N-acetylcysteine as a therapeutic approach to post-COVID-19 pulmonary fibrosis adjunctive treatment.

OBJECTIVE: Growing interest is directed to the outcomes of COVID-19 in survivors, both in the convalescent period and in the long-term, which are responsible for morbidity and quality of life deterioration. This article aims to describe the mechanisms supporting the possible use of NAC as an adjuvant treatment for post-COVID-19 pulmonary fibrosis. MATERIALS AND METHODS: A search was performed in PubMed/MEDLINE. RESULTS: Interstitial changes have been observed in the CT scan of COVID-19 pneumonia. In patients with respiratory outcomes in the post-COVID-19 stage, glutathione (GSH) deficiency was found and interpreted as a reaction to the inflammatory cascade caused by the viral infection, while the pathophysiological process of pulmonary fibrosis involves numerous cytokines, such as TGF-ß, TNF-α, IL-1, PDGF and VEGF. NAC has a good tolerability profile, is easily administered orally and inexpensively, and has antioxidant and anti-inflammatory effects that may target the pathophysiologic mechanisms involved in pulmonary fibrosis. It may revert GSH deficiency, exerts direct and indirect antioxidant activity, anti-inflammatory activity and improves immune T-cell response. CONCLUSIONS: The mechanism of action of NAC suggests a role in the treatment of pulmonary fibrosis induced by COVID-19.

[Development of a PCR for the detection and quantification of parasitism by Demodex folliculorum infestation in biopsies of skin neoplasms periocular area]. / Desarrollo de una PCR para la detección y cuantificación de la parasitación por Demodex folliculorum en biopsias de neoplasias cutáneas del área periocula.

OBJECTIVE: To standardize the relative quantification by mass of tissue parasitism by Demodex folliculorum infestation from neoplastic skin biopsies periocular using molecular amplification to study the possible relationship of the appearance of eyelid basal cell carcinoma with the presence and density of the mite in later works. METHODS: A quantitative PCR was developed real-time probes TaqMan. PCR was tested in a pilot 46 actual biopsy samples nodular basal cell carcinoma series. RESULTS: The sensitivity was placed with a detection limit of between 1 and 10 copies / µl. 50% (23/46) of the biopsies were positive for D. folliculorum. The specificity was 100% confirmed by sequencing. CONCLUSIONS: The technique shows good results for sensitivity and specificity that can make it useful as a tool for studies of cause and effect D. folliculorum and basal cell carcinoma.

Intratracheal infection as an efficient route for testing vaccines against Chlamydia abortus in sheep.

Pregnant ewes have been widely used to test vaccines against Chlamydia abortus. However, this model entails many disadvantages such as high economic costs and long periods of pregnancy. The murine model is very useful for specific studies but cannot replace the natural host for the later stages of vaccine evaluation. Therefore, a non-pregnant model of the natural host might be useful for a vaccine trial to select the best vaccine candidates prior to use of the pregnant model. With this aim, two routes of infection were assessed in young non-pregnant sheep, namely, intranasal (IN) and intratracheal (IT). In addition, groups of non-vaccinated sheep and sheep immunised with an inactivated vaccine were established to investigate the suitability of the model for testing vaccines. After the experimental infection, isolation of the microorganism in several organs, with pathological and immunohistochemical analyses, antibody production assessment and investigation by PCR of the presence of chlamydia in the vagina or rectum were carried out. Experimental IT inoculation of C. abortus induced pneumonia in sheep during the first few days post-infection, confirming the suitability of the IT route for testing vaccines in the natural host. The course of infection and the resulting pathological signs were less severe in vaccinated sheep compared with non-vaccinated animals, demonstrating the success of vaccination. IN infection did not produce evident lesions or demonstrate the presence of chlamydial antigen in the lungs and cannot be considered an appropriate model for testing vaccines.

IFN-γ expression in placenta is associated to resistance to Chlamydia abortus after intragastric infection.

