Approaches for recombinant human factor IX production in serum-free suspension cultures.

OBJECTIVE: To establish a serum-free suspension process for production of recombinant human factor IX (rhFIX) based on the human cell line HEK 293T by evaluating two approaches: (1) serum-free suspension adaptation of previously genetic modified cells (293T-FIX); and (2) genetic modification of cells already adapted to such conditions (293T/SF-FIX). RESULTS: After 10 months, 293T-FIX cells had become adapted to FreeStyle 293 serum-free medium (SFM) in Erlenmeyer flasks. After 48 and 72 h of culture, 2.1 µg rhFIX/ml and 3.3 µg rhFIX/ml were produced, respectively. However, no biological activity was detected. In the second approach, wild-type 293T cells were adapted to the same SFM (adaptation process took only 2 months) and then genetically modified for rhFIX production. After 48 h of culture, rhFIX reached 1.5 µg/ml with a biological activity of 0.2 IU/ml, while after 72 h, the production was 2.4 µg/ml with a biological activity of 0.3 IU/ml. CONCLUSION: The findings demonstrate that the best approach to establish an rhFIX production process in suspension SFM involves the genetic modification of cells already adapted to the final conditions. This approach is time saving and may better ensure the quality of the produced protein.

Antioxidant, analgesic and anti-inflammatory effects of lavender essential oil.

Several studies have investigated the antinociceptive, immunomodulatory and anti-inflammatory properties of compounds found in the lavender essential oil (LEO), however to date, there is still lack of substantial data. The objective of this study was to assess the antioxidant, anti-inflammatory and antinociceptive effects of lavender essential oil. The 1,1-diphenyl-2-picrylhydrazyl radical decolorization assay was used for antioxidant activity evaluation. The anti-inflammatory activity was tested using two models of acute inflammation: carrageenan-induced pleurisy and croton oil-induced ear edema. The antinociceptive activity was tested using the pain model induced by formalin. LEO has antioxidant activity, which is dose-dependent response. The inflammatory response evoked by carrageenan and by croton oil was reduced through the pre-treatment of animals with LEO. In the pleurisy model, the drug used as positive control, dexamethasone, was more efficacious. However, in the ear swelling, the antiedematogenic effect of the oil was similar to that observed for dexamethasone. In the formalin test, LEO consistently inhibited spontaneous nociception and presented a similar effect to that of tramadol. The results of this study reveal (in vivo) the analgesic and anti-inflammatory activities of LEO and demonstrates its important therapeutic potential.

Evaluation of in vivo and in vitro efficacy of solasonine/solamargine-loaded lipid-polymer hybrid nanoparticles against bladder cancer.

Solasonine (SS) and solamargine (SM) are alkaloids known for their antioxidant and anticancer properties, which can be further enhanced by encapsulating them in nanoparticles. This led to a study on the potential therapeutic benefits of SS and SM against bladder cancer when encapsulated in lipid-polymer hybrid nanoparticles (LPHNP). The LPHNP loaded with SS/SM were prepared using the emulsion and sonication method and their physical-chemical properties characterized. The biological effects of these nanoparticles were then tested in both 2D and 3D bladder cancer cell culture models, as well as in a syngeneic orthotopic mouse model based on the MB49 cell line and ethanol epithelial injury. The LPHNP-SS/SM had an average size of 130 nm, a polydispersity index of 0.22 and a positive zeta potential, indicating the presence of chitosan coating on the nanoparticle surface. The dispersion of LPHNP-SS/SM was found to be monodispersed with a span index of 0.539, as measured by nanoparticle tracking analysis (NTA). The recrystallization index, calculated from DSC data, was higher for the LPHNP-SS/SM compared to LPHNPs alone, confirming the presence of alkaloids within the lipid matrix. The encapsulation efficiency (EE%) was also high, with 91.08 % for SS and 88.35 % for SM. Morphological analysis by AFM and Cryo-TEM revealed that the nanoparticles had a spherical shape and core-shell structure. The study showed that the LPHNP-SS/SM exhibited mucoadhesive properties by physically interacting with mucin, suggesting a potential improvement in interaction with mucous membrane. Both the free and nanoencapsulated SS/SM demonstrated dose-dependent cytotoxicity against bladder cancer cell lines after 24 and 72 h of treatment. In 3D bladder cell culture, the nanoencapsulated SS/SM showed an IC50 two-fold lower than free SS/SM. In vivo studies, the LPHNP-SS/SM displayed an antitumoral effect at high doses, leading to a significant reduction in bladder volume compared to the positive control. However, there were observed instances of systemic toxicity and liver damage, indicated by elevated levels of transaminases (TGO and TGP). Overall, these results indicate that the LPHNPs effectively encapsulated SS/SM, showing high encapsulation efficiency and stability, along with promising in vitro and in vivo antitumoral effects against bladder cancer. Further evaluation of its systemic toxicity effects is necessary to ensure its safety and efficacy for potential clinical application.

