Precision Medicine, Artificial Intelligence, and Genomic Markers in Urology. Do we need to Tailor our Clinical Practice? / Medicina de precisión, inteligencia artificial y marcadores genómicos en urología. ¿Debemos cambiar nuestra práctica clínica?

Precision medicine plays a key role in urological oncology practice nowadays, with the breakthrough of the poly (ADP-ribose) polymerase inhibitors (PARPi), which play a critical role in different DNA damage repair (DDR) pathways, the immune checkpoint inhibitors, the genomic expression profiles and current genome manipulation-directed targeted therapy. Information and technology (IT) are set to change the way we assess and treat patients and should be reviewed and discussed. The aim of the present article is to demonstrate a detailed revision on precision medicine, including novel therapeutic targets, genomic markers, genomic stratification of urological patients, and the top-notch technological breakthroughs that could change our clinical practice We performed a review of the literature in four different databases (PubMed, Embase, Lilacs, and Scielo) on any information concerning prostate, bladder, kidney and urothelial cancer novel treatments with PARPi, immune checkpoint inhibitors (ICIs), targeted therapy with fibroblast growth factor receptor inhibitors (FGFRi), and theranostics with prostate-specific membrane antigen (PSMA) targeted monoclonal antibodies. Artificial intelligence, machine learning, and deep learning algorithm in urological practice were also part of the search. We included all articles written in English, published within the past 7 years, that discussed outstanding therapies and genomics in urological cancer and artificial intelligence applied to urology. Meanwhile, we excluded articles with lack of a clear methodology and written in any other language than English. One-hundred and twenty-six articles of interest were found; of these, 65 articles that presented novel treatments of urological neoplasms, discussed precision medicine, genomic expression profiles and biomarkers in urology, and latest deep learning and machine learning algorithms as well as the use of artificial intelligence in urological practice were selected. A critical review of the literature is presented in the present article. Urology is a constantly changing specialty with a wide range of therapeutic breakthroughs, a huge understanding of the genomic expression profiles for each urological cancer and a tendency to use cutting-edge technology to treat our patients. All of these major developments must be analyzed objectively, taking into account costs to the health systems, risks and benefits to the patients, and the legal background that comes with them. A critical analysis of these new technologies and pharmacological breakthroughs should be made before considering changing our clinical practice. Nowadays, research needs to be strengthened to help us improve results in assessing and treating our patients

La medicina de precisión juega un rol fundamental en la práctica clínica de la urologia oncológica en la actualidad, con el desarrollo de los inhibidores de la poli (ADP-ribosa) polimerasa (PARPi), que juegan un papel fundamental en las distintas vías del reparo del ADN dañado (RAD), los inhibidores del punto de chequeo inmune (ICI), los perfiles de expresión genómicos, y la terapia blanco-dirigida a la manipulación genómica. El desarrollo tecnológico y la informática están cambiando la forma como evaluamos y tratamos a los pacientes, y se debe discutir y revisar a detalle. El objetivo de este artículo es hacer una revisión detallada acerca de la medicina de precisión, genómica, y los avances tecnológicos en nuestro campo. Realizamos una revisión de la literatura en cuatro bases de datos diferentes (PubMed, Embase, Lilacs, y Scielo), buscando cualquier información relacionada con cáncer de próstata, vejiga, riñón y carcinoma urotelial, tratamientos novedosos con PARPi, ICI, terapia-blanco con inhibidores del receptor del factor de crecimiento de los fibroblastos (FGFRi) y teragnósticos con anticuerpos monoclonales dirigidos al antígeno de membrana específico de la próstata (AMEP). Inteligencia artificial, aprendizaje de máquinas y algoritmos de aprendizaje profundo en la práctica urológica también fueron revisados. Incluimos artículos escritos en inglés, publicados dentro de los últimos 7 años, que abordaran terapias novedosas y genómica en cáncer urológico e inteligencia artificial aplicada a la urología. Excluimos artículos con falta de una metodología adecuada y escritos en cualquier idioma diferente al inglés. En total, 126 artículos de interés fueron encontrados, y, de estos seleccionamos 65 artículos que reportaban tratamientos novedosos para neoplasias urológicas, discutían medicina de precisión y perfiles de expresión genómica y bio-marcadores en urología, algoritmos de aprendizaje profundo, aprendizaje de máquina, y el uso de inteligencia artificial en la práctica urológica. Se hizo una revisión crítica de la literatura que se presenta en este artículo. La urología es una especialidad constantemente en cambio, con un gran rango de avances terapéuticos, un gran conocimiento de los perfiles de expresión genómica para cada cáncer urológico, y una tendencia a utilizar tecnología de punta para estudiar y tratar a nuestros pacientes. Todos estos desarrollos se deben analizar objetivamente, y hay que tener en cuenta los costos al sistema de salud, los riesgos y beneficios para los pacientes, y el contexto legal que implica cada uno. Hasta la fecha, estos avances tecnológicos y farmacológicos se deben analizar con cautela antes de vernos en la posición de cambiar nuestra práctica clínica. Se debe fortalecer la investigación médica para mejorar los resultados en el tratamiento y abordaje de nuestros pacientes.

