Overexpression of SPARC obliterates the in vivo tumorigenicity of human hepatocellular carcinoma cells

Hepatocellular carcinoma (HCC) is the sixth most common cancer and the third leading cause of cancer-related death worldwide. Current treatments are extremely disappointing. SPARC (Secreted protein, acidic and rich in cysteine) is a matricellular glycoprotein with differential expression in several tumors, including HCC, which significance remains unclear. We infected HCC cells (HepG2, Hep3B and Huh7) with an adenovirus expressing SPARC (AdsSPARC) to examine the role of SPARC expression on HCC cells and its effect on tumor aggressiveness. The in vitro HCC cells substrate-dependent proliferation and cell cycle profile were unaffected; however, SPARC overexpression reduced HCC proliferation when cells were grown in spheroids. A mild induction of cellular apoptosis was observed upon SPARC overexpression. SPARC overexpression resulted in spheroid growth inhibition in vitro while no effects were found when recombinant SPARC was exogenously applied. Moreover, the clonogenic and migratory capabilities were largely decreased in SPARC-overexpressing HCC cells, altogether suggesting a less aggressive HCC cell phenotype. Consistently, AdsSPARC-transduced cells showed increased E-cadherin expression and a concomitant decrease in N-cadherin expression. Furthermore, SPARC overexpression was found to reduce HCC cell viability in response to 5-FU-based chemotherapy in vitro, partially through induction of apoptosis. In vivo experiments revealed that SPARC overexpression in HCC cells inhibited their tumorigenic capacity and increased animal survival through a mechanism that partially involves host macrophages. Our data suggest that SPARC overexpression in HCC cells results in a reduced tumorigenicity partially through the induction of mesenchymal-to-epithelial transition (MET). These evidences point to SPARC as a novel target for HCC treatment.Fil: Atorrasagasti, María Catalina. Universidad Austral. Facultad de Ciencias Biomédicas. Laboratorio de Terapia Genética; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Malvicini, Mariana. Universidad Austral. Facultad de Ciencias Biomédicas. Laboratorio de Terapia Genética; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Aquino, Jorge Benjamin. Universidad Austral. Facultad de Ciencias Biomédicas. Laboratorio de Terapia Genética; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Alaniz, Laura Daniela. Universidad Austral. Facultad de Ciencias Biomédicas. Laboratorio de Terapia Genética; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: García, Mariana Gabriela. Universidad Austral. Facultad de Ciencias Biomédicas. Laboratorio de Terapia Genética; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Bolontrade, Marcela Fabiana. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigaciones Bioquímicas de Buenos Aires. Fundación Instituto Leloir. Instituto de Investigaciones Bioquímicas de Buenos Aires; ArgentinaFil: Rizzo, Manglio Miguel. Universidad Austral. Facultad de Ciencias Biomédicas. Laboratorio de Terapia Genética; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Podhajcer, Osvaldo Luis. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigaciones Bioquímicas de Buenos Aires. Fundación Instituto Leloir. Instituto de Investigaciones Bioquímicas de Buenos Aires; ArgentinaFil: Mazzolini Rizzo, Guillermo Daniel. Universidad Austral. Facultad de Ciencias Biomédicas. Laboratorio de Terapia Genética; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentin

Dual effect of Thymosin α 1 on human monocyte-derived dendritic cellin vitrostimulated with viral and bacterial toll-like receptor agonists

OBJECTIVES: Thymosin α 1 (Tα1) recently gained interest as immune adjuvant for vaccines because of its ability to modulate the T-cell/dendritic cell (DC) axis and to improve antibody production. The objective of this study was to determine whether Tα1 would address in vitro the response of human primary monocyte-derived DC, crucial regulators of vaccine-induced immunity, upon exposure to different toll-like receptor (TLR) agonists or infection with viruses or bacteria. METHODS: DC maturation and production of pro-inflammatory cytokines were analyzed. RESULTS: Our data revealed a dual effect of Tα1 on DC biology upon viral or bacterial stimulation. Interestingly, Tα1 enhanced human leukocyte antigen (HLA)-I and II surface expression and secretion of IL-6, TNF-α and IL-8 when DCs were treated with viral TLR3 and TLR7/8 agonists. Similarly, in pandemic H1N1 influenza A-infected DCs, Tα1 raised the expression of maturation markers and type I and III Interferon (IFN). In contrast, following bacterial TLR2 and 4 stimulation, as well as upon Bacillus Calmette-Guerin infection, the presence of Tα1 in DC cultures drastically lowered the analyzed cellular parameters. CONCLUSION: The knowledge that Tα1 pleiotropic effect might ameliorate anti-viral immune responses and, at the same time, dampen inflammation caused by bacterial infections could lay the groundwork for a more appropriate therapeutic application of this molecule

