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Background: Exclusive breast milk is the recommended nutrition by the World Health Organization (WHO) until six months of age, followed by introducing foods with continual breastfeeding. Bisphenol A (BPA) is a synthetic chemical used as a monomer in polycarbonate plastics and epoxy resins. Human exposure to BPA can cause numerous adverse health effects such as diabetes, obesity and cardiovascular disease, with children and infants as the most susceptible groups. Thus, this study aimed to determine BPA concentration in the colostrum of 64 mothers hospitalized at the University Hospital Professor Polydoro Ernani de São Thiago through the ELISA technique. We also assessed possible associations between breast milk BPA concentration and lactogenesis II. The results showed that all the breast milk samples contained a high concentration of BPA with a median value of 34.18 ng/mL. In addition, exposure to BPA did not alter the nutritional status of the mother; however, BPA concentration in mothers is influenced by the consumption of food packaged in plastic containers. The total daily intake of BPA in breastfed babies was 19.5 µg/kg/day and 28.5 µg/kg/day was recorded in the 95th percentile of the body weight per day, which is higher than the maximum daily intake by the European Food Safety Authority. This is of public health importance as a high concentration of BPA in the breast milk of the lactating mothers in this study could cause severe health problems in these mothers and much more in the infants breastfeeding on these BPA-contaminated breast milk.

This study aimed to describe the prevalence of invasive breast cancer (IBC) in women assisted in a public hospital in Brazil and to establish a correlation between two models of classification by immunophenotypes, one of them based on the 13th St. Gallen Conference classification and the other on biomarker-defined subtypes based on HER2 and oestrogen receptor status, as described in World Health Organization (WHO). We selected IBC of 1335 cases between 1994 and 2018. Univariate frequencies and associations were estimated using chi-square tests. The concordance between the two immunohistochemical analysis models above mentioned using Cohen's kappa coefficients. The most prevalent subtype was luminal B/HER2, and the frequency of tumours with a worse prognosis was 62.7%. Has been identified an association between histological grade 3 (G3) and the worst prognostic subtypes: non-luminal A, Triple Negative Breast Cancer (TNBC), non-ER+/HER2- and ER-/HER2-. A similar association was found in nuclear G3 tumours. The results showed agreement between 99.48% and 100% when we compared the two immunohistochemical analysis models. Furthermore, there was an absolute agreement between the two models of immunohistochemical analysis. These results can contribute to institutions that do not have a molecular investigation, enabling accessible routine practice tools. Among the most important questions addressed in this work is the association between histological and nuclear G3 with the worst prognostic subtypes. Using the St. Gallen Conference classification and HER2 and ER status based on subtypes verified the feasibility of selecting IBC with different prognoses and correlated them with recognized predictive and prognostic factors.

Imidazo[1,2-a]pyridines have been studied regarding drug development. The objective of this work was to evaluate the antileukemic capacity of imidazo[1,2-a]pyridine derivatives by screening its ability as a pro-oxidant. Imidazo[1,2-a]pyridine derivatives were synthesized and oral bioavailability and toxicity were analyzed in silico. Redox screening was performed on human Kasumi, KG-1, K562 and Jurkat leukemia cells. The imidazo[1,2-a]pyridine derivative and the most responsive leukemic cell were selected for cytotoxicity, cell proliferation, cell senescence and oxidative stress assays. The predictive toxicity analysis showed a possible effect on the reproductive system, but without mutagenic, carcinogenic or irritability effects. MRK-107 against K562 cells was the compound that showed the best redox profile. MRK-107 did not induce cell death in K562 and monocyte cells. However, this compound was able to decrease cell proliferation and increase cell senescence after 48 and 72 hours. Furthermore, MRK-107 induced oxidative stress in K562 cells after 72h, increasing lipid peroxidation and decreasing reduced glutathione (GSH) contents. This study demonstrated that MRK-107-induced senescence with the involvement of oxidative stress as a possible mechanism of action, addressing this compound as a potential antitumor drug against chronic myeloid leukemia.

The present study investigated the effects of perinatal exposure to glyphosate-based herbicide (GBH) in offspring's liver. Pregnant Wistar rats were exposed to GBH (70 mg glyphosate/Kg body weight/day) in drinking water from gestation day 5 to postnatal day 15. The perinatal exposure to GBH increased

Acute promyelocytic leukemia is a subtype of acute myeloid leukemia, characterized by the presence of neoplastic promyelocytes, due to the reciprocal balanced translocation between chromosomes 15 and 17. Currently, with the use of agents that act directly on this molecular change, such as all-trans retinoic acid and arsenic trioxide, APL has shifted from a highly mortal to a curable disease. However, some cases are still at high risk of death, especially early death, and acquiring a better understanding of the clinical and biological factors involving APL is needed to correctly identify and treat such cases. The early suspected diagnosis and prompt initiation of the target therapy are important for better response rates. The follow-up and outcomes, using real-life data from 44 consecutive APL patients, were studied between 2001 and 2013. The overall survival rate was 82.7% and early death was 16%. Almost all patient deaths were due to severe bleeding, which was confirmed by multivariate analysis, as the most important prognostic factor leading to death. A better understanding the pathogenesis of the hemorrhagic complications in APL is needed, as well as the risk factors associated with early death in APL patients, as this has become synonymous with overall mortality.

