Phytochemical characterization, antioxidant potential and antibacterial activity of the Croton argyrophylloides Muell. Arg. (Euphorbiaceae) / Caracterização fitoquímica, potencial antioxidante e atividade antibacteriana do Croton argyrophylloides Muell. Arg. (Euphorbiaceae)

Croton argyrophylloides Muell. Arg., from the Euphorbiaceae family, popularly known as marmeleiro prateado or sacatinga, is a plant from the Caatinga biome commonly found in Brazil's northeastern region. The present study aimed to evaluate the antioxidant activity of the species. The phytochemical study was performed through qualitative analysis of chemical constituents and quantitative determination of the total phenol content through the Folin-Ciocalteu test. The qualitative and quantitative antioxidant tests were performed using the DPPH method (2.2 diphenyl-1-picryl hydrazil) and ferric reducing antioxidant power (FRAP). The minimum inhibitory concentration (MIC) was determined by microdilution in 96-well plates. The ethanolic extract of the leaves of C. argyrophylloides manifested antioxidant action in the quantitative DPPH test with a significant bioactivity of 84.70 AAO% in 500 µg/mL, with an EC50 of 236.79. The content of total phenolic compounds was 946.06 mg of gallic acid equivalents/g of sample, and total flavonoids was 58.11 mg of quercetin equivalents/g of sample, the result obtained for FRAP was 15294.44 µM Trolox/g of sample and ABTS was 718 µM Trolox of sample. The prospecting of the chemical constituents of the leaves of C. argyrophylloides revealed the presence of the main compounds that manifests the antioxidant activity and it was proven by the DPPH method that there is antioxidant activity in the analyzed sample, in addition to demonstrating a significant content of phenolic compounds and total flavonoid content in the species, which corroborates the antioxidant activity of the plant sample. The leaf extracts presented growth inhibition halos of 10 and 12 mm upon Staphylococcus aureus ATCC 25923.

Croton argyrophylloides Muell. Arg., pertencente à família Euphorbiaceae, conhecida popularmente como marmeleiro prateado e sacatinga, é um vegetal do bioma caatinga comumente encontrado no Nordeste do Brasil. O presente trabalho teve como objetivo avaliar a atividade antioxidante da espécie. O estudo fitoquímico foi realizado por meio de análise qualitativa dos constituintes químicos e determinação quantitativa do teor de fenóis totais pelo teste de Folin-Ciocalteu. Os testes antioxidantes qualitativos e quantitativos foram realizados pelo método do DPPH (2,2 difenil-1- picril-hidrazila) e redução do ferro (FRAP). A concentração inibitória mínima (CIM) foi determinada por microdiluição em placas de 96 poços. O extrato etanólico das folhas de C. argyrophylloides apresentou ação antioxidante no teste DPPH quantitativo com uma significativa bioatividade de 84.70 AAO% em 500 µg/mL, apresentando um CE50 de 236.79. O teor de compostos fenólicos totais, foi de 946,06 mg equivalentes de ácido gálico/g de amostra, e de flavonoides totais de 58,11 mg equivalentes de quercetina/g da amostra, o valor encontrado para FRAP foi de 15294,44 µM Trolox/g da amostra e de ABTS foi 718 µM Trolox da amostra. A prospecção dos constituintes químicos das folhas de C. argyrophylloides revelou a presença dos principais compostos que caracterizam a atividade antioxidante e foi possível comprovar pelo método de DPPH que há atividade antioxidante na amostra analisada, além de demonstrar um resultado significativo de teor de compostos fenólicos e teor de flavonoides totais na espécie e o que corrobora com a atividade antioxidante da amostra vegetal. Os extratos das folhas apresentaram halos de inibição de crescimento de 10 e 12mm frente a Staphylococcus aureus ATCC 25923.

Phytochemical characterization, antioxidant potential and antibacterial activity of the Croton argyrophylloides Muell. Arg. (Euphorbiaceae)

