Enhanced recovery after fetal spina bifida surgery: what is our global practice?

OBJECTIVE: Prenatal spina bifida closure results in improved outcomes for the child compared to postnatal surgery but is associated with significant maternal morbidity. Optimization of the perioperative care for women who underwent fetal spina bifida surgery could improve maternal and pregnancy outcomes. Enhanced Recovery After Surgery (ERAS) protocols are multimodal, evidence-based care plans that have been adopted for multiple surgical procedures to promote faster and better patient recovery and shorter hospitalization. This study aims to explore if fetal centers have implemented ERAS principles in this setting. Furthermore, we provide recommendations for the perioperative management of patients undergoing fetal spina bifida surgery. METHODS: Fifty-three fetal therapy centers offering prenatal surgery for open spina bifida were identified and invited to complete a digital questionnaire covering their pre-, intra- and postoperative management. An overall score was calculated per center based on the center's compliance with 20 key ERAS principles, extrapolated from ERAS guidelines for cesarean section, gynecologic oncology and colorectal surgery. Each item was scored 1 or 0 when the center did or did not comply with each principle, with a maximum score of 20. RESULTS: The questionnaire was completed by 46 centers in 17 countries (response rate 87%). Twenty-two centers (48%) exclusively perform open fetal surgery (laparotomy and hysterotomy), whereas 14 (30%) offer both open and fetoscopic procedures and 10 (22%) use fetoscopy only. The perioperative management of patients undergoing fetoscopic and open surgery was highly similar. The median ERAS score was 12 (mean 12.5, SD 2.4, range 8-17). Center compliance was the highest for the use of regional anesthesia (98%), avoidance of bowel preparation (96%), and thromboprophylaxis (96%), while the lowest compliance was achieved for preoperative carbohydrate loading (15%), postoperative nausea and vomiting prevention (33%), avoidance of overnight fasting (33%) and a 2-hour fasting period for clear fluids (20%). ERAS scores were similar in centers with a short (2-5 days), medium (6-10 days) and long (≥11 days) hospital stay (12.8 ± 2.4, 12.1 ± 2.0, and 10.3 ± 3.2, respectively, p=0.15). Furthermore, there was no significant association between ERAS score and surgical technique or center volume. CONCLUSION: The perioperative management of fetal spina bifida surgery is highly variable across fetal therapy centers worldwide. Standardizing protocols according to ERAS principles may improve patient recovery, reduce maternal morbidity, and shorten hospital stay after fetal spina bifida surgery. This article is protected by copyright. All rights reserved.

Growth curve of Nile tilapia from different families of the AquaAmérica variety / Curva de crescimento de tilápia-do-Nilo de diferentes famílias da variedade AquaAmérica

Selection can affect growth, changing performance and asymptotic values. However, there is little information about the growth of families in fish breeding programs. The aim of this study was to evaluate the performance and growth of families of Nile tilapia AquaAmérica. Twenty AquaAmérica families cultivated in a net cage (13.5 m3) for 181 days were evaluated. The nonlinear Gompertz regression model was fitted to the data by the weighted least squares method, taking the inverse of the variance of weight in different families and at different ages as the weighting variable. The model was adjusted to describe the growth in weight and morphometric characteristics. Two families showed highest (P<0.05) weights at both 133 days (family AA10: 743.2 g; family AA16: 741.2 g) and 181 days (family AA10: 1,422.1 g; family AA16: 1,393.4 g) of the experiment. In both experimental periods, the males showed a heavier weight, with the greatest contrast between the sexes occurring at 181 days. The analysis of the three most contrasting families (AA1, AA9 and AA14) showed that the asymptotic value for weight was higher (P<0.05) in family AA9 (3,926.3 g) than in family AA14 (3,251.6 g), but specific growth rate and age at the inflection point did not differ significantly between families. In conclusion, two of the 20 families were superior; males exhibited a greater growth, mainly in the period of 181 days; and the growth curve differed between the families, especially for asymptotic weight.