Objective: The aim of this study was to describe the characteristics of blood donors and the serological profile of the blood donations at the blood bank of the University Hospital Polydoro Ernani de São Thiago of the Federal University of Santa Catarina from January 2011 to December 2016. Methods: The characteristics of donors and the serological results of the donated blood were compiled from databases. Only donations with a negative serology or a positive serology confirmed by second-sample testing were included in the study. Results: A total of 14,368 donations were included in the study, of which 118 (0.8%) had a confirmed positive serology. Of the total donations, 94.3% were from spontaneous donations and 5.7% from replacement donation. Donations were predominantly from men (54.1%), individuals aged 18 to 29 years (69.1%), and repeat donors (47.7%). Detection rates were higher for HBV (0.63%), followed by syphilis (0.13%), HIV (0.05%), HCV (0.02%), and Chagas disease (0.01%). With the exception of HIV, positive results were more frequent in the older age groups. Positive results for HBV, HCV, and HIV were more frequent among first-time donors. Replacement donations were more likely to have HBV (OR 7.7; 95% CI 4.9–12.1, p

The present study investigated the effects of perinatal exposure to glyphosate-based herbicide (GBH) on liver of immature Wistar rats. Female rats were exposed to GBH (70 mg glyphosate/Kg body weight/day) in drinking water from gestation day 5 and continually up to lactation day 15. The perinatal exposure to GBH increased 45Ca2+ influx in offspring’s liver Pharmacological tools indicated a role played by oxidative stress, phospholipase C (PLC) and Akt pathways, as well as voltage-dependent Ca2+ channel modulation to GBH-induced Ca2+ influx in liver. In addition, changes in the enzymatic antioxidant defense system, decreased GSH content, lipid peroxidation and protein carbonylation suggest a connection between GBH hepatotoxic mechanism and redox imbalance. The perinatal exposure to GBH also increased the enzymatic activities of transaminases and gamma-glutamyl transferase in offspring's liver and blood, suggesting a pesticide-induced liver injury. Moreover, we detected increased iron levels in liver, blood and bone marrow of GBH-exposed rats, which were accompanied by increased transferrin saturation and decreased transferrin levels in blood. The levels of tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) were increased in the liver of rats exposed to GBH. We also detected increased phospho-p65NFκB immunocontent, corresponding to the active form of this transcription factor. Therefore, we propose that excessive amounts of iron induced by perinatal exposure to GBH in offspring’s liver, blood and bone marrow may account for iron-driven hepatotoxicity, which was associated with Ca2+ influx, oxidative damage and inflammation in immature rats. Further studies will clarify whether these events can ultimately impact on liver function.

Obesity is a global health problem and is associated with a chronic low-grade inflammatory state. Surgical obesity treatment is being increasingly common due to its efficacy. From this, we evaluate the metabolic state improvement and inflammation remission in patients with obesity undergoing bariatric surgery. Methods: The clinical data and serum levels of leptin and adiponectin were assessed in patients with obesity before and one, three and six months after bariatric surgery. Also, serum amyloid A (SAA), monocyte chemoattractant protein-1 (MCP-1) levels were measured during the follow-up surgery and compared with a lean group of individuals. Results: Weight loss decreased body mass index (BMI), comorbidities percentage, drugs use and leptin levels. Adiponectin levels increased after surgery. SAA and MCP-1 showed no difference after surgery, but a trend decrease for MCP-1 and a significant decrease was observed when the patients with obesity were compared to the lean participants. Conclusion: Bariatric surgery alters metabolic status improving obesity-related comorbidities and the adiposity biomarkers leptin and adiponectin, but not inflammatory cytokines SAA and MCP-1.

Liver cirrhosis is often complicated by an immunological imbalance known as cirrhosis-associated immune dysfunction. This study aimed to investigate disturbances in circulating monocytes and dendritic cells in patients with acute decompensation (AD) of cirrhosis. The sample included 39 adult cirrhotic patients hospitalized for AD, 29 patients with stable cirrhosis (SC), and 30 healthy controls (CTR). Flow cytometry was used to analyze monocyte and dendritic cell subsets in whole blood and quantify cytokines in plasma samples. Cirrhotic groups showed higher frequencies of intermediate monocytes (iMo) than CTR. AD patients had lower percentages of nonclassical monocytes than CTR and SC. Cirrhotic patients had a profound reduction in absolute and relative dendritic cell numbers compared with CTR and showed higher plasmacytoid/classical dendritic cell ratios. Increased plasma levels of IL-6, IL-10, and IL-17A, elevated percentages of CD62L+ monocytes, and reduced HLA-DR expression on classical monocytes (cMo) were also observed in cirrhotic patients. Patients with more advanced liver disease showed increased cMo and reduced tissue macrophages (TiMas) frequencies. It was found that cMo percentages greater than 90.0% within the monocyte compartment and iMo and TiMas percentages lower than 5.7% and 8.6%, respectively, were associated with increased 90-day mortality. Monocytes and dendritic cells are deeply altered in cirrhotic patients, and subset profiles differ between stable and advanced liver disease. High cMo and low TiMas frequencies may be useful biomarkers of disease severity and mortality in liver cirrhosis.

