Ki‑67 Expression in Oral Potential Malignant and Malignant Lesions and Correlation of Mitotic Index with MIB‑1 Labeling Index

Introduction: Oral cancers are the most serious health issues in underdeveloped countries such as India and considered as the main cause of death. Among them, oral squamous cell carcinoma is the most common type (90%) of all malignancies. Various oral potential malignant lesions (OPMLs) can transform into malignancies. This study was conducted to determine the significance of Ki‑67 expression in oral potential malignant and malignant lesions (MLs) as well as correlation of mitotic index (MI) with MIB‑1 labeling index (LI) in these lesions. MaterialsandMethods: The study was performed on 60 cases in a tertiary care center over a period of 2 years. Ki‑67 expression, MI and MIB‑1 LI were calculated and correlated. Results: In the studied population, there were 49 (81.7%) males and 11 (18.3%) females. The mean age was 46.60 ± 9.94 (23–68 years), with majority of patients in 41–60 years of age group (46/60 cases). Anterior 2/3rd tongue is the most affected site, presented ulcer as the most common lesion. Smoking, tobacco, and betel nutchewing addiction were presented in 72% of the patients. Among 60 cases, 45 (75%) were OPMLs, while 15 (25%) cases were MLs. MI increases in OPMLs and MLs and comparison was significant (P < 0.01). MIB‑1 LI was significant (P < 0.01) on comparison to dysplasia III and MLs. A positive correlation (0.01) was established between MI and MIB‑1 LI of OPMLs and MLs. Conclusion: Ki‑67 expression was found correlated with the progression of disease from OPMLs to MLs. Therefore, it is considered a proliferative marker that corresponds with disease progression. Both proliferative indices (MI and MIB‑1 LI) are positively correlated

Variables pronósticas de pacientes con melanomas gruesos. Análisis de 362 casos / Prognostic variables in patients with thick melanomas. Analysis of 362 cases

Resumen Antecedentes: Los estudios sobre factores pronóstico de melanoma están basados en poblaciones caucásicas, con predominio de melanomas delgados (Breslow < 3 mm). Los pacientes mexicanos muestran predominio de melanomas gruesos (Breslow ≥ 3 mm). Objetivo: Identificar factores asociados al pronóstico de pacientes con melanomas gruesos. Material y métodos: Se analizó la influencia pronóstica de factores clinicopatológicos en 362 melanomas gruesos. Resultados: La mediana de Breslow fue de 7 mm, 271 (74.9 %) pacientes tuvieron melanoma acral y 49 (13.5 %) melanoma nodular. El 56.6 % de los pacientes se encontró en etapa clínica [EC] III), 269 (74.3 %) tenía ulceración y 15 (4.1 %) márgenes positivos. Las variables asociadas con menor supervivencia global [SG] fueron la EC (p < 0.001), Breslow (p = 0.044), ulceración (p = 0.004), mitosis (p < 0.001) y margen < 2 cm (p < 0.001) . En el análisis multivariante los factores que influyen en SG fueron la EC, mitosis y el margen quirúrgico. Conclusiones: En pacientes con melanomas gruesos la SG es influida por un margen positive, mitosis y EC.

Abstract Background: Studies on prognostic factors in melanoma are based on Caucasian populations, with a predominance of thin melanomas (Breslow <3 mm). Mexican patients show a predominance of thick melanomas (Breslow ≥ 3 mm). Objective: To identify factors associated with the prognosis of patients with thick melanomas. Material and methods: The prognostic influence of clinicopathological factors was analyzed in 362 thick melanomas. Results: The Breslow median was 7 mm, 271 (74.9 %) patients had acral melanoma and 49 (13.5 %) nodular melanoma. The 56.6 % of patients were found in clinical stage [CS] III), 269 (74.3 %) had ulceration, and 15 (4.1 %) had positive margins. The variables associated with lower overall survival [OS] were CS (p < 0.001), Breslow (p = 0.044), ulceration (p = 0.004), mitosis (p < 0.001) and margin < 2 cm (p < 0.001). In the multivariate analysis, the factors influencing OS were CD, mitosis, and the surgical margin. Conclusions: In patients with thick melanomas, OS is influenced by a positive margin, mitosis and CS.

