Associations of plasma lipids, lipoproteins, and cardiovascular outcomes with climatic variations in a large Brazilian population of Campinas, São Paulo state: an eight-year study.

In this eight-year retrospective study, we evaluated the associations between climatic variations and the biological rhythms in plasma lipids and lipoproteins in a large population of Campinas, São Paulo state, Brazil, as well as temporal changes of outcomes of cardiovascular hospitalizations. Climatic variables were obtained at the Center for Meteorological and Climatic Research Applied to Agriculture (University of Campinas - Unicamp, Brazil). The plasma lipid databases surveyed were from 27,543 individuals who had their lipid profiles assessed at the state university referral hospital in Campinas (Unicamp). The frequency of hospitalizations was obtained from the Brazilian Public Health database (DATASUS). Temporal statistical analyses were performed using the methods Cosinor or Friedman (ARIMA) and the temporal series were compared by cross-correlation functions. In normolipidemic cases (n=11,892), significantly different rhythmicity was observed in low-density lipoprotein (LDL)- and high-density lipoprotein (HDL)-cholesterol (C) both higher in winter and lower in summer. Dyslipidemia (n=15,651) increased the number and amplitude of lipid rhythms: LDL-C and HDL-C were higher in winter and lower in summer, and the opposite occurred with triglycerides. The number of hospitalizations showed maximum and minimum frequencies in winter and in summer, respectively. A coincident rhythmicity was observed of lower temperature and humidity rates with higher plasma LDL-C, and their temporal series were inversely cross-correlated. This study shows for the first time that variations of temperature, humidity, and daylight length were strongly associated with LDL-C and HDL-C seasonality, but moderately to lowly associated with rhythmicity of atherosclerotic outcomes. It also indicates unfavorable cardiovascular-related changes during wintertime.

Associations of plasma lipids, lipoproteins, and cardiovascular outcomes with climatic variations in a large Brazilian population of Campinas, São Paulo state: an eight-year study

In this eight-year retrospective study, we evaluated the associations between climatic variations and the biological rhythms in plasma lipids and lipoproteins in a large population of Campinas, São Paulo state, Brazil, as well as temporal changes of outcomes of cardiovascular hospitalizations. Climatic variables were obtained at the Center for Meteorological and Climatic Research Applied to Agriculture (University of Campinas - Unicamp, Brazil). The plasma lipid databases surveyed were from 27,543 individuals who had their lipid profiles assessed at the state university referral hospital in Campinas (Unicamp). The frequency of hospitalizations was obtained from the Brazilian Public Health database (DATASUS). Temporal statistical analyses were performed using the methods Cosinor or Friedman (ARIMA) and the temporal series were compared by cross-correlation functions. In normolipidemic cases (n=11,892), significantly different rhythmicity was observed in low-density lipoprotein (LDL)- and high-density lipoprotein (HDL)-cholesterol (C) both higher in winter and lower in summer. Dyslipidemia (n=15,651) increased the number and amplitude of lipid rhythms: LDL-C and HDL-C were higher in winter and lower in summer, and the opposite occurred with triglycerides. The number of hospitalizations showed maximum and minimum frequencies in winter and in summer, respectively. A coincident rhythmicity was observed of lower temperature and humidity rates with higher plasma LDL-C, and their temporal series were inversely cross-correlated. This study shows for the first time that variations of temperature, humidity, and daylight length were strongly associated with LDL-C and HDL-C seasonality, but moderately to lowly associated with rhythmicity of atherosclerotic outcomes. It also indicates unfavorable cardiovascular-related changes during wintertime.

