Adverse conditions in the life of children with developmental retardation

O objetivo deste estudo foi o de identificar os fatores de risco a que foram expostas as crianças portadoras de atraso no desenvolvimento, encaminhadas ao Centro de Psicologia Aplicada da FFCLRP-USP, nos últimos cinco anos. O grupo ficou constituído por doze meninos e dez meninas, cujas idades variavam de um mês a oito anos. Os dados foram extraídos das entrevistas realizadas com as mães, no momento da inscrição da criança junto ao serviço. Os problemas referidos pelas mães, na queixa, bem como os eventos e as circunstâncias adversas relatados, na história de vida da criança, foram classificados de acordo com os Sistemas de Categorias elaborados pelas autoras desse estudo. Os resultados mostram que, além do atraso, aparecem referidos na queixa, com maior freqüência, os problemas de fala, coordenação motora, aprendizagem e agitação/inquietude. A análise das circunstâncias adversas, presentes na história de vida desse grupo de crianças, evidenciou que todas elas foram expostas a uma ou mais das condições de risco biológico, sendo as mais freqüentes: problemas congênitos, problemas de saúde física da mãe durante a gestação, problemas de saúde física da criança, crises convulsivas, complicações no parto e hospitalização. Além disso, observou-se que 36% das crianças havia sido exposta a fatores de risco ambiental, especialmente aqueles ligados a atitudes e práticas de cuidado e educação inadequadas. Com base nos resultados, discutem-se as implicações da presença do risco biológico como condição importante para o acesso da criança e da família a serviços especializados.The objective of the present study was to identify the risk factors to which children with delayed development referred to the Center of Applied Psychology of FFCLRP-USP over the last five years had been exposed. The group consisted of 12 boys and 10 girls ranging in age from 1 month to 8 years. The data were extracted from the interviews held with the mothers as the time of child enrollment in the service. The problems reported by de mothers on the occasion of the complaint and the adverse events and circumstances that ocurred in the child’s life were classified according to the Category System elaborated by the authors of the present study. The results showed that, in addition to retardation, the most frequent problems reported on the occasion of the complaint were speech, motor coordination, learning and agitation/restlessness problems. Analysis of the adverse circumstances in the life history of this group of children demonstrated that all of them had been exposed to one or more biological risk conditions, the most frequent being congenital problems, mother’s physical health problems during pregnancy, child’s physical health problems, convulsive seizures, complications of delivery, and hospitalization. Furthermore, 36% of the children had been exposed to environmental risk factors, especially those linked to inadequate attitudes and practices of care and education. On the basis of the results obtained, we discuss the implications of the presence of biological risks as an important condition for access to specialized services on the part of the children and their families

Palladium(II) complexes with thiosemicarbazones: syntheses, characterization and cytotoxicity against breast cancer cells and anti-mycobacterium tuberculosis activity

Three PdII complexes were prepared from N(4)-substituted thiosemicarbazones: [Pd(aptsc)(PPh3)](NO3)•H2O, 1, [Pd(apmtsc)(PPh3)](NO3), 2, and [Pd(apptsc)(PPh3)](NO3)•H2O, 3, where PPh3 = triphenylphosphine; Haptsc = 2-acetylpyridine-thiosemicarbazone; Hapmtsc = 2-acetylpyridine-N(4)-methyl-thiosemicarbazone and Happtsc = 2-acetylpyridine-N(4)-phenyl-thiosemicarbazone. All complexes were characterized by elemental analysis, IR, UV-Vis, 1H and 31P{1H} NMR spectroscopies, and had their crystalline structures determined by X-ray diffractometry from single crystals. The monoanionic thiosemicarbazonate ligands act in a tridentate mode, binding to the metal through the pyridine nitrogen, the azomethine nitrogen and the sulfur atoms. The cytotoxic activity against the breast cancer cell line MDA-MB231 and the anti-Mycobacterium tuberculosis H37Rv ATCC 27294 activity were evaluated for the compounds. All PdII complexes were highly active against the studied cell line, presenting similar values of IC50, around 5 µmol L-1, while the clinically applied antitumor agent cisplatin was inactive. The compounds show remarkable anti-M. tuberculosis activities, presenting MIC values comparable or better than some commercial anti-M tuberculosis drugs.FAPESPCAPESCNPqFINE

Palladium(II) complexes with thiosemicarbazones: syntheses, characterization and cytotoxicity against breast cancer cells and Anti-Mycobacterium tuberculosis activity