Evaluation Of Mutagenic And Antimutagenic Activity Of Methanol Extract Of Cousinia Thomsonii Against Cyclophosphamide

The present study was conducted to evaluate the mutagenic and antimutagenic potential of Cousinia thomsonii (CT) extract in bone marrow cells of male wistar rats using some important parameters like micronucleated polychromatic erythrocyte (MnPCE), mitotic index (MI), chromosomal aberrations (CA) and polychromatic erythrocyte to normochromatic erythrocyte ratio (PCE/NCE). 30 male rats of wistar strain were divided into 6 groups with 5 rats each group. Group 1 rats were taken as negative control having free access to distilled water and rat feed. Group 2 rats were taken as positive control treated with mutagen cyclophosphamide (CP) at dose of 60 mg/kg b wt. for 2 days. Group 3 and 4 were treated with CT extract at dose of 100 and 200 mg/kg b wt. for 20 days. Group 5 and 6 were treated with 100 and 200 mg/kg b wt of CT extract for first 18 days and for last 2 days with CP at dose concentration of 60 mg/kg. It was found that rats treated with CT extract alone did not produce any significant changes in MnPCE, PCE/NCE ratio, CA and MI when compared with control treated rats (group 1). However in group 5 and 6 rats treated with CT extract in combination with CP a protective effect was observed against the cyclophosphamide induced cellular mutagenicity. In concluding remark Cousinia thomsonii was found to show antigenotoxic potential and also produce protective antimutagenic effects against CP induced chromosomal damage.

Cytotoxic, clastogenic and genotoxic effects of cis-tetraammine(oxalato)ruthenium(III) dithionate on human peripheral blood lymphocytes

There is a concern about stablishing the clinical risk of drugs used for cancer treatment. In this study, the cytotoxic, clastogenic and genotoxic properties of cis-tetraammine(oxalato)ruthenium(III) dithionite - cis-[Ru(C2O4)(NH3)4]2(S2O6), were evaluated in vitro in human lymphocytes. The mitotic index (MI), chromosomal aberrations (CA) and DNA damage by comet assay were also analyzed. The MTT test revealed that the ruthenium compound showed a slight cytotoxic effect at the highest concentration tested. The IC50 value for the compound after 24 hours of exposure was 185.4 µM. The MI values of human peripheral blood lymphocytes treated with 0.015, 0.15, 1.5 and 150 µM of cis-[Ru(C2O4)(NH3)4]2(S2O6) were 6.1, 3.9, 3.2 and 0.2%, respectively. The lowest concentration, 0.015 µM, did not show any cytotoxic activity. The CA values for the 0.015, 0.15 and 1.5 µM concentrations presented low frequency (1.5, 1.6 and 2.3%, respectively), and did not express clastogenic activity when compared to the negative control, although it was observed clastogenic activity in the highest concentration tested (150 µM). The results obtained by the comet assay suggest that this compound does not present genotoxic activity at lower concentrations. The results show that cis-[Ru(C2O4)(NH3)4]2(S2O6) has no cytotoxic, clastogenic or genotoxic in vitro effects at concentrations less than or equal to 0.015 µM. This information proves as promising in the treatment of cancer and is crucial for future trials.

Análisis de las modificaciones en la estadificación del T según el manual del American Joint Committee on Cancer. ¿Cómo cambia nuestro manejo de los pacientes? Estudio de corte transversal / Analysis of the change in the staging of the T according to the manual of the American Joint Committee on Cancer. How does our patient management change?

Antecedentes: En la octava edición del manual de estadificación del cáncer del American Joint Committee on Cancer (AJCC), se introdujeron cambios importantes en las categorías T, N y M. Al entrar en vigencia la octava guía de la AJCC, se modificó no solo el T, sino también la indicación de biopsia del ganglio centinela (BGC). Entre los cambios más significativos en la estadificación se encuentran: la exclusión del índice mitótico (IM) de la categoría T en los melanomas finos (de hasta 1 mm de espesor) y el cambio del punto de corte para el espesor tumoral para discriminar un T1a (< 0,8 mm sin ulceración) de un T1b (≥ 0,8 mm). Objetivo: Comparar la estadificación inicial de los melanomas finos según el criterio utilizado en la séptima edición, con la que tendrían de acuerdo con la perspectiva actual del AJCC, con especial atención en el índice mitótico. Diseño y métodos: Estudio observacional, de corte transversal, realizado mediante la recolección de datos de las historias clínicas desde el 1 de enero de 2000 hasta el 31 de diciembre de 2017. Resultados: De 131 melanomas finos incluidos, 28 tendrían cambios en su estadificación. Al considerar el nuevo punto de corte para el espesor tumoral, 22 melanomas T1a pasarían a T1b. Asimismo, se detectaron 20 melanomas con un IM ≥ 1 mitosis/mm2, de los cuales solo 6 tuvieron indicación de BGC por este criterio exclusivamente y serían clasificados como T1a en la actualidad. De estos, en 2 no se realizó la BGC por autodeterminación de los pacientes y en los 4 restantes el resultado fue negativo. Conclusiones: Veintiocho de nuestros pacientes tendrían hoy diferencias en la indicación de BGC: 22 serían considerados con mayor riesgo de metástasis ganglionares y serían candidatos a su pesquisa. Los otros 6 pacientes ya no tendrían indicación de ese estudio por la baja posibilidad de encontrar metástasis ocultas, lo cual coincide con el resultado negativo de la BGC en los 4 pacientes que se sometieron al procedimiento. (AU)