Heterogeneity of Human γδ T Cells and Their Role in Cancer Immunity

The γδ T cells are unconventional lymphocytes that function in both innate and adaptive immune responses against various intracellular and infectious stresses. The γδ T cells can be exploited as cancer-killing effector cells since γδ TCRs recognize MHC-like molecules and growth factor receptors that are upregulated in cancer cells, and γδ T cells can differentiate into cytotoxic effector cells. However, γδ T cells may also promote tumor progression by secreting IL-17 or other cytokines. Therefore, it is essential to understand how the differentiation and homeostasis of γδ T cells are regulated and whether distinct γδ T cell subsets have different functions. Human γδ T cells are classified into Vδ2 and non-Vδ2 γδ T cells. The majority of Vδ2 γδ T cells are Vγ9δ2 T cells that recognize pyrophosphorylated isoprenoids generated by the dysregulated mevalonate pathway. In contrast, Vδ1 T cells expand from initially diverse TCR repertoire in patients with infectious diseases and cancers. The ligands of Vδ1 T cells are diverse and include the growth factor receptors such as endothelial protein C receptor. Both Vδ1 and Vδ2 γδ T cells are implicated to have immunotherapeutic potentials for cancers, but the detailed elucidation of the distinct characteristics of 2 populations will be required to enhance the immunotherapeutic potential of γδ T cells. Here, we summarize recent progress regarding cancer immunology of human γδ T cells, including their development, heterogeneity, and plasticity, the putative mechanisms underlying ligand recognition and activation, and their dual effects on tumor progression in the tumor microenvironment.

Efecto reno-protector y reno-reparador del Factor de Crecimiento Epidérmico en biomodelo de Insuficiencia Renal Crónica / Reno-protective and reno-restorative effect of the Epidermal Growth Factor in biomodel of Chronic Renal Failure

Introducción: La Enfermedad Renal es un problema de salud mundial. El Factor de Crecimiento Epidérmico actúa como citoprotector y trófico reparador. Objetivo: Evaluar el efecto reno-protector y reno-reparador del Factor de Crecimiento Epidérmico en biomodelo de Insuficiencia Renal Crónica. Material y Métodos: Se estudiaron 120 ratas, Wistar, en 6 grupos: Control Negativo y positivo, Solución Salina Dosis Única y Múltiple, Factor de Crecimiento Epidérmico Dosis Única y Múltiple. Se aplicó para efecto reno-protector dosis única antes del daño, y para el reno-reparador dosis múltiples posterior al daño, a razón de 100 µg/kg de peso. Resultados: La creatinina, urea y ácido úrico disminuyeron significativamente en los grupos experimentales, con mayor disminución para el grupo experimental dosis única, por lo que el efecto reno-protector fue mayor que el reno-reparador para los esquemas de tratamiento utilizados. Conclusiones: El Factor de Crecimiento Epidérmico mostró efecto reno-protector y reno-reparador al disminuir las variables hematológicas de daño renal(AU)

Introduction: Kidney disease is a world health problem. Epidermic Growth Factor acts as cyto-protector, and trophic restorative. Objective: To assess the reno-protective and reno-restorative effect of the Epidermal Growth Factor in biomodel of Chronic Renal Failure. Material and Methods: 120 Wistar rats were studied in 6 groups: Negative and Positive Control, Saline Solution Single-Dose and Multiple-Dose, Epidermal Growth Factor Single-Dose and Multiple-Dose. A single dose was applied before the damage for the reno-protective effect, and multiple doses after the damage for the reno-restorative effect, at a rate of 100 µg/kg of weight. Results: Creatinine, urea, and uric acid diminished significantly in the experimental groups, with a higher decrease for experimental group with single dose; therefore, the reno-protective effect was higher than reno-restorative one for the treatment patterns used. Conclusions: Epidermal Growth Factor showed reno-protective and reno-restorative effect by diminishing the hematological variables in kidney damage(AU)

Insuficiencia venosa crónica: relación de la funcionalidad de las venas perforantes con los trastornos tróficos

Introducción: La insuficiencia venosa crónica es una patología cuyos síntomas signos son expresión de la hipertensión venosa persistente en los miembros inferiores. Se la describe en la historia de la medicina desde sus comienzos, sin embargo matiene altas prevalencia a nivel mundial y representa un problema sanitario de gran impacto socio-económico, afectando en sus formas severas a más del 25% de la población adulta en países industrializados. Su fisiopatología sigue siendo motivo de investigación, en ese sentido los parámetros para definir la funcionalidad de las venas perforantes se encuentran en constante revisión. El estudio morfológico y hemodinámico de las venas de los miembros inferiores mediante Eco Doppler es considerado el patrón de oro en el diagnóstico de la insuficiencia venosa crónica

Summary: Introduction: Chronic venous insufficiency is a condition with symptoms expressed as presistent venous hypertension in the lower extremities. It has been described throughout the history of medicine, yet today it is still quite prevalent worldwide, and represents a health problem of great socio economic impact. In its severe form, it affects more than 25% of the adult population in industrialized countries. Its pathophysiology remains a matter of inquiry: in this respect, the parameters that define the functionality of the perforatin veins are under constant review. The morphologic and hemodynamic study of veins in the lower extremities via Doppler echocardiography is considered the gold standard in the diagnosis of chronic venous insufficiency

La metformina como una alternativa en la prevención y tratamiento del cáncer / Metformin as an alternative for the prevention and treatment of cancer

