Development of Radiosterilized Porcine Skin Electrosprayed with Silver Nanoparticles Prevents Infections in Deep Burns.

Extensive burns represent a significant challenge in biomedicine due to the multiple systemic and localized complications resulting from the major skin barrier loss. The functionalization of xenografts with nanostructured antibacterial agents proposes a fast and accessible application to restore barrier function and prevent localized bacterial contamination. Based on this, the objective of this work was to functionalize a xenograft by electrospray deposition with silver nanoparticles (AgNPs) and to evaluate its antibiofilm and cytotoxic effects on human fibroblasts. Initially, AgNPs were synthesized by a green microwave route with sizes of 2.1, 6.8, and 12.2 nm and concentrations of 0.055, 0.167, and 0.500 M, respectively. The AgNPs showed a size relationship directly proportional to the concentration of AgNO3, with a spherical and homogeneous distribution determined by high-resolution transmission electron microscopy. The surface functionalization of radiosterilized porcine skin (RPS) via electrospray deposition with the three AgNP concentrations (0.055, 0.167, and 0.500 M) in the epidermis and the dermis showed a uniform distribution on both surfaces by energy-dispersive X-ray spectroscopy. The antibiofilm assays of clinical multidrug-resistant Pseudomonas aeruginosa showed significant effects at the concentrations of 0.167 and 0.500 M, with a log reduction of 1.3 and 2.6, respectively. Additionally, viability experiments with human dermal fibroblasts (HDF) exposed to AgNPs released from functionalized porcine skin showed favorable tolerance, with retention of viability more significant than 90% for concentrations of 0.05 and 0.167 M after 24 h exposure. Antibacterial activity combined with excellent biocompatibility makes this biomaterial a candidate for antibacterial protection by inhibiting bacterial biofilms in deep burns during early stages of development.

Toxicological Evaluations in Macrophages and Mice Acutely and Chronically Exposed to Halloysite Clay Nanotubes Functionalized with Polystyrene.

Halloysite clay nanotubes (HNTs) have been proposed as highly biocompatible for several biomedical applications. Various polymers have been used to functionalize HNTs, but scarce information exists about polystyrene for this purpose. This work evaluated polystyrene-functionalized HNTs (FHNTs) by comparing its effects with non-FHNTs and innocuous talc powder on in vitro and in vivo models. Monocyte-derived human or murine macrophages and the RAW 264.7 cell line were treated with 0.01, 0.1, 1, and 100 µg mL-1 FHNTs, HNTs, or talc to evaluate the cytotoxic and cytokine response. Our results show that nanoclays did not cause cytotoxic damage to macrophages. Only the 100 µg mL-1 concentration induced slight proinflammatory cytokine production at short exposure, followed by an anti-inflammatory response that increases over time. CD1 mice treated with a single dose of 1, 2.5, or 5 mg Kg-1 of FHNTs or HNTs by oral and inhalation routes caused aluminum accumulation in the kidneys and lungs, without bodily signs of distress or histopathological changes in any treated mice, evaluated at 48 h and 30 days post-treatment. Nanoclay administration simultaneously by four different parenteral routes (20 mg Kg-1) or the combination of administration routes (parenteral + oral or parenteral + inhalation; 25 mg Kg-1) showed accumulation on the injection site and slight surrounding inflammation 30 days post-treatment. CD1 mice chronically exposed to HNTs or FHNTs in the bedding material (ca 1 mg) throughout the parental generation and two successive inbred generations for 8 months did not cause any inflammatory process or damage to the abdominal organs and the reproductive system of the mice of any of the generations, did not affect the number of newborn mice and their survival, and did not induce congenital malformations in the offspring. FHNTs showed a slightly less effect than HNTs in all experiments, suggesting that functionalization makes them less cytotoxic. Doses of up to 25 mg Kg-1 by different administration routes and permanent exposure to 1 mg of HNTs or FHNTs for 8 months seem safe for CD1 mice. Our in vivo and in vitro results indicate that nanoclays are highly biocompatible, supporting their possible safe use for future biomedical and general-purpose applications.

Improvement in extracellular secretion of recombinant L-asparaginase II by Escherichia coli BL21 (DE3) using glycine and n-dodecane.