Background: In the 8th edition of the cancer staging manual of the American Joint Committee on Cancer (AJCC), important changes were made in the T, N and M categories. When the 8th guideline of the AJCC came into effect, not only was the T stage modified, but also the indication for sentinel lymph node biopsy (SLNB). The most significant changes in staging included: the exclusion of the mitotic index (MI) as a determinant of the T category and the change of the threshold of tumor thickness to discriminate a T1a (< 0.8 mm without ulceration) from a T1b (≥ 0.8 mm). Objective: To compare the initial staging of thin melanomas according to the criteria used in the 7th edition, with the one that would have been used according to the current AJCC recommendations, with special focus on MI. Design and methods: Observational, cross-sectional study, carried out through the collection of data from medical records from January 1, 2000 to December 31, 2017. Results: There were 131 thin melanomas included, 28 of which would have had changes in their staging. When considering the modified threshold for tumor thickness, 22 T1a melanomas would be classified as T1b. Among 20 thin melanomas with a MI ≥ 1, only 6 had an indication for SLNB solely due to the MI criterion and would be now classified as T1a. Two of these did not undergo SLNB because they rejected the procedure, and in the remaining 4, there were no SLN metastasis. Conclusions: Nowadays, 28 of our patients would have differences in the indication for SLNB: 22 would be considered to be at greater risk of lymph node metastasis and would be candidates for screening. The other 6 patients would no longer have an indication for this procedure due to the low probability of clinically occult metastases, which seems to concur with the negative result of SLNB in the 4 patients who underwent the procedure. (AU)

Acute toxicity of Jatropha curcas oil on plant cell cycle / Toxicidade aguda do óleo de jatropha curcas no ciclo celular de plantas

Today, a great interest in Jatropha-based products exists worldwide, mainly for the production of biofuel.However, the oil obtained from this plant is known to be toxic due to contained curcins andphorbol esters. Bioassays, including plant cytogenetic assays based on cell cycle observation, are useful for determining the toxicity of J. curcas oil. Hence, the aim of this study was to describe the mechanism of action of J. curcas oil by cell cycle analysis using Lactuca sativa as plant testing model. A decrease in root growth was observed, closely related to the reduction in mitotic index, along with an increase in condensed nuclei. J. curcas chemicals act both as aneugenic agents, leading to the formation of lagged, sticky chromosomes and c-metaphase cells, as well as clastogenic agents, inducing the formation of chromosome bridges and fragments. The cytotoxicity and genotoxicity of phorbol esters and other chemical components of J. curcas oil was determined and discussed.

Um grande interesse mundial existe em produtos à base de pinhão manso, principalmente para a produção de biocombustíveis. No entanto, o óleo obtido a partir desta planta é conhecidamente tóxico por conter curcina e ésteres de forbol. Bioensaios, incluindo ensaios citogenéticos em plantas-modelo com base na observação do ciclo celular, são úteis para determinar a toxicidade do óleo de J. curcas. Assim, o objetivo deste estudo foi descrever o mecanismo de ação do óleo de J. curcas por análise do ciclo celular usando Lactuca sativa como modelo de teste em plantas. Foi observada uma redução no crescimento das raízes, intimamente relacionada com a redução do índice mitótico e com um aumento de núcleos condensados. Os constituintes químicos de J. curcas atuam simultaneamente como agentes aneugênicos, levando à formação de cromossomos perdidos e pegajosos e células em c-metáfase, bem como agentes clastogênicos, induzindo a formação de pontes e fragmentos cromossômicos. A citotoxicidade e genotoxicidade do éster de forbol e outros componentes químicos do óleo de J. curcas foram determinados e discutidos.

Relationship between mitotic index and Ki67 expression in meningioma

Background: Meningioma is the most common primary central nervous system (CNS) tumor which covers 36.4% of all CNS tumors. Two important factors determining the prognosis of patients with a diagnosis of meningioma are the percentage of tumor resection and the degree of tumor histopathology. Because there are limitations to routine histopathological examination in predicting tumor progressivity, several examination techniques have been developed including cytogenetics and use of immunohistochemical examination.Method: Observational analytic study was carried out on 68 tumor samples in dr. Wahidin Sudirohusodo Hospital and the Makassar Pathology Diagnostic Center with diagnoses of meningioma from 2012-2018. The Sample size is determined by consecutive sampling method.Results: Sample Size were 68 people from Dr. Wahidin Sudirohusodo Hospital and the Makassar Pathology Diagnostic Center with diagnoses of meningioma which fulfilled the inclusion criteria and consisted of 19(27.9%) men and 49 (72.1%) women with an average age of 42 years. The most common location of the tumor was in the convexity area with an incidence of 29.4%. Grade I meningioma was found 41.2%, grade II of 32.4% and grade III of 26.5%. In grade I, the mean mitotic index was 0.25, grade II was (7.4) and grade III was 22.75. In grade I, the mean Ki67 expression was 1.01%. The highest expression was obtained in grade III with amean of 14.8% and the highest expression was 53%. The Spearman’s rho test results between the mitotic index and Ki67 expression show that there is a positive correlation of 0.490, which means that there is a moderate correlation.Conclusion: IHC expression of Ki67 increases proportionally to the degree of histopathology of meningioma. There is a positive correlation of 0.490 which means that there is a moderate correlation between the mitotic index and Ki67 expression.

