Abstract Introduction The nonspecific hyperreactivity of rhinitis has been attributed to neurotrophins activating sensory nerves and inflammatory cells. The relationship between these markers and the intensity of the symptoms is not well established and few studies have evaluated individuals with idiopathic rhinitis. Objective The present study aims to evaluate whether perivascular innervation and nerve growth factor (NGF) are related to the intensity of the clinical conditions in allergic rhinitis (AR) and idiopathic rhinitis (IR). Methods A total of 15 patients with AR and 15 patients with IR with the indication for inferior turbinectomy (associated or not with septoplasty) were selected. The patients received a score according to their signs and symptoms. After the surgery, we quantified eosinophils, mast cells, NGF, and nerve fibers in the nasal turbinate. Results The score of the signs and symptoms was higher in the AR group. Nerve growth factor was found in the cytoplasm of inflammatory cells in the submucosa in greater quantity in the AR group. The nerve fibers were distributed throughout the tissue, mainly in the subepithelial, glandular, and vascular regions, and there was no difference between the groups. Greater perivascular innervation was associated with a higher signs and symptoms score. Conclusions We concluded that these findings suggest that the NGF produced by submucosal inflammatory cells stimulates increased perivascular innervation in rhinitis, thus directly reflecting in more intense clinical conditions, especially in AR. (NGF (AR IR. . (IR) 1 septoplasty selected surgery eosinophils turbinate group tissue subepithelial glandular regions groups (IR

Discinesia ciliar primária (DCP) é uma doença genética que compromete a estrutura e/ou a função ciliar, causando retenção de muco e bactérias no trato respiratório e levando a infecções crônicas nas vias aéreas superiores e inferiores, defeitos de lateralidade visceral e problemas de fertilidade. Revisamos os sinais e sintomas respiratórios da DCP, os testes de triagem e a investigação diagnóstica, bem como detalhes relacionados ao estudo da função, ultraestrutura e genética ciliar. Descrevemos também as dificuldades em diagnosticar a DCP por meio de microscopia eletrônica de transmissão, bem como o seguimento dos pacientes.

Primary ciliary dyskinesia (PCD) is a genetic disorder of ciliary structure or function. It results in mucus accumulation and bacterial colonization of the respiratory tract which leads to chronic upper and lower airway infections, organ laterality defects, and fertility problems. We review the respiratory signs and symptoms of PCD, as well as the screening tests for and diagnostic investigation of the disease, together with details related to ciliary function, ciliary ultrastructure, and genetic studies. In addition, we describe the difficulties in diagnosing PCD by means of transmission electron microscopy, as well as describing patient follow-up procedures.

<title>Abstract</title><sec><title>Background:</title><p>The choice between heterologous expression versus chemical synthesis for synthesizing short cysteine-rich insecticidal peptides from arthropods may impact the obtainment of yields and well-folded bioactive molecules for scientific research. Therefore, two recombinant expression systems were compared to that of chemical synthesis for producing Ba1, a cysteine-rich spider neurotoxin.</p></sec><sec><title>Methods:</title><p>The transcription of the insecticidal neurotoxin Ba1 was obtained from a cDNA library of venom glands of the spider <italic>Brachypelma albiceps.</italic>It was cloned into the pCR®2.1-TOPO® cloning vector and then introduced in two different expression vectors, pQE40 and pET28a⁺. Each vector was transfected into E. <italic>coli</italic> M15 and BL21 cells, respectively, and expressed under induction with isopropyl thiogalactoside (IPTG). The chemical synthesis of Ba1 was performed in an Applied Biosystems 433A peptide synthesizer.</p></sec><sec><title>Results:</title><p>Both expression systems pQE40 and pET28a⁺ expressed the His-tagged recombinant protein products, HisrDFHRBa1 and HisrBa1, respectively, as inclusion bodies. The recombinant proteins HisrDFHRBa1 and HisrBa1 presented respective molecular masses of 28,289 and 8274.6 Da, and were not biologically active. These results suggested that both HisrDFHRBa1 and HisrBa1 were oxidized after cell extraction, and that their insecticidal activities were affected by their N-terminal pro-peptides and different disulfide bridge arrangements. The respective protein expression yields for HisrDFHRBa1 and HisrBa1 were 100 μg/L and 900 μg/L of culture medium. HisrBa1 was reduced and folded under <italic>in vitro</italic>conditions. The <italic>in vitro</italic> folding of HisrBa1 produced several isoforms, one of which, after removing its N-terminal pro-peptide by enzymatic cleavage, presented elevated insecticidal activities compared to the native Ba1. Furthermore, the His-tagged protein HisrDFHRBa1 underwent enzymatic cleavage to obtain recombinant Ba1 (rBa1). As expected, the molecular mass of rBa1 was 4406.4 Da. On the other hand, Ba1 was chemically synthesized (sBa1) with a yield of 11 mg per 0.1 mmol of amino acid assembly.</p></sec><sec><title>Conclusions:</title><p>The two recombinant insecticidal peptides and the one synthesized chemically were as active as the native Ba1; however, toxin yields differed drastically.</p></sec>

