[Show abstract][Hide abstract] ABSTRACT: Millions of blood products are transfused each year, and many lives are directly affected by transfusion. Platelet concentrate (PC) is one of the main products derived from blood. Even under good storage conditions, PC is likely to suffer cell damage. The shape of platelets changes after 5 to 7 days of storage at 22°C. Taking into consideration that some platelet proteins undergo changes in their shape and functionality during PC storage. Sixteen PC bags were collected and each PC bag tube was cut into six equal pieces to perform experiments with platelets from six different days of storage. Thus, on the first day of storage, 1/6 of the tube was used for miRNA extraction, and the remaining 5/6 was stored under the same conditions until extraction of miRNAs on each the following five days. Samples were sequenced on an Illumina Platform to demonstrate the most highly expressed miRNAs. Three miRNAs, mir127, mir191 and mir320a were validated by real-time quantitative PCR (RQ-PCR) in 100 PC bags tubes. Our method suggests, the use of the miRNAs mir127 and mir320a as biomarkers to assess the "validity period" of PC bags stored in blood banks for long periods. Thus, bags can be tested on the 5th day of storage for the relative expression levels of mir127 and mir320a. Thus, we highlight candidate miRNAs as biomarkers of storage damage that can be used as tools to evaluate the quality of stored PC. The use of miRNAs as biomarkers of damage is unprecedented and will contribute to improved quality of blood products for transfusions.
PLoS ONE 06/2015; 10(6):e0129399. DOI:10.1371/journal.pone.0129399 · 3.23 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: With the aim of discovering new anticancer agents, we have designed and synthesized novel 6-hydroxy-benzo[d][1,3]oxathiol-2-one Schiff bases. The synthesis started with the selective nitration at 5-position of 6-hydroxybenzo[d][1,3]oxathiol-2-one (1) leading to the nitro derivative 2. The nitro group of 2 was reduced to give the amino intermediate 3. Schiff bases 4a-r were obtained from coupling reactions between 3 and various benzaldehydes and heteroaromatic aldehydes. All the new compounds were fully identified and characterized by NMR (1H and 13C) and specifically for 4q by X-ray crystallography. The in vitro cytotoxicity of the compounds was evaluated against cancer cell lines (ACP-03, SKMEL-19 and HCT-116) by using MTT assay. Schiff bases 4b and 4o exhibited promising cytotoxicity against ACP-03 and SKMEL-19, respectively, with IC50 values lower than 5 μM. This class of compounds can be considered as a good starting point for the development of new lead molecules in the fight against cancer.
[Show abstract][Hide abstract] ABSTRACT: Aim:
To investigate frequent quantitative alterations of intestinal-type gastric adenocarcinoma.
Materials and methods:
We analyzed genome-wide DNA copy numbers of 22 samples and using CytoScan® HD Array.
We identified 22 gene alterations that to the best of our knowledge have not been described for gastric cancer, including of v-erb-b2 avian erythroblastic leukemia viral oncogene homolog 4 (ERBB4), SRY (sex determining region Y)-box 6 (SOX6), regulator of telomere elongation helicase 1 (RTEL1) and UDP-Gal:betaGlcNAc beta 1,4- galactosyltransferase, polypeptide 5 (B4GALT5). The most significant alterations related to peritoneal invasion involved the regions 13q21.1 (gain) and 15q15.1, 17q23.1, 19q13.2 and 20q11.22 (loss of heterozygozity; LOH), where we found LOH of erythrocyte membrane protein band 4.1-like 1 (EPB41L1) gene. In relation to early age of onset, the most significant alterations were gains in the regions Xq26 and Xp22.31 and a loss in the region 11p15.4.
These quantitative changes may play a role in the development of this type of neoplasia and may be used as markers in evaluating poor prognosis, as well as act as potential therapeutic targets for gastric cancer.
Anticancer research 11/2014; 34(11):6405-6415. · 1.83 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Background
Helicobacter pylori (HP) and Epstein-Barr virus (EBV) have been associated with cancer development. We evaluated the prevalence of HP, HP CagA+ and EBV infection in gastric cancer (GC) samples from adults and in gastric tissues from patients who underwent upper endoscopy (UE).
Samples from UE and GC were collected to investigate the presence of HP infection and the HP virulence factor CagA by a urease test and PCR. The presence of EBV was detected by Eber-1 in situ hybridization.