Induced genetic variations in Cuminum cyminum through supplemental UV-B radiation

Aim: The present study was conducted to investigate the genotoxic effect of UV-B radiation on morphological, biochemical and cytological parameters of Cuminum cyminum. Methodology: The germinated roots of cumin were exposed to UV-B radiation at different intervals viz., 20 min 40 min and 60 min, respectively, and some treated seeds were sown on pots for morphological and biochemical observation. Results: Chromosomal studies divulged that UV-B radiation has substantial impact on Active Mitotic Index (AMI %). On increasing UV-B radiation dose, the chromosomal aberration rate elevated, thus AMI % moderately decreased and Total Abnormality percentage (TAB %) gradually increased. Different types of chromosomal abnormalities were ascertained, among which scattering was more prominent. The morphological observation showed that survival percentage and plant height decreased at elevated dose of UV-B. Biochemical results indicated that chlorophyll a, chlorophyll b and carotenoid contents decreased as compared to control, but proline content showed significant increment at higher doses of UV-B.Interpretation: The observation elucidates that UV-B causes chromosomal aberrations during cell division and acts as a potent genotoxic agents for roots. Thus, it can be concluded from the above experiment that UV-B rays promote plant growth at low doses but at the higher doses, it subsequently hinders plant growth by damaging important constituents of plant cell.

Efecto antigenotóxico de las semillas de Salvia hispanica "chia" utilizando la prueba de Allium cepa / Antigenotoxic effect of Salvia hispanica "chia" seeds using the Allium cepa test

Objetivo: El objetivo fue evaluar el efecto antigenotóxico de las semillas de Salvia hispanica expuestas al efecto genotóxico de la azida de sodio utilizando la prueba de Allium cepa. Materiales y métodos: Estudio experimental, las unidades de análisis fueron células meristemáticas de 40 bulbos de Allium cepa. Se dispuso 5 grupos de trabajo: control negativo, control positivo, pretratamiento, simultáneo y postratamiento, además de dos concentraciones (10% y 20%). Las raíces se expusieron a los tratamientos por un total de 48 horas. Resultados: Se observó una disminución significativa del índice mitótico (IM) del control negativo (23,7) respecto al control positivo (3,4) confirmando la acción genotóxica de la azida de sodio. En el grupo pretratamiento, la reducción de daño (RD%) fue de -151,2% y -59,2% señalando su nulo papel desmutagénico. En el grupo postratamiento y simultáneo, la RD% fue de 37,6%, 67,2% y 59,2%, 64,8% respectivamente para ambas concentraciones indicando su actividad bioantimutagénica dependiente de la concentración. Sin embargo, a las concentraciones evaluadas no se halló significancia estadística. Conclusión: Las semillas de Salvia hispanica expuestas al efecto genotóxico de la azida de sodio utilizando la prueba de Allium cepa no tienen un efecto antigenotóxico a las concentraciones de 10% y 20%.

Objective: The objective was to assess the antigenotoxic effect of Salvia hispanica seeds exposed to the genotoxic effect of sodium azide using the Allium cepa test. Materials and Methods: This is an experimental study. The analyzed units were meristem cells from forty Allium cepa bulbs Five working groups were available: negative controls, positive controls, pre-treatment, simultaneous, and post-treatment, and two concentrations (10% and 20%) were used. Bulbs were exposed to treatment for a total 48-hour time. Results: A significant reduction in the mitotic index (MI) of negative controls (23.7) was observed, compared to positive controls (3.4), confirming the genotoxic action of sodium azide. In the pre-treatment group, damage reduction values (DR%) were -151.2% and -59.2%, indicating their absent demutagenic role. In both posttreatment and simultaneous groups, DR% values were 37.6 %, 67.2 %, 59.2 %, and 64.8 % for both concentrations, respectively, indicating their concentration-dependent bioantimutagenic activity. However, no statistically significant differences were found between the concentrations tested. Conclusion: Salvia hispanica seeds exposed to the genotoxic effect of sodium azide using the Allium cepa test have no antigenotoxic effect at 10% and 20% concentrations.