Antecedentes: la metformina, fármaco económico y seguro, ha demostrado mejorar el pronóstico de varios tipos de cánceres. Objetivo: revisar los aspectos más relevantes de la relación entre la diabetes mellitus, la metformina y el cáncer. Desarrollo: la diabetes mellitus, en especial la tipo 2, se relaciona con algunos cánceres (mama, hígado, páncreas, ginecológico, vejiga, colon y recto), y en el sexo masculino, aumenta su recurrencia y la mortalidad. Los mecanismos responsables de esta relación no están del todo esclarecidos. La insulina y el factor de crecimiento similar a la insulina en un estado de hiperinsulinismo e insulinorresistencia, pudieran desempeñar un papel fundamental en el desarrollo de cáncer, así como otros factores de riesgo comunes a la diabetes mellitus y al cáncer (alimentación no saludable, sedentarismo, adicciones, edad, sexo, etnia y raza). La proteína liver kinase B1 se ha identificado como una proteína supresora tumoral, y al unirse con la metformina interrumpe el complejo 1 de la cadena respiratoria mitocondrial, y conduce a la disminución de la síntesis de trifosfato de adenosin, y al aumento del cociente proteína activada por mitógenos-trifosfato de adenosin en el espacio intracelular. Los quimioterápicos, esteroides y antiandrógenos, pueden afectar negativamente el metabolismo hidrocarbonado. Algunas drogas antihiperglucemiantes se han relacionado a cánceres específicos, aunque las evidencias son pobres, indirectas y controversiales. Conclusiones: la metformina pudiera utilizarse en la prevención y el tratamiento de algunos cánceres, y reducir su recurrencia y la mortalidad. Parece existir una relación entre cáncer y la diabetes mellitus, aunque muchos aspectos quedan por dilucidar, como el papel desempeñado por los fármacos anticancerígenos y antihiperglucemiantes utilizados en ambas entidades(AU)

Background: metformin, a safe inexpensive drug, has proved to improve the prognosis of several types of cancer. Objective: to review the most relevant aspects of the relationship among diabetes mellitus, metformin and cancer. Development: diabetes mellitus, particularly type 2, is related to some kinds of cancer (breast, liver, pancreas, gynecological, gallbladder, colon and rectum), and its recurrence and mortality increase in men. The mechanism behind this relationship is not fully clarified. Insulin and insulin-like growth factor under hyperinsulinism and insulin resistance conditions may play a fundamental role in developing cancer as well as other common risk factors for diabetes mellitus and cancer (unhealthy feeding, sedentary lifestyle, addictions, age, sex, ethnic group and race). Liver kinase B1 protein has been identified as tumor suppressor protein which binds the metformin to impair the mitochondrial respiratory chain complex I and leads to reduction of adenosine triphosphate synthesis and to the increase of mytogen-activated protein-adenosine triphosphate quotient in the intracellular space. Chemotherapeutic, steroid and anti-androgen drugs may negatively affect the hydrocarbon metabolism. Some antihyperglycemic drugs have been related to specific cancers, although the evidence is still poor, indirect and controversial. Conclusions: metformin may be used to prevent and treat some types of cancer and to reduce recurrence and mortality. There seems to be some relationship between cancer and diabetes mellitus, even when many aspects remain to be ascertained such as the role played by anticancer and antihyperglycemic drugs intended to treat both diseases(AU)

Emancipatory practices of nurses in primary health care: the home visit as an instrument of health needs assessment / Prácticas emancipadoras del enfermero en la atención primaria de salud: la visita domiciliaria como instrumento de reconocimiento de necesidades de la salud / Práticas emancipatórias de enfermeiros na Atenção Básica à Saúde: a visita domiciliar como instrumento de reconhecimento de necessidades de saúde

Objective Identify nurses’ emancipatory practices in primary care, to contribute to the improvement of health care. Method A case study type social research of qualitative nature, in which nurses of a primary health care service unit in São Paulo were interviewed. Results The home visit was identified as a nursing practice possible to be expanded in order to identify social determinants of health, triggering emancipatory practices in the service. This expansion occurred because the design of health care labour intended by the service team changed its focus from the traditional object of health services, the disease. Conclusion First, it is advocated that social policies lead projects with the purpose of improving health needs. On the other hand, the daily labour needs to provide opportunities for reflection and discussion of healthcare projects, leading workers to propose labour-processes targeted to both the social determinants of health and people’s illness. .

Objetivo Identificar las prácticas emancipadoras de enfermeras en Unidad de Salud Familiar fueron el objeto de este estudio. Método La investigación social cualitativa tipo estúdio de caso. Fueron entrevistados enfermeros de una Unidad de Salud Familiar en Sao Paulo. Resultados Se identificó que la Visita Domiciliaria ha ampliado su alcance y identificado determinantes del proceso salud-enfermedad, lo que provocó en la Unidad de Salud Familiar prácticas emancipadoras. Esta expansión se produjo debido a que el diseño de la atención en propósito por la USF amplió el tradicional objeto de los servicios de salud. Conclusión Se aboga que las directrices de las políticas sociales basen proyectos que tengan como fin el mejoramiento de las necesidades de salud y que el trabajo diario proporcione la reflexión y discusión de los proyectos de atención, para proponer prácticas que enfoquen en los determinantes del proceso salud-enfermedad, tanto cuanto en sus resultados - la enfermedad en el cuerpo individual. .

Objetivo Identificar as práticas emancipatórias de enfermeiros da Atenção Primária, com a finalidade de contribuir para o aprimoramento do cuidado em saúde. Método Pesquisa social de natureza qualitativa, do tipo estudo de caso. Foram entrevistados os enfermeiros de uma Unidade de Saúde da Família em São Paulo. Resultados Identificou-se que a visita domiciliária, prática protocolar, ampliou seu escopo e identificou determinantes do processo saúde-doença, desencadeando na Unidade de Saúde da Família práticas emancipatórias. Essa ampliação ocorreu porque o projeto de cuidado intencionalizado ampliou o objeto tradicional dos serviços de saúde. Conclusão Advoga-se que as diretrizes das políticas sociais ancorem projetos que tomem como finalidade o aprimoramento das necessidades de saúde e que o cotidiano do trabalho proporcione reflexão e discussão dos projetos de cuidado, para intencionalizar práticas que incidam nos determinantes do processo saúde-doença, tanto quanto nos resultados - a doença expressa no corpo individual. .