Três complexos de PdII com tiossemicarbazonas N(4)-substituídas foram preparados: [Pd(aptsc)(PPh3)](NO3) H2O, 1, [Pd(apmtsc)(PPh3)](NO3), 2, e [Pd(apptsc)(PPh3)](NO3) H2O, 3, sendo PPh3 = trifenilfosfina; Haptsc = 2-acetilpyridina-tiossemicarbazona; Hapmtsc = 2-acetilpiridina-N(4)-metil-tiossemicarbazona e Happtsc = 2-acetilpiridina-N(4)-fenil-tiossemicarbazona. Os complexos foram caracterizados por análise elementar, IR, UV-Vis, ¹H e 31P{¹H} NMR e tiveram suas estruturas cristalinas determinadas por difratometria de raios X em monocristal. Os ligantes tiossemicarbazonatos monoaniônicos atuam de modo tridentado, ligando-se ao metal pelos átomos de nitrogênio piridínico, nitrogênio azometínico e enxofre. A atividade citotóxica frente à linhagem de células tumorais MDA-MB231 (tumor de mama) e a atividade anti-Mycobacterium tuberculosis H37Rv ATCC 27294 dos compostos foram investigadas. Os complexos de PdII mostraram-se altamente ativos contra as células tumorais, com valores de IC50 em torno de 5 µmol L-1, enquanto o agente antitumoral em uso clínico cisplatina mostrou-se inativo. Os compostos apresentaram atividade anti-M. tuberculosis significante, com valores de CIM comparáveis ou melhores que aqueles referentes a alguns fármacos usados clinicamente contra tuberculose.Three PdII complexes were prepared from N(4)-substituted thiosemicarbazones: [Pd(aptsc)(PPh3)](NO3) H2O, 1, [Pd(apmtsc)(PPh3)](NO3), 2, and [Pd(apptsc)(PPh3)](NO3) H2O, 3, where PPh3 = triphenylphosphine; Haptsc = 2-acetylpyridine-thiosemicarbazone; Hapmtsc = 2-acetylpyridine-N(4)-methyl-thiosemicarbazone and Happtsc = 2-acetylpyridine-N(4)-phenyl-thiosemicarbazone. All complexes were characterized by elemental analysis, IR, UV-Vis, ¹H and 31P{¹H} NMR spectroscopies, and had their crystalline structures determined by X-ray diffractometry from single crystals. The monoanionic thiosemicarbazonate ligands act in a tridentate mode, binding to the metal through the pyridine nitrogen, the azomethine nitrogen and the sulfur atoms. The cytotoxic activity against the breast cancer cell line MDA-MB231 and the anti-Mycobacterium tuberculosis H37Rv ATCC 27294 activity were evaluated for the compounds. All PdII complexes were highly active against the studied cell line, presenting similar values of IC50, around 5 µmol L-1, while the clinically applied antitumor agent cisplatin was inactive. The compounds show remarkable anti-M. tuberculosis activities, presenting MIC values comparable or better than some commercial anti-M tuberculosis drugs

Synthesis, Characterization, Cytotoxic Activity, and Interactions with CT-DNA and BSA of Cationic Ruthenium(II) Complexes Containing Dppm and Quinoline Carboxylates

The complexes cis-[Ru(quin)(dppm)2]PF6 and cis-[Ru(kynu)(dppm)2]PF6 (quin = quinaldate; kynu = kynurenate; dppm = bis(diphenylphosphino)methane) were prepared and characterized by elemental analysis, electronic, FTIR, 1H, and 31P{H1} NMR spectroscopies. Characterization data were consistent with a cis arrangement for the dppm ligands and a bidentate coordination through carboxylate oxygens of the quin and kynu anions. These complexes were not able to intercalate CT-DNA as shown by circular dichroism spectroscopy. On the other hand, bovine serum albumin (BSA) binding constants and thermodynamic parameters suggest spontaneous interactions with this protein by hydrogen bonds and van der Waals forces. Cytotoxicity assays were carried out on a panel of human cancer cell lines including HepG2, MCF-7, and MO59J and one normal cell line GM07492A. In general, the new ruthenium(II) complexes displayed a moderate to high cytotoxicity in all the assayed cell lines with IC50 ranging from 10.1 to 36 µM and were more cytotoxic than the precursor cis-[RuCl2(dppm)2]. The cis-[Ru(quin)(dppm)2]PF6 were two to three times more active than the reference metallodrug cisplatin in the MCF-7 and MO59J cell lines

Cytotoxicity and anti-tumor effects of new ruthenium complexes on triple negative breast cancer cells