Abstract Croton argyrophylloides Muell. Arg., from the Euphorbiaceae family, popularly known as marmeleiro prateado or sacatinga, is a plant from the Caatinga biome commonly found in Brazils northeastern region. The present study aimed to evaluate the antioxidant activity of the species. The phytochemical study was performed through qualitative analysis of chemical constituents and quantitative determination of the total phenol content through the Folin-Ciocalteu test. The qualitative and quantitative antioxidant tests were performed using the DPPH method (2.2 diphenyl-1-picryl hydrazil) and ferric reducing antioxidant power (FRAP). The minimum inhibitory concentration (MIC) was determined by microdilution in 96-well plates. The ethanolic extract of the leaves of C. argyrophylloides manifested antioxidant action in the quantitative DPPH test with a significant bioactivity of 84.70 AAO% in 500 µg/mL, with an EC50 of 236.79. The content of total phenolic compounds was 946.06 mg of gallic acid equivalents/g of sample, and total flavonoids was 58.11 mg of quercetin equivalents/g of sample, the result obtained for FRAP was 15294.44 µM Trolox/g of sample and ABTS was 718 M Trolox of sample. The prospecting of the chemical constituents of the leaves of C. argyrophylloides revealed the presence of the main compounds that manifests the antioxidant activity and it was proven by the DPPH method that there is antioxidant activity in the analyzed sample, in addition to demonstrating a significant content of phenolic compounds and total flavonoid content in the species, which corroborates the antioxidant activity of the plant sample. The leaf extracts presented growth inhibition halos of 10 and 12 mm upon Staphylococcus aureus ATCC 25923.

Resumo Croton argyrophylloides Muell. Arg., pertencente à família Euphorbiaceae, conhecida popularmente como marmeleiro prateado e sacatinga, é um vegetal do bioma caatinga comumente encontrado no Nordeste do Brasil. O presente trabalho teve como objetivo avaliar a atividade antioxidante da espécie. O estudo fitoquímico foi realizado por meio de análise qualitativa dos constituintes químicos e determinação quantitativa do teor de fenóis totais pelo teste de Folin-Ciocalteu. Os testes antioxidantes qualitativos e quantitativos foram realizados pelo método do DPPH (2,2 difenil-1- picril-hidrazila) e redução do ferro (FRAP). A concentração inibitória mínima (CIM) foi determinada por microdiluição em placas de 96 poços. O extrato etanólico das folhas de C. argyrophylloides apresentou ação antioxidante no teste DPPH quantitativo com uma significativa bioatividade de 84.70 AAO% em 500 µg/mL, apresentando um CE50 de 236.79. O teor de compostos fenólicos totais, foi de 946,06 mg equivalentes de ácido gálico/g de amostra, e de flavonoides totais de 58,11 mg equivalentes de quercetina/g da amostra, o valor encontrado para FRAP foi de 15294,44 µM Trolox/g da amostra e de ABTS foi 718 M Trolox da amostra. A prospecção dos constituintes químicos das folhas de C. argyrophylloides revelou a presença dos principais compostos que caracterizam a atividade antioxidante e foi possível comprovar pelo método de DPPH que há atividade antioxidante na amostra analisada, além de demonstrar um resultado significativo de teor de compostos fenólicos e teor de flavonoides totais na espécie e o que corrobora com a atividade antioxidante da amostra vegetal. Os extratos das folhas apresentaram halos de inibição de crescimento de 10 e 12mm frente a Staphylococcus aureus ATCC 25923.

Phytochemical characterization, antioxidant potential and antibacterial activity of the Croton argyrophylloides Muell. Arg. (Euphorbiaceae) / Caracterização fitoquímica, potencial antioxidante e atividade antibacteriana do Croton argyrophylloides Muell. Arg. (Euphorbiaceae)

Abstract Croton argyrophylloides Muell. Arg., from the Euphorbiaceae family, popularly known as marmeleiro prateado or sacatinga, is a plant from the Caatinga biome commonly found in Brazil's northeastern region. The present study aimed to evaluate the antioxidant activity of the species. The phytochemical study was performed through qualitative analysis of chemical constituents and quantitative determination of the total phenol content through the Folin-Ciocalteu test. The qualitative and quantitative antioxidant tests were performed using the DPPH method (2.2 diphenyl-1-picryl hydrazil) and ferric reducing antioxidant power (FRAP). The minimum inhibitory concentration (MIC) was determined by microdilution in 96-well plates. The ethanolic extract of the leaves of C. argyrophylloides manifested antioxidant action in the quantitative DPPH test with a significant bioactivity of 84.70 AAO% in 500 µg/mL, with an EC50 of 236.79. The content of total phenolic compounds was 946.06 mg of gallic acid equivalents/g of sample, and total flavonoids was 58.11 mg of quercetin equivalents/g of sample, the result obtained for FRAP was 15294.44 µM Trolox/g of sample and ABTS was 718 μM Trolox of sample. The prospecting of the chemical constituents of the leaves of C. argyrophylloides revealed the presence of the main compounds that manifests the antioxidant activity and it was proven by the DPPH method that there is antioxidant activity in the analyzed sample, in addition to demonstrating a significant content of phenolic compounds and total flavonoid content in the species, which corroborates the antioxidant activity of the plant sample. The leaf extracts presented growth inhibition halos of 10 and 12 mm upon Staphylococcus aureus ATCC 25923.