A seleção pode impactar a forma de crescimento, mudando o desempenho e os valores assintóticos. No entanto, existem poucas informações sobre o crescimento das famílias em programas de criação de peixes. O objetivo deste estudo foi examinar o desempenho e as curvas de crescimento de famílias de tilápia-do-Nilo AquaAmérica. Foram avaliadas 20 famílias AquaAmérica cultivadas em tanques-rede (13,5 m3) por 181 dias. O modelo de regressão não linear de Gompertz foi ajustado aos dados pelo método dos mínimos quadrados ponderados, tomando o inverso da variância do peso nas diferentes famílias e nas diferentes idades como variável de ponderação. O modelo foi ajustado para descrever o crescimento em peso e características morfométricas. Duas famílias apresentaram pesos maiores (P <0,05) em 133 dias (família AA10: 743,2 g; família AA16: 741,2 g) e 181 dias (família AA10: 1422,1 g; família AA16: 1393,4 g) de experimento em relação a outras famílias. Em ambos os períodos experimentais, os machos apresentaram maior peso, com maior contraste entre os sexos ocorrendo aos 181 dias. A análise das três famílias mais contrastantes (AA1, AA9 e AA14) mostrou que o valor assintótico para o peso foi maior (P <0,05) na família AA9 (3926,3 g) do que na família AA14 (3251,6 g), mas a taxa de crescimento específica e a idade no ponto de inflexão não diferiu significativamente entre as famílias. Em conclusão, duas das 20 famílias eram muito superiores; machos exibiram um maior crescimento, principalmente no período de 181 dias; e a curva de crescimento diferiu entre as famílias, principalmente quanto ao peso assintótico.

Growth curve of Nile tilapia from different families of the AquaAmérica variety / Curva de crescimento de tilápia-do-Nilo de diferentes famílias da variedade AquaAmérica

Selection can affect growth, changing performance and asymptotic values. However, there is little information about the growth of families in fish breeding programs. The aim of this study was to evaluate the performance and growth of families of Nile tilapia AquaAmérica. Twenty AquaAmérica families cultivated in a net cage (13.5 m3) for 181 days were evaluated. The nonlinear Gompertz regression model was fitted to the data by the weighted least squares method, taking the inverse of the variance of weight in different families and at different ages as the weighting variable. The model was adjusted to describe the growth in weight and morphometric characteristics. Two families showed highest (P<0.05) weights at both 133 days (family AA10: 743.2 g; family AA16: 741.2 g) and 181 days (family AA10: 1,422.1 g; family AA16: 1,393.4 g) of the experiment. In both experimental periods, the males showed a heavier weight, with the greatest contrast between the sexes occurring at 181 days. The analysis of the three most contrasting families (AA1, AA9 and AA14) showed that the asymptotic value for weight was higher (P<0.05) in family AA9 (3,926.3 g) than in family AA14 (3,251.6 g), but specific growth rate and age at the inflection point did not differ significantly between families. In conclusion, two of the 20 families were superior; males exhibited a greater growth, mainly in the period of 181 days; and the growth curve differed between the families, especially for asymptotic weight.

A seleção pode impactar a forma de crescimento, mudando o desempenho e os valores assintóticos. No entanto, existem poucas informações sobre o crescimento das famílias em programas de criação de peixes. O objetivo deste estudo foi examinar o desempenho e as curvas de crescimento de famílias de tilápia-do-Nilo AquaAmérica. Foram avaliadas 20 famílias AquaAmérica cultivadas em tanques-rede (13,5 m3) por 181 dias. O modelo de regressão não linear de Gompertz foi ajustado aos dados pelo método dos mínimos quadrados ponderados, tomando o inverso da variância do peso nas diferentes famílias e nas diferentes idades como variável de ponderação. O modelo foi ajustado para descrever o crescimento em peso e características morfométricas. Duas famílias apresentaram pesos maiores (P <0,05) em 133 dias (família AA10: 743,2 g; família AA16: 741,2 g) e 181 dias (família AA10: 1422,1 g; família AA16: 1393,4 g) de experimento em relação a outras famílias. Em ambos os períodos experimentais, os machos apresentaram maior peso, com maior contraste entre os sexos ocorrendo aos 181 dias. A análise das três famílias mais contrastantes (AA1, AA9 e AA14) mostrou que o valor assintótico para o peso foi maior (P <0,05) na família AA9 (3926,3 g) do que na família AA14 (3251,6 g), mas a taxa de crescimento específica e a idade no ponto de inflexão não diferiu significativamente entre as famílias. Em conclusão, duas das 20 famílias eram muito superiores; machos exibiram um maior crescimento, principalmente no período de 181 dias; e a curva de crescimento diferiu entre as famílias, principalmente quanto ao peso assintótico.