Capsiate treatment in liver surgeries may compromise its recovery

Abstract Purpose: To investigate the effects of capsiate treatment on hepatic hyperplasia in partially hepatectomized rats. Methods: The animals were divided into a Capsiate group (CPH), a Capsiate Post-Partial Hepatectomy group (CPPH) and a Partial Hepatectomy Control group (PH). CPH and CPPH animals received 60 mg/kg/day Capsiate for 30 days. Next, the rats underwent partial hepatectomy. CPPH animals continued to receive treatment for 48 h after partial hepatectomy. Liver tissue and intracardiac blood samples were obtained 24 or 48 h after PH. Results: Capsiate treatment interfered with hepatic parameters, reducing the number of mitoses and apoptosis and increasing blood ALT and alkaline phosphatase concentrations. Conclusion: Capsiate treatment preceding hepatic surgery may compromise the initial period of postoperative recovery.

Cytogenotoxicity of rice crop water after application of the tricyclazole fungicide

ABSTRACT Tricyclazole is currently one of the fungicides recommended for the treatment of diseases in irrigated rice. However, there is relatively little information on its cytotoxic and genotoxic potential. The objective of this study was to evaluate the cytotoxicity and genotoxicity of rice crop water after apllication of the tricyclazole fungicide through the Allium cepa L. test. The rice crop water samplings were collected before and 1, 15 and 30 days after application of the fungicide in rice plant shoots. The Allium cepa roots were placed in contact with the rice crop water to check for possible chromosomal abnormalities and mitotic index of the bioindicators meristematic cells. The data obtained by the Allium cepa test indicates that the application of the tricyclazole fungicide leads to an increase in the genotoxic activity in the rice crop water, through the appearance of chromosomal abnormalities, without, however, causing significant effects on the mitotic index. The major chromosomal alterations observed were anaphasic and telophasic bridges and laggard chromosomes.

Lycopene and resveratrol pretreatment did not interfere with the liver of hepatectomized rats

Abstract Purpose: To investigate the effects of lycopene and resveratrol pretreatment on hepatic hyperplasia in partially hepatectomized rats. Methods: The lycopene group and the resveratrol group received 40 mg/kg/day of lycopene or resveratrol, respectively (dissolved in olive oil or in saline solution, respectively) and administered via a gastric tube for 30 days. The partially hepatectomzed (PH) control groups received saline or olive oil via a gastric tube for 30 days, respectively, and the normal control group received no treatment. Liver tissue and intracardiac blood samples were obtained 24, 36 or 48 h after PH. Results: No areas of fibrosis were detected. No significant changes in mitotic index, in the number of apoptosis events or in aspartate aminotransferase and alanine aminotransferase levels were observed. Conclusions: Lycopene and resveratrol pretreatment did not interfere on hepatic hyperplasia in partially hepatectomized rats.

Assessment of the antiproliferative and antigenotoxic activity and phytochemical screening of aqueous extracts of Sambucus australis Cham. & Schltdl. (ADOXACEAE)

ABSTRACT The purpose of this study was to evaluate the antiproliferative and antigenotoxic activity of Sambucus australis Cham. & Schltdl. aqueous extracts on the cell cycle of Allium cepa L. as well as determine the phenolic compounds in such extracts. S. australis inflorescences and leaves of two accessions were used for aqueous extract preparation at concentrations: 0.003 g/ml and 0.012 g/ml. A. cepa bulbs were rooted in distilled water and, subsequently, placed in treatments for 24 hours. Rootlets were collected and fixed in modified Carnoy's solution for 24 hours and kept. The squash technique was performed for slide preparation. Root tips were smashed and stained with 2% acetic orcein, and a total of 4000 cells per treatment were analyzed. The phenolic compounds were determined using high-performance liquid chromatography and data was analyzed using the Scott-Knott test. The results show that S. australis aqueous extracts have antiproliferative potential. Besides, the extracts prepared from S. australis leaves of both accessions at a concentration of 0.012 g/ml have shown antigenotoxic activity. The phytochemical analysis allowed us to determine the presence of flavonoids and phenolic acids, of which kaempferol and chrologenic acid were the most predominant compounds in the extracts from the inflorescences and leaves, respectively.