Intragastric infection mimics the natural route of infection of Chlamydia abortus (etiological agent of ovine enzootic abortion). In the mouse model, intragastric experimental infection induces very mild signs of infection followed by late term abortions, as it is shown by the natural ovine host. In order to evaluate the immune mechanisms associated to the dissemination of the pathogen from the gastrointestinal tract, we have administered an intragastric dose of C. abortus to pregnant mice. Systemic and local expression of cytokines, tissue colonization and excretion of bacteria after parturition were monitored during pregnancy. Susceptible CBA/J mice showed a higher bacterial colonization of the placenta and excretion of live bacteria after parturition that were related to a higher local IL-10 expression. By contrast, resistant C57BL/6 mouse strain had higher local IFN-γ mRNA expression in the placenta just before parturition and a transient bacterial colonization of the reproductive tract, with no excretion of C. abortus after parturition. In summary, intragastric infection not only mimics the natural route of infection of C. abortus, but can also be useful in order to understand the immunopathogenesis of chlamydial abortion in the mouse.

[Utility and validity of indicators from the Nursing Outcomes Classification as a support tool for diagnosing Ineffective Self Health Management in patients with chronic conditions in primary health care]. / Utilidad y validez de un instrumento basado en indicadores de la Nursing Outcomes Classification como ayuda al diagnóstico de pacientes crónicos de At.

BACKGROUND: Self-care and management of therapeutic regime (drugs adherence, preventive behaviours and development of healthy life-styles) are key components for managing chronic diseases. Nursing has standardized languages which describe many of these situations, such as the diagnosis "Ineffective Self Health Management" (ISHM) or many of the Nursing Outcomes Classification (NOC) indicators. The aims of this study were to determine the interobserver reliability of a NOC-based instrument for assessment and aid in diagnosis of the ISHM in patients with chronic conditions in Primary Health Care, to determine its diagnostic validity and to describe the prevalence of patients with this problem. METHODS: Cross-sectional validation study developed in the provinces of Málaga, Cádiz and Almería from 2006 to 2009. Each patient was assessed by 3 independent observers: the first two observers evaluated scoring of the NOC indicators and the third one acted as the "gold-standard". RESULTS: Two hundred and twenty-eight patients were included, 37.7% of them with more than one chronic condition. NOC indicators showed a high interobserver reliability (ICC>0,70) and a consistency (Cronbach's alpha: 0.81). With a cut-point of 10.5, sensitivity was 61% and specificity 85%, and the area under the curve was 0.81 (CI95%: 0.77 to 0.85). The prevalence of patients with ISHM was 36% (CI 95%: 34 to 40). CONCLUSIONS: The use of NOC indicators allows evaluation of management of the therapeutic regime in people with chronic conditions with a satisfactory validity and it provides new approaches for dealing with this problem.

Utilidad y validez de un instrumento basado en indicadores de la Nursing Outcomes Classification como ayuda al diagnóstico de pacientes crónicos de Atención Primaria con gestión ineficiente de la salud propia / Utility and validity of indicators from the Nursing Outcomes Classification as a support tool for diagnosing Ineffective Self Health Management in patients with chronic conditions in Primary Health Care

Fundamento. El autocuidado y el manejo del régimen terapéutico por parte del paciente (adherencia a su medicación, conductas preventivas y desarrollo de estilos de vida saludables) son componentes clave del abordaje de las enfermedades crónicas. La enfermería dispone de lenguajes estandarizados para describir muchas de estas situaciones, como el diagnóstico ”gestión ineficiente dela propia salud” (GIPS) o muchos indicadores de la Clasificación de Resultados de Enfermería (NOC). Los objetivos de este estudio, pretenden conocer la fiabilidad inter observador de un instrumento de valoración y ayuda al diagnóstico de la GIPS en pacientes crónicos que acuden a consultas de Atención Primaria, basado en indicadores NOC, además de determinar su validez diagnóstica y describir la prevalencia de pacientes con este problema. Material y métodos. Estudio observacional transversal de validación, en las provincias de Málaga, Cádiz y Almería durante los años 2006 a 2009. Cada paciente era valorado por 3 observadores de forma independiente: una pareja evaluaba la puntuación de los distintos indicadores y un tercer observador experto actuaba como patrón oro .Resultados. Doscientos veintiocho pacientes fueron incluidos, con un 37,7% de ellos con más de un proceso crónico concomitante. Los indicadores NOC mostraron una alta fiabilidad inter observador (CCI>0,70) y una consistencia interna buena (alfa de Cronbach:0,81). Con un punto de corte en 10,5, se obtuvo una sensibilidad del61% y una especificidad del 85%, con un área bajo la curva de 0,81(IC95%: 0,77 a 0,85). La prevalencia de pacientes con GIPS fue del36% (IC 95%: 34 a 40).Conclusiones. El uso de indicadores de la NOC permite valorar el manejo del régimen terapéutico en pacientes crónicos, con una validez aceptable y aporta nuevos enfoques para el abordaje de este problema(AU)

