside (2OH3MeOBAPR), which has shown interesting anti-cancer activity under in vitro conditions. In order to study the biological activity of this interesting compound in more detail, a rapid and highly efficient method for its purification from complex samples (e.g. blood and plant extracts) is needed. Therefore, we prepared monoclonal antibodies against 2OH3MeOBAPR. The antibody had undetectable cross-reactivity with all natural isoprenoid cytokinins, but relatively high cross-reactivity with aromatic cytokinins as well as some synthetic di- and tri-substituted 6-benzylaminopurines and the corresponding ribosides. The antibody also showed strong responses and specificity in enzyme-linked immunoassays (ELISAs). In addition, it was used to prepare, for the first time, an immunoaffinity sorbent with high specificity and capacity for aromatic cytokinins. A batch immunoextraction method was then developed and optimized for the purification of 2OH3MeOBAPR from murine blood samples. The high efficacy and simplicity of this method (in off-line combination with HPLC-MS) for the isolation of target analytes from biological material is demonstrated in this study.

[Expression of markers of immunocompetent cells, cytokine level, and L-arginine metabolism in complex extremely high frequency and interferon therapy of inflammatory diseases in women of highlands]. / Ekspressiia markerov immunokompetentnykh kletok, uroven' tsitokinov i metabolizm L-arginina pri kompleksnoi krainevysokochastotnoi i interferonoterapii vospalitel'nykh zabolevanii u zhenshchin v vysokogor'e.

Chronic inflammatory gynecological diseases (CIGD) in highlanders (2,100-2,200 m a.s.l.) are accompanied with the following changes in immunoregulatory parameters: strengthening in the expression of CD3+ (TCR), CD4+ (T-helpers), CD16+ (NK-cells), CD25+ (activated IL-2R), increase in CD4+/CD8+ ratio, levels of circulating cytokines--alpha-, beta-, gamma-interferon (IFN) and alpha-, beta-tumor necrotizing factor (TNF), strengthening in immune adhesion of thrombocytes (IAT), and inhibition of the oxidative metabolic way of L-arginine with redistribution of the main metabolites formed via its non-oxidative way. Complex extremely high frequency (EHF) therapy and interferon (with Laferon) therapy of CIGD normalized expression of the differentiated and functional CD-markers of immunocompetent cells (ICC), CD4+/CD8+ ratio, increased expression of CD8+, decreased IAT and levels of circulating cytokines (IFN, TNF), changed L-arginine metabolism with activation of the oxidative way. Full-value of the population signs of immune stabilization due to the treatment was complemented with increase in the ICC functional parameters--markers of mitochondrial, lysosomal and mitotic activity. We first demonstrated a high effectiveness of complex use of EHF and Laferon as an immunocorrective therapy of CIGD in highlanders.