L-asparaginase II (ASNase) is the biopharmaceutical of choice for the treatment of acute lymphoblastic leukaemia. In this study, E. coli BL21 (DE3) transformed with the pET15b + asnB vector which expresses recombinant ASNase was used as a source to obtain this enzyme. The ideal conditions to produce ASNase would be a high level of secretion into the extracellular medium, which depends not only on the application of molecular biology techniques but also on the development of a strategy to modify cell permeability such as the addition of substances to the culture medium that stimulate destabilisation of structural components of the cell. Thus, the growth of E. coli BL21 (DE3) in modified Luria-Bertani broth, supplemented with 0.8% (w/v) glycine and 6% (v/v) n-dodecane, increased the total yield of ASNase by about 50% (15,108 IU L-1) and resulted in a 16-fold increase in extracellular enzymatic productivity (484 IU L-1 h-1), compared to production using the same medium without addition of these substances. Most of the enzyme (89%) was secreted into the culture medium 24 h after the induction step. This proposed approach presents a simple strategy to increase extracellular production of ASNase in E. coli.

Uremic endothelial-derived extracellular vesicles: Mechanisms of formation and their role in cell adhesion, cell migration, inflammation, and oxidative stress.

p-Cresyl sulfate (PCS), indoxyl sulfate (IS), and inorganic phosphate (Pi) are uremic toxins found in chronic kidney disease (CKD) that are closely related to endothelial extracellular vesicles (EVs) formation. The present study aimed to understand the role of EVs and their role in cell adhesion and migration, inflammation, and oxidative stress. Human endothelial cells were treated with PCS, IS, and Pi in pre-established uremic and kinetic recommendations. EVs were characterized using scanning electron microscopy, flow cytometry, and NanoSight assays. The concentrations of EVs were established using Alamar Blue and MTT assays. Cell adhesion to extracellular matrix proteins was analyzed using an adhesion assay. Inflammation and oxidative stress were assessed by vascular cell adhesion molecule-1 expression/monocyte migration and reactive oxygen species production, respectively. The capacity of EVs to stimulate endothelial cell migration was evaluated using a wound-healing assay. Our data showed that endothelial cells stimulated with uremic toxins can induce the formation of EVs of different sizes, quantities, and concentrations, depending on the uremic toxin used. Cell adhesion was significantly (P < 0.01) stimulated in cells exposed to PCS-induced extracellular vesicles (PCSEVs) and inorganic phosphate-induced extracellular vesicles (PiEVs). Cell migration was significantly (P < 0.05) stimulated by PCSEVs. VCAM-1 expression was evident in cells treated with PCSEVs and IS-induced extracellular vesicles (ISEVs). EVs are not able to stimulate monocyte migration or oxidative stress. In conclusion, EVs may be a biomarker of endothelial injury and the inflammatory process, playing an important role in cell-to-cell communication and pathophysiological processes, although more studies are needed to better understand the mechanisms of EVs in uremia.

Obtención de hidrolizados proteicos de leguminosas usando una proteasa recombinante de Pseudomonas aeruginosa M211 / Obtaining protein hydrolysates from leguminous using a recombinant protease from Pseudomonas aeruginosa M211

El género Pseudomonas es una fuente importante de proteasas; sin embargo, su uso está restringido en la industria alimentaria. El clonaje permite aprovechar la capacidad catalítica de estas enzimas mediante su producción en microorganismos inocuos. Por otro lado, las leguminosas son fuentes ricas en proteínas, a partir de las cuales se pueden obtener compuestos con valor agregado mediante procesos de hidrólisis enzimática. En este estudio, se produjo y caracterizó una proteasa recombinante (PT4) alcalina y termoestable de Pseudomonas aeruginosa M211, para la obtención de hidrolizados proteicos de leguminosas. Para ello, el gen de la proteasa se clonó en el vector pJET1.2/blunt utilizando E. coli DHalfa como hospedero. El análisis de la secuencia nucleotídica parcial de la proteasa indicó un 99 % de similitud con Peptidasas de la Familia M4 de Pseudomonas aeruginosa. La enzima recombinante presentó un peso molecular de 80 kDa, demostró ser activa y estable en condiciones alcalinas y termófilas con un pH y temperatura óptimos de 8 y 60 °C, respectivamente, y fue inhibida por EDTA. Además, hidrolizó proteínas de semillas de Glycine max, Phaseolus lunatus, Lupinus mutabilis y Erythrina edulis, obteniéndose fracciones peptídicas menores a 40 kDa. Esta proteasa recombinante se podría utilizar en la elaboración de hidrolizados proteicos funcionales a partir proteínas de distintas fuentes y residuos agroalimentarios.