Introduction: Breast cancer (BC) is the most incident form of cancer in women worldwide. The widespread use of breast screening programs, as well advances in molecular biology, and new drugs in chemotherapy have contributed to the recent survival rate improvement in high income countries. Furthermore, the study of cancer genome led to the elucidation of the intrinsic subtypes of invasive breast cancer (IBC), consequently, the success rate of targeted therapies improved the outcome in patients. However, considering that immunohistochemistry (IHC) is one of the main methods to determine the profile of protein expression in surgical pathology, most antibodies used have a presumed or already established role and represent proteins whose transcription has been previously described in genetic profile studies. Objectives: Describe the prevalence of IBC in women admitted to a public hospital in Brazil from 1994 to 2018, to establish a correlation between two models of immunohistochemical analysis, the 13th St. Gallen Conference classification and the biomarkerdefined subtypes based on HER2 and estrogen receptor (ER) status, and to investigate the profile of these cases. Methods: Retrospective database analysis was performed. 1,335 women with histologic diagnosis of IBC were included in the study from a public hospital in Brazil between 1994 and 2018. Frequencies and univariate associations were estimated by using chi-square tests. Agreement between the immunohistochemical groups were tested by using Cohen’s kappa coefficients. Results: The mean age was 56.1 years. The most prevalent subtype was luminal B/HER2 and the frequency of tumors with worse prognosis was 62.7%. An association was found between histological grade 3 (G3) and the worst prognostic subtypes: non-luminal A (OR=31.18; 95%CI 13.76–70.64), TNBC (OR=8.77; 95%CI 6.20–12.41), non-ER+/HER2- (OR=5.37; 95%CI 4.11–7.04) and ER-/HER2- (OR=8.50; 95%CI 6.10–11.85). A similar association was found for nuclear G3: non-luminal A (OR=6.3; 95%CI 4.29–9.47), TNBC (OR=5.14; 95%CI 3.64–7.31), non-ER+/HER2- (OR=4.83; 95%CI 3.80–6.15) and ER-/HER2- (OR=5.41; 95%CI 3.92–7.50). When the two models of immunohistochemical analysis were compared, the results showed an agreement rate of 99.48% to 100%. Conclusions: Our results show that most cases had worse outcomes, and there was absolute agreement between the two models of immunohistochemical analysis. These results can contribute to institutions that do not have molecular investigation, enabling accessible tools in routine practice.

Chalcones and their derivatives have been described as promising compounds with antiproliferative activity against leukemic cells. This study aimed to investigate the cytotoxic effect of three synthetic chalcones derived from 1-naphthylacetophenone (F07, F09, and F10) in acute leukemia cell lines (K562 and Jurkat) and examine the mechanisms of cell death induced by these compounds. The three compounds were cytotoxic to K562 and Jurkat cells, with IC50 values ranging from 1.03 to 31.66 µM. Chalcones induced intrinsic and extrinsic apoptosis, resulting in activation of caspase-3 and DNA fragmentation. F07, F09, and F10 were not cytotoxic to human peripheral blood mononuclear cells, did not produce any significant hemolytic activity, and did not affect platelet aggregation after ADP stimulation. These results, combined with calculations of molecular properties, suggest that chalcones F07, F09, and F10 are promising molecules for the development of novel antileukemic drugs.

Background: B cell lymphomas' (BCL) current diagnosis is usually based on a combination of morphology, immunophenotype, recurrent cytogenetic aberration and clinical features. However, even with these diagnostic tools, a definitive diagnosis can be difficult to achieve. Therefore, the aim of this study was to assess the profile of CD39, CD43, CD81 and CD95 expression in diffuse large B cell lymphoma (DLBCL), follicular lymphoma (FL) and Burkitt lymphoma (BL) cases. Methods: To address this issue, we investigated the expression of CD39, CD43, CD81 and CD95 by eight-color flow cytometry in retrospective cases from 2014 to 2016. Results: The study included 27 adult patients diagnosed with DLBCL, FL and BL during the study period. Four patients were diagnosed with germinal center B cell-like DLBCL (GCB DLBCL), seven with non-GCB DLBCL, nine with FL, and seven with BL. CD39 seems to be especially relevant to differentiate non-GCB DLBCL from BL and from FL. BL showed stronger expression of CD43 when compared to FL and GCB DLBCL. Moreover, CD43 may help to distinguish non-GCB DLBCL from GCB DLBCL. CD81 expression was much stronger in BL when compared to the other three groups of patients. Lastly, CD95 may also help to distinguish BL from the other subtypes, as BL cells expressed this antigen at low levels. Conclusions: In combination, CD39, CD43, CD81 and CD95 expression appears to be helpful to distinguish CD10+ BCL, particularly BL. Phenotypic distinction between FL and GCB DLBCL remains challenging and requires further studies. This article is protected by copyright. All rights reserved.