Brazilian red propolis extract free and encapsulated into polymeric nanoparticles against ovarian cancer: formulation, characterisation and biological assays in 2D and 3D models.

Cancer incidence worldwide is alarming and among the cancers that affect women ovarian cancer is the most fatal. Many side effects are associated with conventional therapies and none of them are completely effective, so the development of new treatments is necessary. Brazilian red propolis extract is a natural product with complex composition and great potential for cancer treatment. However, its clinical application is harmed due to unfavourable physicochemical characteristics. To enable its application encapsulation in nanoparticles can be used. OBJECTIVES: The aims of this work were to develop polymeric nanoparticles with Brazilian red propolis extract and compare their action with the free extract against ovarian cancer cells. METHODS: Box Behnken design was used and nanoparticles were characterised using the techniques dynamic light scattering, nanoparticle tracking analysis, transmission electron microscopy, differential scanning calorimetry and encapsulation efficiency. Activity against OVCAR-3 was also tested on 2D and 3D models. KEY FINDINGS: Nanoparticles' sizes were ~200 nm with monomodal size distribution, negative zeta potential, spherical shape and with extract molecularly dispersed. Encapsulation efficiency was above 97% for the biomarkers chosen. Nanoparticles had greater efficacy in comparison with free propolis in OVCAR-3. CONCLUSIONS: So far, the nanoparticles here described have the potential to be a chemotherapy treatment in the future.

Phenolic enriched extract of Baccharis trimera presents anti-inflammatory and antioxidant activities.

Baccharis trimera is a plant popularly used as a tea and to treat gastrointestinal diseases and inflammatory processes as well. The total phenolic content was determined and the antioxidant and anti-inflammatory activities of six extracts (dichloromethane, ethyl acetate, butanol, aqueous, saponin and phenolic) from B. trimera were evaluated. Using carrageenan-induced pleurisy as a model of acute inflammation, the phenolic extract at 15 mg/kg decreased significantly the analyzed parameters when compared to the carrageenan group ( p < 0.05), thus showing potential anti-inflammatory activity. The total phenolic content and antioxidant activity were evaluated by the Folin-Ciocalteau and DPPH methods, respectively. Phenolic and ethyl acetate extracts presented higher antioxidant activity ( p < 0.05) than ascorbic acid. The phenolic extract also showed the highest antioxidant potential in relation to the other extracts, thus suggesting that the antioxidant and anti-inflammatory activities were due to the presence of phenolic compounds.

Anti-inflammatory and immunomodulatory effects of RDV-8 [C18H22N2O2S (ethyl 1-butyl-6-methyl-2-phenyl-4-thioxo-1,4-dihydropyrimidine-5-carboxylate)] in a rat model of induced pleurisy and in vitro lymphoproliferation.

RDV-8 [C(18)H(22)N(2)O(2)S (ethyl 1-butyl-6-methyl-2-phenyl-4-thioxo-1,4-dihydropyrimidine-5-carboxylate)] is derived from the 4-thioxopyrimidine, and presents important clinical effects. The present study explored the RDV-8 effects in the proliferation of human peripheral blood mononuclear cells (PBMCs), as well as in a pleurisy-induced rat model. PBMCs were directly plated in four different RDV-8 concentrations (0.0125, 0.025, 0.05 and 0.1 mg/mL). RDV-8 decreased cell proliferation and monocyte chemotactic protein 1 synthesis. The interleukin 1 levels and the cytotoxic effect were not significantly affected by RDV-8 treatment. In the carrageenan-induced pleurisy model, the RDV-8 (3 mg/kg) treatment induced a significant reduction in the exudate volume, in the polymorphonuclear leukocyte migration and in the pleural exudate NO levels. The results indicate that RDV-8 may have an immunomodulatory effect, as well as anti-inflammatory actions suggesting that it could represent a new strategy in the inflammatory response modulation.