Resumo Croton argyrophylloides Muell. Arg., pertencente à família Euphorbiaceae, conhecida popularmente como marmeleiro prateado e sacatinga, é um vegetal do bioma caatinga comumente encontrado no Nordeste do Brasil. O presente trabalho teve como objetivo avaliar a atividade antioxidante da espécie. O estudo fitoquímico foi realizado por meio de análise qualitativa dos constituintes químicos e determinação quantitativa do teor de fenóis totais pelo teste de Folin-Ciocalteu. Os testes antioxidantes qualitativos e quantitativos foram realizados pelo método do DPPH (2,2 difenil-1- picril-hidrazila) e redução do ferro (FRAP). A concentração inibitória mínima (CIM) foi determinada por microdiluição em placas de 96 poços. O extrato etanólico das folhas de C. argyrophylloides apresentou ação antioxidante no teste DPPH quantitativo com uma significativa bioatividade de 84.70 AAO% em 500 µg/mL, apresentando um CE50 de 236.79. O teor de compostos fenólicos totais, foi de 946,06 mg equivalentes de ácido gálico/g de amostra, e de flavonoides totais de 58,11 mg equivalentes de quercetina/g da amostra, o valor encontrado para FRAP foi de 15294,44 µM Trolox/g da amostra e de ABTS foi 718 μM Trolox da amostra. A prospecção dos constituintes químicos das folhas de C. argyrophylloides revelou a presença dos principais compostos que caracterizam a atividade antioxidante e foi possível comprovar pelo método de DPPH que há atividade antioxidante na amostra analisada, além de demonstrar um resultado significativo de teor de compostos fenólicos e teor de flavonoides totais na espécie e o que corrobora com a atividade antioxidante da amostra vegetal. Os extratos das folhas apresentaram halos de inibição de crescimento de 10 e 12mm frente a Staphylococcus aureus ATCC 25923.

Phytochemical characterization, antioxidant potential and antibacterial activity of the Croton argyrophylloides Muell. Arg. (Euphorbiaceae).

Croton argyrophylloides Muell. Arg., from the Euphorbiaceae family, popularly known as marmeleiro prateado or sacatinga, is a plant from the Caatinga biome commonly found in Brazil's northeastern region. The present study aimed to evaluate the antioxidant activity of the species. The phytochemical study was performed through qualitative analysis of chemical constituents and quantitative determination of the total phenol content through the Folin-Ciocalteu test. The qualitative and quantitative antioxidant tests were performed using the DPPH method (2.2 diphenyl-1-picryl hydrazil) and ferric reducing antioxidant power (FRAP). The minimum inhibitory concentration (MIC) was determined by microdilution in 96-well plates. The ethanolic extract of the leaves of C. argyrophylloides manifested antioxidant action in the quantitative DPPH test with a significant bioactivity of 84.70 AAO% in 500 µg/mL, with an EC50 of 236.79. The content of total phenolic compounds was 946.06 mg of gallic acid equivalents/g of sample, and total flavonoids was 58.11 mg of quercetin equivalents/g of sample, the result obtained for FRAP was 15294.44 µM Trolox/g of sample and ABTS was 718 µM Trolox of sample. The prospecting of the chemical constituents of the leaves of C. argyrophylloides revealed the presence of the main compounds that manifests the antioxidant activity and it was proven by the DPPH method that there is antioxidant activity in the analyzed sample, in addition to demonstrating a significant content of phenolic compounds and total flavonoid content in the species, which corroborates the antioxidant activity of the plant sample. The leaf extracts presented growth inhibition halos of 10 and 12 mm upon Staphylococcus aureus ATCC 25923.

Radiologic analysis of hindfoot alignment: Comparison of Méary, long axial, and hindfoot alignment views.

