Daniela Diógenes de Carvalho

Effect of BJcuL (a lectin from the venom of the snake Bothrops jararacussu) on adhesion and growth of tumor and endothelial cells.

Lectins are polyvalent carbohydrate-binding proteins of non-immune origin. Recently, we have isolated and characterized a lectin from the venom of the snake Bothrops jararacussu. This lectin (BJcuL) has been shown to bind to lactose moieties and induce agglutination of erythrocytes. In the present work, we observed that cells from human metastatic breast cancer (MDA-MB-435) and human ovarian carcinoma (OVCAR-5) cell lines adhere, although weakly, to BJcuL. However, BJcuL did not inhibit adhesion of these cells to the extracellular matrix proteins fibronectin, laminin and type I collagen. Importantly, viability of these tumor cells and cells from other human tumor cell lines and a bovine brain endothelial cell line was suppressed by BJcuL. These findings suggest that the lectin BJcuL may serve as an interesting tool for combating tumor progression by inhibiting tumor cell and endothelial cell growth.

Primary Structure Characterization of Bothrops jararacussu Snake Venom Lectin

The complete amino acid sequence of the lectin from Bothrops jararacussu snake venom (BJcuL) is reported. The sequence was determined by Edman degradation and amino acid analysis of the S-carboxymethylated BJcuL derivative (RC-BJcuL) and from its peptides originated from enzymatic digestion. The sequence of amino acid residues showed that this lectin displays the invariant amino acid residues characterized in C-type lectins. Amino acids analysis revealed a high content of acidic amino acids and leucine. These findings suggest that BJcuL, like other snake venom lectins, possesses structural similarities to the carbohydrate recognition domain (CRD) of calcium-dependent animal lectins belonging to the C-type β-galactoside binding lectin family.

Lectins from seeds of Crotalaria pallida (smooth rattlebox)

A lectin from the seeds of Crotalaria pallida (CPL), with an apparent molecular mass of 30 kDa, determined by SDS-polyacrylamide gel electrophoresis, showed human type A and B erythrocytes agglutination activity, which is inhibited by raffinose and galactose. The lectin requirement for divalent cation was demonstrated with EDTA/EGTA blocking hemagglutination activity. Although the N-terminal amino acid sequence of CPL is identical to another lectin from Crotalaria striata, which is taxonomically synonymous to Crotalaria pallida, these lectins differ in amino acid composition and hemagglutination properties.

Isolation and characterization of a new lectin from the venom of the snake Bothrops jararacussu

Bothrops jararacussu venom contains a new potent hemagglutinin (BJcuL), isolated by affinity chromatography on immobilized D‐galactose. Hemagglutination activity of BJcuL against pig erythrocytes was inhibited by galactosides, EDTA or EGTA. The purified lectin is a di sulfide dimer composed of 15 kDa subunits. Association of these subunits is essential for lectin activity, since DTT‐reduced BJcuL showed no hemagglutination activity against pig erythrocytes. Reduced and S‐pyridylethylated BJcuL (RP‐BJcuL) was separated as a single peak by reverse‐phase HPLC on a C‐18/μBondapak column. Results of the application of Edman degradation methodology showed that RP‐BJcuL has a single N‐terminal amino acid sequence, indicating that BJcuL is a homodimer. The N‐terminal 8‐residue (NNCPQDWL) shows homology with other lectins from snake venoms.

Diacerein improves left ventricular remodeling and cardiac function by reducing the inflammatory response after myocardial infarction.

Background The inflammatory response has been implicated in the pathogenesis of left ventricular (LV) remodeling after myocardial infarction (MI). An anthraquinone compound with anti-inflammatory properties, diacerein inhibits the synthesis and activity of pro-inflammatory cytokines, such as tumor necrosis factor and interleukins 1 and 6. The purpose of this study was to investigate the effects of diacerein on ventricular remodeling in vivo. Methods and Results Ligation of the left anterior descending artery was used to induce MI in an experimental rat model. Rats were divided into two groups: a control group that received saline solution (n = 16) and a group that received diacerein (80 mg/kg) daily (n = 10). After 4 weeks, the LV volume, cellular signaling, caspase 3 activity, and nuclear factor kappa B (NF-κB) transcription were compared between the two groups. After 4 weeks, end-diastolic and end-systolic LV volumes were reduced in the treatment group compared to the control group (p < .01 and p < .01, respectively). Compared to control rats, diacerein-treated rats exhibited less fibrosis in the LV (14.65%± 7.27% vs. 22.57%± 8.94%; p < .01), lower levels of caspase-3 activity, and lower levels of NF-κB p65 transcription. Conclusions Treatment with diacerein once a day for 4 weeks after MI improved ventricular remodeling by promoting lower end-systolic and end-diastolic LV volumes. Diacerein also reduced fibrosis in the LV. These effects might be associated with partial blockage of the NF-κB pathway.