<div><p>Triple-negative breast cancer (TNBC) is a highly aggressive breast cancer subtype. The high rate of metastasis associated to the fact that these cells frequently display multidrug resistance, make the treatment of metastatic disease difficult. Development of antitumor metal-based drugs was started with the discovery of cisplatin, however, the severe side effects represent a limitation for its clinical use. Ruthenium (Ru) complexes with different ligands have been successfully studied as prospective antitumor drugs. In this work, we demonstrated the activity of a series of biphosphine bipyridine Ru complexes <b>(1)</b> [Ru(SO₄)(dppb)(bipy)], <b>(2)</b> [Ru(CO₃)(dppb)(bipy)], <b>(3)</b> [Ru(C₂O₄)(dppb)(bipy)] and <b>(4)</b> [Ru(CH₃CO₂)(dppb)(bipy)]PF₆ [where dppb = 1,4-bis(diphenylphosphino)butane and bipy = 2,2’-bipyridine], on proliferation of TNBC (MDA-MB-231), estrogen-dependent breast tumor cells (MCF-7) and a non-tumor breast cell line (MCF-10A). Complex <b>(4)</b> was most effective among the complexes and was selected to be further investigated on effects on tumor cell adhesion, migration, invasion and in apoptosis. Moreover, DNA and HSA binding properties of this complex were also investigated. Results show that complex <b>(4)</b> was more efficient inhibiting proliferation of MDA-MB-231 cells over non-tumor cells. In addition, complex <b>(4)</b> was able to inhibit MDA-MB231 cells adhesion, migration and invasion and to induce apoptosis and inhibit MMP-9 secretion in TNBC cells. Complex <b>(4)</b> should be further investigated <i>in vivo</i> in order to stablish its potential to improve breast cancer treatment.</p></div

Effect of complex 4 on MDA-MB-231, MCF-7 and MCF-10A cells.

<p><b>(A)</b> Cell morphology was examined after 2 h and 24 h of treatment. <b>(B)</b> Clonogenic assay of untreated MDA-MB-231 cells (control) or cells treated with complex <b>(4)</b>. A photograph of a representative experiment is shown along with graph quantifications of colony number and size. Significant at the *<i>p</i><0.005, **<i>p</i><0.001 levels using ANOVA and Bonferroni tests.</p

Interactions with DNA and HSA.

<p>Binding constants for the interaction between Ru(II) complexes <b>(1–4)</b> and calf thymus <i>ct</i>-DNA and stern-Volmer quenching constant (K_sv, M^-1), biomolecular quenching rate constant (Kq, M^-1s^-1), binding constant (Kb, M^-1), the number of binding sites (n), ΔG(KJ.mol^-1), ΔH (KJ.mol^-1) and ΔS (J.mol^-1K) values for the complex-HSA system at different temperatures.</p

Absorption spectral titration.

<p><b>(A)</b> Absorption spectral titration of complex <b>(4)</b> in the presence of increasing concentrations of <i>ct</i>-DNA at 298 K and fluorescence emission spectra of HSA at 37°C. Inset graph represents the plots of (ε_a-ε_f)/(ε_b-ε_f) versus [DNA] for the titration of DNA with Ru(II) complexes. <b>(B)</b> CD spectrum of <i>ct</i>-DNA (100 μM) in the presence of complexes <b>(1–4)</b> in Tris-HCl buffer after incubating by 18 h at 37°C. <b>(C)</b> Electrophoretic mobility pattern of pBR322 plasmid DNA incubated with metal complexes <b>(1–4)</b> by 18 h at 37°C, at indicated concentrations (μM). Ri = ratio complex/DNA; MM = molecular marker. <b>(D)</b> Fluorescence quenching spectra of HSA with different concentrations of complex (4) with the excitation wavelength at 270 nm at 37°C in a Trizma buffer, pH 7.4. The arrow shows the intensity changes upon increasing the concentration of the quencher (0 to 50 μM, orange line to light blue line, respectively).</p

Effects of complex (4) on MDA-MB-231 cell adhesion and cytoskeleton structure.

<p><b>(A)</b> Type I collagen, fibronectin, laminin and vitronectin were coated on the wells of a 96-well plate. Complex <b>(4)</b> was incubated with MDA-MB-231 cells, which were subsequently plated on the ECM coatings. After washes, remaining cells were quantified by colorimetry. <b>(B)</b> MDA-MB-231 cells were incubated with complex <b>(4)</b> and then with Phalloidin and DAPI. Significant at the *<i>p</i><0.005, **<i>p</i><0.001 levels using ANOVA and Bonferroni tests.</p

Effect of complex (4) on apoptosis in MDA-MB-231 breast tumor cells and MCF-10A non-tumor breast cells.

<p><b>(A)</b> After treatment with the indicated concentrations of complex <b>(4)</b>, cells were incubated with PE-Annexin-V and 7AAD for 15min, harvested and then analyzed by cytometry. <b>(B)</b> The percentage of apoptotic and necrotic cells was plotted in a graph for MDA-MB-231 and MCF-10A cells. The fluorescence of 7AAD is detected in the FL3-A channel and the fluorescence of PE-Annexin-V is detected in the FL2-A channel. Camptothecin (campto) was used as a positive control for apoptosis. <b>(C)</b> Nuclear fragmentation promoted by complex <b>(4)</b> in MDA-MB-231 cells was investigated using DAPI staining. Staurosporine was used as a positive control for nuclear fragmentation. White arrows show fragmented nuclei. The expression of apoptotic and anti-apoptotic molecules was investigated by <b>(D)</b> qRT-PCR and <b>(E)</b> Western blotting analysis. Significant at the *<i>p</i><0.005, **<i>p</i><0.001, ***<i>p</i><0.0001 levels using ANOVA and Bonferroni tests.</p