The genus Pseudomonas is an important source of proteases; however, in the food industry the use of this bacterium is restricted. Cloning allows for the use of the proteolytic activity of Pseudomonas proteases through their production in innocuous microorganisms. Leguminous are protein-rich sources from which value-added compounds can be obtained through enzymatic hydrolysis. In this study, an alkaline and thermostable recombinant protease (PT4) from Pseudomonas aeruginosa M211 was cloned and characterized in order to obtain protein hydrolysates from leguminous. Therefore, protease gene was cloned into the pJET1.2 / blunt vector using E. coli DHalpha as a host. Analysis of protease partial nucleotide sequence showed 99% homology with Peptidases M4 Family from Pseudomonas aeruginosa. The molecular weight of the recombinant enzyme was 80 kDa, it was active and stable under alkaline and thermophilic conditions, presented an optimum pH and temperature of 8 and 60 °C, respectively, and was inhibited by EDTA. In addition, it hydrolysed Glycine max, Phaseolus lunatus, Lupinus mutabilis y Erythrina edulis proteins, obtaining peptide fractions less than 40 kDa. This recombinant protease could be used in the elaboration of functional hydrolysates using protein from different sources and agricultural waste.

Hands Up, Now What?: Black Families' Reactions to Racial Socialization Interventions.

Given the heightened national attention to negative race-related issues and the subsequent community solution-oriented outcry (e.g., Black Lives Matter movement), it is crucial to address healing from racial discrimination for Black Americans. Clinical and community psychologists have responded by developing and implementing programs that focus on racial socialization and psychological wellness, particularly given disproportionate issues with utilization, access, and the provision of quality services within urban and predominantly Black communities. The aim of this article is to describe 2 applied programs (Engaging, Managing, and Bonding through Race and Family Learning Villages), which seek to address and heal racial stress through crucial proximal systems-families and schools-and to highlight participant reactions. These programs offer solutions through strengths-based and participatory approaches which draw from Black Americans' own protective mechanisms related to improved mental health. We conclude with a discussion on practice, assessments, and models specific to racial stress for researchers, practitioners, and consumers of mental health services.

Production and Characterization of Extremophilic Proteinases From a New Enzyme Source, Barrientosiimonas sp. V9.

Microbial proteases are widely used as commercial enzymes, which have an active role in several industrial processes. The aim of this study was to investigate the production and properties of extracellular proteases from Barrientosiimonas sp. strain V9. The cultivation conditions for protease production were studied using different carbon and nitrogen sources. Maximum protease production was obtained in medium containing 25 g L-1 sucrose, 7 g L-1 KNO3, and initial pH 7.0 at 35 °C and 150 rpm during 72 h. Under these conditions, maximum proteolytic activity reached 1200 U mL-1. The enzyme extract showed optimum activity at 60 °C, pH 9.0, and was stable from 30 to 50 °C within a pH range from 4.0 to 10.0 and NaCl concentration up to 2.5 M. The enzyme was stable in the presence of EDTA, urea, Triton X-100 and laundry detergent (sodium lauryl sulfate as main component). The addition of 1% sodium dodecyl sulfate, Tween-80, or Tween-20 increased the activity by 183% and 119% respectively, while 2-mercaptoethanol reduced the activity to 71%. Casein zymogram analysis revealed three hydrolysis zones suggesting that Barrientosiimonas sp. V9 expresses proteases with molecular weights about 60, 45, and 35 kDa, which were inhibited in the presence of phenylmethylsulfonyl fluoride. Barrientosiimonas sp. V9 produces halotolerant serine proteases with great biotechnological potential.

Arthroscopic treatment of lateral ankle instability. Is there a safe zone? An anatomic study.

BACKGROUND: The purpose of this study is to describe the relative location of superficial anatomic landmarks and likely location of structures at risk in order to predict the proximity of the later and avoid their injury during the arthroscopic treatment of lateral ankle instability. METHODS: Fifteen cadaver ankles were dissected. Based on superficial anatomic landmarks, the location and distances to the structures at risk (extensor tendons, peroneus tertius, peroneal tendons, main branch or intermediate branch of the superficial peroneal nerve, and the sural nerve) were measured. RESULTS: The distance from the lateral malleolus along the peroneus brevis to its intersection by the sural nerve was 38.5±10.5mm and from it to the superficial peroneal nerve was 32.0±7.4mm. Based on the minimum distances, a rectangular area of 25mm×22mm was obtained. The anterior talofibular ligament and the proximal border of the inferior extensor retinaculum were within this area. CONCLUSIONS: Our study suggests that based on superficial anatomic landmarks, it is possible to define an anatomic area in order to avoid structures at risk. In addition to the usual precautions, these anatomical references may contribute to lower the complication rate associated to the arthroscopic treatment of lateral ankle instability.