Background Red blood cell genes are highly polymorphic with the distribution of alleles varying between different populations and ethnic groups. The objective of this study was to investigate gene polymorphisms of blood groups in the state of Santa Catarina, Southern Brazil. Methods Three hundred and seventy-three unrelated blood donors and 31 transfusion-dependent patients were evaluated to investigate polymorphisms of the Rh, Kell, Duffy, Kidd, and Diego blood group systems in a population from the state of Santa Catarina. The subjects, from seven regions that comprise the blood-banking network of the state, were assessed between August 2011 and March 2014. The genotypes of the Rh, Kell, Duffy, Kidd, and Diego systems were determined using the restriction fragment length polymorphism-polymerase chain reaction and allele-specific polymerase chain reaction techniques. Results The genotype frequencies in this study were significantly different when populations from different regions of Santa Catarina were compared. Furthermore, there were also significant differences in the genetic frequencies compared to other Brazilian states. The genotype frequencies of the Kell and Kidd blood groups are similar to European populations from Naples, Italy and Zurich, Switzerland. Conclusion This article reports for the first time the frequency of polymorphisms of blood group systems in blood donors from Santa Catarina, Southern Brazil.

Flow cytometry has emerged as a useful screening approach to evaluate whether specific cell populations are present or absent. Previous studies have shown different reference ranges in several countries. The aim of this study was to determine reference ranges of lymphocyte subsets in peripheral blood by flow cytometric method in Brazilian adults. In this study, relative and absolute reference ranges of lymphocyte subsets were: CD3+: 51.3-83.5%, 718-2494cells/μl; CD4+: 24.4-54.2%, 456-1492cells/μl; CD8+: 12.8-40.2%, 272-1144cells/μl; CD4+CD8+: double-positive 0.01-3.6%, 2-88cells/μl; TCR γδ: 1.0-15.9%, 19-345cells/μl; CD3+CD4-CD8-: 1.2-13.3%, 28-292cells/μl; TCR αβ+: 44.3-77.0%, 855-2384cells/μl; CD4/CD8 ratio: 0.68-3.61; CD19+: 6.3-20.8%, 112-622cells/μl; mature NK cells: 3.1-27.4%, 70-745cells/μl; immature NK cells: 0.08-1.1%, 1-23cells/μl; total NK cells: 3.7-28.5%, 82-760cells/μl; and NKT cells: 0.9-21.4%, 18-488cells/μl. Comparison with other studies showed differences among some of them. This suggests that there are differences among lymphocyte subsets in the worldwide population and also it is important to determine reference ranges in different populations in order to better assess and monitor patients.

In 2010, new monoclonal antibodies were submitted to the 9th International Workshop on Human Leukocyte Differentiation Antigens, and there are few studies demonstrating normal expression patterns of these markers. Thus, the objective of this study was to determine the normal patterns of cell expression of CD86, CD210a, CD261, CD262, CD264, CD358, and CD361 in peripheral blood (PB) and bone marrow (BM) samples by flow cytometry. In the present study, CD86 was expressed only in monocytes and B lymphocytes in PB and in monocytes and plasma cells in BM. Regarding CD210a expression, in PB samples, monocytes and NK cells showed weak expression, while neutrophils, B and T lymphocytes, and basophils showed weak and partial expression. In BM samples, expression of CD210a was observed in eosinophils, monocytes, and B and T/NK lymphocytes. Weak expression of CD210a was also observed in neutrophilic cells and plasma cells. All B cell maturation stages had weak expression of CD210a except for immature B cells, which did not express this marker. In the present study, no cell type in PB samples showed positivity for CD261 and, in BM samples, there was very weak expression in neutrophilic series, monocytes, and B lymphocytes. Conversely, plasma cells showed positivity for CD261 with a homogeneous expression. For CD262, there was weak expression in monocytes, neutrophils, and B lymphocytes in PB samples and weak expression in monocytes, B lymphocytes, and plasma cells in BM samples. The evaluation of CD264 showed very weak expression in B cells in PB samples and no expression in BM cells. Very weak expression of CD358 was observed in neutrophils, monocytes, and B lymphocytes in PB and BM samples. In addition, in BM samples, plasma cells and T lymphocytes showed weak expression of CD358. In relation to the maturation stages of B cells, there was weak expression in pro-B cel, pre-B cell, and mature B cell. In the present study, it was possible to observe expression of CD361 in all cell types analyzed in PB and BM samples. The analyzed markers presented varied profiles of expression and, in some cases, these profiles were different from those observed in other studies. Further studies are needed to evaluate these molecules, mainly in relation to a possible application in the diagnosis of hematological malignancies or as new therapeutic targets for the treatment of hematological neoplasms or autoimmune diseases.

When the FLT3 gene is mutated, it originates a modified receptor with structural changes, which give survival advantage and malignant hematopoietic cell proliferation. Thus, the presence of mutations in this gene is considered an unfavorable prognostic factor. A total of 85 consecutive samples of newly diagnosed untreated patients with AL were included in the study after they provided their informed consent. FLT3 gene mutations were detected by PCR. For the pediatric group, a positive correlation was observed between WBC count and the presence of FLT3-ITD in patients with AML and ALL. Furthermore, children with AML who had the FLT3-ITD mutation showed a tendency to express CD34 in blast cells. In the adult group, the AML patients with FLT3-ITD who expressed CD34 in blast cells had a tendency to worse progression. The present data indicate no association between the prognostic factors evaluated and FLT3 gene mutations in adult with AL. Yet, the presence of FLT3-ITD mutation was significantly related with WBC count in the pediatric group. These findings demonstrate that FLT3 gene mutations can be considered as independent poor prognostic factors.