BACKGROUND: Among radiographic views available for assessing hindfoot alignment, the antero-posterior weight-bearing view with metal cerclage of the hindfoot (Méary view) is the most widely used in France. Internationally, the long axial view (LAV) and hindfoot alignment view (HAV) are used also. The objective of this study was to compare the reliability of these three views. HYPOTHESIS: The Méary view with cerclage of the hindfoot is as reliable as the LAV and HAV for assessing hindfoot alignment. MATERIAL AND METHODS: All three views were obtained in each of 22 prospectively included patients. Intra-observer and inter-observer reliabilities were assessed by having two observers collect the radiographic measurements then computing the intra-class correlation coefficients (ICCs). RESULTS: The intra-observer and inter-observer ICCs were 0.956 and 0.988 with the Méary view, 0.990 and 0.765 with the HAV, and 0.997 and 0.991 with the LAV, respectively. Correlations were far stronger between the LAV and HAV than between each of these and the Méary view. Compared to the LAV and HAV, the Méary view indicated a greater degree of hindfoot valgus. DISCUSSION: Intra-observer reliability was excellent with both the LAV and HAV, whereas inter-observer reliability was better with the LAV. Excellent reliability was also obtained with the Méary view. Combining the Méary view to obtain a radiographic image of the clinical deformity with the LAV to measure the angular deviation of the hindfoot axis may be useful when assessing hindfoot malalignment. A comparison of the three views in a larger population is needed before clinical recommendations can be made. LEVEL OF EVIDENCE: II, prospective study.

Prognostic factors for all-inside meniscal repair. A 87-case series.

INTRODUCTION: Repair is indicated for tears in non-degenerative menisci. The literature reports a 15% failure rate for all-inside repair. The aim of the present study was to determine prognostic factors for failure of all-inside meniscal repair. The study hypothesis was that epidemiological, clinical and surgical factors affect success. MATERIAL AND METHODS: A retrospective study included 87 meniscal repair procedures, with or without anterior cruciate ligament (ACL) tear. Lesions were located in red-red or red-white zones. After freshening, repair comprised an all-inside arthroscopic technique using the FasT-Fix® system (Smith & Nephew), with (70.1%) or without ligament reconstruction; all ACL tears were reconstructed. Preoperative data comprised: age, gender, smoking status, sports activity, trauma-to-surgery time, body mass index (BMI), frontal morphotype, and IKDC score. Intra- and postoperative data comprised: meniscal lesion characteristics, location, number of sutures, type of ACL reconstruction, presence of chondropathy, authorized postoperative ranges of motion, and IKDC score. Failure was defined by secondary meniscectomy. RESULTS: At 31 months' follow-up, there were 13 failures (15%). Mean postoperative IKDC score was 88.19 (range: 64.37-98.95). Bucket-handle lesion (P=0.006) and BMI>25 (P=0.014) emerged as significant factors of poor prognosis. DISCUSSION: The present failure rate matched those reported in the literature. The more extensive the lesion, the higher the risk of failure. High BMI incurs mechanical stresses that increase the risk of failure. LEVEL OF EVIDENCE: IV, retrospective cohort study.

Medial patellofemoral ligament reconstruction for patellar instability following total knee arthroplasty: A review of 6 cases.

INTRODUCTION: Patellar instability is a frequent cause of total knee arthroplasty (TKA) failure. In cases of post-arthroplasty patellar instability, the medial structures may be damaged. The objective of this study was to study the effectiveness of medial patellofemoral ligament (MPFL) reconstruction. We hypothesized that MPFL reconstruction will effectively realign the patella, making it a viable treatment option for managing post-arthroplasty patellar instability. MATERIAL AND METHODS: In this retrospective study of six patients treated by four different surgeons, patients were included if they had a recurring or permanent patellar dislocation after undergoing TKA. Patients were excluded if the patellar instability was painful but did not result in dislocation. Each patient underwent MPFL reconstruction using the gracilis; additional procedures could be performed depending on the diagnosis. One patient required TKA revision because of an abnormally rotated femoral implant. The main outcome measure was the non-recurrence of the dislocation. The IKDC and Kujala functional scores, joint range of motion and patellar tilt on X-rays were analyzed preoperatively and at the last follow-up. RESULTS: At a mean follow-up of 23 months (6-46), none of the patients experienced a recurrence of the patellar dislocation. Only one patient had no improvements in the functional outcome scores. The patellar tilt was reduced in all patients. CONCLUSION: MPFL reconstruction-in isolation or with femoral component revision-is effective at treating post-arthroplasty patellar instability. It has its place in the treatment of patellar dislocation following TKA and its indications must be based on exact analysis of the reasons for the instability. LEVEL OF EVIDENCE: IV - Retrospective cohort study.

Results of anatomic gracilis MPFL reconstruction with precise tensioning.