Fed-Batch Production of Saccharomyces cerevisiae L-Asparaginase II by Recombinant Pichia pastoris MUT s Strain.

L-Asparaginase (ASNase) is used in the treatment of acute lymphoblastic leukemia, being produced and commercialized only from bacterial sources. Alternative Saccharomyces cerevisiae ASNase II coded by the ASP3 gene was biosynthesized by recombinant Pichia pastoris MUT s under the control of the AOX1 promoter, using different cultivation strategies. In particular, we applied multistage fed-batch cultivation divided in four distinct phases to produce ASNase II and determine the fermentation parameters, namely specific growth rate, biomass yield, and enzyme activity. Cultivation of recombinant P. pastoris under favorable conditions in a modified defined medium ensured a dry biomass concentration of 31 gdcw.L-1 during glycerol batch phase, corresponding to a biomass yield of 0.77 gdcw.g glycerol - 1 and a specific growth rate of 0.21 h-1. After 12 h of glycerol feeding under limiting conditions, cell concentration achieved 65 gdcw.L-1 while ethanol concentration was very low. During the phase of methanol induction, biomass concentration achieved 91 gdcw.L-1, periplasmic specific enzyme activity 37.1 U.g dcw - 1 , volumetric enzyme activity 3,315 U.L-1, overall enzyme volumetric productivity 31 U.L-1.h-1, while the specific growth rate fell to 0.039 h-1. Our results showed that the best strategy employed for the ASNase II production was using glycerol fed-batch phase with pseudo exponential feeding plus induction with continuous methanol feeding.

Heavy Metal Enrichment Factors in Fluvial Sediments of an Amazonian Basin Impacted by Gold Mining.

Artisanal and small-scale gold mining (ASGM) has been performed in the southern Ecuadorian Amazon since the colonial period. However, its effects on fluvial systems have been poorly investigated. Thus, in order to calculate the normalized enrichment factors (NEF) of several heavy metals in fluvial sediments of the Zamora River basin (Ecuadorian Amazon), we analyzed bottom sediments along ASGM-affected and unaffected river sections. The results indicated that sediments of the Congüime River have NEF between 2.2 and 2.3 for Cu (moderate contamination) and higher than 3 for Mn, Zn, Pb, and Hg (severe contamination). Similarly, a severe contamination is also observed in the lower Nangaritza River, due to sediments of this sector have NEF > 3 for Zn, Pb, and Hg. Bottom sediments from the Nambija and Zamora rivers showed a severe contamination with Hg (NEF > 3), suggesting the existence of ASGM activities in the upper Zamora River basin.

Microbial cell disruption methods for efficient release of enzyme L-asparaginase.

The efficacy of a simple laboratory method for cell disruption based on the glass bead stirring, sonication, osmotic shock, freezing and grinding, or use of solvents and detergents was assessed in this study, via measurements of the release of total protein and L-asparaginase activity. Three different microbial sources of L-asparaginase were used: Escherichia coli BL21 (DE3), Leucosporidium muscorum, and Aspergillus terreus (CCT 7693). This study adjusted and identified the best procedure for each kind of microorganism. Sonication and glass bead stirring led to obtaining filamentous fungus cell-free extracts containing high concentrations of soluble proteins and specific activity; however, sonication was the best since it obtained 4.61 ± 0.12 IU mg-1 after 3 min of operation time. Mechanical methods were also the most effective for yeast cell disruption, but sonication was the technique which yielded a higher efficiency releasing 7.3 IUtotal compared to glass bead stirring releasing 2.7 IUtotal at the same operation time. For bacterium, sonication proved to be the best procedure due to getting the highest specific activity (9.01 IU mg-1) and total enzyme activity (61.7 IU). The data presented lead to conclude that the mechanical methods appeared to be the most effective for the disintegration of the all microbial cells studies. This is the first report related to the experimental comparison of L-ASNase extraction procedures from different microorganisms, which can also be used for extracting periplasm located enzymes from other organisms.

Pharmacological and toxicological study of a chemical-standardized ethanol extract of the branches and leaves from Eysenhardtia polystachya (Ortega) Sarg. (Fabaceae).