Composição Química e Atividade Antioxidante de Agaricus blazei e seu Efeito Sobre a Modulação da Mitose / Chemical Composition and Antioxidant Activity of Agaricus blazei and its Effect on Mitosis Modulation

Este estudo teve como objetivo verificar a composição química e atividade antioxidante do Cogumelo do Sol (Agaricus blazei) e seu efeito sobre o ciclo celular de Allium cepa. A análise da composição química revelou uma elevada concentração em compostos fenólicos (65,36 mg/ EAG), ácido ascórbico (1,74mg/g), ß-caroteno (384,28µg/mg) e licopeno (33,61µg/mg). A eficiência no sequestro de radicais livres do DPPH foi considerada primária (68,15%) e a análise do ciclo celular mostrou que o tratamento com Agaricus blazei promove atraso na fase de anáfase da mitose em células meristemáticas de Allium cepa. Os resultados sugerem que a composição química e antioxidante de Agaricus blazei pode influenciar na modulação do ciclo celular de Allium cepa e possivelmente reduz a taxa de mutação espontânea, por diminuir a velocidade de disjunção e migração dos cromossomos durante a mitose. (AU)

The chemical composition and antioxidant activity of mushrooms Agaricus blazei and its effect on the Allium cepa cell cycle was investigated. Chemical analysis showed high concentration of phenolic compounds (65.36 mg/GAE), ascorbic acid (1.74 mg/g), ß-carotene (384.28 µg/mg) and lycopene (33.61 µg/mg). The efficiency free radical DPPH scavenging acitvity was primary (68.15%). Analysis of cell cycle demonstrated that treatment with Agaricus blazei delayed the mitosis anaphase phase in meristem cells of Allium cepa. The results suggest that the chemical composition and antioxidant activity of Agaricus blazei may affect the modulation of the Allium cepa cell cycle, and possibly reduce the spontaneous mutation rate, by decreasing the disjunction and migration speed of chromosomes during mitosis. (AU)

The Prognostic Role of Mitotic Index in Hepatocellular Carcinoma Patients after Curative Hepatectomy / Journal of the Korean Cancer Association, 대한암학회지

PURPOSE: High proliferation rate is a hallmark of cancer. The mitotic index is a useful and simple method for analysis of cell proliferation. However, the practical utility of mitotic index as a predictor of prognosis in patients with hepatocellular carcinoma (HCC) has not been determined. Therefore, we examined mitotic index as a prognostic marker in HCC patients. MATERIALS AND METHODS: We counted the number of mitotic cells in 10 high-power fields of the tumor area on hematoxylin and eosin-stained slides representing 282 surgically resected HCCs. The highest number of mitotic cells was defined as the mitotic index. RESULTS: High mitotic index was observed in 127 of 282 HCCs. High mitotic index showed significant association with younger age, larger tumor size, higher Edmondson grade, microvascular invasion, major portal vein invasion, intrahepatic metastasis, higher American Joint Committee on Cancer (AJCC) T-stage, higher Barcelona Clinic Liver Cancer (BCLC) stage, higher alpha-fetoprotein level, hepatitis B virus etiology, and liver cirrhosis. Patients with high mitotic index had shorter disease-specific survival (DSS) (p < 0.001) and tended to have shorter recurrence-free survival (p=0.112). In subgroup analysis among patients with a larger tumor size, microvascular invasion, intrahepatic metastasis, higher AJCC T-stage, and higher BLCL stage, high mitotic index showed unfavorable influences on DSS (p=0.001, p=0.008, p=0.003, p=0.012, and p < 0.001, respectively). In addition, high mitotic index was an independent predictor of shorter DSS (p=0.004). CONCLUSION: High mitotic index may be a novel predictor of DSS in patients with HCC and may have utility as an auxiliary prognostic factor in HCC.

T. thermophilus Rhamnolipids Induce Cytogenetic Damage on Human Lymphocytes and Bind DNA in vitro.

Aims: Bacteria including Pseudomonas and T. thermophilus secretes rhamnose–containing glycolipid biosurfactants called rhamnolipids (RLs), known as bacterial virulence factors. The aim of this investigation was the evaluation of DNA damage induced on human lymphocytes by both RLs itself, secreted in a host organism by pathogens during a bacterial attack or symbiosis and in combination with the camptothecin (CPT), and on calf thymus DNA. Study Design: Human lymphocytes and calf thymus DNA were treated with isolated T. thermophilus RLs for studying DNA damage in vitro. Methodology: RLs DNA damaging action was evaluated by the Sister Chromatid Exchanges (SCEs) methodology, a method for estimating genotoxicity of human exposure to different chemicals or other mutagenic agents and by DNA electrophoretic mobility experiments. Results: RLs at concentrations of 100 and 150 μg/mL reveal significant toxicity. The highest concentration of 200 μg/mL reveals higher genotoxicity. The frequency of SCEs/cell was increased two times over the control level. When CPT, an antineoplastic drug with DNA damaging action, was tested together with RLs the genotoxic activity was reduced significantly (P<0.01) compared to the action caused by CPT itself. Sequential increase in the concentration of RLs results in the proportional reduction of Proliferation Rate Index (PRI) which is a cytostatic index. Also, Mitotic Index (MI), a cytotoxic index, was also significantly decreased at concentration of 200 μg/mL RLs. Addition of RLs in the same concentration together with CPT doesn’t affect the MI so much. Moreover, RLs are obviously capable for strong binding to plasmid or calf thymus DNA in vitro. Conclusion: RLs exert genotoxicity, cytotoxicity and cytostaticity in human lymphocytes and play probably a protective role for cells against CPT due to RLs’ detergent capability to enrobe CPT and DNA, providing a significant property that might support its possible involvement in DNA horizontal transfer phenomena.