Background. Self-care and management of therapeutic regime(drugs adherence, preventive behave ours and development of healthy life-styles) are key components for managing chronic diseases. Nursing has standardized languages which describe many of these situations, such as the diagnosis “Ineffective Self Health Management” (ISHM) or many of the Nursing Outcomes Classification (NOC) indicators. The aims of this study were to determine the inter observer reliability of a NOC-based instrument for assessment and aid in diagnosis of the ISHM in patients with chronic conditions in Primary Health Care, to determine its diagnostic validity and to describe the prevalence of patients with this problem. Methods. Cross-sectional validation study developed in the provinces of Málaga, Cádiz and Almería from 2006 to 2009. Each patient was assessed by 3 independent observers: the first twoobservers evaluated scoring of the NOC indicators and the third one acted as the “gold-standard”. Results. Two hundred and twenty-eight patients were included,37.7% of them with more than one chronic condition. NOC indicators showed a high inter observer reliability (ICC>0,70) and a consistency (Cronbach’s alpha: 0.81). With a cut-point of 10.5,sensitivity was 61% and specificity 85%, and the area under the curve was 0.81 (CI95%: 0.77 to 0.85). The prevalence of patients with ISHM was 36% (CI 95%: 34 to 40).Conclusions. The use of NOC indicators allows evaluation of management of the therapeutic regime in people with chronic conditions with a satisfactory validity and it provides new approaches for dealing with this problem(AU)

Tratamiento de las malformaciones linfáticas mediante esclerosis con OK-432. Nuestra experiencia / Treatment of lymphatic malformations with ok-432 sclerosis. Our experience

Introducción. Las malformaciones linfáticas se han tratado clásicamente mediante exéresis quirúrgica. En los últimos años se ha extendido el tratamiento mediante esclerosis con diferentes sustancias, entre las que destaca el OK-432. El objetivo de este trabajo es presentar nuestra experiencia con este tratamiento .Material y Métodos. Revisión de los pacientes con diagnóstico de malformación linfática, tratados mediante esclerosis con OK-432 en nuestro centro. Los procedimientos se realizaron bajo anestesia general mediante control ecográfico y quistografía previa por parte del radió-logo intervencionista pediátrico y un cirujano de la sección. Resultados. 15 pacientes han sido tratados. En total se han realizado 36 procedimientos, 2,4 infiltraciones de media por paciente con una cantidad media de 1,5 miligramos (mg) de preparado diluido inyectado en cada una. Los resultados han sido satisfactorios en el 73% de (..) (AU)

Introduction. The elective treatment of lymphatic malformations has always been surgical resection. However sclerotherapy with a variety of substances is being used as an alternative to surgery special yin the last years. The aim of this essay is to present our results usings clerosis with OK-432.Materials and methods. We have reviewed all patients affected by lymphatic malformation that have been treated by sclerosis with OK-432. Each procedure was performed under general anesthesia by a radiologist and a pediatric surgeon. Results. Fifteen patients have been treated. On the whole, 36 procedures have been performed, with a mean of 2.4 for each patient. The mean amount of OK-432 injected was 1.5 milligrams (mg) in each procedure. 73% of the patients showed nearly complete or complete (AU)

[Treatment of lymphatic malformations with OK-432 sclerosis. Our experience]. / Tratamiento de las malformaciones linfáticas mediante esclerosis con OK-432. Nuestra experiencia.

INTRODUCTION: The elective treatment of lymphatic malformations has always been surgical resection. However sclerotherapy with a variety of substances is being used as an alternative to surgery specially in the last years. The aim of this essay is to present our results using sclerosis with OK-432. MATERIALS AND METHODS: We have reviewed all patients affected by lymphatic malformation that have been treated by sclerosis with OK-432. Each procedure was performed under general anesthesia by a radiologist and a pediatric surgeon. RESULTS: Fifteen patients have been treated. On the whole, 36 procedures have been performed, with a mean of 2.4 for each patient. The mean amount of OK-432 injected was 1.5 milligrams (mg) in each procedure. 73% of the patients showed nearly complete or complete response. There were two complications; one of them was a limited bleeding and the other one, a case of high fever resolved by antipyretics and antibiotics. Microcystic or mixed malformations required a higher number of procedures and presented poorer response. CONCLUSION: Sclerotherapy with OK-432 is an effective and safe treatment for lymphatic malformation specially the macrocystic ones, therefore it should be considered as first line therapeutic option.