Growth curve of Nile tilapia from different families of the AquaAmérica variety

Abstract Selection can affect growth, changing performance and asymptotic values. However, there is little information about the growth of families in fish breeding programs. The aim of this study was to evaluate the performance and growth of families of Nile tilapia AquaAmérica. Twenty AquaAmérica families cultivated in a net cage (13.5 m3) for 181 days were evaluated. The nonlinear Gompertz regression model was fitted to the data by the weighted least squares method, taking the inverse of the variance of weight in different families and at different ages as the weighting variable. The model was adjusted to describe the growth in weight and morphometric characteristics. Two families showed highest (P 0.05) weights at both 133 days (family AA10: 743.2 g; family AA16: 741.2 g) and 181 days (family AA10: 1,422.1 g; family AA16: 1,393.4 g) of the experiment. In both experimental periods, the males showed a heavier weight, with the greatest contrast between the sexes occurring at 181 days. The analysis of the three most contrasting families (AA1, AA9 and AA14) showed that the asymptotic value for weight was higher (P 0.05) in family AA9 (3,926.3 g) than in family AA14 (3,251.6 g), but specific growth rate and age at the inflection point did not differ significantly between families. In conclusion, two of the 20 families were superior; males exhibited a greater growth, mainly in the period of 181 days; and the growth curve differed between the families, especially for asymptotic weight.

Resumo A seleção pode impactar a forma de crescimento, mudando o desempenho e os valores assintóticos. No entanto, existem poucas informações sobre o crescimento das famílias em programas de criação de peixes. O objetivo deste estudo foi examinar o desempenho e as curvas de crescimento de famílias de tilápia-do-Nilo AquaAmérica. Foram avaliadas 20 famílias AquaAmérica cultivadas em tanques-rede (13,5 m3) por 181 dias. O modelo de regressão não linear de Gompertz foi ajustado aos dados pelo método dos mínimos quadrados ponderados, tomando o inverso da variância do peso nas diferentes famílias e nas diferentes idades como variável de ponderação. O modelo foi ajustado para descrever o crescimento em peso e características morfométricas. Duas famílias apresentaram pesos maiores (P 0,05) em 133 dias (família AA10: 743,2 g; família AA16: 741,2 g) e 181 dias (família AA10: 1422,1 g; família AA16: 1393,4 g) de experimento em relação a outras famílias. Em ambos os períodos experimentais, os machos apresentaram maior peso, com maior contraste entre os sexos ocorrendo aos 181 dias. A análise das três famílias mais contrastantes (AA1, AA9 e AA14) mostrou que o valor assintótico para o peso foi maior (P 0,05) na família AA9 (3926,3 g) do que na família AA14 (3251,6 g), mas a taxa de crescimento específica e a idade no ponto de inflexão não diferiu significativamente entre as famílias. Em conclusão, duas das 20 famílias eram muito superiores; machos exibiram um maior crescimento, principalmente no período de 181 dias; e a curva de crescimento diferiu entre as famílias, principalmente quanto ao peso assintótico.

Effects of a nanoemulsion with Copaifera officinalis oleoresin against monogenean parasites of Colossoma macropomum: A Neotropical Serrasalmidae.

Monogeneans are ectoparasites that may cause losses in production and productivity in the aquaculture of Colossoma macropomum. Chemotherapeutics used in aquaculture usually have major adverse effects on fish; hence, the use of essential oils has been considered advantageous, but these are not soluble in water. Thus, the use of nanostructures to enhance water solubility of compounds and improve bioactivity may be very promising. This study investigated the antiparasitic activity of nanoemulsion prepared with Copaifera officinalis oleoresin (50, 100, 150, 200 and 300 mg/L), against monogenean parasites from the gills of C. macropomum. The particle size distribution and zeta potential suggested that a potentially kinetic stable system was generated. The nanoemulsion from C. officinalis oleoresin achieved high efficacy (100%) at low concentrations (200 and 300 mg/L) after 15 min of exposure. This was the first time that a nanoemulsion was generated from C. officinalis oleoresin using a solvent-free, non-heating and low-energy method. Moreover, this was the first time that an antiparasitic against monogeneans on fish gills, based on nanoemulsion of C. officinalis oleoresin, was tested.

Neonatal tobacco smoke reduces thermogenesis capacity in brown adipose tissue in adult rats.