Association Study of Fibroblast Growth Factor 2 and Fibroblast Growth Factor Receptors Gene Polymorphism in Korean Ossification of the Posterior Longitudinal Ligament Patients

OBJECTIVE: The aim of this study was to determine whether single nucleotide polymorphisms (SNPs) of fibroblast growth factor (FGF) 2 gene and fibroblast growth factor receptor (FGFR) genes are associated with ossification of the posterior longitudinal ligament (OPLL). METHODS: A total of 157 patients with OPLL and 222 controls were recruited for a case control association study investigating the relationship between SNPs of FGF2, FGFR1, FGFR2 and OPLL. To identify the association among polymorphisms of FGF2 gene, FGFR1, FGFR2 genes and OPLL, the authors genotyped 9 SNPs of the genes (FGF2 : rs1476217, rs308395, rs308397, and rs3747676; FGFR1 : rs13317 and rs2467531; FGFR2 : rs755793, rs1047100, and rs3135831) using direct sequencing method. SNPs data were analyzed using the SNPStats, SNPAnalyzer, Haploview, and Helixtree programs. RESULTS: Of the SNPs, a SNP (rs13317) in FGFR1 was significantly associated with the susceptibility of OPLL in the codominant (odds ratio=1.35, 95% confidence interval=1.01-1.81, p=0.048) and recessive model (odds ratio=2.00, 95% confidence interval=1.11-3.59, p=0.020). The analysis adjusted for associated condition showed that the SNP of rs1476217 (p=0.03), rs3747676 (p=0.01) polymorphisms in the FGF2 were associated with diffuse idiopathic skeletal hyperostosis (DISH) and rs1476217 (p=0.01) in the FGF2 was associated with ossification of the ligament flavum (OLF). CONCLUSION: The results of the present study revealed that an FGFR1 SNP was significantly associated with OPLL and that a SNP in FGF2 was associated with conditions that were comorbid with OPLL (DISH and OLF).

Estudo comparativo do uso isolado de plasma rico em plaquetas e combinado com cimento de alfa-fosfato tricálcico no reparo ósseo em ratos / Comparative study on use of platelet-rich plasma alone and in combination with alpha-tricalcium phosphate cement for bone repair in rats

OBJETIVOS: Avaliar o efeito do cimento α-TCP combinado com PRP sobre a osteogênese, comparando os resultados com PRP utilizado isoladamente. MÉTODOS: Foi confeccionado defeito bilateral no fêmur de ratos e preenchido com um dos dois tipos de tratamentos (PRP ou α-TCP+PRP), sendo avaliado em quatro e oito semanas. As imagens radiográficas forneceram valores da área da lesão, e a histologia (coloração Picrosirius) indicou a área de neoformação óssea. RESULTADOS: As médias referentes à área de lesão do grupo α-TCP+PRP (2,64mm² ± 2,07 e 1,91mm² ± 0,93; quatro e oito semanas, respectivamente) demonstraram numericamente melhores resultados, porém não significativos (p > 0,05), em comparação com aqueles observados no grupo PRP (5,59mm² ± 2,69 e 3,23mm² ± 1,46; quatro e oito semanas, respectivamente). As médias de neoformação óssea foram de 62,7% ± 12,1% e 79,01% ± 6,25 no grupo PRP, e 73,3% ± 12,7 e 85,86% ± 10,45 no grupo α-TCP+PRP, em quatro e oito semanas, respectivamente (p > 0,05). CONCLUSÃO: Os dados deste estudo sugerem que o tratamento com cimento α-TCP combinado com PRP não demonstra diferença significativa quando comparado ao PRP isolado. Entretanto, há um possível efeito precoce sobre a regeneração óssea quando os dois biomateriais são aplicados em conjunto.

OBJECTIVES: To evaluate the effect of alpha-tricalcium phosphate (α-TCP) cement combined with platelet-rich plasma (PRP) on osteogenesis, and to compare the results with use of PRP alone. METHODS: A bilateral defect was produced in rat femurs and was filled with one of two types of treatments (PRP or α-TCP + PRP). The outcomes were evaluated after four and eight weeks. Radiographic images provided values for the lesion area, and histology (picrosirius staining) indicated the area of ​​new bone formation. RESULTS: The means relating to the lesion area of the α-TCP + PRP group (2.64 ± 2.07 and 1.91 ± 0.93 mm², after four and eight weeks, respectively) showed numerically better but non-significant results (p > 0.05) than those seen in the PRP group (5.59 mm ² ± 2.69 and 3.23 ± 1.46 mm ², after four and eight weeks, respectively). The mean new bone formation rates were 62.7% ± 12.1 and 79.01% ± 6.25 in the PRP group, and 73.3% ± 12.7 and 85.86% ± 10.45 in α-TCP + PRP group, after four and eight weeks, respectively (p > 0.05). CONCLUSION: The data from this study suggest that treatment with α-TCP cement combined with PRP does not show any significant difference in comparison with PRP alone. However, there is a possible early effect on bone regeneration when the two biomaterials are applied together.