Cytotoxic and chemosensitizing effects of glycoalkaloidic extract on 2D and 3D models using RT4 and patient derived xenografts bladder cancer cells.

Glycoalkaloids have been widely demonstrated as potential anticancer agents. However, the chemosensitizing effect of these compounds with traditional chemotherapeutic agents has not been explored yet. In a quest for novel effective therapies to treat bladder cancer (BC), we evaluated the chemosensitizing potential of glycoalkaloidic extract (GE) with cisplatin (cDDP) in RT4 and PDX cells using 2D and 3D cell culture models. Additionally, we also investigated the underlying molecular mechanism behind this effect in RT4 cells. Herein, we observed that PDX cells were highly resistant to cisplatin when compared to RT4 cells. IC50 values showed at least 2.16-folds and 1.4-folds higher in 3D cultures when compared to 2D monolayers in RT4 cells and PDX cells, respectively. GE + cDDP inhibited colony formation (40%) and migration (28.38%) and induced apoptosis (57%) in RT4 cells. Combination therapy induced apoptosis by down-regulating the expression of Bcl-2 (p < 0.001), Bcl-xL (p < 0.001) and survivin (p < 0.01), and activating the caspase cascade in RT4 cells. Moreover, decreased expression of MMP-2 and 9 (p < 0.01) were observed with combination therapy, implying its effect on cell invasion/migration. Furthermore, we used 3D bioprinting to grow RT4 spheroids using sodium alginate-gelatin as a bioink and evaluated the effect of GE + cDDP on this system. Cell viability assay showed the chemosensitizing effect of GE with cDDP on bio-printed spheroids. In summary, we showed the cytotoxicity effect of GE on BC cells and also demonstrated that GE could sensitize BC cells to chemotherapy.

Cell spheroids as a versatile research platform: formation mechanisms, high throughput production, characterization and applications.

Three-dimensional (3D) cell culture has tremendous advantages to closely mimic thein vivoarchitecture and microenvironment of healthy tissue and organs, as well as of solid tumors. Spheroids are currently the most attractive 3D model to produce uniform reproducible cell structures as well as a potential basis for engineering large tissues and complex organs. In this review we discuss, from an engineering perspective, processes to obtain uniform 3D cell spheroids, comparing dynamic and static cultures and considering aspects such as mass transfer and shear stress. In addition, computational and mathematical modeling of complex cell spheroid systems are discussed. The non-cell-adhesive hydrogel-based method and dynamic cell culture in bioreactors are focused in detail and the myriad of developed spheroid characterization techniques is presented. The main bottlenecks and weaknesses are discussed, especially regarding the analysis of morphological parameters, cell quantification and viability, gene expression profiles, metabolic behavior and high-content analysis. Finally, a vast set of applications of spheroids as tools forin vitrostudy model systems is examined, including drug screening, tissue formation, pathologies development, tissue engineering and biofabrication, 3D bioprinting and microfluidics, together with their use in high-throughput platforms.

A Simple Three-Dimensional In Vitro Culture Mimicking the In Vivo-Like Cell Behavior of Bladder Patient-Derived Xenograft Models.

Patient-derived xenograft (PDX) models allow for personalized drug selection and the identification of drug resistance mechanisms in cancer cells. However, PDX models present technical disadvantages, such as long engraftment time, low success rate, and high maintenance cost. On the other hand, tumor spheroids are emerging as an in vitro alternative model that can maintain the phenotype of cancer cells long enough to perform all assays and predict a patient's outcome. The present work aimed to describe a simple, reproducible, and low-cost 3D in vitro culture method to generate bladder tumor spheroids using human cells from PDX mice. Cancer cells from PDX BL0293 and BL0808 models, previously established from advanced bladder cancer, were cultured in 96-well round-bottom ultra-low attachment (ULA) plates with 5% Matrigel and generated regular and round-shaped spheroids (roundness > 0.8) with a diameter larger than 400 µm and a hypoxic core (a feature related to drug resistance in solid tumors). The responses of the tumor spheroids to the antineoplastic drugs cisplatin, gemcitabine, and their combination were similar to tumor responses in in vivo studies with PDX BL0293 and BL0808 mice. Therefore, the in vitro 3D model using PDX tumor spheroids appears as a valuable tool that may predict the outcome of in vivo drug-screening assays and represents a low-cost strategy for such purpose.

Multiple stab wounds on the left side of the chest in a patient with Situs Inversus Totalis: A lifesaving coincidence.