Atorvastatin improves ventricular remodeling after myocardial infarction by interfering with collagen metabolism

Purpose Therapeutic strategies that modulate ventricular remodeling can be useful after acute myocardial infarction (MI). In particular, statins may exert effects on molecular pathways involved in collagen metabolism. The aim of this study was to determine whether treatment with atorvastatin for 4 weeks would lead to changes in collagen metabolism and ventricular remodeling in a rat model of MI. Methods Male Wistar rats were used in this study. MI was induced in rats by ligation of the left anterior descending coronary artery (LAD). Animals were randomized into three groups, according to treatment: sham surgery without LAD ligation (sham group, n = 14), LAD ligation followed by 10mg atorvastatin/kg/day for 4 weeks (atorvastatin group, n = 24), or LAD ligation followed by saline solution for 4 weeks (control group, n = 27). After 4 weeks, hemodynamic characteristics were obtained by a pressure-volume catheter. Hearts were removed, and the left ventricles were subjected to histologic analysis of the extents of fibrosis and collagen deposition, as well as the myocyte cross-sectional area. Expression levels of mediators involved in collagen metabolism and inflammation were also assessed. Results End-diastolic volume, fibrotic content, and myocyte cross-sectional area were significantly reduced in the atorvastatin compared to the control group. Atorvastatin modulated expression levels of proteins related to collagen metabolism, including MMP1, TIMP1, COL I, PCPE, and SPARC, in remote infarct regions. Atorvastatin had anti-inflammatory effects, as indicated by lower expression levels of TLR4, IL-1, and NF-kB p50. Conclusion Treatment with atorvastatin for 4 weeks was able to attenuate ventricular dysfunction, fibrosis, and left ventricular hypertrophy after MI in rats, perhaps in part through effects on collagen metabolism and inflammation. Atorvastatin may be useful for limiting ventricular remodeling after myocardial ischemic events.

Lidocaine and Pinacidil Added to Blood versus Crystalloid Cardioplegic Solutions: Study in Isolated Hearts

Objective The present study aimed the functional recovery evaluation after long term of cardiac arrest induced by Custodiol (crystalloid-based) versus del Nido (blood-based) solutions, both added lidocaine and pinacidil as cardioplegic agents. Experiments were performed in isolated rat heart perfusion models. Methods Male rat heart perfusions, according to Langendorff technique, were induced to cause 3 hours of cardiac arrest with a single dose. The hearts were assigned to one of the following three groups: (I) control; (II) Custodiol-LP; and (III) del Nido-LP. They were evaluated after ischemia throughout 90 minutes of reperfusion. Left ventricular contractility function was reported as percentage of recovery, expressed by developed pressure, maximum dP/dt, minimum dP/dt, and rate pressure product variables. In addition, coronary resistance and myocardial injury marker by alpha-fodrin degradation were also evaluated. Results At 90 minutes of reperfusion, both solutions had superior left ventricular contractile recovery function than the control group. Del Nido-LP was superior to Custodiol-LP in maximum dP/dt (46%±8 vs. 67%±7, P<0.05) and minimum dP/dt (31%±4 vs. 51%±9, P<0.05) variables. Coronary resistance was lower in del Nido-LP group than in Custodiol-LP (395%±50 vs. 307%±13, P<0.05), as well as alpha-fodrin degradation, with lower levels in del Nido-LP group (P<0.05). Conclusion Del Nido-LP cardioplegia showed higher functional recovery after 3 hours of ischemia. The analysis of alpha-fodrin degradation showed del Nido-LP solution provided greater protection against myocardial ischemia and reperfusion (IR) in this experimental model.

A Simpler and Shorter Neuromuscular Electrical Stimulation Protocol Improves Functional Status and Modulates Inflammatory Profile in Patients with End-Stage Congestive Heart Failure

Background: Neuromuscular electrical stimulation (NMES) using a stimulation wave for 5 days/week over 8 weeks has been used as a treatment option for congestive heart failure (CHF) patients who are unable to tolerate aerobic exercise. Objective: We assessed the impact of a shorter NMES protocol using a Russian stimulation wave on the functional status, quality of life (QoL) and inflammatory profile of end-stage CHF patients. Methods: Twenty-eight patients with end-stage CHF (53 ± 11 years) were randomized to the NMES or control group. Treatment was an NMES training program with Russian stimulation wave, applied for 50 minutes to both quadriceps femoral muscles twice weekly over seven weeks. The stimulation intensity was chosen to elicit muscle contractions in the NMES group and current input up to sensory threshold in the control group. Distance in the 6-minute walk test (6MWD) and QoL score by the Minnesota Living with Heart Failure Questionnaire were evaluated before, immediately after and one month after NMES protocol completion. Peripheral leukocytes were obtained to measure the gene expression levels of inflammatory cytokines. Results: The NMES group showed increases in the 6MWD (324 ± 117 vs. 445 ± 100 m; p = 0.02) and QoL score (64 ± 22 vs. 45 ± 17; p < 0.01) immediately but not 1 month after protocol completion, as well as increased gene expression levels of IL-1β, IL-6 and IL-8 after protocol completion. Conclusion: Using a shorter and fewer sessions NMES protocol improved the QoL score and functional class of severe CHF patients, and modulated the gene expression levels of some cytokines. This protocol might be a good alternative for patients with severe CHF and limitations in protocol adherence. (Int J Cardiovasc Sci. 2017;30(6)484-495)Mailing Address: Maria Carolina Basso Sacilotto Avenida Arlindo Joaquim de Lemos, 865, Apto: 32. Postal Code: 13100450, Vila Lemos, Campinas, SP – Brazil E-mail: mariacarolinasacilotto@gmail.com; orlandopetrucci@gmail.com A Simpler and Shorter Neuromuscular Electrical Stimulation Protocol Improves Functional Status and Modulates Inflammatory Profile in Patients with End-Stage Congestive Heart Failure Maria Carolina Basso Sacilotto, Carlos Fernando Ramos Lavagnoli, Lindemberg Mota Silveira-Filho, Karlos Alexandre de Souza Vilarinho, Elaine Soraya Barbosa de Oliveira, Daniela Diógenes de Carvalho, Pedro Paulo Martins de Oliveira, Otávio Rizzi Coelho-Filho, Orlando Petrucci Junior