Eysenhardtia polystachya is used for the empirical treatment of cancer, infections, diarrhea, inflammation, and pain. This study identified, using GC-MS, the main chemical components in an ethanol extract of E. polystachya branches and leaves (EPE) and tested its cytotoxic, antimicrobial, anti-diarrheal, anti-inflammatory, and antinociceptive effects. The in vitro and in vivo toxicity of EPE was evaluated using the comet assay in human peripheral blood mononuclear cells (PBMC) and the acute toxicity test in mice, respectively. The cytotoxic and the antimicrobial effects were performed using the MTT assay and the minimum inhibitory concentration (MIC) test, respectively. The levels of pro-inflammatory mediators in LPS-stimulated macrophages were measured to evaluate the in vitro anti-inflammatory effects of EPE. The antidiarrheal (castor oil test, small intestine transit, and castor oil-induced enteropooling), and anti-inflammatory activities (TPA and carrageenan) of EPE were also performed. The antinociceptive actions of EPE were carried out with the following tests: acetic acid, formalin, and hot plate. The hypnotic and locomotor effects were analyzed using pentobarbital and a rotarod system, respectively. The main component in EPE was D-pinitol (26.93%). The antidiarrheal and antinociceptive effects of D-pinitol were also evaluated. EPE showed low in vitro toxicity (DNA damage in PBMC at concentrations higher than 200 µg/ml), and low in vivo toxicity (LD50 > 2000 mg/kg i.p. and p.o.). Furthermore, EPE lacked cytotoxic activity (IC50 > 300 µg/ml) on human cancer cells, but showed good antimicrobial effects in E. coli (MIC=1.56 µg/ml) and S. aureus (MIC = 0.78 µg/ml). In multi-drug resistant microorganisms, EPE showed MIC> 100 µg/ml. EPE exerted in vitro anti-inflammatory effects, mainly, by the decrease in the production of H2O2 (IC50 = 43.9 ±â€¯3.8 µg/ml), and IL-6 (73.3 ±â€¯6.9 µg/ml). EPE (ED50 =7.5 ±â€¯0.9 mg/kg) and D-pinitol (ED50 = 0.1 ±â€¯0.03 mg/kg) showed antidiarrheal activity, and antinociceptive effects in the acetic acid test with ED50 = 117 ±â€¯14.5 mg/kg for EPE and 33 ±â€¯3.2 mg/kg for D-pinitol. EPE showed also antinociceptive activity in the phase 2 of the formalin test (ED50 = 48.9 ±â€¯3.9 mg/kg), without inducing hypnotic effects or altering the locomotor activity in mice. The results here presented corroborate the folk medicinal use of Eysenhardtia polystachya in the treatment of infections, diarrhea, inflammation, and pain. D-pinitol, the main metabolite of EPE, showed antinociceptive and antidiarrheal effects with similar potency compared to standard drugs.

Manganese and Mercury Levels in Water, Sediments, and Children Living Near Gold-Mining Areas of the Nangaritza River Basin, Ecuadorian Amazon.

Artisanal and small-scale gold-mining activities performed in mountain areas of the Southern Ecuadorian Amazon have incorporated several heavy metals into the aquatic systems, thus increasing the risk of exposure in populations living in adjacent zones. Therefore, the objective of this study was to evaluate the contamination levels of mercury (Hg) and manganese (Mn) in several rivers of the Nangaritza River basin and assess the exposure in school-aged children residing near the gold-mining zones. River water and sediment samples were collected from a highly contaminated (HEx) and a moderately contaminated (MEx) zones. Hair Mn (MnH) and urinary Hg (HgU) levels were determined in school-aged children living in both zones. High concentrations of dissolved Mn were found in river waters of the HEx zone (between 2660 and 3990 µg l(-1)); however, Hg levels, in general, were lower than the detection limit (DL; <1.0 µg l(-1)). Similarly, Mn levels in sediments were also increased (3090 to 4086 µg g(-1)). Median values of MnH in children of the HEx and MEx zones were 5.5 and 3.4 µg g(-1), respectively, whereas the median values of HgU concentrations in children living in the HEx and MEx zones were 4.4 and 0.62 µg g-creat(-1), respectively. Statistically significant differences were observed between both biomarkers in children from the HEx and MEx zones. In addition, boys presented significantly greater MnH levels in both zones. The greater MnH values were found in children living in alluvial areas, whereas children living in the high mountain areas, where some ore-processing plants are located close to or inside houses and schools, had the greater HgU concentrations. In summary, the data reported in this paper highlights that artisanal and small-scale gold-mining activities can not only produce mercurial contamination, that can also release other heavy metals (such as Mn) that may pose a risk to human health.