Tumor associated fibroblast: impact on osteosarcoma primary and metastatic tumoral microenvironment and treatment response

Tumor associated fibroblast (TAF) have been implicated in almost every aspect of tumoral biology. Of relevance TAF could be modulating response to treatment and overall microenvironment development. Given that Osteosarcoma (OS) have the same 5-year survival rate for metastatic and treatment resistant patients since 1970 ́s, we dediced to investigate the role of TAF in OS primary and metastatic niches. Aim: Evaluate TAF and human OS cell lines interaction in primary and pulmonary metastatic environments. To analyze if metastatic OS cell line has a higher inducing power than non-metastatic OS cell line analyzing the expression of different ABC transporters im-plicated in chemoresistance and the ability to exclude doxorubicin and rhodamine. Methods: The expression of ABC transporters was analyzed by RT-qPCR on conditioned fibroblast. Rhodamine 123 exclution assay was used to determine the activity of P-glycoprotein (P-gp) mediated transport and doxorubicin (DOX) exclution was performed to analysis the overall ABC-related chemoresistant capacity. To evaluate the interaction of fibroblast with metastatic (LM7) and non-metastatic (SAOS2) OS human cells hetero – spheroid formation assays were performed. Results: LM7 conditioned medium (CM) induced an overall upregulation of ABC transporters in comparison with SAOS2 CM. Conditioned fibroblast with LM7 CM showed lower levels of intracellular DOX and Rhodomine in comparison with SAOS2 CM fibroblast. Mixed spheroid compose of fibroblast ans OS cell lines display a lower area and more compact than single type aggregates. OS has not changed the 5-year rate survival for metastatic patients since the 70’, so the need to understand aspects of OS metastatic biology and chemoresistance could be helpful to develop new treat-ments to this group. Knowing aspects of the associated stroma and in particular TAF, could allow the development of new therapeutic possibilities targeting the tumoral associated stroma.Fil: Valenzuela Alvarez, Matias Juan Pablo. Consejo Nacional de Investigaciones Cientificas y Tecnicas. Oficina de Coordinacion Administrativa Houssay. Instituto de Medicina Traslacional E Ingenieria Biomedica. - Hospital Italiano. Instituto de Medicina Traslacional E Ingenieria Biomedica. - Instituto Universitario Hospital Italiano de Buenos Aires. Instituto de Medicina Traslacional E Ingenieria Biomedica.; ArgentinaFil: Rizzo, Matias Eduardo. Consejo Nacional de Investigaciones Cientificas y Tecnicas. Oficina de Coordinacion Administrativa Houssay. Instituto de Medicina Traslacional E Ingenieria Biomedica. - Hospital Italiano. Instituto de Medicina Traslacional E Ingenieria Biomedica. - Instituto Universitario Hospital Italiano de Buenos Aires. Instituto de Medicina Traslacional E Ingenieria Biomedica.; ArgentinaFil: Auzmendi, Jeronimo. Universidad de Buenos Aires. Facultad de Medicina; ArgentinaFil: Lazarowski, Alberto. Universidad de Buenos Aires; ArgentinaFil: Bolontrade, Marcela Fabiana. Consejo Nacional de Investigaciones Cientificas y Tecnicas. Oficina de Coordinacion Administrativa Houssay. Instituto de Medicina Traslacional E Ingenieria Biomedica. - Hospital Italiano. Instituto de Medicina Traslacional E Ingenieria Biomedica. - Instituto Universitario Hospital Italiano de Buenos Aires. Instituto de Medicina Traslacional E Ingenieria Biomedica.; ArgentinaLXVI Reunión Anual de la Sociedad Argentina de Investigación Clínica: LXIX Reunión Anual de la Sociedad Argentina de Inmunología; LIII Reunión Anual de la Asociación Argentina de Farmacología Experimental y XI Reunión Anual de la Asociación Argentina de NanomedicinasArgentinaSociedad Argentina de Investigación ClínicaSociedad Argentina de InmunologíaAsociación Argentina de Farmacología Experimenta

Impact of two different exercise programs on persistent cancer-related fatigue and physical fitness