BACKGROUND: The medial patellofemoral ligament (MPFL) is the most commonly injured structure in patients with objective patellar instability. The objective of this study was to prospectively evaluate the clinical and radiographic results of MPFL reconstruction in 50 patients with chronic patellar instability. METHODS: Fifty patients with chronic patellar instability, aged 15-39 years, were included. The MPFL was reconstructed using a free gracilis autograft tendon. Two anchors were used for patellar fixation, and femoral fixation was achieved with an interference screw placed into a tunnel between the adductor tubercle and medial epicondyle. The graft was tensioned to 10 N with the knee in 30° flexion. IKDC and Kujala scores were assessed pre- and post-operatively. Patellar tilt was measured from CT scans with the quadriceps relaxed and contracted, both pre- and post-operatively. RESULTS: The follow-up period was 7 to 44 months (mean: 25 months, SD 10.3). The mean raw IKDC score increased from 51.5 preoperatively to 71.7 at last follow-up, the mean overall IKDC score increased from 38.5 to 61.7 and the Kujala score increased from 48.3 to 82.4. On CT scans, the mean patellar tilt went from 24° to 16.2° with the quadriceps relaxed and 27.7° to 18.1° in contraction. No recurrent dislocation was observed. CONCLUSION: This technique of MPFL reconstruction provided significant improvements in IKDC and Kujala scores and significant reduction in patellar tilt. No recurrent dislocations were observed during the study period.

Medial patellofemoral ligament reconstruction: clinical and radiographic results in a series of 90 cases.

INTRODUCTION: Among the numerous techniques available, medial patellofemoral ligament (MPFL) reconstruction is increasingly used for the surgical treatment of objective patellar instability. The main objective of the present study was to assess efficacy in preventing recurrence of patellar dislocation and in correcting radiographic patellar tilt. The study hypothesis was that MPFL reconstruction, isolated or with associated bone surgery, by restoring "favorable" graft anisometry, provides a good trade-off between patellar stability and absence of postoperative stiffness. MATERIALS AND METHODS: Eighty-seven patients (90 reconstructions) presenting with objective patellar instability were prospectively included. The standardized procedure comprised MPFL reconstruction using the gracilis tendon. Femoral fixation used an interference screw in a blind tunnel between the adductor magnus tubercle and the medial epicondyle; patellar fixation used 2 anchors. Complementary distal bone graft was associated in 21 patients due to a preoperative tibial tubercle-trochlear groove (TT-TG) distance exceeding 20mm or to patella alta. Functional IKDC and Kujala scores and radiographic measurement of patellar tilt and femoral tunnel position were assessed preoperatively and at end of follow-up. RESULTS: Mean follow-up was 24.3months (range, 6-49months). Three patients showed recurrence of patellar dislocation. Mean Kujala score rose from 53.88 preoperatively to 86.24 postoperatively, and mean real IKDC score from 45.15 to 73.92 (P<0.001). Patellar tilt decreased significantly between pre- and postoperative X-ray (P<0.001). DISCUSSION: MPFL gracilis reconstruction provides good clinical results and good radiologic correction of patellar tilt, making it a technique of choice in the treatment of objective patellar instability. LEVEL OF EVIDENCE: Level IV. Retrospective case series study.

Role of immunoglobulin G fragment C receptor polymorphism-mediated antibody-dependant cellular cytotoxicity in colorectal cancer treated with cetuximab therapy.

Antibody-dependent cellular cytotoxicity (ADCC), which is activated by effector cells via immunoglobulin G (IgG) fragment C receptors (FcRs), was proposed as a mechanism of cetuximab efficacy. Peripheral blood mononuclear cells (PBMCs) from 23 healthy donors and 13 patients with metastatic colorectal cancer (mCRC) treated with cetuximab were tested for FcγR polymorphisms and cetuximab-mediated ADCC. ADCC was measured by chromium-51 release on a epidermal growth factor receptor (EGFR)-positive human colon cancer cell line. Overall, 86 mCRC patients were genotyped for study purposes. PBMCs harbouring the FcγRIIIa 158 V/V genotype had a significantly higher cetuximab-mediated ADCC. No correlation was found between FcγR polymorphisms and response rate or time to progression after cetuximab-based therapy. Despite the in vitro analysis showing that the FcγRIIIa 158 V/V genotype is associated with higher ADCC, clinical data do not support a predictive role of FcγRIIIa polymorphisms in mCRC treated with cetuximab.

Associations between two single nucleotide polymorphisms of the adiponectin gene, its circulating concentrations and cardiometabolic risk factors in prepubertal children with and without abdominal obesity.