Effect of Clinoptilolite and Sepiolite Nanoclays on Human and Parasitic Highly Phagocytic Cells.

Nanoclays have potential applications in biomedicine raising the need to evaluate their toxicity in in vitro models as a first approach to its biocompatibility. In this study, in vitro toxicity of clinoptilolite and sepiolite nanoclays (NC) was analyzed in highly phagocytic cultures of amoebas and human and mice macrophages. While amebic viability was significantly affected only by sepiolite NC at concentrations higher than 0.1 mg/mL, the effect on macrophage cultures was dependent on the origin of the cells. Macrophages derived from human peripheral blood monocytes were less affected in viability (25% decrease at 48 h), followed by the RAW 264.7 cell line (40%), and finally, macrophages derived from mice bone marrow monocytes (98%). Moreover, the cell line and mice macrophages die mainly by necrosis, whereas human macrophages exhibit increased apoptosis. Cytokine expression analysis in media of sepiolite NC treated cultures showed a proinflammatory profile (INFγ, IL-1α, IL-8, and IL-6), in contrast with clinoptilolite NC that induced lees cytokines with concomitant production of IL-10. The results show that sepiolite NC is more toxic to amoebas and macrophages than clinoptilolite NC, mostly in a time and dose-dependent manner. However, the effect of sepiolite NC was comparable with talc powder suggesting that both NC have low cytotoxicity in vitro.

Large foot schwannoma with bone invasion - a case report.

A rare case of a large foot schwannoma with an intraosseous component is presented. Due to suspicion of malignancy, an amputation had previously been proposed. When the patient presented to us, the prolonged clinical course as well as some elements of the imaging exams suggested benignancy and we decided for complete tumor excision. Intraoperative findings supported the benign character of the neoplasm and pathology would later confirm the diagnosis of a benign schwannoma. The outcome, at 18 months of follow-up was a fully functional limb. Recurrence must still be considered but malignant transformation is very rare in a solitary schwannoma and can be discounted. Solitary benign schwannomas have a negligible malignization potential independently of their size or osseous component. When benignancy is considered, primary total tumor excision, as part of a staged procedure, is a safe approach. It allows for a thorough pathologic examination and eventually for a definitive treatment.

[Prenatal diagnosis of choledochal cyst]. / Quiste de colédoco detectado prenatalmente.

The choledocal cyst is a defect of the biliary extrahepatic route, the incidence is 1 in 100-150,000 newborns. This paper reports the case of a female newborn with choledocal cyst detected prenatally, from a 32-year-old mother, 2nd term pregnancy, who was diagnosed in the routine obstetric ultrasound as an abdominal fetal cyst. An anatomic obstetric ultrasound confirmed the diagnosis that was defined of a probable hepatic origin. Newborn was delivered at 39 weeks by caesarean section, with weight of 3,980 g and Apgar score 9-9 in conventional time. Newborn presented a maxim bilirubin level of 16 mg, controlled with phototherapy; the hepatic function was normal. Ultrasound showed a choledocal cyst which measured 50 x 49 x 48 mm, with dilatation of the hepatic common conduct, the gall bladder was normal. The abdominal scan reported a cystic mass in the middle abdominal region of 44 x 53 x 52 mm confirming a choledocal cyst. The diagnosis was confirmed after surgery. Patient had a satisfactory post-surgical evolution.

[Atypical femoral fractures associated with long-term treatment with bisphosphonates]. / Fraturas Atípicas do Fémur Associadas a Terapêutica Prolongada com Bisfosfonatos.

INTRODUCTION: Long-term treatment with bisphosphonates has been associated to atypical femoral fractures whose features are now clearly defined. CLINICAL CASES: We present two cases of female patients under bisphosphonate treatment for over 10 years who were admitted to our institution for subtrochanteric and femoral shaft fractures after low-energy trauma. They presented, respectively, a transverse and a short oblique femoral fracture, with thickening of the lateral cortex. They underwent surgical treatment obtaining good functional and imaging result. DISCUSSION: These cases fulfill the established criteria for atypical femoral fracture, thereby illustrating a serious adverse event of long-term treatment with bisphosphonates. Such relationship has still not been clearly established by scientific evidence. However, its effectiveness in preventing osteoporotic fractures is well proven. CONCLUSION: The atypical femoral fractures are possibly a serious adverse effect of the long-term treatment with bisphosphonates. Scientific evidence still supports its use, however, the physician must be aware of these events and closely follow-up these patients.