In UE, 85.5% of juvenile patients showed some degree of gastritis (45.3% of patients with mild gastritis and 54.7% with moderate/severe gastritis) and patients with mild gastritis were younger than patients with moderate/severe gastritis. Among adults, 48.7% presented mild gastritis and 51.3% moderate/severe gastritis. HP infection was detected in 0% of normal mucosa, 58.5% of juvenile gastritis patients, 69.2% of adult gastritis patients and 88% of GC patients. In these same groups, HP CagA+ was detected in 0%, 37.7%, 61.5% and 67.2% of tissue samples, respectively. In juvenile patients, HP infection was more common in those with gastritis than in normal samples (p = 0.004). The patients with either HP or HP CagA+ were older than patients without these pathogens (p < 0.05). In juvenile patients, HP infection was more frequent in cases of moderate/severe gastritis than in cases of mild gastritis (p = 0.026). Moreover, in patients with GC, HP infection was more frequent in males than in females (p = 0.023). GC patients with HP CagA+ were older than patients with HP CagA- (p = 0.027). HP CagA+ was more common in intestinal-type than diffuse-type GC (p = 0.012). HP CagA+ was also associated with lymph-node (p = 0.024) and distal (p = 0.005) metastasis. No association between EBV infection and HP infection or any clinicopathological variable was detected.
Our results suggest that HP is involved in the pathophysiology of severe gastric lesions and in the development of GC, particularly when CagA+ is present. EBV was not the primary pathogenic factor in our samples.
[Show abstract][Hide abstract] ABSTRACT: The immune response modifier Canova® is a homeopathic remedy indicated for patients with depressed immune system, since this drug appears to increase adaptive immunity and induce an immune response against multiple and severe pathological conditions, including cancer. We evaluated the pattern of immune cellular response in non-human primates of the species Cebus apella exposed to N-methyl-N-nitrosourea (MNU) with and without Canova®. Twelve animals were divided into four groups, with three animals each: negative control and three experimental groups, MNU-alone (35 days); MNU (35 days)-plus-Canova® (3 days) and Canova®-alone (3 days). The animals received MNU orally and Canova® by three intravenous injections. Evaluation of the cellular immune response was performed by immunophenotyping of T-lymphocytes (CD4(+), CD8(+)), B-lymphocytes and natural killer cells. Analysis was also performed of the cell cycle. Our results suggest an increase of T-lymphocytes (CD4(+)CD3(+)) only in the Canova® group, while in the MNU-plus-Canova® group only B-lymphocytes increased.
In vivo (Athens, Greece) 09/2014; 28(5):837-841. · 0.97 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Background
Hereditary diffuse gastric cancer (HDGC) is a hereditary autosomal inherited syndrome associated with CDH1 germline mutations. In Brazil, gastrointestinal tumors are among the most prevalent tumor types and constitute a serious public health problem, especially in the northern and northeastern regions. This study aimed to investigate germline mutations, methylation pattern and genomic rearrangements in the CDH1 gene and quantitative changes in the DNA of HDGC patients in northern and northeastern Brazil.
Twenty-seven DNA samples from the members of four families affected by HDGC were analyzed using array comparative genomic hybridization (aCGH), DNA sequencing and methylation pattern.
No evidence of gain and loss events or any rearrangements were found in any of the samples tested using aCGH. No promoter region hypermethylation was observed either. Two of the four families presented different types of germline mutations. The 185G > T and 1018A > G germline mutations detected in this study have been described in Asian and European families, respectively. The ancestors of the two families carrying these mutations had originated from those continents.
This is the first study to evaluate CDH1 gene germline mutations in Brazilian families with HDGC. In our study, 50% of the families showed no CDH1 gene alterations, and it is possible that in regions with a high incidence of gastric cancer, such as northern and northeastern Brazil, environmental factors might have induced the different genetic alterations analyzed in this study.
Hereditary Cancer in Clinical Practice 08/2014; 12(1):18. DOI:10.1186/1897-4287-12-18 · 1.47 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Benzothiazole hydrazones have been synthesized and evaluated for their in vitro antiproliferative activity against three human cancer cell lines: HL-60 (leukemia), MDAMB-435 (breast) and HCT-8 (colon). The good cytotoxicity for the three cancer cell lines and theoretical profile of compounds 3o and 3p pointed them as promising lead molecules for anticancer drug design.