Avaliação das técnicas de imunodifusão em gel de ágar, ensaio imunoenzimático indireto e reação em cadeia da polimerase no diagnóstico da brucelose ovina / Evaluation of technical, immunodiffusion agar gel, enzyme-linked immunosorbent assay and polymerase chain reaction in the diagnosis of ovine brucellosis

A brucelose na espécie ovina tem recebido destaque, uma vez que se trata de uma enfermidade que acomete o sistema reprodutivo dos animais, provocando sério comprometimento no setor produtivo. Dessa forma, objetivou-se a avaliação de três métodos para o diagnóstico da brucelose ovina: o ensaio imunoenzimático indireto (ELISAi), a técnica imunodifusão em gel de ágar (IDGA) e a reação em cadeia da polimerase (PCR). Para tanto, utilizaram-se 211 amostras de sangue de ovinos oriundos de propriedades de nove municípios da microrregião homogênea de Teresina, Piauí. As 211 amostras de sangue foram submetidas aos testes sorológicos e à PCR, visando detectar anticorpos anti-B. ovis e DNA de Brucella ovis, respectivamente. Foram obtidos resultados positivos nos testes sorológicos, sendo 36 (17,06%) positivos no teste IDGA e sete (3,31%) positivos no teste ELISAi, contudo não houve resultados positivos na técnica de PCR. Dos métodos de diagnóstico utilizados neste estudo, o teste IDGA foi o que apresentou melhor desempenho na detecção de animais reagentes, quando comparado ao teste ELISAi e à PCR em amostras de sangue, e o percentual de animais soropositivos sugere uma ampla distribuição de ovinos infectados por Brucella ovis na região em estudo, o que pode causar prejuízos aos produtores.(AU)

Brucellosis in sheep has received a major focus, since it is a disease that affects the reproductive system of animals, causing serious impairment in the productive sector. Thus, three methods for the diagnosis of ovine brucellosis were evaluated as goal, the indirect Linked Immunosorbent Assay (ELISAi) test, the Immunodiffusion Agar Gel (AGID) technique and the Polymerase Chain Reaction (PCR). Therefore, we used 211 sheep blood samples from properties of nine municipalities of the homogeneous micro-region of Teresina, Piaui. The 211 blood samples were subjected to serologic testing and PCR to detect anti-B. ovis antibodies, and Brucella ovis DNA, respectively. Positive results in serological tests were obtained, 36 (17%) positive in the AGID test and seven (3.3%) positive to the ELISAi test, however, there were no positive results in the PCR technique. Of the diagnostic methods used in this study, the AGID test was the one that presented the best performance in the detection of reactive animals, when compared to ELISAi and PCR in blood samples and, the percentage of seropositive animals suggests a wide distribution of Brucella ovis infected sheep in the study region and could cause loss to producers.(AU)

Bioinspired seeding of biomaterials using three dimensional microtissues induces chondrogenic stem cell differentiation and cartilage formation under growth factor free conditions.

Cell laden biomaterials are archetypically seeded with individual cells and steered into the desired behavior using exogenous stimuli to control growth and differentiation. In contrast, direct cell-cell contact is instructive and even essential for natural tissue formation. Namely, microaggregation and condensation of mesenchymal progenitor cells triggers chondrogenesis and thereby drives limb formation. Yet a biomimetic strategy translating this approach into a cell laden biomaterial-based therapy has remained largely unexplored. Here, we integrate the microenvironment of cellular condensation into biomaterials by encapsulating microaggregates of a hundred human periosteum-derived stem cells. This resulted in decreased stemness-related markers, up regulation of chondrogenic genes and improved in vivo cartilage tissue formation, as compared to single cell seeded biomaterials. Importantly, even in the absence of exogenous growth factors, the microaggregate laden hydrogels outperformed conventional single cell laden hydrogels containing supraphysiological levels of the chondrogenic growth factor TGFB. Overall, the bioinspired seeding strategy described herein represents an efficient and growth factor-free approach to efficiently steer cell fate and drive tissue formation for biomaterial-based tissue engineering strategies.

High throughput generated micro-aggregates of chondrocytes stimulate cartilage formation in vitro and in vivo.