Cancer-related fatigue is a common symptom in patients with cancer, which is experienced by 70% to 100% of these patients and brings some impairment of physical and mental performance, hinders their working or carrying out regular daily activities, and hence results in a substantial reduction of the quality of life. Physical exercise has consistently been identified as a central element of rehabilitation for many chronic diseases like cancer, and increasing evidence supports the contention that physical activity is a valuable intervention that can be utilized in conjunction with conventional therapies during CRF treatment. Objective: The aim of this study was to assess the impact of a program of physical exercise on fatigue levels and physical performance of cancer patients. Method: A consecutive series of 44 adult patients with neoplastic disease (solid or hematological), with a medical diagnosis of fatigue, who were enrolled in an oncological treatment, with the ability to walk and willing to enter a rehabilitation program of exercise for at least 4 consecutive months. The exercise program was performed two times per week, each session lasting one hour and consisting of aerobic, resistance, and flexibility exercises. The protocol was divided into aerobic exercise and resistance training combined with aerobic exercise. The patients were evaluated with two assessments: one prior to their beginning the exercise program and other at the end of the four-month program. In both assessments the patients completed the Revised Piper Fatigue Scale and the six-minute walk test. The primary outcome of change over baseline and after 16 weeks in PFS-R score and six-minute walk test were compared using a two sample two-sided t-test for both groups. Alpha level was set at p < 0.05. Results: After 16 weeks, the patients who participated in the aerobic or the combined exercise program reported significantly higher levels of physical functioning (6-minute walking test, p = 0.0009 and p = 0.001, respectively) and significantly lower fatigue (PFS-R, p = 0.003 and p = 0.002, respectively) than at the beginning the exercise program. Conclusion: The results of patients who underwent aerobic or aerobic + anaerobic exercise showed statistically significant improvement of physical performance and of fatigue. Data from this study corroborates with the literature showing that exercise programs with aerobic or resistance exercises are an effective strategy for the treatment of fatigue. The results of this study confirm that physical exercise could be useful in rehabilitation of cancer survivors, especially for fatigued patients.A fadiga relacionada ao câncer é um dos sintomas mais comuns entre pacientes com câncer, relatada em 70% a 100% desses pacientes resultando em uma reduçao significativa da qualidade de vida, funcionalidade e independência. O exercício físico tem sido identificado como um elemento central de reabilitaçao de muitas doenças crônicas como câncer, e cada vez mais evidências apoiam a tese de que a atividade física é uma intervençao útil, que pode ser utilizada em conjunto com terapias convencionais durante o tratamento da fadiga relacionada ao câncer. Objetivo: Avaliar o impacto de dois programas de exercício físico sobre os níveis de fadiga e desempenho físico de pacientes com câncer. Método: Relato de uma série consecutiva de 44 doentes adultos com doença neoplásica (sólido ou hematológicas), e diagnóstico médico de fadiga, submetidos a dois diferentes programas de exercício físico. Todos os doentes foram avaliados quanto a desempenho físico com o uso do teste de caminhada de 6 minutos e avaliados quanto aos níveis de fadiga com o teste de Piper, antes e depois de 4 meses de atividade física supervisionada (exercícios aeróbicos isolados e treino de resistência combinado ao exercícios aeróbicos). Resultados: Após 16 semanas, os doentes que participaram do programa de exercícios aeróbicos ou que participaram do protocolo de exercício aeróbico combinado com anaeróbio, relataram níveis significativamente mais elevados do desempenho físico (6 minutos teste de caminhada, p = 0,0009 e p = 0,001, respectivamente) e níveis de fadiga significativamente menor (PFS- R, p = 0,003 e p = 0,002, respectivamente) do que no início do programa de exercícios. Conclusão: Estes resultados demonstram que tanto um protocolo de exercício aeróbico quanto de exercício aeróbico combinado com exercício anaeróbio apresentam melhora significativa do desempenho físico e dos níveis de fadiga de doentes oncológicos. Os dados deste estudo corroboram a literatura mostrando que a atividade física é uma estratégia eficaz para o tratamento da fadiga. Os resultados deste estudo confirmam que o exercício físico pode ser útil na reabilitaçao de sobreviventes de câncer, especialmente para pacientes com fadiga oncológica