Diagnosis of placental pathogens in small ruminants by immunohistochemistry and PCR on paraffin-embedded samples.

A histological study was carried out on 58 formalin-fixed and paraffin-embedded samples of placenta from sheep and goats that had aborted, and the placental lesions were graded. Sequential histological sections of each cotyledon were then immunostained with specific antibodies and used for PCR detection of Chlamydophila abortus, Coxiella burnetii, Salmonella Abortusovis, Brucella melitensis, Listeria monocytogenes and Toxoplasma gondii. Most of the cotyledons showed different degrees of placentitis. The proportional agreement between the two techniques was 0.879 (kappa value 0.746). C abortus was the most prevalent pathogen. Mixed infections were common.

Chlamydophila abortus infection in the mouse: a useful model of the ovine disease.

Chlamydophila (C.) abortus is an obligate intracellular bacterium able to colonize the placenta of several species of mammals, which may induce abortion in the last third of pregnancy. The infection affects mainly small ruminants resulting in major economic losses in farming industries worldwide. Furthermore, its zoonotic risk has been reported in pregnant farmers or abattoir workers. Mouse models have been widely used to study both the pathology of the disease and the role of immune cells in controlling infection. Moreover, this animal experimental model has been considered a useful tool to evaluate new vaccine candidates and adjuvants that could prevent abortion and reduce fetal death. Future studies using these models will provide and reveal information about the precise mechanisms in the immune response against C. abortus and will increase the knowledge about poorly understood issues such as chlamydial persistence.

Characterization of a murine model of intranasal infection suitable for testing vaccines against C. abortus.

Mouse models have been widely used to test candidate vaccines against Chlamydophila abortus infection in mice. Although the induction of a systemic infection by endogenous or intraperitoneal inoculation is a useful tool for understanding the immune mechanism involved in the protection conferred by the vaccination, a different approach is necessary to understand other factors of the infection, such as mucosal immunity or the colonization of target organs. To test whether C. abortus intranasal model of infection in mice is a useful tool for testing vaccines in a first group of experiments mice, were infected intranasally with C. abortus to characterize the model of infection. When this model was used to test vaccines, two inactivated experimental vaccines, one of them adjuvated with QS-21 and another with aluminium hydroxide, and a live attenuated vaccine (strain 1B) were used. Non-vaccinated control mice died within the first 8 days, after displaying substantial loss of weight. Histologically, the mice showed lobar fibrinopurulent bronchointerstitial pneumonia. Prior immunization with QS-21 adjuvated vaccine or 1B vaccine presented mortality and the recipients showed a greater number of T cells in the lesions, especially CD8(+) T cells, than the control mice and mice immunized with vaccine adjuvated with aluminium hydroxide. The results confirm that the C. abortus intranasal model of infection in mice is a useful tool for testing vaccines.

Role of polymorphonuclear neutrophils (PMNs) and NK cells in the protection conferred by different vaccines against Chlamydophila abortus infection.

Ovine enzootic abortion (OEA) is caused by Chlamydophila abortus, an intracellular bacterium which acts by infecting the placenta, causing abortion in the last term of gestation. The main prevention strategy against OEA is the vaccination of flocks. An effective vaccine against C. abortus must induce a Th1-like specific immune response, which is characterized by the early production of IFN-gamma and the activation of CD8(+)T cells. Moreover, vaccine effectiveness could be modulated by the functioning of the innate immunity. The purpose of this study was to ascertain how polymorphonuclear neutrophils (PMNs) and NK cells might influence vaccine-induced protection. The live attenuated 1B vaccine and two inactivated experimental vaccines, adjuvated with aluminium hydroxide (AH) or QS-21 (QS), were used in PMN-depleted or NK cell-depleted mice. For PMN depletion, RB6-8C5 monoclonal antibody, which recognizes GR1(+) receptors (Robben, P.M., LaRegina, M., Kuziel, W.A., Sibley, L.D. 2005. Recruitment of Gr-1(+) monocytes is essential for control of acute toxoplasmosis. The Journal of Experimental Medicine 201, 1761-1769.) was used, while for NK cell-depletion the anti-asialo GM1 polyclonal antibody was used. The depletion of PMNs caused 100% mortality in non-vaccinated mice (NV) and 60% mortality in the AH-vaccinated mice by day 10 p.i., while both groups showed a significant increase in their bacterial burden in the liver by day 4 p.i. The depletion of NK cells caused mortality only in the NV group (50% by day 10 p.i.), although this group and the 1B vaccinated mice showed an increased bacterial burden in the liver at day 4 p.i. Our results suggest that the importance of PMNs in inactivated vaccines depends on the adjuvant chosen. The results also demonstrated that the importance of NK cells is greater in live vaccines than in inactivated vaccines.