Chromatographic analysis, antiproliferative effect and genotoxicity of aqueous extracts of Citrus sinensis (L) osbeck on the Allium cepa L test system / Análise cromatográfica, efeito antiproliferativo e genotoxicidade de extratos aquosos de Citrus sinensis (L) osbeck sobre o sistema teste de Allium cepa L

The species Citrus sinensis (L.) Osbeck (orange) is used in folk medicine as an infusion (tea) for the treatment of nerve irritation, insomnia, spasms, whooping cough, flue, and gastrointestinal problems. The aim of this study was to evaluate the antiproliferative and genotoxic potential of aqueous extracts of C. sinensis testing by Allium cepa L., as well as to determine the components in the complex mixture (tea). Leaves from two populations of this species were used in the preparation of infusions at two concentrations: 6g.L-1 and 24g.L-1, besides using distilled water as a negative control and 9.6% glyphosate as a positive control. Six groups of four bulbs of A. cepa were used, each group corresponding to one of the treatments. The bulbs were rooted in distilled water and then transferred to the respective treatments where they remained for 24 hours (the bulbs of the negative control remained in distilled water). The rootlets were collected, fixated in ethanol-acetic acid (3:1) for 24 hours and stored in 70% alcohol. 4,000 cells were analyzed per group of bulbs, and the calculated mitotic indices were submitted to statistical analysis using the c 2 test. The aqueous extracts of C. sinensis have as main components flavonoids and phenolic acids. The values of the mitotic index decreased with increasing concentration of the extracts relative to the control in water. The results indicated that the orange aqueous extracts in the studied concentrations in both populations have antiproliferative and genotoxic effects on the cell cycle of A. cepa.

A espécie Citrus sinensis (laranjeira) (L.) Osbeck é usada na medicina popular como infusão (chá) para o tratamento de irritações nervosas, insônia, espasmos, tosse convulsa, gripes e problemas gastrointestinais. O objetivo desse estudo foi avaliar o potencial antiproliferativo e genotóxico dos extratos aquosos de Citrus sinensis pelo teste de Allium cepa L., bem como determinar os componentes na mistura complexa (chá). Folhas de duas populações dessa espécie foram utilizadas no preparo das infusões em duas concentrações: 6g.L-1 e 24g.L-1, além de serem usados água destilada como controle negativo e glifosato 9,6% como controle positivo. Utilizaram-se seis grupos de quatro bulbos de A. cepa, cada grupo correspondendo a um dos tratamentos. Os bulbos foram enraizados em água destilada e então transferidos para os respectivos tratamentos onde permaneceram por 24 horas (os bulbos do controle negativo permaneceram em água destilada). As radículas foram coletadas, fixadas em etanol-ácido acético (3:1) por 24 horas e armazenadas em álcool 70%. Foram analisadas 4,000 células por grupo de bulbos, e os índices mitóticos calculados submetidos à análise estatística pelo teste c 2. Os extratos aquosos de C. sinensis têm como componentes principais flavonoides e ácidos fenólicos. Os valores dos índices mitóticos diminuíram com o aumento das concentrações dos extratos, em relação ao controle em água. Os resultados indicaram que os extratos aquosos de laranjeira nas concentrações estudadas das duas populações possuem ação antiproliferativa e genotóxica sobre o ciclo celular de A. cepa.

Comparative study of the different degrees of risk of gastrointestinal stromal tumor / Estudo comparativo dos diferentes graus de risco no tumor estromal gastrointestinal

OBJECTIVE: To evaluate the applicability of the main categories of risk and morphological factors in the prognosis of gastrointestinal stromal tumors. METHODS: we retrospectively studied fifty-four cases of GIST, assessing the main prognostic factors of this neoplasis: risk levels, topography, size, mitotic index, necrosis, histological subtype and immunophenotype. We also verified their association and the reduction of overall survival. RESULTS: Univariate analysis showed that tumors with mitoses number greater than 5 per 50CGA (high-power fields), the presence of necrosis and a high risk for both the systems proposed by Fletcher and Miettinen had a significant association with reduced survival (p = 0.00001, 0.0056, 0.03 and 0.009, respectively). The remaining analyzed factors (size, histological subtype, topography and immunophenotype) had no such association. Multivariate analysis (Jacard index) showed that the Miettinen degree of risk was the one that best correlated with prognosis. CONCLUSION: the risk criteria of Fletcher and Miettinen are important in assessing the prognosis of patients with gastrointestinal stromal tumors, especially the latter, which adds to the mitotic index and the presence of tumor necrosis.