Epigallocatechin-3-gallate inhibits paracrine and autocrine hepatocyte growth factor/scatter factor-induced tumor cell migration and invasion

Aberrant activation of hepatocyte growth factor/scatter factor (HGF/SF) and its receptor, Met, is involved in the development and progression of many human cancers. In the cell-based screening assay, (-)epigallocatechin-3-gallate (EGCG) inhibited HGF/SF-Met signaling as indicated by its inhibitory activity on HGF/SF-induced cell scattering and uPA activation (IC50 = 15.8 microg/ml). Further analysis revealed that EGCG at low doses specifically inhibited HGF/SF-induced tyrosine phosphorylation of Met but not epidermal growth factor (EGF)-induced phosphorylation of EGF receptor (EGFR). On the other hand, high-dose EGCG decreased both Met and EGFR proteins. We also found that EGCG did not act on the intracellular portion of Met receptor tyrosine kinase, i.e., it inhibited InlB-dependent activation of Met but not NGF-induced activation of Trk-Met hybrid receptor. This inhibition decreased HGF-induced migration and invasion by parental or HGF/SF-transfected B16F10 melanoma cells in vitro in either a paracrine or autocrine manner. Furthermore, EGCG inhibited the invasion/metastasis of HGF/SF-transfected B16F10 melanoma cells in mice. Our data suggest the possible use of EGCG in human cancers associated with dysregulated paracrine or autocrine HGF/SF-Met signaling.

In vitro migration capacity of human adipose tissue-derived mesenchymal stem cells reflects their expression of receptors for chemokines and growth factors

The homing properties of adipose tissue-derived mesenchymal stem cells (AdMSCs) have stimulated intravenous applications for their use in stem cell therapy. However, the soluble factors and corresponding cellular receptors responsible for inducing chemotaxis of AdMSCs have not yet been reported. In the present study, the migration capacity of human AdMSCs (hAdMSCs) toward various cytokines or growth factors (GFs) and the expression of their receptors were determined. In a conventional migration assay, PDGF-AB, TGF-beta1, and TNF-alpha showed the most effective chemoattractant activity. When AdMSCs were preincubated with various chemokines or GF, and then allowed to migrate toward medium containing 10% FBS, those preincubated with TNF-alpha showed the highest migratory activity. Next, hAdMSCs were either preincubated or not with TNF-alpha, and allowed to migrate in response to various GFs or chemokines. Prestimulation with TNF-alpha increased the migration activity of hAdMSCs compared to unstimulated hAdMSCs. When analyzed by FACS and RT-PCR methods, hAdMSCs were found to express C-C chemokine receptor type 1 (CCR1), CCR7, C-X-C chemokine receptor type 4 (CXCR4), CXCR5, CXCR6, EGF receptor, fibroblast growth factor receptor 1, TGF-beta receptor 2, TNF receptor superfamily member 1A, PDGF receptor A and PDGF receptor B at both the protein and the mRNA levels. These results indicate that the migration capacity of hAdMSCs is controlled by various GFs and chemokines. Prior in vitro modulation of the homing capacity of hAdMSCs could stimulate their movement into injured sites in vivo when administered intravenously, thereby improving their therapeutic potential.

In vitro migration capacity of human adipose tissue-derived mesenchymal stem cells reflects their expression of receptors for chemokines and growth factors

The homing properties of adipose tissue-derived mesenchymal stem cells (AdMSCs) have stimulated intravenous applications for their use in stem cell therapy. However, the soluble factors and corresponding cellular receptors responsible for inducing chemotaxis of AdMSCs have not yet been reported. In the present study, the migration capacity of human AdMSCs (hAdMSCs) toward various cytokines or growth factors (GFs) and the expression of their receptors were determined. In a conventional migration assay, PDGF-AB, TGF-beta1, and TNF-alpha showed the most effective chemoattractant activity. When AdMSCs were preincubated with various chemokines or GF, and then allowed to migrate toward medium containing 10% FBS, those preincubated with TNF-alpha showed the highest migratory activity. Next, hAdMSCs were either preincubated or not with TNF-alpha, and allowed to migrate in response to various GFs or chemokines. Prestimulation with TNF-alpha increased the migration activity of hAdMSCs compared to unstimulated hAdMSCs. When analyzed by FACS and RT-PCR methods, hAdMSCs were found to express C-C chemokine receptor type 1 (CCR1), CCR7, C-X-C chemokine receptor type 4 (CXCR4), CXCR5, CXCR6, EGF receptor, fibroblast growth factor receptor 1, TGF-beta receptor 2, TNF receptor superfamily member 1A, PDGF receptor A and PDGF receptor B at both the protein and the mRNA levels. These results indicate that the migration capacity of hAdMSCs is controlled by various GFs and chemokines. Prior in vitro modulation of the homing capacity of hAdMSCs could stimulate their movement into injured sites in vivo when administered intravenously, thereby improving their therapeutic potential.