Pyrazoline is an important 5-membered nitrogen heterocycle that has been extensively researched. Ten derivatives were synthesized and tested for antileukemic effects on 2 human acute leukemia cell lines, K562 and Jurkat. The most cytotoxic of these derivatives, compound 21, was chosen for investigation of cytotoxicity mechanisms. The results obtained with selectivity calculations revealed that compound 21 is more selective for acute leukemia (K562 and Jurkat cell lines) than for other tumor cell lines. Moreover, compound 21 was not cytotoxic to normal cell lines, indicating a potential use in clinical tests. Compound 21 caused a significant cell cycle arrest in the S-phase in Jurkat cells and increased the proportion of cells in the sub G0/G1 phase in both cell lines. Cells treated with compound 21 demonstrated morphological changes characteristic of apoptosis in the EB/AO assay, confirmed by externalization of phosphatidylserine by the annexin V – fluorescein isothiocyanate method and by DNA fragmentation. An investigation of cytotoxicity mechanisms suggests the involvement of an intrinsic apoptosis pathway due to mitochondrial damage and an increase in the ratio of mitochondrial Bax/Bcl2. Pyrazoline 21 obeyed Lipinski’s “rule of five” for drug-likeness. Based on these preliminary results, the antileukemic activity of compound 21 makes it a potential anticancer agent.

B cell lymphomas' (BCL) current diagnosis is usually based on a combination of morphology, immunophenotype, recurrent cytogenetic aberration and clinical features. However, even with these diagnostic tools, a definitive diagnosis can be difficult to achieve. Therefore, the aim of this study was to assess the profile of CD39, CD43, CD81, and CD95 expressions in diffuse large B cell lymphoma (DLBCL), follicular lymphoma (FL), and Burkitt lymphoma (BL) cases.To address this issue, we investigated the expression of CD39, CD43, CD81, and CD95 by eight-color flow cytometry in retrospective cases from 2014 to 2016.The study included 27 adult patients diagnosed with DLBCL, FL, and BL during the study period. Four patients were diagnosed with germinal center B cell-like DLBCL (GCB DLBCL), seven with non-GCB DLBCL, nine with FL, and seven with BL. CD39 seems to be especially relevant to differentiate non-GCB DLBCL from BL and from FL. BL showed stronger expression of CD43 when compared to FL and GCB DLBCL. Moreover, CD43 may help to distinguish non-GCB DLBCL from GCB DLBCL. CD81 expression was much stronger in BL when compared to the other three groups of patients. Lastly, CD95 may also help to distinguish BL from the other subtypes, as BL cells expressed this antigen at low levels.In combination, CD39, CD43, CD81, and CD95 expressions appear to be helpful to distinguish CD10

In this study, we assessed the cytotoxic effect of synthetic 2′,4′,5′-trimethoxychalcones on the human K562 acute myeloid leukemia cell and human Jurkat acute lymphoid leukemia cell. Compounds 13, 16, 19, and 26 showed low IC50 values (4.10–8.56 μM at 72 h) for both cell lines and did not have a cytotoxic effect on normal human lymphocytes. The mechanism of cell death induced by these compounds involves a decrease in the expression of cell proliferation marker Ki67, suggesting inhibition of cell proliferation. Furthermore, these chalcones reduced mitochondrial potential, decreased Bcl-2 expression, and increased Bax expression, indicating that the mechanism of apoptosis induced by them involves the intrinsic apoptosis pathway. The mechanism of action also involves increase in active caspase-3 and decrease in survivin expression. These results support the chalcones as potential antitumoral agents for further optimization.

Cyclic imides are known for their antitumor activity, especially the naphthalimide derivatives, such as Mitonafide and Amonafide. Recently, we have demonstrated the cytotoxic effect of a series of naphthalimide derivatives against B16F10 melanoma cells. On the basis of this fact, we have developed a study starting from the synthesis of different cyclic imides and the evaluation of their cytotoxic properties on human acute leukemia cells (K562 and Jurkat). Initially, a screening test was conducted to select the compound with the best cytotoxic effect, using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. After this selection, structural modifications were performed in the most active compound to obtain five more derivatives. All compounds presented a good cytotoxic effect. The results of cell cycle analysis, fluorescence microscopy, and Annexin V-FITC assay confirmed that the cells observed in the sub-G0/G1 phase were undergoing apoptosis. From this set of results, cyclic imides 8, 10, and 12 were selected for the evaluation of the mechanisms involved in the apoptotic process. The results demonstrate the involvement of the intrinsic pathway of apoptosis, evidenced by the reduction in mitochondrial potential, an increase in the level of AIF protein expression, a decreased level of expression of anti-apoptotic Bcl-2 protein, and an increased level of expression of pro-apoptotic protein Bax in both K562 and Jurkat cells treated with cyclic imides (8, 10, and 12). Furthermore, cyclic imides 8 and 10 caused an increase in the level of Fas expression in Jurkat cells, indicating the additional involvement of the extrinsic apoptosis pathway. The compounds (8, 10, and 12) also caused a decreased level of expression of anti-apoptotic protein survivin. The biological effects observed with these cyclic imide derivatives in this study suggest promising applications against acute leukemia.