Cell-based cartilage repair strategies such as matrix-induced autologous chondrocyte implantation (MACI) could be improved by enhancing cell performance. We hypothesised that micro-aggregates of chondrocytes generated in high-throughput prior to implantation in a defect could stimulate cartilaginous matrix deposition and remodelling. To address this issue, we designed a micro-mould to enable controlled high-throughput formation of micro-aggregates. Morphology, stability, gene expression profiles and chondrogenic potential of micro-aggregates of human and bovine chondrocytes were evaluated and compared to single-cells cultured in micro-wells and in 3D after encapsulation in Dextran-Tyramine (Dex-TA) hydrogels in vitro and in vivo. We successfully formed micro-aggregates of human and bovine chondrocytes with highly controlled size, stability and viability within 24 hours. Micro-aggregates of 100 cells presented a superior balance in Collagen type I and Collagen type II gene expression over single cells and micro-aggregates of 50 and 200 cells. Matrix metalloproteinases 1, 9 and 13 mRNA levels were decreased in micro-aggregates compared to single-cells. Histological and biochemical analysis demonstrated enhanced matrix deposition in constructs seeded with micro-aggregates cultured in vitro and in vivo, compared to single-cell seeded constructs. Whole genome microarray analysis and single gene expression profiles using human chondrocytes confirmed increased expression of cartilage-related genes when chondrocytes were cultured in micro-aggregates. In conclusion, we succeeded in controlled high-throughput formation of micro-aggregates of chondrocytes. Compared to single cell-seeded constructs, seeding of constructs with micro-aggregates greatly improved neo-cartilage formation. Therefore, micro-aggregation prior to chondrocyte implantation in current MACI procedures, may effectively accelerate hyaline cartilage formation.

Enzymatically-crosslinked injectable hydrogels based on biomimetic dextran-hyaluronic acid conjugates for cartilage tissue engineering.

Polysaccharide hybrids consisting of hyaluronic acid (HA) grafted with a dextran-tyramine conjugate (Dex-TA) were synthesized and investigated as injectable biomimetic hydrogels for cartilage tissue engineering. The design of these hybrids (denoted as HA-g-Dex-TA) is based on the molecular structure of proteoglycans present in the extracellular matrix of native cartilage. Hydrogels of HA-g-Dex-TA were rapidly formed within 2 min via enzymatic crosslinking of the tyramine residues in the presence of horseradish peroxidase and hydrogen peroxide. The gelation time, equilibrium swelling and storage modulus could be adjusted by varying the degree of substitution of tyramine residues and polymer concentration. Bovine chondrocytes incorporated in the HA-g-Dex-TA hydrogels remained viable, as shown by the Live-dead assay. Moreover, enhanced chondrocyte proliferation and matrix production were observed in the HA-g-Dex-TA hydrogels compared to Dex-TA hydrogels. These results suggest that HA-g-Dex-TA hydrogels have a high potential as injectable scaffolds for cartilage tissue engineering.

Synthesis and characterization of hyaluronic acid-poly(ethylene glycol) hydrogels via Michael addition: An injectable biomaterial for cartilage repair.

Injectable hydrogels based on hyaluronic acid (HA) and poly(ethylene glycol) (PEG) were designed as biodegradable matrices for cartilage tissue engineering. Solutions of HA conjugates containing thiol functional groups (HA-SH) and PEG vinylsulfone (PEG-VS) macromers were cross-linked via Michael addition to form a three-dimensional network under physiological conditions. Gelation times varied from 14min to less than 1min, depending on the molecular weights of HA-SH and PEG-VS, degree of substitution (DS) of HA-SH and total polymer concentration. When the polymer concentration was increased from 2% to 6% (w/v) in the presence of 100Uml(-1) hyaluronidase the degradation time increased from 3 to 15days. Hydrogels with a homogeneous distribution of cells were obtained when chondrocytes were mixed with the precursor solutions. Culturing cell-hydrogel constructs prepared from HA185k-SH with a DS of 28 and cross-linked with PEG5k-4VS for 3weeks in vitro revealed that the cells were viable and that cell division took place. Gel-cell matrices degraded in approximately 3weeks, as shown by a significant decrease in dry gel mass. At day 21 glycosaminoglycans and collagen type II were found to have accumulated in hydrogels. These results indicate that these injectable hydrogels have a high potential for cartilage tissue engineering.

