Morphological characterization of a human glioma cell l ine

A human malignant continuous cell line, named NG97, was recently established in our laboratory. This cell line has been serially subcultured over 100 times in standard culture media presenting no sign of cell senescence. The NG97 cell line has a doubling time of about 24 h. Immunocytochemical analysis of glial markers demonstrated that cells are positive for glial fibrillary acidic protein (GFAP) and S-100 protein, and negative for vimentin. Under phase-contrast microscope, cultures of NG97 showed cells with variable morphological features, such as small rounded cells, fusiform cells (fibroblastic-like cells), and dendritic-like cells. However, at confluence just small rounded and fusiform cells can be observed. At scanning electron microscopy (SEM) small rounded cells showed heterogeneous microextentions, including blebs and filopodia. Dendritic-like cells were flat and presented extensive prolongations, making several contacts with small rounded cells, while fusiform cells presented their surfaces dominated by microvilli. We believe that the knowledge about NG97 cell line may be useful for a deeper understanding of biological and immunological characteristics of gliomas

Total hepatic warm ischemia and reperfusion associated with controlled hemorrhagic shock: effects of neutrophil sequestration in rat liver

BACKGROUND:The purpose of this experimental study was to evaluate the effects of total hepatic ischemia and reperfusion on the accumulation of neutrophils in the liver of rats, under normal conditions and in rats submitted to controlled hemorrhagic shock . METHODS: Thirty two adult male Wistar rats, were divided into four groups: the Control group, was submitted to the standard procedures for a period of 60 min of observation; Shock group, was submitted to controlled hemorrhagic shock (mean arterial blood pressure = 40 mmHg, 20 min) followed by volume resuscitation (lactated Ringer's solution + blood, 3:1) and reperfusion for 60 min; Pringle group, was submitted to total hepatic ischemia for 15 min and reperfusion for 60 min; The Total group, was submitted to controlled hemorrhagic shock for 15 min followed by volume resuscitation (lactated Ringer's solution + blood, 3:1), total hepatic ischemia for 15 min and reperfusion for 60 min. Measurements of serum lactate and base excess were used to characterize the hemorrhagic shock state with low tissue perfusion. The counting of neutrophils on the liver tissue was performed after the euthanasia of animals. RESULTS: Values for the counting of neutrophils on the liver indicate that, the animals from Pringle group differed from Shock and Total groups (Control 10.30±3.20, Shock 13.94±2.84, Pringle 7.00±3.40, Total 12.45±3.65) but did not differ from Control group. CONCLUSIONS: Rats submitted to controlled hemorrhagic shock state associated to total hepatic ischemia for 15 minutes, followed by 60 minutes of reperfusion, did not present significant neutrophils accumulation on liver tissue.O propósito deste trabalho experimental foi estudar os efeitos da isquemia e reperfusão hepática total sobre o acúmulo de neutrófilos no fígado de ratos, em condições de normalidade e submetidos ao estado de choque hemorrágico controlado. MÉTODO: Trinta e dois ratos Wistar, machos, foram divididos em quatro grupos de oito animais cada: grupo Controle, submetido à laparotomia com um período de 60 minutos de observação; grupo Choque, submetido a choque hemorrágico controlado (PAM = 40 mmHg, 20 min.) seguido de reposição volêmica (Ringer lactato + sangue, 3:1) e reperfusão (60 min.); grupo Pringle, submetido a isquemia hepática total (15 min.) e reperfusão (60 min.); grupo Total submetido a choque hemorrágico controlado (15 min.) seguido de reposição volêmica (Ringer lactato + sangue, 3:1) mais isquemia hepática total (15 min.) e reperfusão (60 min.). A dosagem do lactato arterial e déficit de base foram utilizados para caracterizar o estado de choque hemorrágico com baixa perfusão tecidual. Após a morte dos animais, procedeu-se à contagem de neutrófilos no tecido hepático. RESULTADOS: Na contagem de neutrófilos no fígado o grupo Pringle diferiu dos grupos Choque e Total, os quais não diferiram entre si (Controle 10,30±3,20; Choque 13,94±2,84; Pringle 7,00±3,40; Total 12,45±3,65). CONCLUSÃO: Em ratos submetidos a estado de choque hemorrágico controlado, associado à isquemia hepática total de 15 minutos, seguido de 60 minutos de reperfusão, não ocorreu acúmulo significativo de neutrófilos no fígado304275281The purpose of this experimental study was to evaluate the effects of total hepatic ischemia and reperfusion on the accumulation of neutrophils in the liver of rats, under normal conditions and in rats submitted to controlled hemorrhagic shock . METHODS: Thirty two adult male Wistar rats, were divided into four groups: the Control group, was submitted to the standard procedures for a period of 60 min of observation; Shock group, was submitted to controlled hemorrhagic shock (mean arterial blood pressure = 40 mmHg, 20 min) followed by volume resuscitation (lactated Ringer's solution + blood, 3:1) and reperfusion for 60 min; Pringle group, was submitted to total hepatic ischemia for 15 min and reperfusion for 60 min; The Total group, was submitted to controlled hemorrhagic shock for 15 min followed by volume resuscitation (lactated Ringer's solution + blood, 3:1), total hepatic ischemia for 15 min and reperfusion for 60 min. Measurements of serum lactate and base excess were used to characterize the hemorrhagic shock state with low tissue perfusion. The counting of neutrophils on the liver tissue was performed after the euthanasia of animals. RESULTS: Values for the counting of neutrophils on the liver indicate that, the animals from Pringle group differed from Shock and Total groups (Control 10.30±3.20, Shock 13.94±2.84, Pringle 7.00±3.40, Total 12.45±3.65) but did not differ from Control group. CONCLUSIONS: Rats submitted to controlled hemorrhagic shock state associated to total hepatic ischemia for 15 minutes, followed by 60 minutes of reperfusion, did not present significant neutrophils accumulation on liver tissu