INTRODUCTION: Situs Inversus Totalis (SIT) is a rare finding of complete reversal of the thoracic and abdominal organs with an estimated incidence of 0.005%-0.01% in the population. Severe trauma has not been reported in this population. We present a case of multiple chest stab wounds in a patient with previously unknown SIT. PRESENTATION OF CASE: A 39-year-old male was admitted to the emergency room with multiple stab wounds on the left side of the chest. Upon admission the patient was hypotensive, with miosis and intubated. Significant ECG findings were an inverted P wave, inverted QRS complex and inverted T wave in V1. A chest CT scan showed SIT, hemopneumothorax on the left side and, despite multiple stab wounds on the left side of the chest, no cardiac damage. The surgical team decided for a conservative approach and the patient remained in the ICU for two days. After five days he was discharged in good clinical conditions. DISCUSSION: SIT generally does not have a clinical relevance throughout the patients life and most diagnoses are coincidental. However, when discovered in acute surgical cases, it requires an accurate evaluation by the surgical team due to anatomical differences that may produce undesirable outcomes in emergency cases such as appendicitis and general trauma. CONCLUSION: There have been very few reports of SIT and trauma in the medical literature. This might be the first ever reported case of a patient with SIT who suffered multiple stab wounds on the left side of the chest and was saved because of his condition.

Development, characterization and biological in vitro assays of paclitaxel-loaded PCL polymeric nanoparticles.

Adenocarcinoma is the most lethal gynecologic tumor and treatment usually consists in surgery followed by chemotherapy. However, the chemotherapy benefits are eventually limited due to drug toxicity to normal tissues and cells, which cause several and harsh side effects. Paclitaxel (PCX) is the drug of first choice for ovarian cancer treatment, but it has low aqueous solubility, which reduces its bioavailability. Thus, in the commercial drug, Taxol®, PCX is solubilized in a mixture of toxic surfactants. The development of drug nanocarriers has been investigated to promote the reduction of toxic effects and increase the safety and therapeutic efficacy of PCX. The aim of this work was the development and characterization of PCX loaded nanoparticles (PNPCX) and evaluation of in vitro efficacy of developed system using adenocarcinoma cell line. The nanocarrier was successfully obtained using nanoprecipitation technique. The results showed that the PNPCX-A had a particle size distribution around 140 nm and polydispersity index smaller than 0.1, with high PCX encapsulation efficiency. The results obtained were suitable for the intravenous administration route and promotion of passive targeting in the tumor microenvironment. The in vitro cytotoxicity assays of SKOV-3 cell line demonstrated that PNPCX-A was able to release PCX and reduce cell viability. The flow cytometry assays first reported that a nanostructured system with such composition (PNPCX-A) presented a time dependent cellular uptake, showing the ability of nanocarrier to be internalized. PNPCX-A present a distinguish potential for ovarian cancer therapy optimization. In vivo studies are needed to confirm the in vitro results and provide additional data regarding safety and efficacy of ovarian cancer treatment.

Assessing the cytotoxic potential of glycoalkaloidic extract in nanoparticles against bladder cancer cells.

OBJECTIVE: This study proposed to use the nanotechnology to deliver glycoalkaloidic extract (AE) to bladder cancer cells, evaluating their activity in 2D and 3D models and the biological mechanism of cell death. METHODS: NPs were prepared by nanoprecipitation method using polylactic acid (PLA) and characterized considering their size, charge, particle concentration and stability. The cytotoxicity was evaluated in 2D and 3D model, and the apoptosis and cell cycle were investigated using flow cytometry. KEY FINDINGS: NPs loading AE (NP-AE) had diameter around 125 ± 6 nm (PdI <0.1) and negative charge. The encapsulation efficiency of SM and SS was higher than 85% for both compounds. The obtained formulation showed a significant in-vitro cytotoxic effect against RT4 cells in a dose-dependent manner with IC50 two fold lower than the free AE. The cytotoxic effect of NP-AE was mediated by apoptosis and cell cycle arrested in the S phase. RT4 cells cultured under 3D conditions exhibited a higher resistance to the treatments (IC50 ~ three fold higher than in 2D cell culture). CONCLUSION: The NP-AE might be a promising nanocarrier to load and deliver glycoalkaloids against bladder cancer.

Serological profile of potential solid organ donors in Santa Catarina, Brazil.