Injectable chitosan-based hydrogels for cartilage tissue engineering.

Water-soluble chitosan derivatives, chitosan-graft-glycolic acid (GA) and phloretic acid (PA) (CH-GA/PA), were designed to obtain biodegradable injectable chitosan hydrogels through enzymatic crosslinking with horseradish peroxidase (HRP) and H2O2. CH-GA/PA polymers were synthesized by first conjugating glycolic acid (GA) to native chitosan to render the polymer soluble at pH 7.4, and subsequent modification with phloretic acid (PA). The CH-GA43/PA10 with a degree of substitution (DS, defined as the number of substituted NH2 groups per 100 glucopyranose rings of chitosan) of GA of 43 and DS of PA of 10 showed a good solubility at pH values up to 10. Short gelation times (e.g. 10 s at a polymer concentration of 3 wt%), as recorded by the vial tilting method, were observed for the CH-GA43/PA10 hydrogels using HRP and H2O2. It was shown that these hydrogels can be readily degraded by lysozyme. In vitro culturing of chondrocytes in CH-GA43/PA10 hydrogels revealed that after 2 weeks the cells were viable and retained their round shape. These features indicate that CH-GA/PA hydrogels are promising as an artificial extracellular matrix for cartilage tissue engineering.

Postoperative cardiorespiratory instability following ligation of the preterm ductus arteriosus is related to early need for intervention.

OBJECTIVE: To investigate the effect of postnatal age at patent ductus arteriosus (PDA) ligation on postoperative need for cardiotropic support. STUDY DESIGN: A significant proportion of premature infants with a hemodynamically significant ductus arteriosus (HSDA) require surgical intervention. The relationship of postnatal maturation to postoperative cardiorespiratory stability is poorly understood. All preterm neonates who underwent PDA ligation between October 2002 and September 2004 were identified and divided according to postnatal age at ductal ligation, into early (4 weeks, n=36) groups. Neonatal birth demographics and morbidities, characteristics of the HSDA and reason for ligation were reviewed. Indices of cardiorespiratory stability (for example heart rate, mean arterial pressure) were recorded before and 1, 4, 8, 12 and 24 h following ligation. Details of perioperative cardiovascular interventions (for example crystalloid boluses, cardiotropic agents) were recorded. RESULT: There were no differences in baseline neonatal demographics between the groups although a greater number of babies requiring early ligation had contraindication to indomethacin as the main reason for ligation. Early PDA ligation was associated with increased postoperative cardiotrope requirement (27.6 vs 5.6%, P=0.02) and decreased systolic blood pressure 8 h after intervention (P<0.05). Although increased oxygenation index was observed in both groups during the first 12 h it was only sustained to 24 h in the early group (P<0.05). Age at ligation remained a significant risk factor for postoperative cardiotrope requirement on multiple logistic regression analysis. CONCLUSION: Surgical ligation of the ductus arteriosus in preterm infants less than 28 days old is associated with oxygenation difficulties, decreased systolic arterial pressure and an increased need for cardiotropic support in the initial 24 postoperative hours. These data suggest the need to prospectively characterize perioperative cardiovascular physiology and identify predictive factors of clinical deterioration.

Non-influence of fetal gender on ductus venosus Doppler flow in the first trimester.