Composting process evaluation of two mixtures of poultry manures

Los residuos del sector avícola, principalmente guano (aves ponedoras) y cama de parrilleros (aves de engorde), pueden generar un impacto negativo en el ambiente contribuyendo a la contaminación de suelo, agua y aire. La estabilización aeróbica a través del compostaje es una alternativa de tratamiento para reducir la contaminación. El objetivo de este trabajo fue evaluar el proceso de compostaje en dos mezclas con diferentes porcentajes de residuos avícolas (guano de aves ponedoras y cama de pollos parrilleros). Se compostaron dos mezclas que contenían 81% y 70% de residuos avícolas durante 16 semanas. Las variables analizadas fueron: temperatura (T°), pH, conductividad eléctrica (CE), humedad (H), capacidad de intercambio catiónico (CIC), carbono orgánico total (COT), amonio (NH4+), nitrato (NO3 - ), nitrógeno total (NT ) y carbono soluble (CS). Las características finales de los compost A y B fueron: pH 7,1 - 6,8, CE 3,3 - 2,9 (mS. cm- 1), COT 14,8 - 17,9 %, NT 0,97 - 0,88 %, NH4 + 501 - 144,9 mg kg-1, NO3-552,3 - 543,0 mg kg-1 respectivamente. El proceso de compostaje podría ser una herramienta para estabilizar los residuos avícolas minimizando su impacto en el ambiente.Chicken production wastes, principally poultry manure (layers) and litter (from chickens for fattening), may impact negatively on the environment, contributing to soil, water and air pollution. Aerobic stabilization through composting is an alternative to reduce contamination. This study aims to evaluate the composting process in two mixtures with different percentages of poultry manure of laying chickens and broiler poultry litter. Two mixtures were composted containing 81% and 70% of poultry wastes for 16 weeks. The analytical parameters were: temperature (T°), pH, electrical conductivity (EC), humidity (H), cationic exchange capacity (CEC), total organic carbon (TOC), amonium (NH4+), nitrate (NO3- ), total nitrogen (TN), soluble carbon soluble (SC) were determined in each treatment. The final compost A and B characteristics were 7,1 - 6,8, CE 3,3 - 2,9 (mS.cm-1), COT 14,8 - 17,9%, NT 0.97 - 0.88%, NH4+ 501 - 144,9 mg kg-1, NO3 - 552,3 -543,0 mg kg-1 respectively. The composting process may be a tool to stabilize poultry wastes, minimizing its environmental damage.Fil: Riera, Nicolás I.. Instituto Nacional de Tecnología Agropecuaria Castelar (Buenos Aires). Laboratorio de Transformación de Residuos, Instituto de Microbiología y Zoología Agrícola.Fil: Della Torre, Virginia. Instituto Nacional de Tecnología Agropecuaria Castelar (Buenos Aires). Laboratorio de Transformación de Residuos, Instituto de Microbiología y Zoología Agrícola.Fil: Rizzo, Pedro F.. Instituto Nacional de Tecnología Agropecuaria Castelar (Buenos Aires). Laboratorio de Transformación de Residuos, Instituto de Microbiología y Zoología Agrícola.Fil: Butti, Mariano. Instituto Nacional de Tecnología Agropecuaria Castelar (Buenos Aires). Laboratorio de Transformación de Residuos, Instituto de Microbiología y Zoología Agrícola.Fil: Bressan, Fabiana M.. Instituto Nacional de Tecnología Agropecuaria Castelar (Buenos Aires). Laboratorio de Transformación de Residuos, Instituto de Microbiología y Zoología Agrícola.Fil: Zarate, Natalia. Instituto Nacional de Tecnología Agropecuaria Castelar (Buenos Aires). Laboratorio de Transformación de Residuos, Instituto de Microbiología y Zoología Agrícola.Fil: Weigandt, Cristian. Instituto Nacional de Tecnología Agropecuaria Castelar (Buenos Aires). Laboratorio de Transformación de Residuos, Instituto de Microbiología y Zoología Agrícola.Fil: Crespo, Diana Elvira. Instituto Nacional de Tecnología Agropecuaria Castelar (Buenos Aires). Laboratorio de Transformación de Residuos, Instituto de Microbiología y Zoología Agrícola

Apoptotic epitope-specific CD8+ T cells and interferon signaling intersect in chronic hepatitis C virus infection

CD8(+) T cells specific to caspase-cleaved antigens derived from apoptotic T cells represent a principal player in chronic immune activation (CIA). Here, we found that both apoptotic epitope (AE)-specific and hepatitis C virus (HCV)-specific CD8(+) T cells were mostly confined within the effector memory (EM) or terminally differentiated EM CD45RA(+) cell subsets expressing a dysfunctional T-helper-1-like signature program in chronic (c)HCV infection. However, AE-specific CD8(+) T cells produced tumor necrosis factor (TNF)-α and interleukin-2 at the intrahepatic level significantly more than HCV-specific CD8(+) T cells, despite both populations acquiring high levels of programmed death-1 receptor expression. Contextually, only AE-specific CD8(+) T cells correlated with both interferon-stimulated gene levels in T cells and hepatic fibrosis score. Taken together, these data suggest that AE-specific CD8(+) T cells can sustain CIA by their capacity to produce TNF-α and be resistant to inhibitory signals more than HCV-specific CD8(+) T cells in cHCV infection