OBJECTIVES: Serological tests are essential for the donation process. We performed a study to identify the seroprevalence of cytomegalovirus (CMV), toxoplasmosis, HIV, Chagas disease, HTLV, hepatitis B virus (HBV), hepatitis C virus (HCV) and Lues among our potential donors. METHODS: Among sera of 233 potential donors tested between January 2006 and April 2007, only 97 resulted in effective donation. RESULTS: The seroprevalence of CMV immunoglobulin G (IgG) was 89.3%. Anti-HBc was positive in 63 samples (27%) and just three people were HBsAg antigen-positive. HIV, HCV, HTLV, and Chagas disease showed low prevalence among the potential donors. Toxoplasmosis IgG antibody had a high prevalence in the tested group. CONCLUSION: CMV and toxoplasmosis were prevalent in the whole sample.

Comparative Analysis of 3D Bladder Tumor Spheroids Obtained by Forced Floating and Hanging Drop Methods for Drug Screening.

Introduction: Cell-based assays using three-dimensional (3D) cell cultures may reflect the antitumor activity of compounds more accurately, since these models reproduce the tumor microenvironment better. Methods: Here, we report a comparative analysis of cell behavior in the two most widely employed methods for 3D spheroid culture, forced floating (Ultra-low Attachment, ULA, plates), and hanging drop (HD) methods, using the RT4 human bladder cancer cell line as a model. The morphology parameters and growth/metabolism of the spheroids generated were first characterized, using four different cell-seeding concentrations (0.5, 1.25, 2.5, and 3.75 × 104 cells/mL), and then, subjected to drug resistance evaluation. Results: Both methods generated spheroids with a smooth surface and round shape in a spheroidization time of about 48 h, regardless of the cell-seeding concentration used. Reduced cell growth and metabolism was observed in 3D cultures compared to two-dimensional (2D) cultures. The optimal range of spheroid diameter (300-500 µm) was obtained using cultures initiated with 0.5 and 1.25 × 104 cells/mL for the ULA method and 2.5 and 3.75 × 104 cells/mL for the HD method. RT4 cells cultured under 3D conditions also exhibited a higher resistance to doxorubicin (IC50 of 1.00 and 0.83 µg/mL for the ULA and HD methods, respectively) compared to 2D cultures (IC50 ranging from 0.39 to 0.43). Conclusions: Comparing the results, we concluded that the forced floating method using ULA plates was considered more suitable and straightforward to generate RT4 spheroids for drug screening/cytotoxicity assays. The results presented here also contribute to the improvement in the standardization of the 3D cultures required for widespread application.

Safety and patient subjective efficacy of using galvanopuncture for the treatment of striae distensae.

BACKGROUND: Striae distensae are linear atrophic dermal scars with associated epidermal atrophy. This recurrent skin disorder causes a significant cosmetic and psychologic concern and remains a therapeutic challenge, especially when they are mature and hypopigmented (striae alba). AIMS: In this prospective single-center study, we evaluated the efficacy, safety, and patient's satisfaction of galvanopuncture for the treatment of striae alba. PATIENTS/METHODS: Thirty-two female subjects with striae alba present on the buttocks were treated with galvanopuncture once a week over a period of 10 weeks. Photographs and a percentage category scale were used to assess striae improvement and patient's satisfaction. Biochemical analyses were performed to assess possible systemic inflammatory effects or oxidative stress induction by the treatment. RESULTS: All patients achieved a substantial increase in clinical improvement in their striae within 10 treatment sessions. Galvanopuncture did not induce any inflammatory effect; however, it reduced oxidative injury. CONCLUSION: The use of galvanopuncture for the treatment of striae alba demonstrated a significant improvement in the lesions with visible results. This study supports the high degree of patient's satisfaction and demonstrate the safe and effective use of galvanopuncture in the treatment of striae alba on several skin types.