OBJETIVO: Avaliar a aplicabilidade das principais categorias de risco e de fatores morfológicos no prognóstico tumor estromal gastrointestinal. MÉTODOS: cinquenta e quatro casos de GIST foram estudados retrospectivamente considerando-se os principais fatores prognósticos da neoplasia: graus de risco, topografia, tamanho, índice mitótico, necrose, subtipo histológico e imunofenótipo. Foi também verificada a sua associação e a redução da sobrevida global dos pacientes. RESULTADOS: a análise univariada mostrou que os tumores com número de mitoses maior que 5/50CGA (campos de grande aumento), a presença de necrose, de alto risco tanto para os sistemas propostos por Fletcher, quanto para Miettinen tiveram associação significativa com redução da sobrevida (p=0,00001, 0,0056, 0,03 e 0,009, respectivamente). Enquanto que os demais fatores analisados (tamanho, subtipo histológico, topografia e imunofenótipo) não tiveram tal associação. A análise multivariada (índice de Jacard) demonstrou que o grau de risco de Miettinen foi aquele que melhor se relacionou com o prognóstico. CONCLUSÃO: os critérios de risco de Fletcher e de Miettinen são importantes na avaliação do prognóstico de pacientes com tumor estromal gastrointestinal, principalmente este último, que se soma ao índice mitótico e a necrose tumoral.

Evaluation of allelopathic potential of leaf extract of Kielmeyera coriacea on Lactuca sativa L / Avaliação do potencial alelopático de extrato de folhas de Kielmeyera coriácea sobre Lactuca sativa L

Kielmeyera coriacea Mart. (Clusiaceae), popularly known as "pau-santo", is a typical Brazilian cerrado tree known due to its varied secondary metabolites. This study aimed to determine the allelopathic potential of the hydroalcoholic extract of leaves of K. coriacea through bioassays of seed germination, seedling growth and mitotic index of Lactuca sativa L. (lettuce). In addition it was done the tetrazolium assay and a phytochemical screening. The extract concentrations caused alterations in germination parameters, in root growth and in the mitotic index. The phytochemical screening showed the presence of triterpenes, coumarins, steroids, flavonoids and condensed tannins, compounds known to confer allelopathic characteristics upon other species. These data indicate that K. coriacea presents an allelopathic potential because its leaf extracts interfere with germination and growth without any interference of pH and osmotic potential in the results.

Kielmeyera coriacea Mart. (Clusiaceae), conhecida popularmente como "pau-santo", é uma árvore típica do cerrado brasileiro, conhecida pelos seus variados metabólitos secundários. O estudo teve como objetivo determinar o potencial alelopático do extrato hidroalcoólico de folhas de K. coriacea por meio dos bioensaios de germinação, crescimento de plântulas e índice mitótico de Lactuca sativa L. (alface). Além disso, foi feito o teste de tetrazólio e uma triagem fitoquímica. As concentrações de extrato causaram alterações nos parâmetros de germinação, no crescimento radicular e no índice mitótico. A triagem fitoquímica mostrou a presença de triterpenos, cumarinas, esteroides, flavonoides e taninos condensados, compostos conhecidos por conferir características alelopáticas sobre outras espécies. Estes dados indicam que K. coriacea apresenta potencial alelopático pois o extrato de sua folha interfere na germinação e no crescimento da planta alvo, sem qualquer interferência do pH e do potencial osmótico.

Physiological and Cytogenetic Responses of Wheat and Barley to Silver Nanopriming Treatment.

This experiment was carried out to study the effect of silver-nanopriming on germination percentage, seedling growth and mitotic cell division. The results showed that silver nanoparticles (AgNPs) significantly increased the germination percentage. Mean comparison showed that the highest germination percentages (98, 92, 96 %) were observed in the pretreated wheat (Beni Sweif 1 and Gemmieza 9) and barley (Giza 130) seeds. The results clearly revealed that shoot length, fresh and dry weights were slightly promoted by AgNPs while in roots corresponding parameters were reduced compared with those of control. Photosynthetic pigments and chlorophyll fluorescence were affected to some extent by AgNPs priming. Cytological changes in root tips of 72h and 120h germinated seeds were observed by disturbed chromosomes at metaphase and anaphase. The main types of chromosomal aberrations are: chromosomes aneuploidy, binucleate cells, deletion chromosomes, deform nuclei, micronuclei, chromosomes fragment, and stickiness chromosomes. It was found that the mitotic index significantly increased in the three pretreated tested plants compared to the control. This study infers that AgNPs could penetrate plant system and might impair stages of cell division causing chromosomal aberrations.