Maternal smoking is a risk factor for progeny obesity. We have previously shown, in a rat model of neonatal tobacco smoke exposure, a mild increase in food intake and a considerable increase in visceral adiposity in the adult offspring. Males also had secondary hyperthyroidism, while females had only higher T4. Since brown adipose tissue (BAT) hypofunction is related to obesity, here we tested the hypothesis that higher levels of thyroid hormones are not functional in BAT, suggesting a lower metabolic rate. We evaluated autonomic nerve activity in BAT and its function in adult rats that were exposed to tobacco smoke during lactation. At birth, litters were adjusted to 3 male and 3 female pups/litter. From postnatal day (PND) 3 to 21, Wistar lactating rats and their pups were divided into SE group, smoke-exposed in a cigarette smoking machine (4 times/day) and C group, exposed to filtered air. Offspring were sacrificed at PND180. Adult SE rats of both genders had lower interscapular BAT autonomic nervous system activity, with higher BAT mass but no change in morphology. BAT UCP1 and CPT1a protein levels were decreased in the SE groups of both genders. Male SE rats had lower ß3-AR, TRα1, and TRß1 expression while females showed lower PGC1α expression. BAT Dio2 mRNA and hypothalamic POMC and MC4R levels were similar between groups. Hypothalamic pAMPK level was higher in SE males and lower in SE females. Thus, neonatal cigarette smoke exposure induces lower BAT thermogenic capacity, which can be obesogenic at adulthood.

Nanoemulsion from essential oil of Pterodon emarginatus (Fabaceae) shows in vitro efficacy against monogeneans of Colossoma macropomum (Pisces: Serrasalmidae).

Outbreaks of diseases pose a major threat to sustainable aquaculture development worldwide. Application of herbal products to combat parasitic diseases provides an alternative approach for sustainable aquaculture. This study investigated the in vitro antiparasitic effects of an oil-in-water nanoemulsion prepared using the essential oil from Pterodon emarginatus, against monogeneans infesting Colossoma macropomum. Gill arches from C. macropomum (47.6 ± 14.5 g and 13.5 ± 1.4 cm) that were naturally parasitized by Anacanthorus spathulatus, Notozothecium janauachensis and Mymarothecium boegeri were immersed in different dispersions of the P. emarginatus nanoemulsions (0, 50, 100, 200, 400 and 600 mg/L). The major compounds presented in the essential oil of P. emarginatus were ß-elemene, ß-caryophyllene and α-humulene. Characterization of these nanoemulsions showed that they have a small mean droplet size and low polydispersity index, which is concordant with stable systems. In this in vitro trial, the P. emarginatus nanoemulsion concentrations of 100, 200, 400 and 600 mg/L presented 100% helminthic efficacy against monogeneans of the gills of C. macropomum. The highest two concentrations used (400 and 600 mg/L) were seen to immobilize the parasites after only 15 min. Therefore, it would be worthwhile testing these concentrations in therapeutic baths against monogeneans of C. macropomum.

First cytotoxic, genotoxic, and antigenotoxic assessment of Euterpe oleracea fruit oil (açaí) in cultured human cells.

Euterpe oleracea Mart., popularly known as "açaí", is a tropical fruit from the Amazon region where it has considerable economic importance. Açaí has been used as food and for several medicinal purposes. Despite the widespread use of this fruit, there is a lack of data regarding the safety of using this fruit oil exclusively. Therefore, we evaluated the in vitro cytotoxic, genotoxic, and antigenotoxic effects of E. oleracea fruit oil (EOO) in cultured human lymphocytes (non-metabolizing cells) and HepG2 cell line (human hepatoma) (metabolizing cells) by using MTT, comet, and micronucleus assays. A wide range of EOO concentrations was tested with a preliminary MTT assay, which allowed selecting five concentrations for comet and micronucleus assays: 2.5, 10, 100, 500, and 1000 µg/mL. The results showed that none of the EOO tested concentrations presented cytotoxic effects. The genotoxic assessment revealed an absence of significant DNA and chromosome damage in human lymphocytes and HepG2 cells but did not show chemoprotection against the DNA damage induced by methyl methanesulfonate and benzo[a]pyrene, used as DNA-damaging agents.

In vitro study of the cytotoxic and genotoxic effects of indomethacin-loaded Eudragit(®) L 100 nanocapsules.