Introdução: A terapêutica prolongada com bisfosfonatos tem sido associada a fraturas atípicas do fémur cujas características estão neste momento definidas.Casos Clínicos: Apresentam-se dois casos clínicos de doentes do género feminino sob terapêutica com bisfosfonatos há mais de 10 anos e que foram admitidas na nossa instituição por fraturas dos fémures resultantes de traumatismos de baixa energia. Estas fraturas localizavam-se na região subtrocantérica e mediodiafisária do fémur apresentando, respectivamente, traço simples transversal e oblíquo curto, com espessamento da cortical externa. Ambas foram submetidas a tratamento cirúrgico com bom resultado clínico e radiológico.Discussão: Os casos apresentados cumprem os critérios aceites para o diagnóstico de fratura atípica do fémur, ilustrando um efeito adverso grave da terapêutica prolongada com bisfosfonatos. A evidência científica ainda não estabeleceu esta associação de forma inequívoca. Por outro lado, a eficácia destes fármacos na prevenção de fraturas osteoporóticas está comprovada.Conclusão: A fratura atípica do fémur pode constituir um efeito adverso grave da terapêutica prolongada com bisfosfonatos. A evidência científica continua a suportar a sua utilização, mas o clínico deverá estar alerta e acompanhar atentamente estes doentes.

[Pituitary tumor detected prenatally]. / Tumor hipofisiario detectado prenatalmente.

INTRODUCTION: Brain tumors are present in 2.9 per 100,000 newborn. Craniopharyngioma is a benign and slow growing brain tumor, frequently localized in the sellar and suprasellar region. There are few reports of pituitary tumor detected prenatally. CASE REPORT: We report a neonate with a craniopharyngioma detected prenatally as a pituitary tumor. In a 23 year old mother, second gestation, with no important history, was detected a sellar tumor at 31 gestation weeks, the obstetric ultrasound reported a suprasellar tumor of 2 per 3 cm diameter. Pregnancy ended in a vaginal delivery at 39 weeks, and obtained a 3.9 kg female, with cephalic diameter of 37.5 cm, the Apgar score was 8-9 at 1st and 5th minutes. In early neonatal period was scanned and confirmed a 3.2/2.3/2.9 cm suprasellar tumor with calcium deposits. The Paediatric Oncology department suggested a surgery and was realized a craniotomy at 3rd week of age. The surgery allowed to obtain 30% of the tumor and confirmed by histology craniopharyngioma. Patient had favourable evolution and was discharged at 3 months of age. CONCLUSIONS: We report a neonate in who was detected by prenatal ultrasound the presence of a suprasellar solid tumor, scan and magnetic resonance images in neonatal period defined its size and location and a craniopharyngioma was confirmed by histology. Patient had a satisfactory postsurgical evolution and was discharged at 3 months of age.

Infarto cerebral espontáneo en un recién nacido de término / Spontaneous cerebral infarction in a full-term neonate

Introducción. Un accidente cerebro-vascular de origen isquémico y/o hemorrágico aumenta la morbi-mortalidad neonatal. Se estima que ocurre en 1:4,000 recién nacidos. En este trabajo se reporta un recién nacido de término con un accidente cerebro-vascular aparentemente espontáneo. Caso Clínico. Se trata de un recién nacido, madre de 31 años, segunda gesta de término que cursó con diabetes gestacional y fue tratada sólo con dieta. Inició con trabajo de parto a las 41.5 semanas de gestación por FUM; se efectuó cesárea por desproporción céfalo pélvica. Se obtuvo producto femenino de 3,640 g calificado con Apgar 8-9, se le aplicaron sólo pasos iniciales de reanimación. En el primer día de vida presentó desviación de la comisura labial a la derecha y movimientos tónico-clónicos en hemicuerpo derecho en 2 ocasiones; se inició fenobarbital. Los resultados de laboratorio para glucemia, sodio, calcio, potasio, creatinina, tiempos de coagulación y biometría hemática fueron normales; los cultivos de líquido cefalo-raquídeo fueron negativos, la punción lumbar se consideró traumática. El ultrasonido fontanelar no mostró alteraciones sugestivas de un evento hemorrágico ni alteraciones a nivel ventricular; sin embargo, al tercer día de vida se practicó tomografía axial computarizada de cráneo simple visualizándose una zona hipodensa con un patrón geográfico hacia la región tempo-parietal izquierda con aspecto de evento vascular isquémico en el territorio de la arteria cerebral media. Conclusiones. Se continuó con fenobarbital y ácido acetilsalicílico. Permaneció asintomática por lo que se egresó al séptimo día de vida. Se citó a consulta externa en Neurología Pediátrica.