Acute leukemia is a disorder of the hematopoietic system characterized by the expansion of a clonal population of cells blocked from differentiating into mature cells. Recent studies have shown that chalcones and their derivatives induce apoptosis in different cell lines. Since new compounds with biological activity are needed, the aim of this study was to evaluate the cytotoxic effect of three synthetic chalcones, derived from 1-naphthaldehyde and 2-naphthaldehyde, on human acute myeloid leukemia K562 cells and on human acute lymphoblastic leukemia Jurkat cells. Based on the results, the most cytotoxic compound (A1) was chosen for further analysis in six human acute leukemia cells and in a human colon adenocarcinoma cell line (HT-29). Chalcone A1 significantly reduced the cell viability of K562, Jurkat, Kasumi, U937, CEM and NB4 cells in a concentration and time-dependent manner when compared with the control group (IC50 values between ∼1.5 μM and 40 μM). It was also cytotoxic to HL-29 cells. To further examine its effect on normal cells, peripheral blood lymphocytes collected from healthy volunteers were incubated with the compound. It has also been incubated with human fibroblasts cultured from bone marrow (JMA). Chalcone A1 is non-cytotoxic to PBL cells and to JMA cells. A1 caused significant cell cycle arrest in all phases according to the cell line, and increased the proportion of cells in the sub G0/G1 phase. To evaluate whether this chalcone induced cell death via an apoptotic or necrotic pathway, cell morphology was examined using fluorescence microscopy. Cells treated with A1 at IC50 demonstrated the morphological characteristic of apoptosis, such as chromatin condensation and formation of apoptotic bodies. Apoptosis was confirmed by externalization of phosphatidylserine, which was detected by the Annexin V-FITC method, and by DNA fragmentation. The results suggest that chalcone A1 has potential as a new lead compound for cancer therapy.

Multidrug resistance (MDR) is a multifactorial phenomenon and the role of these proteins in generating the MDR phenotype is controversial. With this in mind, this review compiled the current data on the role of ABCB1, ABCC1, and LRP proteins in the prognosis of hematologic neoplasms and their influence on the choice of therapy. Literature showed that the detection of these proteins, mainly ABCB1, is important in the AL prognosis. However, there is controversy regarding the methodology used for their detection. In summary, the expression and activity profiles of ABCB1, ABCC1, and LRP, proteins capable of promoting the efflux of a variety of chemotherapeutic agents from the cell cytoplasm represent one of the greatest causes of failure in AL treatment.

Since complement system has been recently associated with metabolic and cardiovascular diseases, and closely related to insulin resistance, we investigated the association of plasma complement factor 3 (C3) and factor 4 (C4) with insulin sensibility and weight loss after bariatric surgery.Serum levels of C3, C4, total cholesterol, triacylglycerol, HDL-cholesterol, LDL-cholesterol and homeostatic model assessment (HOMA-IR) measurements were assessed in morbidly obese patients before and after bariatric surgery, including a 6-month follow-up period, as well as a comparison with a lean group.Weight loss decreased body mass index (BMI), serum triacylglycerol, and increased serum HDL-cholesterol and insulin sensitivity, as expected. C3 and C4 were significantly higher in obese individuals when compared to lean subjects (p0.001). In addition, C3 and C4 positively correlated with BMI and HOMA-IR, however, only C3 were significantly decreased 6months after surgery.C3 was strongly associated with insulin sensitivity after bariatric surgery.

We have previously reported the cytotoxic effects of chalcone A1, derived from 1-naphthaldehyde, in leukemia cell lines. On the basis of these findings, the main aim of this study was to elucidate some of the molecular mechanisms involved in apoptosis induced by chalcone A1 toward K562 and Jurkat cells. In both cell lines, chalcone A1 decreased the mitochondrial membrane potential, increased the expression of Bax proapoptotic protein, and decreased the expression of Bcl-2 antiapoptotic protein (resulting in the inversion of the Bcl-2/Bax ratio), which indicates the involvement of the intrinsic pathway. In addition, chalcone A1 increased the expression of FasR in Jurkat cells, which also indicates the involvement of the extrinsic pathway in this cell line. The results also showed an increased expression of effector caspase-3 and cleaved PARP-1 and a decreased expression of IAP protein survivin, which are consistent with apoptotic cell death. The decreased expression of Ki67 suggests that the mechanism involved in cell death induced by chalcone A1 also involves a decrease in cell proliferation. In ex-vivo experiments, chalcone A1 reduced the cell viability of blast cells collected from eight patients with different types of acute leukemia, confirming the cytotoxicity results found in vitro. The results obtained so far are very promising and further studies need to be carried out so that chalcone A1 can be used as a prototype for the development of new antileukemia agents.

ObjectivesThe aim of the present study was to assess the expression profile of multidrug resistance (MDR)-related proteins ABCB1, ABCC1 and LRP in 46 patients with acute leukemia (AL).MethodsThe levels of MDR gene mRNA expression and protein expression at diagnosis were analyzed by semi-quantitative PCR and flow cytometry, respectively.ResultsIn the adult group, higher expression levels of abcc1 gene were associated with older age and lower levels of lactate dehydrogenase (LDH). In the pediatric group, abcc1 gene expression levels were associated with higher CD34 expression and a higher ABCB1 protein expression was correlated with high WBC counts.Discussion/ConclusionThe present data indicate that abcb1 gene overexpression may be associated with a poor prognosis in adults with AL and that ABCB1 and abcc1 expression correlates with different prognostic factors in pediatric patients with AL. Our findings demonstrate that the method of choice to evaluate chemotherapy resistance-related proteins is a ...