OBJETIVO: Comparar a técnica neurocirúrgica clássica a uma nova técnica simplificada, para correção de mielomeningocele, em fetos de ovelhas. MÉTODOS: Em 9 fetos, foi criado um defeito semelhante à mielomeningocele (laminectomia e excisão de dura-máter) no 90º dia de gestação. O tipo de correção foi randomizado. No Grupo 1, o defeito foi corrigido usando a técnica neurocirúrgica clássica, com a sutura de três camadas (dura-máter, músculo e pele), realizada por um neurocirurgião. No Grupo 2, um especialista em Medicina Fetal utilizou a técnica simplificada, colocando um fragmento de celulose biossintética sobre a medula e suturando apenas da pele sobre a celulose. Próximo ao termo da gestação (132 dias), os fetos foram sacrificados para análise anatomopatológica. RESULTADOS: Ocorreram dois casos de aborto e uma morte materna, restando seis casos para avaliação - três em cada grupo. No Grupo 1, todos os casos mostraram aderência da medula à cicatriz (meningoadesão) e perda da arquitetura medular, com destruição do funículo posterior e perda da visualização da substância cinzenta. No Grupo 2, observou-se, em todos os casos, a formação de uma neodura-máter, separando o tecido nervoso do músculo adjacente, sendo que o funículo posterior e a substância cinzenta estavam preservados. CONCLUSÃO: A técnica simplificada foi superior à neurocirúrgica, com maior preservação da medula e evitando as aderências do tecido nervoso. Os presentes achados sugerem que a técnica utilizada atualmente na correção de mielomeningocele em fetos humanos deva ser reavaliada.

OBJECTIVE: To compare the classical neurosurgical technique with a new simplified technique for prenatal repair of a myelomeningocelelike defect in sheep. METHODS: A myelomeningocele-like defect (laminectomy and dural excision) was created in the lumbar region on day 90 of gestation in 9 pregnant sheep. Correction technique was randomized. In Group 1 the defect was corrected using the classic neurosurgical technique of three-layer suture (dura mater, muscle and skin closure) performed by a neurosurgeon. In Group 2, a fetal medicine specialist used a biosynthetic cellulose patch to protect the spinal cord and only the skin was sutured above it. Near term (day 132 of gestation) fetuses were sacrificed for pathological analysis. RESULTS: There were two miscarriages and one maternal death. In total, six cases were available for pathological analysis, three in each group. In Group 1, there were adherence of the spinal cord to the scar (meningo-neural adhesion) and spinal cord architecture loss with posterior funiculus destruction and no visualization of grey matter. In Group 2, we observed in all cases formation of a neo-dura mater, separating the nervous tissue from adjacent muscles, and preserving the posterior funiculus and grey matter. CONCLUSION: The new simplified technique was better than the classic neurosurgical technique. It preserved the nervous tissue and prevented the adherence of the spinal cord to the scar. This suggests the current technique used for the correction of spina bifida in humans may need to be reassessed.

Durante las últimas décadas las anomalías térmicas han sido más frecuentes y severas en el Caribe, quedando pocos arrecifes exentos de eventos masivos de blanqueamiento (EMB). En el Parque Nacional Los Roques, Venezuela, un archipiélago poco afectado previamente por EMB, la temperatura del agua a 5m de profundidad alcanzó 30,85°C en septiembre 2010, y fue >29,0°C entre mediados de agosto y la primera semana de noviembre en cuatro arrecifes. El 72% de 563 colonias de escleractinios estaban blanqueadas o pálidas para octubre de 2010, mientras que para febrero 2011, el 46% de las colonias aún estaban afectadas. Para febrero 2011, la cobertura béntica coralina promedio disminuyó de 45 a 31%. Además, los arrecifes mostraron una alta prevalencia (de hasta 16%) de enfermedad de banda negra en Octubre 2010, y de plaga blanca (11%) en Febrero 2011. Como consecuencia, es probable que la mortalidad coralina resulte mayor a la reportada acá. Sin embargo, Los Roques es poco afectado por perturbaciones antropogénicas y cuenta con un alto reclutamiento de corales, lo cual podría contribuir a su recuperación