European Journal of Medicinal Chemistry 08/2014; 86C:12-16. DOI:10.1016/j.ejmech.2014.08.039 · 3.45 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: The hydrodistilled oils of Piper hispidum, Piper aleyreanum, and Piper anonifolium, collected in the Carajás National Forest, Pará state, Brazil, were analyzed by GC and GC-MS and then, evaluated their antioxidant, antifungal, anticholinesterase and cytotoxic activities. In total, 87 constituents were identified in the Piper oils. The sesquiterpenes were the most highly represented classes and the main compounds found in the Piper oils were selin-11-en-4-α-ol, β-elemene, β-selinene, α-selinene, bicyclogermacrene, β-caryophyllene, α-humulene, and δ-elemene. All analyzed oils showed powerful antifungal activity, with the detection limit (DL) from 0.1 to 1.0 μg against the Cladosporium cladosporioides and Cladosporium sphareospermum fungi, as well they have no lytic effect against the mice erythrocytes. In the anticholinesterase evaluation, the oils of P. anonifolium (DL = 0.01 ng) and P. hispidum (DL = 0.01 ng) were hundred times more potent than the standard physostigmine (DL = 1.0 ng). Moreover, the oil of P. aleyreanum showed high in vitro cytotoxic activity against the human melanoma cell line SKMEL-19 (IC50 = 7.4 μg/mL) and significant antioxidant activity (DPPH = 412.2 mg TE/mL). The higher cell growth inhibition induced by the oil of P. aleyreanum is probably due to elemene (β-, δ-, and γ-elemene), as well as other structurally related compounds, which were previously reported in the proliferation inhibition, stimulation of apoptosis and induction of cell cycle arrest in malignant cells.
[Show abstract][Hide abstract] ABSTRACT: As part of a continuing search for new potential anticancer candidates, we describe the synthesis, cytotoxicity and mechanistic evaluation of a series of 4-oxoquinoline-3-carboxamide derivatives as novel anticancer agents. The inhibitory activity of compounds 10–18 was determined against three cancer cell lines using the MTT colorimetric assay. The screening revealed that derivatives 16b and 17b exhibited significant cytotoxic activity against the gastric cancer cell line but was not active against a normal cell line, in contrast to doxorubicin, a standard chemotherapeutic drug in clinical use. Interestingly, no hemolytical activity was observed when the toxicity of 16b and 17b was tested against blood cells. The in silico and in vitro mechanistic evaluation indicated OPEN ACCESS Molecules 2014, 19 6652 the potential of 16b as a lead for the development of novel anticancer agents against gastric cancer cells.
[Show abstract][Hide abstract] ABSTRACT: Oral bacteria have been detected in atherosclerotic plaques at a variable frequency; however, the connection between oral health and vascular and oral bacterial profiles of patients with vascular disease is not clearly established. The aim of this study was to evaluate the presence of oral bacterial DNA in the mouth and atherosclerotic plaques, in addition to assessing the patients' caries and periodontal disease history.
Thirty samples of supragingival and subgingival plaque, saliva and atherosclerotic plaques of 13 patients with carotid stenosis or aortic aneurysm were evaluated, through real-time polymerase chain reaction, for the presence of Streptococcus mutans (SM), Prevotella intermedia (PI), Porphyromonas gingivalis (PG) and Treponema denticola (TD). All patients were submitted to oral examination using the DMFT (decayed, missing and filled teeth) and PSR (Periodontal Screening and Recording) indexes. Histopathological analysis of the atherosclerotic plaques was performed.
Most of the patients were edentulous (76.9%). SM, PI, PG and TD were detected in 100.0%, 92.0%, 15.3% and 30.7% of the oral samples, respectively. SM was the most prevalent targeted bacteria in atherosclerotic plaques, detected in 100% of the samples, followed by PI (7.1%). The vascular samples were negative for PG and TD. There was a statistically significant difference (p<0.05) between the presence of PG and TD in the oral cavity and vascular samples.
SM was found at a high frequency in oral and vascular samples, even in edentulous patients, and its presence in atherosclerotic plaques suggests the possible involvement of this bacterium in the disease progression.
International journal of cardiology 04/2014; 174(3). DOI:10.1016/j.ijcard.2014.04.201 · 4.04 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: We herein report the antitumor activity of several substituted alpha- and beta-dihydrofuran naphthoquinones against 4 human tumor cell lines, HL-60 (leukemia), SF-295 (CNS), HCT-8 (colon) and MDA-MB435 (melanoma), and their electrochemical parameters, in the absence and presence of oxygen, in comparison with their non-substituted precursors. These compounds were prepared from readily available lawsone and olefins in the presence of cerium (IV) ammonium nitrate. The beta-dihydrofuran naphthoquinones were shown to be highly cytotoxic, while their positional alpha-isomers were considered less active. The level of intracellular ROS release and the first wave redox potentials were also analyzed and compared with the kinetic constants of the reactivity of quinones with oxygen (k(app)) obtained through cyclic voltammetry. Significantly positive correlations between ROS release and oxygen reactivity were obtained, while IC50 vs. ROS release; -E-plc vs. k(app) or ROS values correlated in an inverse manner, i.e., the less negative the potential, higher the activities. These findings reinforce the effectiveness of the combination of pharmacology and electrochemistry in medicinal chemistry, in the search of lead anticancer compounds.