Immunostaining with D2–40 improves evaluation of lymphovascular invasion, but may not predict sentinel lymph node status in early breast cancer

<p>Abstract</p> <p>Background</p> <p>Sentinel lymph node (SLN) biopsy is a widely used diagnostic procedure in the management of early breast cancer. When SLN is free of metastasis, complete axillary dissection may be skipped for staging in clinically N0 patients, allowing a more conservative procedure. Histological tumor features that could reliably predict SLN status have not yet been established. Since the degree of tumor lymphangiogenesis and vascularization may theoretically be related to the risk of lymph node metastasis, we sought to evaluate the relationship between lymph vessel invasion (LVI), lymphatic microvascular density (LVD), microvascular density (MVD) and VEGF-A expression, with SLN status and other known adverse clinical risk factors.</p> <p>Methods</p> <p>Protein expression of D2–40, CD34, and VEGF-A was assessed by immunohistochemistry on paraffin-embedded sections of primary breast cancer specimens from 92 patients submitted to SLN investigation. The presence of LVI, the highest number of micro vessels stained for D2–40 and CD34, and the protein expression of VEGF-A were compared to SLN status, clinicopathological features and risk groups.</p> <p>Results</p> <p>LVI was detected in higher ratios by immunostaining with D2–40 (p < 0.0001), what would have changed the risk category from low to intermediate in four cases (4.3%). There was no association between LVI and other angiogenic parameters determined by immunohistochemistry with SLN macrometastases, clinical features or risk categories.</p> <p>Conclusion</p> <p>Assessment of LVI in breast carcinoma may be significantly increased by immunostaining with D2–40, but the clinical relevance of altering the risk category using this parameter may not be advocated according to our results, neither can the use of LVI and LVD as predictors of SLN macrometastasis in early breast cancer.</p

Cancer Stem and Progenitor-Like Cells as Pharmacological Targets in Breast Cancer Treatment

The present review is focused on the current role of neoplastic stem and progenitor-like cells as primary targets in the pharmacotherapy of cancer as well as in the development of new anticancer drugs. We begin by summarizing the main characteristics of these tumor-initiating cells and key concepts that support their participation in therapeutic failure. In particular, we discuss the differences between the major carcinogenesis models (ie, clonal evolution vs cancer stem cell (CSC) model) with emphasis on breast cancer (given its importance to the study of CSCs) and their implications for the development of new treatment strategies. In addition, we describe the main ways to target these cells, including the main signaling pathways that are more activated or altered in CSCs. Finally, we provide a comprehensive compilation of the most recently tested drugs

Recrutamento de neutrófilos através da barreira hematoencefálica: importância da isquemia hepática pós-traumática