Apendicectomia aberta versus apendicectomia videolaparoscópica: uma análise a curto prazo

Introdução: Apendicectomia é uma cirurgia frequente nas situações de emergência, podendo ser realizada por via aberta ou videolaparoscópica. Objetivo: Analisar resultados e possíveis diferenças entre apendicectomias aberta e videolaparoscópica. Almeja-se caracterizar epidemiologicamente pacientes operados e avaliar os desfechos de ambas as técnicas a curto prazo. Métodos: Análise de coorte histórica de 238 pacientes diagnosticados com apendicite aguda e submetidos à apendicectomia aberta ou videolaparoscópica no Hospital Regional de São José Dr. Homero de Miranda Gomes, em Santa Catarina, Brasil, no período de 01 de Maio de 2017 a 30 de Abril de 2018. Resultados: Dentre os pacientes analisados, 209 (87,8%) foram submetidos à apendicectomia aberta e 29 (12,2%) à apendicectomia videolaparoscópica. O número total de homens operados superou o de mulheres (141 versus 97), entretanto, houve mais mulheres operadas pela via videolaparoscópica (58,7%; p = 0,04). Nas duas abordagens, a apresentação transoperatória mais comum da apendicite aguda foi a fase supurativa e houve maior prevalência de doentes entre 21 e 30 anos. A via videolaparoscópica apresentou menor taxa de infecção do sítio operatório (3,4% versus 14,8% na via aberta), apesar dessa diferença não ser estatisticamente significativa. O tempo médio do ato operatório foi mais longo (p<0,01) nas cirurgias videolaparoscópicas (86 ± 27,37 minutos versus 62,7 ± 23,80 minutos). Nos quesitos tempo de internação hospitalar e dor abdominal pós-operatória, ambas as abordagens se demonstraram semelhantes. Conclusão: No hospital avaliado, a maioria das apendicectomias foram realizadas por via aberta e a maior parte das apendicectomias videolaparoscópicas foram realizadas em mulheres. A técnica videolaparoscópica apresentou maior tempo médio de execução. Não houve diferenças significativas em relação a taxas de infecção do sítio operatório, dor pós-operatória e tempo de internação hospitalar.

Riscos e ocorrências de eventos adversos na percepção de enfermeiros assistenciais / Risks and occurrences of adverse events in the perception of health care nurses

Objetivo analisar os riscos e ocorrências de eventos adversos em pacientes hospitalizados na perspectiva de enfermeiros. Métodos estudo de corte transversal, desenvolvido com 41 enfermeiros assistenciais. Utilizou-se instrumento autoaplicável intitulado Eventos Adversos Associados às Práticas de Enfermagem validado em Portugal e adaptado à realidade brasileira. Resultados a quantidade de vínculos empregatícios (p=0,019) e a carga horária semanal (p=0,002) se mostraram potencializadores de falhas nos cuidados assistenciais. Houve correlação positiva entre lesões por pressão e quedas (p<0,001), erros de medicamentos e lesões por pressão (p=0,004) e infecções relacionadas à assistência à saúde e erros de medicamentos (p=0,006). Conclusão a análise evidenciou que a ocorrência de eventos adversos nos cuidados assistenciais como lesões por pressão e infecções relacionadas à assistência à saúde foram as mais frequentes na percepção de enfermeiros. Dos domínios explorados, erros de medicamentos apontou entre os resultados com menor incidência.

Objective to analyze the risks and occurrences of adverse events in hospitalized patients from the perspective of nurses. Methods cross-sectional study, developed with 41 nurses. We used a self-administered instrument titled Adverse Events Associated with Nursing Practices validated in Portugal and adapted to the Brazilian reality. Results the amount of employment bonds (p=0.019) and weekly workload (p=0.002) foster failures in care assistance. There was a positive correlation between pressure and falls injuries (p<0.001), medication errors and pressure injuries (p=0.004) and infections related to health care and medication errors (p=0.006). Conclusion the analysis showed that the occurrence of adverse events in care such as pressure injuries and infections related to health care were the most frequent in the perception of nurses. Drug errors had the lowest incidence among the results of the explored domains.

Treatment with N-methyl-D-aspartate receptor antagonist (MK-801) protects against oxidative stress in lipopolysaccharide-induced acute lung injury in the rat.

Acute lung injury (ALI) and the acute respiratory distress syndrome (ARDS) are common syndromes that affect both clinical and surgical patients. This study describes the effects of a potent and specific N-methyl-d-aspartate receptor antagonist (MK-801) against oxidative stress in acute lung injury induced by intratracheal lipopolysaccharide (LPS) injection. This study was performed using male Wistar rats weighing 200-250g. Rats were randomly divided into four groups: control with isotonic saline instillation (n=6); LPS (100µg/100g of body weight) treated with saline (n=6); LPS treated with MK-801 (0.3mg/kg, intraperitoneally; n=6); LPS treated with MK-801 (0.3mg/kg, intratracheally; n=6). Twelve hours after the LPS instillation, rats were anesthetized and a bronchoalveolar lavage (BAL) was performed in order to determine the alveolar-capillary membrane alterations and the inflammatory infiltrate level. Blood and lung samples were isolated and assayed for oxidative stress variables and histopathologic analysis. The use of MK-801 decreased bronchoalveolar lavage fluid protein, LDH activity and inflammatory cells. Indeed, the treatment with MK-801 significantly attenuated lung oxidative damage and histopathologic alterations after LPS instillation. Our data provide the first experimental demonstration that MK-801 decreases oxidative stress and limits inflammatory response and alveolar disarray in lipopolysaccharide-induced acute lung injury.