BACKGROUND: The adiponectin gene has been identified as a susceptibility locus for metabolic syndrome, diabetes and cardiovascular disease. AIM: To examine the influence of two single nucleotide polymorphisms (SNPs) of this gene (+276G>T and +45T>G) on circulating adiponectin concentrations, and to evaluate their relationship with adiposity and cardiometabolic risk factors in prepubertal children with and without abdominal obesity. MATERIAL AND METHODS: 168 children (78M, 6-10 yr) were examined, divided into three groups based on waist circumference (WC). Auxological and biochemical parameters were measured by standard procedures. Adiponectin SNPs were genotyped using TaqMan allelic discrimination assays. RESULTS: Adiponectin concentration correlated inversely with measures of adiposity (rBMIz-score=-0.211, pBMIz-score=0.007; rwc=-0.210, pwc=0.008; rwc/height=-0.215, pwc/height=0.006), and was significantly influenced by blood glucose, insulin and systolic blood pressure (SBP). The +276T-allele carriers had higher SBP and diastolic BP compared to GG-homozygotes (p<0.05), and expressed higher obesity-related measures and lower adiponectin concentrations. As to the +45T>G SNP, the GGsubject had higher total cholesterol and LDL-C concentrations compared to the T-allele carriers (p<0.05), showing worse obesity measures, higher triglyceride, glucose and insulin and lower serum adiponectin values. CONCLUSION: Genetic variants of the adiponectin gene had an impact on adiposity, adiponectin concentrations and some cardiometabolic variables among prepubertal children.

The mechanisms of nadroparin-mediated inhibition of proliferation of two human lung cancer cell lines.

OBJECTIVES: Clinical data suggest that heparin treatment improves survival of lung cancer patients, but the mechanisms involved are not fully understood. We investigated whether low molecular weight heparin nadroparin, directly affects lung cancer cell population growth in conventionally cultured cell lines. MATERIALS AND METHODS: A549 and CALU1 cells' viability was assessed by MTT and trypan blue exclusion assays. Cell proliferation was assessed using 5-bromo-2-deoxyuridine incorporation. Apoptosis and cell-cycle distribution were analysed by flow cytometry; cyclin B1, Cdk1, p-Cdk1 Cdc25C, p-Cdc25C and p21 expressions were analysed by western blotting. mRNA levels were analysed by real time RT-PCR. RESULTS: Nadroparin inhibited cell proliferation by 30% in both cell lines; it affected the cell cycle in A549, but not in CALU-1 cells, inducing arrest in the G(2) /M phase. Nadroparin in A549 culture inhibited cyclin B1, Cdk1, Cdc25C and p-Cdc25C, while levels of p-Cdk1 were elevated; p21 expression was not altered. Dalteparin caused a similar reduction in A549 cell population growth; however, it did not alter cyclin B1 expression as expected, based on previous reports. Fondaparinux caused minimal inhibition of A549 cell population growth and no effect on either cell cycle or cyclin B1 expression. CONCLUSIONS: Nadroparin inhibited proliferation of A549 cells by inducing G(2) /M phase cell-cycle arrest that was dependent on the Cdc25C pathway, whereas CALU-1 cell proliferation was halted by as yet not elucidated modes.

Determination of arsenic in chicken feed by hydride generation atomic absorption spectrometry after pre-concentration with polyurethane foam.

A pre-concentration procedure with solid-phase extraction was developed for the determination of arsenic (As) in chicken feed using hydride generation atomic absorption spectrometry (HG-AAS). The procedure was based on the sorption of As(III) ions as complexes with ammonium pyrrolidine dithiocarbamate onto a mini-column packed with polyurethane foam. After pre-concentration, the As was removed from the mini-column by acid solution, and the analyte content in the eluate was measured by HG-AAS. The following main experimental conditions were established: adjustment of the As solution pH with 0.05 mol l⁻¹ HCl, 2.88 × 10⁻³ mol l⁻¹ complexing agent concentration and 6.0 mol l⁻¹ eluting hydrochloric acid concentration. The proposed method produced an enrichment factor of 67, with 0.050 and 0.165 µg g⁻¹ limits of detection and quantification, respectively. The procedure was applied to the determination of As content in two types of chicken feed using the proposed procedure and atomic absorption spectrometry with electrothermal atomisation (ETAAS). The t-test indicated that the results were not significantly different at a confidence level of 95%.

Autocrine activation of human monocyte/macrophages by monocyte-derived microparticles and modulation by PPARγ ligands.