Inhibition of eNOS/sGC/PKG Pathway Decreases Akt Phosphorylation Induced by Kainic Acid in Mouse Hippocampus

The serine/threonine kinase Akt has been shown to play a role of multiple cellular signaling pathways and act as a transducer of many functions initiated by growth factor receptors that activate phosphatidylinositol 3-kinase (PI3K). It has been reported that phosphorylated Akt activates eNOS resulting in the production of NO and that NO stimulates soluble guanylate cyclase (sGC), which results in accumulation of cGMP and subsequent activation of the protein kinase G (PKG). It has been also reported that PKG activates PI3K/Akt signaling. Therefore, it is possible that PI3K, Akt, eNOS, sGC, and PKG form a loop to exert enhanced and sustained activation of Akt. However, the existence of this loop in eNOS-expressing cells, such as endothelial cells or astrocytes, has not been reported. Thus, we examined a possibility that Akt phosphorylation might be enhanced via eNOS/sGC/PKG/PI3K pathway in astrocytes in vivo and in vitro. Phosphorylation of Akt was detected in astrocytes after KA treatment and was maintained up to 72 h in mouse hippocampus. 2 weeks after KA treatment, astrocytic Akt phosphorylation was normalized to control. The inhibition of eNOS, sGC, and PKG significantly decreased Akt and eNOS phosphorylation induced by KA in astrocytes. In contrast, the decreased phosphorylation of Akt and eNOS by eNOS inhibition was significantly reversed with PKG activation. The above findings in mouse hippocampus were also observed in primary astrocytes. These data suggest that Akt/eNOS/sGC/PKG/PI3K pathway may constitute a loop, resulting in enhanced and sustained Akt activation in astrocytes.

Receptores de factores de crecimiento un ejemplo de blanco molecular / Receptors of growth factors in the cancer treatment

EI bloqueo de los receptores de factores de crecimiento constituye una nueva arma terapéutica en el manejo del cáncer. Esta acción se ha realizado mediante dos tipos de medicamentos, anticuerpos monoclonales que se unen a la porción extra membranosa del receptor y con bloqueadores de la actividad de tirosinquinasa que actúan en la parte intracitoplasmática del receptor. Los mejores resultados se han logrado en el cáncer de mama con el empleo de un anticuerpo monoclonal denominado trastuzumab, que actúa sobre el receptor HER-2 y un inhibidor de tirosinquinasa, lapatinib, que es un bloqueador dual ya que actúa sobre HER-l y HER-2. Ambos potencian la acciónde la quimioterapia. Por otro lado, otro anticuerpo monoclonal, el cetuximab el cual actúa sobre el receptor HER-l ha demostrado actividad en el cáncer de colon metastásico, asociado a la quimioterapia y en el cáncer de cabeza y cuello localmente avanzado, asociado a la radioterapia. Finalmente, dos inhibidores de tirosinquinasa de HER-l, elerlotinib y gefitinib tienen actividad en el cáncer pulmonar de células no pequeñas metastásico, especialmente cuando los tumores tienen mutaciones en el dominio de tirosinquinasa del receptor.

The blockage of the growth factor receptors is the new treatment technique in the cancer treatment. This action has been done by two types of medicament, monoclonal antibo-dies, binding to the receptor of the extramembranose portion, and with blockaders of the thyroxinequinasa activity in the intracytoplasmatic receptor. The best results has been achieved in breast cancer treatment, using monoclonal antibody named trastuzumab, blocking HER-2 receptor, and a thyroxinequinasa inhibitor, lapatinib, with a double blockage action, over HER-l and HER-2. Both of them helps the chemotherapy effect. On the other hand, another monoclonal antibody, the cetuximab, acts over the HER-l receptor, has demonstrate activity in the metastasic colon cancer, when is associated to chemotherapy, and in the head and neck cancer locally advanced, associated to radiotherapy. Finally, two thyroxinequinasa inhibitors of HER-l, the erlotinib and gefitinib, have activity over the metastatic lung cancer of not small cells, especially when the tumors have mutations in the thyroxinequinasa receptor domain.

TGFbeta, activina e sinalização SMAD em câncer de tiróide / TGFbeta, activin and SMAD signalling in thyroid cancer

TGFbeta e activina são membros da superfamília TGFbeta e desempenham um amplo papel no desenvolvimento, proliferação e apoptose. Estes fatores de crescimento exercem seus efeitos biológicos ligando-se a receptores de membrana do tipo I e do tipo II que transduzem a sinalização até o núcleo através da fosforilação das proteínas R-SMADs (SMAD 2/3) e co-SMADs (SMAD4). O controle apropriado da via de TGFbeta/activina ainda depende da regulação negativa exercida pelo SMAD inibitório (SMAD7) e pelas enzimas E3 de ubiquitinação (Smurfs). Fisiologicamente, TGFbeta e activina atuam como potentes inibidores da proliferação na célula folicular tiroidiana. Desta forma, alterações de receptores e componentes da via de sinalização SMAD estão associadas a diferentes tipos de tumores. Desde que TGFbeta e activina geram sua sinalização intracelular utilizando os mesmos componentes da via SMAD, o desequilíbrio desta via prejudica dois processos anti-mitogênicos da célula. Nesta revisão, enfocamos aspectos que indicam o mecanismo de resistência ao efeito inibitório de TGFbeta e activina ocasionado pelo desequilíbrio da via de sinalização SMAD nas neoplasias da tiróide.

TGFbeta and activin are members of the TGFbeta superfamily and play a wide role in development, proliferation and apoptosis. These growth factors exert their biological effects by binding to the type I and II membrane receptors to transduce their signalling through the nucleus by phosphorylation of R-SMADs (SMAD 2/3) and co-SMADs (Smad 4). The proper control of TGFbeta/activin pathway is negatively regulated by inhibitory SMAD (SMAD7) and by E3 ubiquitination enzymes (Smurfs). Physiologically, TGFbeta and activin act as potent growth inhibitors in thyroid follicular cell. Thus, alterations in the receptors and components of SMAD signalling pathway are associated with several types of tumors. Since TGFbeta and activin generate their intracellular signalling through the same components of the SMAD pathway, the unbalance of this pathway impairs both of anti-mitogenic signals in the cell. This review addresses aspects of the molecular mechanisms in the understanding of resistance to the growth inhibitory effects of TGFbeta and activin due to the disequilibrium in the SMAD inhibitory pathway in thyroid neoplasia.

