Perivascular Innervation in the Nasal Mucosa and Clinical Findings in Patients with Allergic Rhinitis and Idiopathic Rhinitis.

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.

Perivascular Innervation in the Nasal Mucosa and Clinical Findings in Patients with Allergic Rhinitis and Idiopathic Rhinitis

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.

Protoporphyrin fluorescence induced by methyl-ALA in skin healing.

BACKGROUND: Estimation of the time period that precedes an injury is critical in forensic medicine. However, there is no reliable method that can be used to evaluate the oldness of a lesion. The aim of this work is to develop a fluorimetric method that can be used to follow the aging process of lesions by applying methyl-ALA (MAL) on wounds and by quantifying protoporphyrin IX (PPIX) fluorescence during the healing process. We also aim to understand the changes in PPIX fluorescence by establishing a correlation with histological evaluations during the healing process. METHODS: Standardized linear wounds were made on the dorsum of 72 mice, which were divided in control (MAL -) and experimental (MAL +) groups. In vivo fluorescence spectra (FS) were collected from normal and wound skin sites of control and experimental groups, corresponding to four groups of FS spectra: (a) FS of skin wound after MAL (+/+); (b) FS of normal skin after MAL (-/+); (c) FS of skin wound without MAL (+/-) and (d) FS of normal skin without MAL (-/-). Animals were monitored periodically for 3 months and euthanized. Tissue specimens were processed for histological analysis using design-based stereological methods. Serial cross-sections were analyzed to evaluate the organization of the dermis and epidermis, collagen deposition and cellular proliferation. RESULTS: FS of skin wound with MAL (+/+) showed an expressive intensity increase from the beginning of the experiment to the 34th day, with maximum fluorescence being observed on the ≈ 11 th day after wounding. There was preferential PPIX accumulation in healing sites as compared to adjacent normal skin (+/-) in the early stage of healing. Histological findings allowed correlation of the fluorescence increase mainly with cell proliferation. The drastic decrease in the FS intensity observed in the end of the healing process was correlated with the decrease in the proliferation rate as well as with the presence of new extracellular fibrous materials. CONCLUSIONS: In the mice wound-healing model tested here, it was possible to distinguish whether the injury was in early or advanced stages by using PPIX fluorescence induced by MAL. We conclude that this method is a promising approach to evaluate the age of skin wounding and we hope this work will stimulate human studies to allow this technique to become standardized in forensic medicine.

Effects of time on ultrastructural integrity of parathyroid tissue before cryopreservation.

BACKGROUND: Cryopreservation of parathyroid tissue is used in the surgical treatment of secondary hyperparathyroidism. After surgical resection, the tissue is temporarily maintained in a cell culture solution until it arrives at the specialized laboratory where the cryopreservation process will take place. The present study evaluates the time that the human hyperplastic parathyroid gland tissue can wait before cryopreservation, based on parathyroid cell ultrastructural integrity. METHODS: This prospective study included 11 patients who underwent total parathyroidectomy with heterotopic autotransplantation and cryopreservation of parathyroid tissue fragments. Part of the tissue was kept in cell culture solution at 4 °C. Five time periods between 2 and 24 h were defined, and parathyroid fragments were kept in the solution for that length of time. At the end of each period, the fragments were removed from the transport solution, fixed, and prepared for ultrathin sections. RESULTS: Of the 11 cases studied, 10 showed ultrastructural findings consistent with cellular viability in tissue fragments that remained in the transport solution up to 12 h. Electron microscopy revealed that cell adhesion and the integrity of plasma membranes, nuclei, and mitochondria were preserved in one case for up to 24 h. Changes in mitochondrial structure represented the most constant ultrastructural damage seen in the cases studied, in addition to the presence of edema and cell vacuoles. CONCLUSIONS: Analysis of the ultrastructure of hyperplastic parathyroid gland tissue showed that ultrastructural integrity was in most cases properly maintained in fragments stored up to 12 h in a cell culture solution at 4 °C.

Neoskin development in the fetus with the use of a three-layer graft: an animal model for in utero closure of large skin defects.

OBJECTIVE: To assess the ability of a three-layer graft in the closuse of large fetal skin defects. METHODS: Ovine fetuses underwent a large (4 × 3 cm) full-thickness skin defect over the lumbar region at 105 days' gestation (term = 140 days). A bilaminar artificial skin was placed over a cellulose interface to cover the defect (3-layer graft). The skin was partially reapproximated with a continuous nylon suture. Pregnancy was allowed to continue and the surgical site was submitted to histopathological analysis at different post-operative intervals. RESULTS: Seven fetuses underwent surgery. One maternal/fetal death occurred, and the remaining 6 fetuses were analyzed. Artificial skin adherence to the wound edges was observed in cases that remained in utero for at least 15 days. Neoskin was present beneath the silicone layer of the bilaminar artificial skin. CONCLUSIONS: Our study shows that neoskin can develop in the fetus using a 3-layer graft, including epidermal growth beneath the silicone layer of the bilaminar skin graft. These findings suggest that the fetus is able to reepithelialise even large skin defects. Further experience is necessary to assess the quality of this repair.

Effects of plastic stenting in commom bile duct of rats: a quantitative reaction analysis using collagen and elastin morphometry / Efeitos de prótese plástica em hepatocolédoco de ratos: uma análise quantitativa utilizando morfometria do colágeno e elastina

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.

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.

Effects of plastic stenting in common bile duct of rats. A quantitative reaction analysis using collagen and elastin morphometry.

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.