Thermal anomalies have become more severe, frequent and well-documented across the Caribbean for the past 30 years. This increase in temperature has caused coral bleaching resulting in reef decline. At Los Roques National Park, Venezuela, temperature has been monitored at four reef sites. In mid-September 2010, seawater temperature reached 30.85°C at 5 m depth in Los Roques, an archipelago only slightly affected by previous bleaching events. For example, bleaching in Los Roques in 2005 was mild compared to the rest of the Caribbean and to the results in this study. In 2010, seawater temperatures remained above 29.0°C from mid-August until the first week of November, resulting in +16 Degree Heating Weeks by that time. Our annual survey of four reef sites indicated that 72% of 563 scleractinian colonies were partial or totally bleached (white) or pale (discolored) in October 2010. In February 2011, there were still 46% of coral colonies affected; but most of them were pale and only 2% were bleached. By February, coral cover had declined 4 to 30% per transect, with a mean of 14.3%. Thus, mean coral cover dropped significantly from 45 to 31% cover (a 34% reduction). In addition to bleaching, corals showed a high prevalence (up to 16%) of black band disease in October 2010 and of white plague (11%) in February 2011. As a consequence, coral mortality is expected to be larger than reported here. Reef surveys since 2002 and personal observations for more than 20 years indicated that this bleaching event and its consequences in Los Roques have no precedent. Our results suggest that reef sites with no previous record of significant deterioration are more likely to become affected by thermal anomalies. However, this archipelago is relatively unaffected by local anthropogenic disturbance and has a high coral recruitment, which may contribute to its recovery

A tendinite calcária aguda pré-vertebral é uma condição benigna e rara que apresenta calcificação das fibras do músculo longo do pescoço com reação inflamatória local, sendo esta uma das formas de apresentação menos frequentes da doença por deposição de hidroxiapatita de cálcio. Manifesta-se com dor cervical aguda e/ou odinofagia, podendo ser erroneamente diagnosticada como abscesso retrofaríngeo, espondilodiscite ou alteração decorrente de trauma. Os achados radiológicos na tendinite calcária pré-vertebral são patognomônicos. O conhecimento de tais achados é muito importante, pois o correto diagnóstico possibilita a resolução precoce dos sintomas e evita intervenções desnecessárias em um paciente que apresenta afecção com boa resposta ao tratamento conservador.

Acute calcific prevertebral tendinitis is a benign and rare condition that presents calcification of the superior oblique fibers of longus colli muscle with local inflammatory reaction. Such condition is one of the less common presentations of calcium hydroxyapatite deposition disease. Clinical signs are usually acute neck pain and odynophagia, and it may be misdiagnosed as retropharyngeal abscess, spondylodiscitis or traumatic injury. The imaging findings in calcific prevertebral tendinitis are pathognomonic. The knowledge of such findings is extremely important to avoid unnecessary interventions in a patient presenting a condition with a good response to conservative treatment.

OBJETIVO: Com o intuito de contornar diversas dificuldades encontradas no uso rotineiro de queratinócitos cultivados in vitro pela técnica descrita por Rheinwald e Green, e obter um substituto cutâneo mais resistente e o menos imunogênico possível para futuras aplicações clínicas, este trabalho avaliou um novo sistema de cultura de queratinócitos que prevê a utilização de um substrato de fibrina em associação com queratinócitos humanos em alta densidade. MÉTODOS: Através de microscopia óptica e eletrônica e análise imunohistoquímica, foram avaliadas as características proliferativas e de diferenciação em longo prazo de queratinócitos cultivados em condição imersa e na interface ar-líquido. RESULTADOS: Apesar da ausência de um substituto dérmico, foi demonstrado que o composto proposto constituiu-se de um substrato de fibrina transparente e elástico coberto por epitélio multi-estratificado, bem aderido, com características morfológicas semelhantes à epiderme humana, incluindo a neo-formação, embora incompleta, da membrana basal. CONCLUSÕES: A maior resistência mecânica com a presença de um substrato de fácil manuseio, a possível liberação de queratinócitos não-confluentes, e a remoção de células com origem animal dos sistemas de cultura sugerem que o composto proposto neste estudo apresenta grande potencial para uso clínico futuro.