Objectives New compounds with biological targets and less cytotoxicity to normal cells are necessary for cancer therapy. In this work ten synthetic chalcones derived from 2-naphtaldehyde were evaluated for their cytotoxic effect in murine acute lymphoblastic leukemia cells L-1210. Methods A series of ten chalcones derived from 2-naphtaldehyde and corresponding acetophenones were prepared by aldolic condensation, using methanol as solvent under basic conditions, at room temperature for 24 h. The cell viability was determined by MTT colorimeter method. The cell cycle phase analysis was carried out by flow cytometry after propidium iodide staining. The apoptosis induction was assessed by exposure to phosphatidylserine (ANNEXIN V-FITC). Cytometric analysis was performed to evaluate the expression of p53, Bcl-2 and Bax protein. The caspase-3 expression was studied by immunoblotting analysis. Key findings A preliminary screening of a series of ten chalcones derived from 2-naphtaldehyde showed that chalcone 8, (2E)-3-(2-naphtyl)-1-(3′-methoxy-4′-hydroxy-phenyl)-2-propen-1-one, had the highest cytotoxic effect (IC50 of 54 µm), but not in normal human lymphocytes. To better understand the cytotoxic mechanism of chalcone 8, its effect on cell cycle and apoptosis was assessed. Our results showed that chalcone 8 caused cell cycle arrest in the G2/M phase and a significant increase in the proportion of cells in the subG0/G1 phase. Our results also demonstrated that chalcone 8 promoted a modification in Bax : Bcl-2 ratio and increased p53 expression and caspase-3 activation. Conclusions The studied chalcone 8 has cytotoxic effect against L-1210 lymphoblastic leukaemic cells, and this effect is associated with increase of p-53 and Bax expression.

Croton celtidifolius Baill is a tree found in the Atlantic Forest South of Brazil, mainly in Santa Catarina. The bark and leaf infusions of this medicinal plant have been popularly used for the treatment of inflammatory diseases. The anti-aggregant activity of C. celtidifolius crude extract (CE) and the column chromatography (CC) isolated compounds flavonoids, catechin and gallocatechin were evaluated in human blood platelets. The platelet-rich plasma (PRP) was incubated with different concentrations of flavonóides (50 - 200 µg/mL) to be tested before platelet aggregation was induced by the agonists adenosine 5'diphosphate (ADP) and collagen. At 200 µg/mL the CE, catechin and gallocatechin markedly inhibited platelet aggregation with the aggregant agents. Using ATP production as an index of platelet secretory capacity, we observed a decreased production of ATP in platelets treated with flavonoids when stimulated by collagen. On the other hand, the flavonoids did not promote inhibitory effect on prothrombin time (PT), thromboplastin time (APTT) and thrombin time (TT). In conclusion, these observations suggest that C. celtidifolius is likely to exert an inhibitory action on platelets in vitro by suppressing secretion and platelet aggregation.Croton celtidifolius Baill é uma árvore encontrada na Mata Atlântica, no sul do Brasil, principalmente no estado de Santa Catarina. A infusão da casca e folhas dessa planta medicinal é utilizada na medicina popular para o tratamento de doenças inflamatórias. A atividade antiagregante do extrato bruto de C. celtidifolius (CE) e de seus flavonóides isolados por coluna cromatográfica (CC), catequina e galocatequina, foi avaliada em plaquetas humanas. O plasma rico em plaquetas (PRP) foi incubado com diferentes concentrações dos flavonóides testados (50 - 200 µg/mL) e posteriormente a agregação foi induzida pelos agonistas adenosina 5'difosfato (ADP) e colágeno. Na concentração de 200 µg/mL o CE, a catequina e a galocatequina inibiram a agregação plaquetária induzida pelos agonistas. A produção de ATP foi utilizada como um índice de capacidade de secreção plaquetária e observamos uma diminuição na produção de ATP nas plaquetas tratadas com os flavonóides e estimuladas com o colágeno. Por outro lado, os flavonóides não promoveram um efeito inibitório no tempo de protrombina (PT), tempo de tromboplastina parcial ativada (APTT) e tempo de trombina (TT). Essas observações sugerem que o C. celtidifolius exerce, in vitro, uma ação inibitória nas plaquetas através da inibição da secreção e agregação plaquetária.

NO (nitric oxide) donating drugs have been investigated for their important role in the sensitization of neoplastic cells to chemotherapy drugs. The goal of this work was to investigate the involvement of NO in the resistance of K562 cells to DNR (daunorubicin). Only simultaneous addition of DNR and SNAP (S-nitroso-N-acetyl-dl-penicillamine) caused significant cell death by apoptosis. Combination of the compounds decreased Bcl-2 and survivin, and increased Bax and active-caspase 3 expression. Fluorescence microscope and cytometric analysis showed that DNR and SNAP together caused DNR intracellular accumulation in K562 cells. RT-PCR (reverse transcription-PCR) analysis showed that DNR and SNAP, alone or in association, produced significant decreases in lrp expression. abcc1 gene expression was unaffected by the presence of SNAP, but when treated with DNR there was a small reduction that was intensified by DNR and SNAP in combination. The transport mechanism involved in the resistance to DNR in K562 cells involves ABCC1 and LRP (lung resistance protein) resistance proteins. DNR and SNAP inhibition of the expression of these proteins occurs by distinct mechanisms, and this disrupts the K562 resistance to DNR.