Relative importance of CD4+ and CD8+ T cells in the resolution of Chlamydophila abortus primary infection in mice.

The role of the specific cellular immune response is well established in Chlamydiaceae infections, but the importance of each T-cell subset seems to be species-dependent. This study was designed to clarify the role of T-cell subsets in the response to Chlamydophila abortus primary infection. C57BL/6 mice were depleted of CD4+ or CD8+, or both, by monoclonal antibody injections and subsequently infected with C. abortus. Mice were killed at intervals and samples were collected for bacteriological and histopathological analysis. Also carried out were spleen cell culture, cytokine quantification, immunolabelling for C. abortus antigen, and a TUNEL assay for apoptosis. CD8+ T cell-depleted mice all died within 12 days of C. abortus infection, while no mortality was observed in the other groups; surprisingly, CD4+ T cell-depleted mice showed lower morbidity (expressed as weight loss) than did a non-depleted (control) group. CD8+ T cell-depleted mice also differed from the other groups in showing a significantly higher chlamydial burden in the liver. CD8+ T cell-depleted mice also had a higher number of apoptotic cells in hepatic inflammatory foci and showed exacerbated IFN-gamma production by spleen cells after specific stimulation. Simultaneous depletion of both T-cell subpopulations led to a chronic infection, but not to early mortality. It is concluded that CD8+ T cells may play a role in the regulatory control of the CD4+ T-cell response and may have a direct cytotoxic or IFN-gamma-mediated effect on infected cells.

Influence of the Th2 immune response established by Nippostrongylus brasiliensis infection on the protection offered by different vaccines against Chlamydophila abortus infection.

Chlamydophila abortus is the aetiological agent of enzootic abortion in small ruminants in which it infects the placenta to cause abortion during the last trimester of gestation. In a mouse model, a Th1 immune response involving IFN-gamma production and CD8+ T cells is necessary for the infection to be resolved. The authors previously demonstrated that infection with Nippostrongylus brasiliensis, a rodent gastrointestinal nematode extensively used in experimental models to induce Th2 responses, alters the specific immune response against C. abortus infection, increasing bacterial multiplication in liver and reducing specific IFN-gamma production. The aim of the present work was to clarify whether a Th2 immune response has any influence on the success of vaccination using both inactivated and attenuated vaccines. The results showed that the Th2 response established prior to vaccination did not influence the induction of protection offered by the vaccines. However, the effectiveness of this protective response can be altered, depending on the adjuvant employed in the inactivated vaccines, when the Th2 response is established after vaccination, just before challenge with C. abortus.

Natural killer (NK) cells play a critical role in the early innate immune response to Chlamydophila abortus infection in mice.

Chlamydophila abortus, the aetiological agent of ovine enzootic abortion, induces a strong inflammatory reaction that leads to the T helper cell (Th1) specific immune response necessary for the clearance of infection. Because the role of natural killer (NK) cells during the first stages of this response has received little attention, this study focused on determining the function of these cells in a mouse model of infection. The location of NK cells in the liver and spleen of infected mice was examined immunohistochemically with an anti-Ly49G monoclonal antibody. The number of NK cells increased during the infection both in spleen and liver. In subsequent experiments, an anti-asialo GM1 polyclonal antibody was injected to deplete the NK cells. NK-depleted mice showed a substantial increase in their susceptibility to C. abortus infection, with high mortality rates and an increased burden of bacteria in the liver. Histopathological studies showed that inflammatory foci, composed mainly of neutrophils, were greater in size and number in depleted mice, while numerous chlamydial inclusions were associated with the foci. Serum concentrations of IFN-gamma, a key cytokine in the control of C. abortus infection, were substantially reduced in the NK-depleted mice. To establish the relationship between NK cells and other components of the innate immune response, neutrophils were depleted with the RB6-8C5 antibody. These cells were shown to be crucial in the recruitment of NK cells to the inflammatory foci.