PURPOSE: In order to circumvent several difficulties that have been met in the routine use of the in vitro keratinocyte cultures using the standard procedure described by Rheinwald and Green, and obtain a more resilient and the least possible immunogeneic skin substitute for a future clinical application, this work studied a new keratinocyte culture system, which envisages the utilization of a fibrin substrate in association with high densities of human keratinocytes. METHODS: Through light and transmission electron microscopy and immunohistochemical assays, long-term proliferative and differentiative characteristics of keratinocytes cultured onto a fibrin gel under immerse and air-liquid interface culture conditions were evaluated. RESULTS: Despite the absence of a dermal substitute, the results demonstrated that the proposed composite was constituted of a transparent and elastic fibrin film covered by a well-attached, multistratified epithelium with morphological characteristics that resemble human epidermis, including the neoformation, albeit incomplete, of the basement membrane. CONCLUSIONS: Increased mechanical resistance due to the presence of an easy handling substrate, the delivery of nonclonfluent keratinocytes as well as the removal of animal-derived cells from the culture system suggest its potential use for future transplantation purposes.

A ventilação mecânica é uma terapia importante, mas pode resultar em complicações. Uma das mais relevantes é a lesão pulmonar induzida por ventilador. Devido à hiperdistensão alveolar, o pulmão inicia um processo inflamatório, com infiltrado neutrofílico, formação de membrana hialina, fibrogênese e prejuízo de troca gasosa. Nesse processo, a mecanotransdução da hiperdistensão celular é mediada através do citoesqueleto da célula e de suas interações com a matriz extracelular e com as células vizinhas, de modo que o estímulo mecânico da ventilação se traduz em sinalização bioquímica intracelular, desencadeando ativação endotelial, permeabilidade vascular pulmonar, quimiotaxia leucocitária, produção de citocinas e, possivelmente, lesão de órgãos à distância. Estudos clínicos demonstram essa relação entre distensão pulmonar e mortalidade em pacientes com lesão pulmonar induzida por ventilador. Entretanto, apesar de o citoesqueleto ter um papel fundamental na patogênese da lesão pulmonar induzida por ventilador, a literatura carece de estudos utilizando modelos in vivo sobre as alterações do citoesqueleto e de suas proteínas associadas durante esse processo patológico.

Although mechanical ventilation is an important therapy, it can result in complications. One major complication is ventilator-induced lung injury, which is caused by alveolar hyperdistension, leading to an inflammatory process, with neutrophilic infiltration, hyaline membrane formation, fibrogenesis and impaired gas exchange. In this process, cellular mechanotransduction of the overstretching stimulus is mediated by means of the cytoskeleton and its cell-cell and cell-extracellular matrix interactions, in such a way that the mechanical stimulus of ventilation is translated into an intracellular biochemical signal, inducing endothelial activation, pulmonary vascular permeability, leukocyte chemotaxis, cytokine production and, possibly, distal organ failure. Clinical studies have shown the relationship between pulmonary distension and mortality in patients with ventilator-induced lung injury. However, although the cytoskeleton plays a fundamental role in the pathogenesis of ventilator-induced lung injury, there have been few in vivo studies of alterations in the cytoskeleton and in cytoskeleton-associated proteins during this pathological process.

OBJETIVO: Avaliar os efeitos da colocação de prótese biliar em ratos, analisando a deposição de colágeno e elastina no hepatocolédoco. MÉTODOS: Vinte ratos machos foram submetidos à laparotomia mediana, duodenotomia e colocação transpapilar de uma prótese plástica 22Fr no hepatocolédoco. Os animais foram randomizados em 4 grupos, com cinco componentes cada: (I) controle, (II) prótese biliar por 7 dias, (III) prótese biliar durante 14 dias e (IV) prótese biliar por 30 dias. Cortes do hepatocolédoco foram corados com Hematoxilina Eosina; Sirius Red para quantificar o colágeno; e Resorcina-fucsina de Weigert para quantificar a elastina. RESULTADOS: A porcentagem de área corada para colágeno foi de 13,4; 21,5; 29,5 e 32,8 para os grupos I a IV, respectivamente. A porcentagem de área corada para fibras de elastina foi de 7,0; 5,2; 4,0 e 2,9, para os grupos I a IV, respectivamente. A razão colágeno / elastina foi de 2,4, 5,1, 11,0 e 14,4 para os grupos I a IV, respectivamente. CONCLUSÕES: O implante da prótese biliar leva à deposição de colágeno e elastina nos ductos biliares; e a deposição de colágeno e a relação colágeno / elastina são proporcionais ao tempo de permanência da prótese.