Indomethacin is a non-steroidal anti-inflammatory agent included in one of the most commonly used drug classes worldwide. The use of this drug results in certain side effects, including gastrointestinal complications. Therefore, there exists a need to develop better methods for the delivery of such drugs into the body, such as those employing nanoparticles. The aim of the present study was to evaluate the cytotoxic and genotoxic effects of indomethacin-loaded Eudragit(®) L 100 nanocapsules (NI; based on methacrylic acid and methyl methacrylate) on cells unable (lymphocytes) and able to metabolize drugs (HepG2 cells), using comet and cytokinesis-block micronucleus (CBMN) assays in vitro. Cells were exposed to NI at concentrations of 5, 10, 50, 125, 250, and 500 µg/mL. The comet assay showed that NI induced no significant DNA damage in either cell type at any of the concentrations tested. The CBMN test confirmed these results; however, the highest concentration of 500 µg/mL resulted in a small but statistically significant clastogenic/aneugenic effect in HepG2 cells. These findings should encourage the development of new investigations of this nanomaterial as a delivery vehicle for anti-inflammatory drugs, such as indomethacin.

Evaluation of the genotoxicity of Euterpe oleraceae Mart. (Arecaceae) fruit oil (açaí), in mammalian cells in vivo.

E. oleracea is a tropical plant from the Amazon region, with its fruit used for food, and traditionally, as an antioxidant, anti-inflammatory, hypocholesterolemic, for atherosclerotic disease, and has anticancer properties. The oil of the fruit has antidiarrheic, anti-inflammatory and antinociceptive activities, but without genotoxicity evaluation. Therefore, the aim of this study was to evaluate the genotoxic potential of E. oleracea fruit oil (EOO), in rat cells. Male Wistar rats were treated with EOO by gavage at doses of 30, 100 and 300 mg/kg, for 14 days, within a 24 h interval. The DNA damage in the leukocytes, liver, bone marrow and testicular cells, was assessed by the comet assay, and the clastogenic/aneugenic effects in the bone marrow cells, by the micronucleus test. Our phytochemicals characterization of the EOO showed the presence of vanillic, palmitic, γ-linolenic, linoleic, oleic, cinnamic, caffeic, protocatechuic, ferulic, syringic acids, and flavonoids quercetin and kaempferol rutinoside as the main constituents. Both cytogenetic tests performed showed that EOO presented no significant genotoxic effects in the analyzed cells, at the three tested doses. These results indicate that, under our experimental conditions, E. oleracea fruit oil did not reveal genetic toxicity in rat cells.

Neonatal Nicotine Exposure Leads to Hypothalamic Gliosis in Adult Overweight Rats.

Astrocytes and microglia, the immune competent cells of central nercous system, can be activated in response to metabolic signals such as obesity and hyperleptinaemia. In rats, maternal exposure to nicotine during lactation leads to central obesity, hyperleptinaemia, leptin resistance and alterations in hypothalamic neuropeptides in the offspring during adulthood. In the present study, we studied the activation of astrocytes and microglia, as well as the pattern of inflammatory mediators, in adult offspring of this experimental model. On postnatal day 2 (P2), osmotic minipumps releasing nicotine (NIC) (-6 mg/kg/day) or saline for 14 days were s.c. implanted in dams. Male offspring were killed on P180 and hypothalamic immunohistochemistry, retroperitoneal white adipose tissue (WAT) polymerase chain reaction analysis and multiplex analysis for plasma inflammatory mediators were carried out. At P180, NIC astrocyte cell number was higher in the arcuate nucleus (ARC) (medial: +82%; lateral: +110%), in the paraventricular nucleus (PVN) (+144%) and in the lateral hypothalamus (+121%). NIC glial fibrillary acidic protein fibre density was higher in the lateral ARC (+178%) and in the PVN (+183%). Interleukin-6 was not affected in the hypothalamus. NIC monocyte chemotactic protein 1 was only higher in the periventricular nucleus (+287%). NIC microglia (iba-1-positive) cell number was higher (+68%) only in the PVN, as was the chemokine (C-X3-C motif) receptor 1 density (+93%). NIC interleukin-10 was lower in the WAT (-58%) and plasma (-50%). Thus, offspring of mothers exposed to nicotine during lactation present hypothalamic astrogliosis at adulthood and microgliosis in the PVN.

Early redox imbalance is associated with liver dysfunction at weaning in overfed rats.