Attitudes, knowledge and Practices of Nurses towards HIV/AIDS Patients. An Observational, Cross Sectional, Multicenter study

Background. Attitudes, knowledge and practices of nurses towards HIV/AIDS patients are of continuous interest, especially in developing countries. However, in Italy, this topic is still scarcely debated. Materials and methods. An observational, cross sectional, multicentre study was conducted on a sample of 144 nurses in two Italian Hospitals of the Puglia Region ("Vito Fazzi" Hospital in Lecce and "San Giuseppe da Copertino" Hospital in Copertino (Le)). Results. A large part of the sample (97.2%, n=140) stated that they never refused to take care of a patient with AIDS. Only 22.9%, (n=33), of the sample had attended a training course and almost half (63.2%, n=91) used the gloves regularly when performing blood samples or when administering therapy to patients. With regard to the assessment of HIV knowledge, the percentage of nurses who know the meaning of the terms " seropositive" (83.3%, n=120), "HIV-positive person" (91.7%, n=132) and "window period" (47.9%, n=69) decreased. Conclusions. The results of the study show that a non-negligible percentage of nurses could be considered at risk of infection due to non-routine use of gloves, incorrect handling of the patient's biological samples and not knowing how to decontaminate a surface with potentially infected blood

Impact of Mycobacterium tuberculosis RD1-locus on human primary dendritic cell immune functions

Modern strategies to develop vaccines against Mycobacterium tuberculosis (Mtb) aim to improve the current Bacillus Calmette-Guerin (BCG) vaccine or to attenuate the virulence of Mtb vaccine candidates. In the present study, the impact of wild type or mutated region of difference 1 (RD1) variants on the immunogenicity of Mtb and BCG recombinants was investigated in human primary dendritic cells (DC). A comparative analysis of transcriptome, signalling pathway activation, maturation, apoptosis, cytokine production and capacity to promote Th1 responses demonstrated that DC sense quantitative and qualitative differences in the expression of RD1-encoded factors - ESAT6 and CFP10 - within BCG or Mtb backgrounds. Expansion of IFN-γ producing T cells was promoted by BCG::RD1-challenged DC, as compared to their BCG-infected counterparts. Although Mtb recombinants acted as a strong Th-1 promoting stimulus, even with RD1 deletion, the attenuated Mtb strain carrying a C-terminus truncated ESAT-6 elicited a robust Th1 promoting phenotype in DC. Collectively, these studies indicate a necessary but not sufficient role for the RD1 locus in promoting DC immune-regulatory functions. Additional mycobacterial factors are likely required to endow DC with a high Th1 polarizing capacity, a desirable attribute for a successful control of Mtb infection

Role of cannabinoids in the treatment of Tinnitus

Tinnitus is a frequent symptom in audiological clinical practice characterized by an abnormal noise perceived in one or both ears or in the head, in which a patient has a conscious hearing percept in absence of external sound. Tinnitus might be caused by a homeostatic response of central dorsal cochlear nucleus auditory neurons that makes them hyperactive in compensation to auditory input loss. One hypothesis suggests that tinnitus is a sensory form of epilepsy that involves the cochlear nucleus and the inferior colliculus, which display impairment in the electrical activity in the auditory system. This alteration determines a synaptic plasticity in the dorsal cochlear nucleus that becomes a target for pharmacological compounds able to treat tinnitus. There is no effective drug treatment for tinnitus, but different studies propose the use of cannabinoid receptors agonist for their anti-epileptic activity, although their practical effects are still unclear. In this review, we want to analyze the emerging pharmacological approaches of cannabinoid receptor agonists to the therapy of tinnitus

V-Edge: Virtual Edge Computing as an Enabler for Novel Microservices and Cooperative Computing

As we move from 5G to 6G, edge computing is one of the concepts that needs revisiting. Its core idea is still intriguing: instead of sending all data and tasks from an end user's device to the cloud, possibly covering thousands of kilometers and introducing delays that are just owed to limited propagation speed, edge servers deployed in close proximity to the user, e.g., at some 5G gNB, serve as proxy for the cloud. Yet this promising idea is hampered by the limited availability of such edge servers. In this paper, we discuss a way forward, namely the virtual edge computing (V-Edge) concept. V-Edge bridges the gap between cloud, edge, and fog by virtualizing all available resources including the end users' devices and making these resources widely available using well-defined interfaces. V-Edge also acts as an enabler for novel microservices as well as cooperative computing solutions. We introduce the general V-Edge architecture and we characterize some of the key research challenges to overcome, in order to enable wide-spread and even more powerful edge services