OBJECTIVES: Recent findings have suggested that ductus venosus blood flow may be influenced by fetal gender. The aim of this study was to investigate further the influence of fetal gender on ductus venosus Doppler flow in the first trimester. METHODS: This was a cross-sectional and retrospective study performed between January 1998 and January 2003. A total of 932 fetuses at between 10 and 14 weeks' gestation were included. The following inclusion criteria were used: singleton gestation; crown-rump length between 39 and 84 mm; and absence of fetal anomalies. The following variables of the ductus venosus were evaluated: peak velocity during ventricular systole (S-wave) and diastole (D-wave); nadir during atrial contraction in late diastole (A-wave); pulsatility index for veins (PIV); peak velocity index for veins (PVIV); and time-averaged maximum velocity (TAMXV). RESULTS: Four hundred and forty-eight (48.1%) female and 484 (51.9%) male fetuses were included in the study. Comparing males and females at between 10 and 14 weeks' gestation, there was no statistically significant difference in S-wave, D-wave, A-wave, PIV, PVIV or TAMXV. CONCLUSIONS: Our study suggests that fetal gender does not influence ductus venosus blood flow in the first trimester.

Ductus venosus Doppler velocimetry in the first trimester: a new finding.

OBJECTIVES: To establish reference curves for ductus venosus blood flow velocities during the first trimester and compare them with previously published curves. METHODS: This was a cross-sectional and retrospective study performed between January 1998 and January 2003. The following inclusion criteria were used: singleton pregnancy, velocity measurements taken when the crown-rump length (CRL) was between 34 and 84 mm, absence of fetal anomalies, full-term pregnancy and newborn birth weight appropriate for gestational age. The following variables of the ductus venosus were measured: peak velocity during ventricular systole (S-wave) and diastole (D-wave), nadir during atrial contraction in late diastole (A-wave), time-averaged maximum velocity (TAMXV) and pulsatility index for veins (PIV). RESULTS: A total of 843 fetuses were included. The mean CRL was 62 (range, 34-84) mm. The S-wave, D-wave, TAMXV and PIV were normally distributed, and logarithmic transformation was performed to achieve a normal distribution for the A-wave. S-wave, D-wave and A-wave and TAMXV increased with CRL. PIV increased up to a CRL of 63 mm and decreased thereafter. Regression analysis revealed a significant quadratic relationship between PIV and CRL. CONCLUSIONS: S-wave, D-wave, A-wave velocities and TAMXV in the ductus venosus increase with CRL between 34 and 84 mm. The reference range for PIV has a biphasic pattern, with an initial non-significant increase up to a CRL of 63 mm and a fall thereafter.

Clinical and radiographic evaluation of pulpotomies performed under intrapulpal injection of anaesthetic solution.

AIM: The purpose of this study was to evaluate clinically and radiographically pulpotomies carried out under intrapulpal injection of anaesthetic solution. METHODOLOGY: Forty-one permanent mandibular molar teeth presenting with deep carious lesions and/or exposed pulps, with or without periapical changes on radiographic examination, were treated with pulpotomy and dressed with calcium hydroxide. The teeth were divided into three groups. Group A consisted of 15 teeth, where intrapulpal anaesthesia was administered by a slow injection of lidocaine hydrochloride 2%. Group B, with 14 teeth, where intrapulpal anaesthesia was obtained with lidocaine hydrochloride 2% with adrenaline 1:100,000. Group C consisted of 12 teeth in which anaesthesia was performed with a mandibular block using prilocaine hydrochloride 3% with felypressin 1: 100,000. Healing was evaluated using clinical and radiographic criteria: dentine barrier formation, absence of clinical symptoms and resolution of periapical involvement. RESULTS: After an observation time of 6-8 weeks (postoperative control) and 24-32 weeks (intermediate control), healing occurred in 13 teeth from group A (87%), in 11 teeth from group B (79%) and in 10 teeth from group C (83%). No statistical difference was demonstrated between the three groups (Fisher's exact test). CONCLUSIONS: Based on the methodology adopted, intrapulpal injection of anaesthetic solution did not impair healing in pulpotomized teeth.