Neonatal overfeeding induced by litter size reduction leads to further obesity and other metabolic disorders, such as liver oxidative stress and microsteatosis at adulthood. We hypothesized that overfeeding causes an early redox imbalance at weaning, which could programme the animals to future liver dysfunction. Thus, we studied lipogenesis, adipogenesis, catecholamine status and oxidative balance in weaned overfed pups. To induce early overfeeding, litters were adjusted to three pups at the 3rd day of lactation (SL group). The control group contained 10 pups per litter until weaning (NL group). Peripheral autonomic nerve function was determined in vivo at 21 days old. Thereafter, pups were killed for further analysis. Differences were considered significant when P < 0.05. The SL pups presented with a higher visceral adipocyte area, higher content of lipogenic enzymes (ACC, FAS) and with a lower content of adipogenic factors (CEBP, PPARγ) in visceral adipose tissue (VAT). Although autonomic nerve activity and adrenal catecholamine production were not significantly altered, catecholamine receptor (ß3ADR) content was lower in VAT. The SL pups also presented with higher triglyceride, PPARγ, PPARα and PGC1α contents in liver. In plasma and liver, the SL pups showed an oxidative imbalance, with higher lipid peroxidation and protein oxidation. The SL group presented with a higher serum alanine aminotransferase (ALT). The early increase in lipogenesis in adipose tissue and liver in weaned overfed rats suggests that the higher oxidative stress and lower catecholamine content in VAT are associated with the early development of liver dysfunction and adipocyte hypertrophy.

Production of L1 protein from different types of HPV in Pichia pastoris using an integrative vector

Human papillomavirus (HPV) infection is the most common sexually transmitted disease in the world and is related to the etiology of cervical cancer. The most common high-risk HPV types are 16 and 18; however, the second most prevalent type in the Midwestern region of Brazil is HPV-33. New vaccine strategies against HPV have shown that virus-like particles (VLP) of the major capsid protein (L1) induce efficient production of antibodies, which confer protection against the same viral type. The methylotrophic yeast Pichia pastoris is an efficient and inexpensive expression system for the production of high levels of heterologous proteins stably using a wild-type gene in combination with an integrative vector. It was recently demonstrated that P. pastoris can produce the HPV-16 L1 protein by using an episomal vector associated with the optimized L1 gene. However, the use of an episomal vector is not appropriate for protein production on an industrial scale. In the present study, the vectors were integrated into the Pichia genome and the results were positive for L1 gene transcription and protein production, both intracellularly and in the extracellular environment. Despite the great potential for expression by the P. pastoris system, our results suggest a low yield of L1 recombinant protein, which, however, does not make this system unworkable. The achievement of stable clones containing the expression cassettes integrated in the genome may permit optimizations that could enable the establishment of a platform for the production of VLP-based vaccines.

Production of L1 protein from different types of HPV in Pichia pastoris using an integrative vector.

Human papillomavirus (HPV) infection is the most common sexually transmitted disease in the world and is related to the etiology of cervical cancer. The most common high-risk HPV types are 16 and 18; however, the second most prevalent type in the Midwestern region of Brazil is HPV-33. New vaccine strategies against HPV have shown that virus-like particles (VLP) of the major capsid protein (L1) induce efficient production of antibodies, which confer protection against the same viral type. The methylotrophic yeast Pichia pastoris is an efficient and inexpensive expression system for the production of high levels of heterologous proteins stably using a wild-type gene in combination with an integrative vector. It was recently demonstrated that P. pastoris can produce the HPV-16 L1 protein by using an episomal vector associated with the optimized L1 gene. However, the use of an episomal vector is not appropriate for protein production on an industrial scale. In the present study, the vectors were integrated into the Pichia genome and the results were positive for L1 gene transcription and protein production, both intracellularly and in the extracellular environment. Despite the great potential for expression by the P. pastoris system, our results suggest a low yield of L1 recombinant protein, which, however, does not make this system unworkable. The achievement of stable clones containing the expression cassettes integrated in the genome may permit optimizations that could enable the establishment of a platform for the production of VLP-based vaccines.

Pre-clinical validation of a vaginal cream containing copaiba oil (reproductive toxicology study).