Expressão protéica no endométrio durante a fase lútea do ciclo menstrual / Endometrial protein expression during the luteal phase of the menstrual cycle

Introdução: O objetivo foi avaliar a expressão de algumas proteínas no endométrio durante a fase lútea do ciclo menstrual de mulheres férteis e inférteis, por meio imunoistoquímica de micro-arranjos teciduais (TMA). Métodos: Analisou-se a expressão de dez proteínas em 52 amostras de endométrio obtidas nas fases lútea inicial, intermediária (janela de implantação) e final. Resultados: As proteínas, fator inibidor de leucemia (LIF), fator de crescimento insulinóide tipo 1 (IGF-1), receptor de progesterona (PR), claudina-4, receptor de fator de crescimento vascular endotelial 3 (VEGFR-3) e citoqueratina 7 (CK-7) mostraram-se expressas no endométrio nas fases lútea inicial, intermediária e final. A proteína morfogenética óssea 4 (BMP-4) expressou-se no endométrio nas fases lútea inicial e intermediária. As proteínas citoqueratina 17 (CK-17), substância solúvel 100 (S100) e calretinina não se expressaram no endométrio durante os três períodos avaliados. Houve correlação entre as expressões protéicas de LIF, IGF-1 e PR. As proteínas LIF e BMP-4 foram diferencialmente expressos no endométrio nas fases lútea inicial, intermediária e final. As proteínas claudina-4 e PR não se expressam simultâneamente no endométrio durante a fase lútea. Conclusão: Baseados nos resultados deste estudo podemos sugerir que a presença das proteínas LIF, IGF-1 e PR durante a janela implantacional teria relevância como preditor do adequado desenvolvimento do endométrio.

Introduction: The objective of this study was to evaluate endometrial protein expressions from fertile and infertile women during the luteal phase of the menstrual cycle by immunohistochemistry in tissue microarrays (TMA). Method: The expression of ten proteins obtained from 52 endometrial samples in the initial, mid (window of implantation) and late (premenstrual) phases of the menstrual cycle were evaluated. Results: The proteins leukemia inhibitory factor (LIF), insulin like growth factor 1 (IGF-1), progesterone receptor (PR), claudin-4, vascular endothelial growth factor receptor 3 (VEGFR-3), and cytokeratin 7 (CK-7) were expressed in the endometrium in the three intervals of the luteal phase. Endometrial expression of the morphogenetic bone protein 4 (BMP-4) occurred during the initial and mid luteal phases. Cytokeratin 17, substance 100 and calretinin were not expressed in the luteal phase. There were positive correlations among endometrial expressions of LIF, IGF- 1, and PR. LIF and BMP-4 were differently expressed in the initial, mid and late phases of the luteal phase. Claudin-4 and PR did not express simultaneously during the different intervals of the luteal phase. Conclusion: These findings suggest that positively correlated endometrial expressions of LIF, IGF-1 and PR at the window of implantation could characterize an adequately developed and receptive endometrium.

cDNA microarray-based study of gene expression profile changes in human esophageal squamous cell carcinoma / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the differentially expressed genes between human esophageal squamous cell carcinoma (ESCC) and normal esophageal mucosa and explore an effective method with high throughput for screening the molecular markers closely correlated with the development, invasion and metastasis of ESCC.</p><p><b>METHODS</b>With cDNA microarray and laser capture microdissection, T7-based amplification were used to detect the mRNA from both the primary carcinoma and the corresponding esophageal epithelium in 15 ESCC cases, and the results were analyzed by bioinformatics methods.</p><p><b>RESULTS</b>Among the 886 target genes, 110 (12.42%) genes were differentially expressed commonly at least twice in all the 15 samples, including 56 (6.32%) up-regulated by at least 2 folds and 54 (6.09%) down-regulated by at least 0.5 folds.</p><p><b>CONCLUSION</b>Many ESCC-associated genes were screened by the high-throughput gene chip method, and functional study of these genes may help to identify the key genes or pathways involved in the pathogenesis and development of ESCC.</p>

MET Expression in Sporadic Renal Cell Carcinomas

Although germline mutations of met proto-oncogene on human chromosome 7q31-34 have been known as useful molecular markers of hereditary papillary renal cell carcinoma (RCC), the expression of MET, a product of met proto-oncogene, has not been fully studied in sporadic RCC, along with its clinical significance. We investigated the expression of MET by immunohistochemistry in 182 cases of renal neoplasm encompassing 145 RCC, 25 urothelial carcinomas of renal pelvis, and 12 oncocytomas. MET was diffusely and strongly expressed in 90% of papillary RCC, all collecting duct carcinomas, and 92% of urothelial carcinomas of renal pelvis. On the contrary, clear cell RCC, chromophobe RCC, and oncocytomas were negative or focally positive for MET expression. In clear cell RCC, MET expression was positively correlated with high nuclear grade, presence of infiltrative growth, tumoral necrosis, papillary architecture, sarcomatoid component, tumoral involvement of the renal pelvis or ureter, involvement of the calyx, and lymphatic invasion. In conclusion, diffuse and strong expression of MET in papillary RCC and collecting duct carcinoma might be helpful in discriminating from the other subtypes of RCC with tubular or papillary growth. In case of MET expression observed in clear cell RCC, it might correlate with those clinicopathological parameters implying aggressive behavior.