BACKGROUND AND PURPOSE: Microparticles (MPs), small membrane-bound particles originating from different cell types during activation or apoptosis, mediate intercellular communication, exert pro-coagulant activity and affect inflammation and other pathophysiological conditions. Monocyte-derived MPs have undergone little investigation and, to our knowledge, have never been evaluated for their possible autocrine effects. Therefore, we assessed the ability of monocyte-derived MPs to stimulate human monocytes and monocyte-derived macrophages (MDM). EXPERIMENTAL APPROACH: MPs were generated from supernatants of human monocytes stimulated by the calcium ionophore A23187 (12 µM), and then characterized. Human monocytes and MDM of healthy donors were isolated by standard procedures. Cells were challenged by MPs or phorbol 12-myristate 13-acetate (PMA, used as standard stimulus), in the absence or presence of PPARγ agonists and antagonists. Superoxide anion production (measured spectrophotometrically), cytokine release (elisa), PPARγ protein expression (immunoblotting) and NF-κB activation (EMSA assay) were evaluated. KEY RESULTS: Monocyte-derived MPs induced, in a concentration-dependent manner, oxygen radical production, cytokine release and NF-κB activation in human monocytes and macrophages, with lower effects than PMA. In both cell types, the PPARγ agonists rosiglitazone and 15-deoxy-Δ(12,14) -prostaglandin J(2) (15d-PGJ(2) ) inhibited MPs-induced stimulation and this inhibition was reversed by a PPARγ antagonist. In human monocyte/macrophages, MPs as well as rosiglitazone and 15d-PGJ(2) induced PPARγ protein expression. CONCLUSION AND IMPLICATIONS: In human monocyte/macrophages, monocyte-derived MPs exert an autocrine activation that was modulated by PPARγ ligands, inducing both pro-inflammatory (superoxide anion production, cytokine release and NF-κB activation) and anti-inflammatory (PPARγ expression) effects.

Evidence for epigenetic abnormalities of the androgen receptor gene in foreskin from children with hypospadias.

CONTEXT: Hypospadias is a malformation of the penis due to an incomplete development of the male urethra, the exact etiology of which in the majority of cases remains unknown. OBJECTIVE: The objective of the study was to assess whether defects of the androgen receptor (AR) gene (CAG repeats and methylation pattern) and DNA methyltransferases (DNMT) family are present in hypospadic patients. DESIGN: CAG repeats length, methylation status, and expression of the AR gene were analyzed. The DNMT family was studied at the protein level and the DNMT3A sequenced. SETTING: The study was performed at a pediatric endocrinology referral clinic. PATIENTS OR OTHER PARTICIPANTS: Twenty boys with isolated glandular hypospadias and 20 age-matched control children undergoing a surgical procedure for circumcision were studied. MAIN OUTCOME MEASURE(S): CAG repeats length and AR methylation pattern in PBLs and foreskin tissue, DNMT expression and sequencing in patients and controls, and in vitro studies in cultured fibroblasts were measured. RESULTS: AR gene methylation in foreskin tissues from patients with hypospadias was higher than in normal children. AR expression in foreskin tissue of hypospadic patients was lower than in controls, whereas the DNMT3A protein level was significantly higher in patients than controls. In cultured fibroblasts, both dihydrotestosterone and testosterone significantly reduced AR gene methylation and DNMT3A expression in a dose-dependent fashion and increased AR expression. CONCLUSION: The AR gene in target tissues from patients with hypospadias is more methylated than in control children, resulting in a decreased expression of the AR. The mechanism underlying the modulation of the AR gene expression seems to be mediated by DNMT3A. This epigenetic alteration of the AR gene might be involved in the pathogenesis of hypospadias.

Role of NF-kappaB and PPAR-gamma in lung inflammation induced by monocyte-derived microparticles.