Background. Cerebral vascular accident (CVA) is an important cause of hemorrhagic or ischemic cerebral injury and increases neonatal morbidity and mortality. It occurs in 1/4000 term neonates. We report a case of a neonate with a spontaneous CVA. Case report. We present the case of a newborn (NB) who was delivered from a 31-year old mother. It was the second pregnancy with 41.5 gestation weeks. The mother presented gestational diabetes controlled only by dietary therapy. A 3640 g, apparently healthy female was obtained by cesarean delivery that was indicated due to cephalopelvic disproportion. Apgar scores were 8-9 according to the conventional time points. The pediatrician used only initial steps of reanimation. During the first day of life, the infant presented a deviation of the right mouth commissure and tonic-clonic movements on the right half of the body two times. The newborn was treated with phenobarbital intravenous infusion. Laboratory tests were all normal, and cultures of cerebrospinal fluid and blood were considered negative. A head sonogram showed no evidence of hemorrhage or ventricular distortion but a cranial CT reported a low-density zone suggesting a cerebral infarction in the left parietal and temporal regions. Conclusions. We continued to treat with phenobarbital and acetylsalicylic acid and the patient remained asymptomatic prior to discharge at the 7th day of life, recommending follow-up with a pediatric neurologist.

Retardo grave en el crecimiento intrauterino en un neonato de 480 g al nacer / Intrauterine growth retardation in a 480 g neonate at birth

Introducción. Existen pocos reportes de neonatos vivos de <500g con retardo en el crecimiento intrauterino (RCIU). Caso clínico. Se reporta el caso de un recién nacido con RCIU grave, madre de 30 años, primera gesta, 31 semanas de gestación y oligohidramnios grave, obtenido por cesárea, femenino, de 480 g, talla 30 cm, Apgar de 7-7, recibió oxígeno al 80% por 48 horas y, a partir de entonces, se inicia vía oral y nutrición parenteral. Las IgG e IgM para citomegalovirus y cuerpos de inclusión citomegálica fueron negativos. A los 20 días de vida pesó 500 g, cursó asintomático y toleró bien su fórmula, aumentando hasta los 1 700 g antes de su egreso a los 5 meses. A los 12 meses, con peso de 2.8 kg y retraso en el desarrollo psicomotor de 6 meses para la edad cronológica, no desarrolló retinopatía del prematuro, leucomalacia periventricular ni hemorragia intrancraneal. Conclusión. Se reporta el caso de un neonato de 480 g al nacer, de 31 semanas de gestación, que presenta RCIU grave y retraso en el desarrollo psicomotor. Sin embargo, no presenta leucomacia paraventricular ni hemorragia paraventricular.

Background. There are few cases of live neonates with intrauterine growth retardation (IGR) <500 g with a satisfactory outcome, including apparent normal neurodevelopment. Clinical case. We report the case of a female 480 g neonate diagnosed with severe IGR who was the product of a 30-year-old mother. This was the first gestation and delivery was at 31 gestation weeks. The neonate demonstrated severe oligoamnios and was delivered by Cesarean section. Apgar score was 7-7, oxygen was administered at 48 h, and parenteral nutrition and formula for premature infants was initiated. Serum cytomegalovirus IgG and IgM and cytomegalic urine inclusion were all negative. The infant accepted formula and was asymptomatic. She reached 500 g at 20 days of life. At 5 months old, her weight increased to 1 700 g before hospital discharge. Patient was followed-up until 1 year of age. She weighed 2.8 kg and demonstrated a 6-month delay in psychomotor neurodevelopment. It is important to mention that she did not develop retinopathy of prematurity, periventricular leukomalacia or intracranial hemorrhage. Conclusion. The patient presented here was born after 31 weeks gestation with a birth weight of 480 g. She showed severe IGR with neurodevelopmental delay; however, no retinopathy of prematurity, periventricular leukomalacia or intracranial hemorrage was demonstrated.