The aims of this study was to evaluate the effects of oil-resin of Copaiba (Copaifera duckei Dwyer), aired in vaginal cream on the reproductive performance of female rats (Rattus norvegicus). To determine the components of the C. duckei oleoresin, gas chromatography coupled with mass spectrometry (CG-MS) was used, and considering the trans-caryophyllene sesquiterpene as a phytochemical marker in the oleoresin. Due to the extensive use of copaiba oleoresin in the suppository form for gynecological infections, an evaluation was carried out on the effects of copaiba oleoresin (Copaifera duckei Dwyer), delivered in a vaginal cream, on the reproductive performance of female Wistar rats. For this purpose, three groups (n=5-6/group) of female rats were treated as follows: 1--vaginal cream of copaiba oleoresin (28.6 mg/kg), 2--base vaginal cream and 3--control (physiological saline 0.9%), administered intravaginally, for 30 days before pregnancy, and from day zero to day 20 during pregnancy. Laparotomy was performed on the 21st day of pregnancy, followed by the determination of reproductive variables: number of live and dead fetuses, mass of the fetuses and placentas, number of implantations and resorptions, number of corpora lutea, pre- and post-implantation loss, and analyses of the fetuses with regard to external and internal anomalies and/or malformations (skeletal and visceral). The trans-caryophyllene present in the sample is suggested as a phytochemical marker and the results of this study demonstrate an absence of maternal toxicity and foetotoxicity embryofoetotoxicity at the dose administered, corresponding to ten times the recommended dose for use in humans. Accordingly, no significant statistical difference was observed between the treated and control groups, for the variables analyzed. Thus, it is concluded that the vaginal cream containing 2.5% copaiba oleoresin is safe during gestation, in female rats (Rattus norvegicus) of the Wistar strain.

Unintentional subdural catheter placement during labor analgesia shows typical radiological pattern but atypical response to the Tsui test.

Subdural injection may be associated with abnormally extensive or limited spread of local anesthetics during neuraxial anesthesia. This complication is difficult to diagnose clinically. Radiological imaging is the gold standard for confirming the location of subdural catheter, but electrical stimulation of the catheter has also been described as a useful diagnostic tool. We present the case of an obstetric patient with unintentional subdural catheter placement that presented as a failed epidural block associated with severe upper back and scapular pain on catheter injection. Electrical stimulation of the catheter did not elicit muscle contractions until a current of 4 mAmp was attained, which is the response pattern of epidural placement. Subdural location of the catheter was subsequently confirmed by contrast radiography. This case adds to the evidence that subdural catheters are difficult to identify clinically, and that electrical stimulation may not differentiate them from epidural catheters.

Effect of nicotine in matrix mineralization by human bone marrow and Saos-2 cells cultured on the surface of plasma-sprayed titanium implants.

Smoking has an established negative impact in the clinical outcome of dental implants. This work analyses the response of human osteoblastic cells to nicotine, at the surface of plasma-sprayed commercial titanium implants. Human bone marrow (HBM) and Saos-2 cells, seeded on the surface of titanium implants and cultured in experimental conditions favoring osteoblastic differentiation, were exposed continuously to nicotine (0.0001 to 0.5 mg mL(-1)) and characterized for cell proliferation and function. Exposure of HBM cells resulted in increased cell proliferation, higher alkaline phosphatase (ALP) activity, and earlier onset of matrix mineralization at levels up to 0.2 mg mL(-1), an initial inhibitory effect in cell growth and functional activity followed by a recovery in the presence of 0.3 mg mL(-1) and a dose-dependent deleterious effect at higher levels. By contrast, exposure to nicotine did not affect cell proliferation of Saos-2 cells at levels up to 0.2 mg mL(-1), and caused only a small positive effect in ALP activity in the presence of 0.05 and 0.1 mg mL(-1); however, matrix mineralization by Saos-2 cells also occurred earlier in the cultures exposed to levels of nicotine up to 0.1 mg mL(-1). Higher concentrations caused dose-dependent inhibitory effects. Considering the high diffusion potential of nicotine, results suggest a local role of nicotine in modulating bone formation events at the implant surface.

Anti-inflammatory and antinociceptive activities of extract, fractions and populnoic acid from bark wood of Austroplenckia populnea.