Microparticles (MP) are phospholipid vesicles shed by cells upon activation or apoptosis. Monocyte-derived MP upregulate the synthesis of proinflammatory mediators by lung epithelial cells; the molecular bases of such activity are unknown. Peroxisome proliferator-activated receptors (PPAR) have been demonstrated to be involved in the modulation of nuclear factor (NF)-κB transcriptional activity and inflammation. We investigated whether the upregulation of the synthesis of proinflammatory cytokines by human lung epithelial cells induced by monocyte/macrophage-derived MP involves NF-κB activation and is modulated by PPAR-γ. MP were generated by stimulation of human monocytes/macrophages with the calcium ionophore, A23187. MP were incubated with human lung epithelial cells. NF-κB translocation was assessed by electrophoretic mobility shift assay. Interleukin (IL)-8 and monocyte chemotactic protein (MCP)-1 synthesis was assessed by ELISA and RT-PCR. Stimulation of A549 alveolar cells with monocyte/macrophage-derived MP caused an increase in NF-κB activation and IL-8 and MCP-1 synthesis that was inhibited by pre-incubation with the PPAR-γ agonists, rosiglitazone and 15-deoxy-Δ12,14-prostaglandin-J2. Parallel experiments with normal human bronchial epithelial cells largely confirmed the results. The effects of PPAR-γ agonists were reversed by the specific antagonist, GW9662. Upregulation of the synthesis of proinflammatory mediators by human lung epithelial cells induced by monocyte/macrophage-derived MP is mediated by NF-κB activation through a PPAR-γ dependent pathway.

Spectrum of F8 gene mutations in haemophilia A patients from a region of Italy: identification of 23 new mutations.

SUMMARY: Haemophilia A (HA) is an X-linked recessive bleeding disorder caused by a lack or decrease of coagulation factor VIII activity. The molecular diagnosis of HA is challenging and a variety of different mutations have been identified throughout the F8 gene. Our aim was to detect the causative mutation in 266 HA patients from Emilia-Romagna region (Italy) and in all suspected carriers. Molecular analysis of F8 in 201 HA patients (152 index cases) was performed with a combination of several indirect and direct molecular approaches, such as long distance polymerase chain reaction, multiplex ligation-dependent probe amplification, denaturing high performance liquid chromatography and direct sequencing. The analysis revealed 78 different mutations, 23 of which were novel, not having been reported in national or international databases. The detection rate was 100%, 86% and 89% in patients with severe, moderate and mild HA, respectively. The information provided by this registry will be helpful for monitoring the treatment of HA patients in Emilia-Romagna and also for reliable genetic counselling of affected families in the future.

[Intestinal perforation due to accidental ingestion of foreign bodies]. / Perforazione intestinale da ingestione accidentale di corpo estraneo.

Usually the ingested foreign bodies (IFB) pass the gastrointestinal tract making no lesions. Sometimes IFB could lead to a gastrointestinal perforation. The most frequently perforating IFB are chicken or fish bones and toothpicks, while risks factors are mental retardation, alcohol or drug abuse, denture usage, quick eating or habitual chewing of toothpicks. The accidentally ingestion in a high risk patient with unclear symptoms, added to a low sensitive diagnostic imaging, lead to intraoperative diagnosis in one half cases of gastrointestinal perforation by IFB. Furthermore the surgical treatment range between the less minimal invasive laparoscopic IFB extraction and intraabdominal hole suture to a laparotomic bowel or colic resection. Herein we describe our experience in 3 cases of gastrointestinal perforation by IFB have been diagnosed at surgery and treated by IFB extraction and hole suture (in 2 patients; 1 laparoscopy, 1 laparotomy) or open right emicolectomy (1 patient).

Karyotype analysis of the euploid cell population of a mouse embryonic stem cell line revealed a high incidence of chromosome abnormalities that varied during culture.

It is common knowledge that mouse embryonic stem cell (mESC) lines accumulate chromosomal changes during culture. Despite the wide use of mESCs as a model of early mammalian development and cell differentiation, there is a lack of systematic studies aimed at characterizing their karyological changes during culture. We cultured an mESC line, derived in our laboratory, for a period of 3 months investigating its chromosome complement at different times. About 60% of the metaphases analysed were euploid throughout the culture period but, from passage 13, only 50% of the euploid metaphases had a proper chromosome complement. The remaining 50% showed chromosome abnormalities, mainly gain or loss of entire chromosomes, both within the same passage and among different passages analysed. The very heterogeneous spectrum of abnormalities indicates a high frequency of chromosome mutations that arise continuously during culture. The heterogeneity of the aberrant chromosome constitution of 2n = 40 metaphases, observed at different passages of culture, might be due either to their elimination or to a shift towards the hypoeu- or hypereuploid population of those metaphases that accumulate further chromosome abnormalities. The stability of the frequency of eu-, hypoeu- and hypereuploid populations during culture might, however, be due to the elimination of those cells that carry a high mutational burden. Based on our results, we suggest that karyotype analysis of the euploid cell population of mESC lines is necessary when such lines are used in the production of chimeric mice, for their contribution to the germ line, or when they are differentiated into specific cell types.