PURPOSE: To study the effects of total hepatic ischemia, and reperfusion on the accumulation of neutrophils in the brain of rats submitted to normovolemic conditions as well as to controlled hemorrhagic shock state. METHODS: Thirty two adult male Wistar rats, were divided into four groups: the Control group, was submitted to the standard procedures for a period of 60 min of observation; Shock group, was submitted to controlled hemorrhagic shock (mean arterial blood pressure=40mmHg, 20min) followed by volemic resuscitation (lactated Ringer's solution + blood, 3:1) and reperfusion for 60min; Pringle group, was submitted to total hepatic ischemia for 15min and reperfusion for 60min. The total group was submitted to controlled hemorrhagic shock for 20min followed by volemic resuscitation (lactated Ringer's solution + blood, 3:1), total hepatic ischemia for 15min and reperfusion for 60min. Measurements of serum lactate and base excess were used to characterize the hemorrhagic shock state with low tissue perfusion. The counting of neutrophils on the brain was performed after the euthanasia of animals. RESULTS: The values for the counting of neutrophils on the brain indicate that did not occur difference among studied groups (p=0.196) (Control 0.12± 0.11, Shock 0.12± 0.13, Pringle 0.02± 0.04, Total 0.14± 0.16). CONCLUSION: Hemorrhagic shock associated to total hepatic ischemia for 15 minutes, followed by 60 minutes of reperfusion, did not causes significant neutrophils accumulation in the brain of rats.Estudar o efeito da isquemia e reperfusão hepática total sobre acúmulo de neutrófilos no cérebro de ratos, em condições de normalidade e submetidos ao estado de choque hemorrágico controlado. MÉTODOS: Foram utilizados 32 ratos Wistar, machos, distribuídos em quatro grupos de oito animais cada: Grupo Controle, submetido aos procedimentos padrões com um período de 60 minutos de observação; Grupo Choque, submetido a choque hemorrágico controlado (PAM=40mmHg, 20 min) seguido de reposição volêmica (Ringer lactato + sangue, 3:1) e reperfusão (60 min); Grupo Pringle, submetido à isquemia hepática total (15 min) e reperfusão (60 min); Grupo Total submetido a choque hemorrágico controlado (15 min) seguido de reposição volêmica (Ringer lactato + sangue, 3:1) mais isquemia hepática total (15 min) e reperfusão (60 min). A dosagem do lactato arterial e déficit de base foram utilizados para caracterizar o estado de choque hemorrágico com baixa perfusão tecidual. Após a eutanásia dos animais, procedeu-se à contagem de neutrófilos no cérebro. RESULTADOS: a contagem de neutrófilos mostrou que não houve diferença estatística entre os grupos (p=0.196). Grupo Controle 0.12± 0.11, Choque 0.12± 0.13, Pringle 0.02± 0.04 e Total 0.14± 0.16. CONCLUSÃO: Em ratos submetidos a estado de choque hemorrágico controlado associado à isquemia hepática total de 15 minutos, seguido de 60 minutos de reperfusão, não ocorreu acúmulo significativo de neutrófilos no cérebro185392397To study the effects of total hepatic ischemia, and reperfusion on the accumulation of neutrophils in the brain of rats submitted to normovolemic conditions as well as to controlled hemorrhagic shock state. METHODS: Thirty two adult male Wistar rats, were divided into four groups: the Control group, was submitted to the standard procedures for a period of 60 min of observation; Shock group, was submitted to controlled hemorrhagic shock (mean arterial blood pressure=40mmHg, 20min) followed by volemic resuscitation (lactated Ringer's solution + blood, 3:1) and reperfusion for 60min; Pringle group, was submitted to total hepatic ischemia for 15min and reperfusion for 60min. The total group was submitted to controlled hemorrhagic shock for 20min followed by volemic resuscitation (lactated Ringer's solution + blood, 3:1), total hepatic ischemia for 15min and reperfusion for 60min. Measurements of serum lactate and base excess were used to characterize the hemorrhagic shock state with low tissue perfusion. The counting of neutrophils on the brain was performed after the euthanasia of animals. RESULTS: The values for the counting of neutrophils on the brain indicate that did not occur difference among studied groups (p=0.196) (Control 0.12± 0.11, Shock 0.12± 0.13, Pringle 0.02± 0.04, Total 0.14± 0.16). CONCLUSION: Hemorrhagic shock associated to total hepatic ischemia for 15 minutes, followed by 60 minutes of reperfusion, did not causes significant neutrophils accumulation in the brain of rats

Modelo experimental de carcinoma mamário em ratas induzidas com 7,12-dimetilbenz(a)antraceno.

Objective: To test an experimental model of chemical mammary carcinogenesis induction in SpragueDawley rats. Methods: Thirty virgin Sprague-Dawley female rats, aged 50 days, received 20 mg of 7,12-dimethylbenz(a)anthracene (DMBA) intragastrically by gavage. At 12 week their mammary glands were examined. Brain, liver, bone and lung tissue were also analyzed. Results: Twelve weeks after DMBA injection, 85% rats presented at least one breast tumor. Conclusin: This experimental animal model of chemical mammary induced carcinogenesis is feasible and can be used in further experiments on the role of tumorigenic biomodulator substances.Objetivo: Testar um modelo experimental de indução de carcinogênese em ratas Sprague-Dawley. Métodos: Foram estudadas 30 ratas fêmeas virgens Sprague-Dawley, induzidas ao carcinogênese mamário. Com 50 dias de vida, foi injetado 7,12-dimetilbenz(a)antrace no ventre por gavage. Com 12 semanas, as glândulas mamárias foram examinadas, assim como os tecidos cerebrais, pulmonares, ossos do fêmur e fígado. Resultados: Doze semanas após a injeção de DMBA, 85% das ratas apresentaram pelo menos um tumor mamário visível. Conclusão: O modelo experimental de carcinoma mamário induzido por DMBA mostrou-se efetivo e de fácil reprodução