Austroplenckia populnea (Reiss) Lund is a Brazilian plant from "cerrado", which belongs to Celastraceae family, popularity know as "marmelinho-do campo, mangabeira-brava, mangabarana, vime and maria-mole". This plant is used in folk medicine to treat dysenteries and inflammatory disorders, such as rheumatism. Austroplenckia populnea bark hydroalcoholic crude extract, and its hexane, chloroform and ethyl acetate fractions, obtained by partition, as well as the isolated populnoic acid were investigated for their anti-inflammatory (carrageenan, dextran and histamine-induced rat paw oedema, histamine-induced increase in vascular permeability, and granulomatous tissue induction) and analgesic activities (writhing and hot plate tests). The ED(50) (oral) of the crude extract for the inhibition of carragenan-induced rat paw oedema assay was determined to be 200 mg/kg, which was also used in the assays with the extract and its fractions in all other experiments. Populnoic acid was administered in the dose of 50 mg/kg. Crude extract, hexane and chloroform fractions (200 mg/kg), and indomethacin (10 mg/kg) inhibited significantly (p<0.05) the formation of the carrageenan-induced rat paw oedema, measured in third hour of experiment (peak of oedema formation) by 43.2%, 37.3%, 31.1% and 59.3%, respectively. There was a significant reduction (p<0.05) in dextran-induced rat paw oedema in all groups, while in the assay using histamine as the oedematogenic agent, only the groups treated with populnoic acid (50 mg/kg) and cyproheptadine (10 mg/kg) displayed significant reduction (p<0.05). The populnoic acid and cyproheptadine reduced the peak of oedema formation (1st hour) by 41.3% and 34.7%, respectively. Only for the groups treated with populnoic acid (50 mg/kg) and cyproheptadine (10 mg/kg) it was observed a significant (p<0.05) reduction in histamine-induced increase in vascular permeability (44.8% and 80.3%, respectively). Granulomatous tissue formation was significantly inhibited (p<0.05) by both hexane fraction (46.0%) and dexamethasone (66.2%). In the analgesic assays, the crude extract and its hexane and chloroform fractions, as well as indomethacin diminished significantly the number of writhings (p<0.05) by 69.6%, 47.2%, 44.8% and 62.8%, respectively. On the other hand, none assayed sample displayed significant result in the hot plate test. Based on the obtained results it is suggested that extracts of Austroplenckia populnea bark and populnoic acid display anti-inflammatory activity, supporting its folkloric use to treat inflammatory disorders.

Topical antiinflammatory and analgesic activities of Copaifera duckei dwyer.

The oleoresin of several Copaifera species is used widely in the Amazonian Region mainly as a topical antiinflammatory and healing agent. The topical analgesic and antiinflammatory activities of Copaifera duckei oleoresin, whose terpenoidal chemical composition has been characterized, are now examined. Antiinflammatory activity was evaluated in rats using the carrageenin-induced paw edema and the granuloma tests, and in mice by the croton oil-induced dermatitis test. Analgesic activity was determined in mice using the writhing test method. In the carrageenin-induced edema and granuloma tests the oleoresin in a dose of 1,802 mg/kg inhibited the edema by 18% and granuloma by 42% (p < 0.05), this last result similar to that observed with dexamethasone. Topical doses of 517 mg/kg, 1,035 mg/kg and 1,802 mg/kg produced 52%, 58% and 62% (p < 0.05) reduction of the edema induced by croton oil, respectively, and 48%, 56% and 65% inhibition of the writhing process (p < 0.05). These results suggest that the Copaifera duckei oleoresin has topical antiinflammatory and analgesic activities.

Anti-inflammatory properties of kefir and its polysaccharide extract.

Kefir is a fermented beverage originating form the Caucasian regions composed of a number of bacteria and yeasts living together in polysaccharide grains secreted by them. Kefir can be considered a probiotic source as it presents anti-bacterial, anti-mycotic, anti-neoplasic and immunomodulatory properties. Aiming to appraise a possible anti-inflammatory effect of kefir we conducted cotton-induced granuloma and paw oedema assays in rats, the latter using carrageenan, dextran and histamine as stimuli. Kefir samples were thawed and continuously cultured during 15 days both in a molasses solution (50 g/l) and in cow's milk. A polysaccharide extract isolated from the grains (kefiran) was also tested in cotton-pellet experiments. The results showed significant inhibition in the formation of granuloma tissue for all the test groups, as compared to the blank group. Kefir suspensions in molasses presented an inhibition of 41 +/- 3% for the inflammatory process, fermented milk prepared from kefir showed 44 +/- 6% inhibition and kefiran extract 34 +/- 15%. Rat paw oedema also showed significant decreases with the mediators. Dextran-induced oedema was completely inhibited at 1 h after input, with a 76% inhibition after 2 h. Carrageenan stimulus was inhibited 62% after the 3rd hour, and histamine by 52% after the 2nd hour. These results points to the existence of anti-inflammatory prebiotic compounds present in symbiotic cultures of kefir growing in both aqueous and milky suspensions.