PURPOSE: To evaluate the effects of biliary stenting in rats through analysis of collagen and elastin deposition in the bile ducts. METHODS: Twenty male rats underwent midline laparotomy, duodenotomy and transampullary stenting of the common bile duct with a 22Fr plastic stent. Animals were randomized in 4 groups, with 5 components in each: (I) control, (II) biliary stenting for 7 days, (III) biliary stenting for 14 days, and (IV) biliary stenting for 30 days. Sections of the common bile duct were stained using Hematoxylin Eosin; Sirius Red to quantitate the amount of collagen present; and Weigert's Resorcin-fuchsin to quantitate the amount of elastin present. RESULTS: The percentage of area stained for collagen was 13.4; 21.5; 29.5 and 32.8, for groups I to IV, respectively. The percentage of area stained for elastin fibers was 7.0; 5.2; 4.0 and 2.9, for groups I to IV, respectively. Collagen/Elastin ratio was 2.4; 5.1; 11.0 and 14.4 for groups I to IV, respectively. CONCLUSIONS: The biliary stenting leads to collagen and elastin deposition in the bile ducts; and collagen deposition and collagen/elastin ratio are proportional to the period of stenting.

RESUMO Objetivo: Modificar a técnica de fetoscopia de suspensão para redução do calibre dos orifícios de entrada no miométrio. Métodos: Sete ovelhas grávidas foram submetidas à fetoscopia para correção de um grande defeito de pele medindo 4,0 × 3,0 cm, criado na região lombar fetal com 100 dias de gestação. O defeito era corrigido através de sutura contínua da pele, aproximando-se as suas bordas. Para realizar a sutura, foi realizada a fetoscopia de suspensão, utilizando-se três orifícios para entrada de trocarte no miométrio. O trocarte da ótica era de 5,0 mm e das pinças de apreensão, dissecção e sutura eram de 3,5 mm. Após a cirurgia, a gestação era mantida até o sacrifício, realizado no 133º dia de gestação, quando os fetos eram avaliados. Resultados: Sete ovelhas prenhes foram operadas. Os dois primeiros casos constituíram o Grupo Piloto, no qual a técnica endoscópica foi modificada e a redução do calibre foi possível em dois dos três orifícios de entrada no miométrio. Nos cinco demais casos (Grupo de Estudo) a correção foi realizada com sucesso em todos os fetos e o tempo médio de duração da fetoscopia foi de 98 minutos. Houve um caso de morte materna atribuído à infecção intrauterina. A média de permanência intraútero após a cirurgia foi de 12 dias. Conclusões: A técnica foi modificada com sucesso, permitindo a redução do calibre dos orifícios uterinos necessários para realizar a correção de um defeito de pele na região lombar do feto através de uma nova técnica de fetoscopia. O impacto desta modificação na correção da meningomielocele em fetos humanos deve ser estudado.

ABSTRACT Objective: To change the gasless fetoscopy technique in order to reduce the diameter of entry orifices in the myometrium. Methods: Seven pregnant ewes were submitted to fetoscopy for repairing a large skin defect measuring 4.0 × 3.0 cm, created in the fetal lumbar region at the gestational age of 100 days. The defect was repaired through continuous suture of the skin with approximation of borders. Gasless fetoscopy was used for performing the suture with three orifices to allow entry of the trocar into the myometrium. A 5.0-mm optical trocar, and 3.5-mm grasping, dissecting and suturing forceps were used. After surgery, pregnancy was maintained until the animals were euthanized on the 133rd day of gestation, and the fetuses were evaluated. Results: Seven pregnant ewes underwent surgery; the first two cases were characterized as the Pilot Group, in which the endoscopic technique was modified and caliber reduction was possible in two out of three entry orifices in the myometrium. In the five remaining cases (Study Group), the repair was successfully carried out in all the fetuses, and the mean duration of fetoscopy was 98 minutes. There was a case of maternal death attributed to intrauterine infection. Mean intrauterine permanence after surgery was 12 days. Conclusions: The technique was successfully modified, allowing reduction of the uterine orifices necessary to perform the repair of a skin defect in the fetal lumbar region through a new fetoscopy technique. The impact of this modification in repair of myelomeningocele in human fetuses should be studied.

The purpose of this study was to compare esophageal infusion with 0.1 N hydrochloridric acid (HCl) to esophageal infusion with saline in patients presenting with typical gastroesophageal reflux symptoms and erosive esophagitis. METHODS: Upper gastrointestinal endoscopy was performed on 44 prospective subjects, 29 of whom were included in the study. Eighteen patients presented with normal esophagi (Control Group "C"), nine of whom were infused with HCl and nine with saline. Eleven patients presented with erosive esophagitis (Lesion Group "L"), five of whom were infused with HCl and six with saline. Biopsies of the esophageal mucosa were collected before and after infusions. RESULTS: No statistically significant difference was found between the two types of infusions in terms of the dilation of the intercellular space of the esophageal epithelium, regardless of the status of the patient. CONCLUSIONS: Response to HCl infusion cannot be used as a marker for gastroesophageal reflux disease.