Hepatocyte Growth Factor/c-Met Signaling in Regulating Urokinase Plasminogen Activator in Human Stomach Cancer: A Potential Therapeutic Target for Human Stomach Cancer

BACKGROUND: Up-regulation of the hepatocyte growth factor (HGF), its transmembrane tyrosine kinase receptor (c-Met), and urokinase type plasminogen activator (uPA), is associated with the development and metastasis of various types of cancers. However, the mechanisms by which HGF/c-Met signaling mediates cancer progression and metastasis are unclear. METHODS: We investigated the roles of HGF/c-Met in tumor progression and metastasis in NUGC-3 and MKN-28 stomach cancer cell lines. RESULTS: Treatment with HGF increased c-Met phosphorylation in a dose-dependent manner, as well as increasing cell proliferation. HGF treatment also increased the protein level and the activity of uPA in NUGC-3 and MKN-28 cells. A monoclonal antibody against human uPA receptor (uPAR), mAb 3936, inhibited HGF-mediated tumor cell invasion in a dose-dependent manner. Down-regulation of uPA using uPA-shRNA induced a decrease in in vitro cell invasion in NUGC-3 cells. CONCLUSIONS: These results suggest that NUGC-3 and MKN-28 cells express functional c-Met, which may provide a therapeutic target for interfering with metastases of cancer cells by inhibiting uPA and uPAR-mediated proteolysis.

Expression of the mRNA of Growth Factor Receptors and Neurotrophic Factors in Porcine Vesical Submucosa and Acellular Matrix / 대한비뇨기과학회지

PURPOSE: Several naturally-derived biopolymers have been introduced as implantable biomaterial for tissue reconstruction, and some of them are already being applied in various clinical fields. They serve as backbones for the regeneration of damaged tissue. It is well known that growth factors play an important role in this process and some of the naturally- derived biopolymers were known to contain several kinds of growth factors, but there are few reports about any growth factors in the various biopolymers. We tried to confirm the existence of growth factor receptors and neurogenic factors through the expression of the mRNA from the biopolymers. MATERIALS AND METHODS: Bladders were harvested from adult pigs. Vesical submucosa was obtained with using mechanical decellulization technique. Acellular matrices were made from pig bladder with using a cell lysis technique, and they were examined by performing scanning electron microscopy. Expression of the mRNA of vascular endothelial growth factor (VEGF) receptor, fibroblast growth factor (FGF)-1 receptor, neuregulin and brain-derived neurotrophic factor (BDNF) in the acellular matrix and the submucosa of porcine bladder were examined for via reverse transcription- polymerase chain reaction (RT-PCR). RESULTS: The ultrastructure of the acelluar matrix shows collagen fibers with many pores. The mRNA's of VEGF receptor, FGF-1 receptor and neuregulin were expressed in the porcine vesical acellular matrix, whereas BDNF was not expressed. On the other hand, none of the mRNA's examined was expressed in the porcine vesical submucosa. CONCLUSIONS: This study's results shows that several growth factors are detected in the acellular matrix of the porcine bladder. These growth factors may play an important role for the growth of tissue when it is implanted in vivo. It is also assumed that there are more growth factors in naturally-derived biopolymers, and it would be useful to investigate the undefined components of naturally-derived biopolymers for developing new and advanced polymers.

Relationship of overexpression of angiogenesis factors and their receptors with invasive growth of keloid / 中华整形外科杂志

<p><b>OBJECTIVE</b>To study the relationship between invasive growth and the angiogenesis factors and their receptors in keloid.</p><p><b>METHODS</b>Biopsies from 17 keloid (Ke) were divided into atrophy group (Ke-A, n = 9), proliferating group (Ke-P, n = 13), infiltrating group (Ke-I, n = 9), normal skin around Ke (Ke-N, n = 10) and normal skin (NS, n = 10). The histology, immunohistochemistry and computerized imaging analysis were used for the study. The levels of basic fibroblast growth factor (bFGF) and its receptor-Flg, vascular endothelial growth factor (VEGF) and VEGF/KDR complex (11B5), and platelet derived growth factor (PDGF-A) and its receptor-PDGFR-alpha, and alpha-smooth muscle actin (alpha-SMA) were determined in specimens with immuneohistochemical staining.</p><p><b>RESULTS</b>In all 5 groups, bFGF, Flg, VEGF, 11B5, PDGF-A, and PDGFR-alpha were all expressed in fibroblasts (Fb), monocyte-phagocytes, vascular endothelial cells, adventitial cells, epidermal (cells and epithelial cells in appendage. The intensities of staining ranked as follows: Ke-I > Ke-N approximately equal to Ke-P > Ke-A approximately equal to NS, Flg > hFGF approximately equal to PDGFR-alpha > PDGF-A approximately equal to 11B5 > VEGF (P < 0.05 to approximately 0.01). 11B5 and VEGF were expressed (intensively in alpha-SMA positive myofibroblasts only in Ke-I group. The histological observation showed hyperplasia of endothelial cells and obliteration of microvessels.</p><p><b>CONCLUSION</b>The invasive growth of keloid may be related to the overexpression of angiogenesis factors and their receptors. The abnormal expression of 11B5 in myofibroblasts may be one of the important factors associated with tumor-like growth feature in the invasive parts sites of keloid. The results suggest that inhibition of these biological activities would be of significance in clinical therapy.</p>