Prevalência de incidentes relacionados à medicação em unidade de terapia intensiva / Prevalence of medication-related incidents in an intensive care unit

Objetivo Estimar a prevalência de incidentes relacionados à medicação em uma Unidade de Terapia Intensiva.Métodos Estudo transversal que incluiu 116 registros de internações hospitalares no período de 12 meses. O instrumento de pesquisa foi elaborado com base nas variáveis de estudo e validado por dois experts. A prevalência foi calculada considerando o número de internações expostas como numerador e o total de internações investigadas como denominador, calculando intervalo de confiança de 95%. Para a verificação de associação significativa entre as variáveis, utilizou-se o Teste Exato de Fisher, assumindo nível de significância máximo de 5% (p<0,05).Resultados Verificou-se que 113 internações foram expostas a pelo menos um tipo de incidente, totalizando 2.869 ocorrências, sendo 1.437 circunstâncias notificáveis, 1.418 incidentes sem dano, nove potenciais eventos adversos e cinco eventos adversos. Os incidentes aconteceram durante a fase da prescrição (45,4%) e a ausência de conduta dos profissionais de saúde frente aos incidentes foi identificada em 99% dos registros.Conclusão Estimou-se prevalência de 97,4% incidentes relacionados à medicação.

Objective To estimate the prevalence of medication-related incidents in an intensive care unit.Methods Cross-sectional study that included 116 records of hospitalizations within a 12-month period. The survey instrument was developed based on the study variables and was validated by two experts. The prevalence was calculated by considering the number of exposed hospitalizations as the numerator and the total of investigated hospitalizations as the denominator, calculating a 95% confidence interval. Fisher’s exact test assuming maximum significance level of 5% (p<0.05) was used to verify significant association.Results It was observed that 113 hospitalizations had been exposed to at least one type of incident, totaling 2,869 occurrences: 1,437 reportable circumstances, 1,418 no-harm incidents, 9 near-miss incidents and 5 adverse events. The incidents occurred during the prescription stage (45.4%) and the absence of information on the actions taken by the health professionals in relation to the incidents was identified in 99% of the records.Conclusion Prevalence of 97.4% of medication-related incidents was estimated.

Contusões cerebrais devido a trauma craniencefálico: princípios fisiopatológicos e conduta / Brain contusion due to cranio-encefalic trauma: conduct and physiopathologic principles

O trauma cranioencefálico (TCE) grave é uma das mais prevalecentes causas de morte em países industrializados. Os pacientes com TCE grave freqüentemente são vítimas de traumatismos multissistêmicos. Em conseqüência de trauma de crânio grave e moderado verifica-se lesão de parênquima cerebral em cerca de 55 por cento dos casos. As contusões cerebrais perfazem 45 por centro e 60 por cento das lesões traumáticas primárias e podem estar associadas a outras lesões intracranianas que envolvem mecanismo fisiopatológico semelhante, como hematoma subdural agudo e hemorragia meníngea traumática. Atualmente, a tendência dos neurotraumatologistas é atuar na prevenção de lesão neuronal secundária, que é a principal responsável pela deterioração clínica em portadores de contusões cerebrais. Normalmente o quadro neurológico está correlacioinado com o tamanho e a localização da contusão. Pacientes com contusão podem apresentar deterioração progressiva ou rápida. Deterioração rápida é geralmente encontrada em contusões bifrontais grandes e ponta de temporal. Inicialmente o tratamento clínico engloba uma série de medidas, como ventilação adequada, medidas posturais, sedação e analgesia, controle hidreletrolítico rigoroso, aporte nutricional, monitorização da PIC e hopotermia convencional. A conduta cirúrgica visa atenuar o desenvolvimento de edema citotóxico, evitável se o foco lesional for operado, reduzir o efeito de massa em lesões em região temporal e evitar ou reduzir lesões secundárias