Nickel and zinc determination by flow-injection solid-phase spectrophotometry exploiting different sorption rates.

Flow-injection solid-phase spectrophotometry is applied for sequential determination of nickel and zinc, exploiting their different sorption rates on 1-(2-thiazolylazo)-2-naphthol (TAN) immobilized on C(18)-bonded silica. The Zn(II) sorption rate on the solid support is constant for flow rates ranging from 0.70 to 2.2 ml min(-1), but for Ni(II) the sorption rate decreases with increasing flow rate. A flow system was designed to perform sequential measurements at two different flow rates (0.85 and 1.9 ml min(-1)). The absorbance was measured at 595 nm, where both TAN-immobilized complexes showed maximum absorption. The coefficients of variation were estimated (n=10) as 1.1 and 1.7% (at 1.9 ml min(-1)) and 1.2 and 2.1% (at 0.85 ml min(-1)) for zinc and nickel, respectively. This strategy was applied to determine zinc and nickel in copper-based alloys and the results agreed with certified values at the 95% confidence level. The sample throughput was estimated as 36 h(-1).

ICP-AES determination of small amounts of zinc in copper-base alloys after separation by adsorption of the zinc-TAN complex on Sep Pak C18 cartridges.

The use of ICP/AES for the determination of zinc, in low concentration levels, in matrices containing high levels of copper is difficult because copper interferes in the zinc main emission wavelength (213.856 nm). In the present work, a separation of zinc from copper matrices was possible, using the reaction of zinc(II) cation with 1-(2-tiazolylazo)-2-naphthol (TAN), in the pH range of 6.5-8.0, resulting in a stable red complex. Copper also reacts with TAN but its interference was avoided by the addition of ascorbic acid and thiosulphate in the reaction medium. In this way, the aqueous solution was passed through a SEP PAK C18 cartridge, in which the zinc(II)-TAN complex was quantitatively retained, but it did not occur with copper which passes through the cartridge, as [Cu(2)(S(2)O(3))(2)](2-), with the aqueous solution. The cartridge was washed with water and the complex eluted with ethanol. Then, the alcohol was evaporated and the complex decomposed by nitric acid. It results in both zinc pre-concentration and separation from copper. The zinc quantification was carried out by ICP/AES at 213.856 nm. The relative standard deviations, for ten different aliquots, were 5.7% and the average recovery found for zinc was 96%, even when the concentration ratio Cu/Zn was up to 500/1 (mg l(-1):mg l(-1)). Other metals, like nickel, for example, can react with TAN in the same way as zinc but they do not interfere in the emission wavelength 213.856 nm.

Spectrophotometric determination of tin in copper-based alloys using pyrocatechol violet.

In the present paper, a new procedure using Pyrocatechol Violet (PCV) for the determination of tin in copper-based alloys is proposed. The use of HEDTA as masking agent allowed tin to be determined in the presence of large amounts of copper, without any separation procedure. The method is more selective than previous methods. Cetyltrimethylammonium bromide (CTAB) and Tween-20 are used to increase the stability of the system. The method can be applied directly to an acidic solution of Sn(IV) in the range 2.0-60.0 mug with a final volume of 50 ml. The pH is adjusted to 2.0 +/- 0.2 with glycine buffer and, after 30 min, the absorbance is measured at 660 nm. Al(III), Cd(II), Co(II), Mg(II), Ca(II), Mn(II), Ni(II) and Pb(II) do not interfere at the 500 mg level; 20 000 mug of Cu(II) and 400 mug of NaCl can be present. The interference at 100 mug of Fe(III) can be masked with ascorbic acid. Bi(III), Sb(V), Ti(IV), Mo(VI), EDTA, tartrate, citrate and iodide interfere. The proposed method was used for tin determination in several copper-based alloys and a comparison of the analytical results with certified values indicates that the procedure provides accurate and precise results.