As síndromes mielodisplásicas (SMDs) são caracterizadas por uma desordem clonal de células primordiais (stem cell) e hematopoese ineficaz que levam à displasia de uma ou mais linhagens celulares da medula óssea, citopenias periféricas e instabilidade genética, as quais aumentam o risco de transformação à leucemia mieloide aguda. Esse grupo heterogêneo de doenças hematopoéticas pode surgir como doença primária, que possui etiologia variada e não completamente definida, ou secundária ao tratamento quimioterápico ou radioterápico para outras neoplasias. O surgimento e aprimoramento de tecnologias de diagnóstico geraram uma melhor compreensão dos processos envolvidos na gênese e evolução das SMDs, o que possibilitou o desenvolvimento de marcadores de diagnóstico e acompanhamentos cada vez mais precoces e específicos. No ano de 2008, a Organização Mundial da Saúde (OMS) redefiniu os critérios para classificação das SMDs, dividindo-as em sete subgrupos. Nessa classificação foram incluídos novos aspectos imunofenotípicos, genéticos, citomorfológicos e moleculares, o que tornou o domínio e o acesso a tecnologias de ponta imprescindíveis para a realização do diagnóstico das SMDs. Apesar dos avanços tecnológicos, alguns pontos, como as bases moleculares da transformação de SMD para LMA, ainda não estão bem esclarecidos, fazendo necessária a continuação de estudos nessa área. Diante disso, essa revisão busca compilar dados atuais dos aspectos moleculares e laboratoriais das SMDs. Myelodysplastic syndromes (MDSs) are characterized by a stem cell clonal disorder and ineffective hematopoiesis which causes dysplasia in one or more bone marrow hematopoietic cell lineages, peripheral cytopenia and genetic instability with enhanced risk to transform into acute myeloid leukemia (AML). This heterogeneous group of hematopoietic diseases can develop as primary diseases, which posses a variable and not completely defined etiology, or as secondary to chemotherapy or radiotherapy for other neoplasias. The evolution of diagnostic tests has improved comprehension of the process involved in the genesis and evolution of MDSs, making the development of earlier and more specific tests for diagnosis and follow ups possible. In 2008, the World Health Organization (WHO) redefined the criteria for the classification of MDSs, dividing them into seven subgroups. This classification included new immunophenotypic, genetic, cytomorphologic and molecular features, which are essential for the diagnosis of MDSs and for a better comprehension of the disease. Despite technological advances, some details, such as the molecular basis of the transformation of MDS to AML, are still not completely understood, which makes further studies in this field necessary. Hence, the objective of this review is to make a compilation of recent molecular and laboratory aspects of MDS.

Croton celtidifolius Baill is a tree found in the Atlantic Forest South of Brazil, mainly in Santa Catarina. The bark and leaf infusions of this medicinal plant have been popularly used for the treatment of inflammatory diseases. The anti-aggregant activity of C. celtidifolius crude extract (CE) and the column chromatography (CC) isolated compounds flavonoids, catechin and gallocatechin were evaluated in human blood platelets. The platelet-rich plasma (PRP) was incubated with different concentrations of flavonóides (50 - 200 µg/mL) to be tested before platelet aggregation was induced by the agonists adenosine 5'diphosphate (ADP) and collagen. At 200 µg/mL the CE, catechin and gallocatechin markedly inhibited platelet aggregation with the aggregant agents. Using ATP production as an index of platelet secretory capacity, we observed a decreased production of ATP in platelets treated with flavonoids when stimulated by collagen. On the other hand, the flavonoids did not promote inhibitory effect on prothrombin time (PT), thromboplastin time (APTT) and thrombin time (TT). In conclusion, these observations suggest that C. celtidifolius is likely to exert an inhibitory action on platelets in vitro by suppressing secretion and platelet aggregation. Croton celtidifolius Baill é uma árvore encontrada na Mata Atlântica, no sul do Brasil, principalmente no estado de Santa Catarina. A infusão da casca e folhas dessa planta medicinal é utilizada na medicina popular para o tratamento de doenças inflamatórias. A atividade antiagregante do extrato bruto de C. celtidifolius (CE) e de seus flavonóides isolados por coluna cromatográfica (CC), catequina e galocatequina, foi avaliada em plaquetas humanas. O plasma rico em plaquetas (PRP) foi incubado com diferentes concentrações dos flavonóides testados (50 - 200 µg/mL) e posteriormente a agregação foi induzida pelos agonistas adenosina 5'difosfato (ADP) e colágeno. Na concentração de 200 µg/mL o CE, a catequina e a galocatequina inibiram a agregação plaquetária induzida pelos agonistas. A produção de ATP foi utilizada como um índice de capacidade de secreção plaquetária e observamos uma diminuição na produção de ATP nas plaquetas tratadas com os flavonóides e estimuladas com o colágeno. Por outro lado, os flavonóides não promoveram um efeito inibitório no tempo de protrombina (PT), tempo de tromboplastina parcial ativada (APTT) e tempo de trombina (TT). Essas observações sugerem que o C. celtidifolius exerce, in vitro, uma ação inibitória nas plaquetas através da inibição da secreção e agregação plaquetária.