Skin Ultrastructure and the Changes of HIF-2α, H-FABP Expression in the Myocardium of Electric Shock Death Rats / 法医学杂志

Objective To observe the skin ultrastructure change of electric shock death rats and to test the expression changes of hypoxia-inducible factor-2α （HIF-2α） and heart type-fatty acid-binding protein （H-FABP） of myocardial cells, in order to provide basis for forensic identification of electric shock death. Methods The electric shock model of rats was established. The 72 rats were randomly divided into control group, electric shock death group and postmortem electric shock group. Each group was divided into three subgroups, immediate （0 min）, 30 min and 60 min after death. The skin changes of rats were observed by HE staining, the changes of skin ultrastructure were observed by scanning electron microscopy, and the expression of HIF-2α and H-FABP in rats myocardium was tested by immunohistochemical staining. Results The skin in the electric shock death group and postmortem electric shock group had no significant difference through the naked eye or by HE staining. Under the scanning electron microscope, a large number of cellular debris, cells with unclear boundaries, withered cracks, circular or elliptical holes scattered on the cell surface and irregular edges were observed. A large number of spherical foreign body particles were observed. Compared with the control group, the expression of HIF-2α in all electric shock death subgroups increased, reaching the peak immediately after death. In the postmortem electric shock group, HIF-2α expression only increased immediately after death, but was lower than that of electric shock death group （P<0.05）. Compared with the control group, the expression of H-FABP in all subgroups of electric shock death group and postmortem electric shock group significantly decreased. The expression of H-FABP in all subgroups of electric shock death group was lower than that of the postmortem electric shock group （P<0.05）. Conclusion Electric shock can increase HIF-2α expression and decrease H-FABP expression in the myocardium, which may be of forensic significance for the determination of electric shock death and identification of antemortem and postmortem electric shock.

Comparación entre suturas convencional coadyuvada con ulmoplus® y sin ulmoplus® en el cierre de incisiones de piel de conejo ( Oryctolagus cuniculus ) / Comparison between conventional sutures with ulmoplus® and without ulmoplus® in the incision closure of rabbit skin ( Oryctolagus cuniculus )

Considerando los avances en las investigaciones que relacionan los efectos terapéuticos de la miel en el proceso de cicatrización de heridas, la presente investigación tuvo como objetivo evaluar y comparar cualitativamente, a través de una análisis morfológico, la respuesta tisular local entre sutura convencional coadyuvada con Ulmoplus® y sin Ulmoplus® en el cierre de la incisión de piel de conejo. Se utilizaron 10 conejos adultos, divididos en grupo S (sutura) y grupo S+U (sutura más Ulmoplus®). Se les realizó una incisión de 10 mm de largo, en la región anterior de la pierna, a través de la epidermis y dermis, hasta llegar a la fascia profunda que cubría al músculo tibial anterior. Posteriormente, solo al grupo S+U se le aplicó Ulmoplus®. Luego, en ambos grupos, la incisión fue suturada con dos puntos intradérmicos de ácido poliglicólico (vicryl 2/0). Después de 10 días post incisión fueron tomadas las biopsias de la piel y procesadas para su observación en microscopio de luz. El estudio se llevó a cabo considerando las recomendaciones de la Guía para el Cuidado y Uso de Animales de Laboratorio. Ambos grupos presentaron una incisión cerrada al día 10, con características microscópicas propias de una fase proliferativa. No obstante, el grupo S+U mostró una cicatriz de menor longitud y de aspecto más plano, cuya epidermis estaba mejor organizada y era de menor grosor, con células basales diferenciadas. A nivel dérmico se observó contracción de la herida, evidenciándose una zona cicatrizal de menor tamaño. Ulmoplus®, es una buena alternativa como coadyuvante en el proceso de cierre de heridas quirúrgicas utilizando sutura convencional, ya que acelera el proceso de cicatrización y permite mejores resultados estéticos de la cicatriz.

Considering the advances in research that relate honey therapeutic effects in wound healing process, the present investigation aimed to evaluate and compare qualitatively, through morphological analysis, the local tissue response between conventional suture with Ulmoplus ® and without Ulmoplus ® in the closure of the rabbit skin incision. We used 10 adult rabbits, divided into group S (suture) and group S + U (suture plus Ulmoplus®). An 10 mm long incision was made in the anterior region of the leg, passing through epidermis and dermis, until reaching the deep fascia that covered the anterior tibial muscle. Subsequently, only the S + U group was given Ulmoplus®. Then, in both groups, the incision was sutured with two intradermal points of polyglycolic acid (vicryl 2/0). After 10 days post incision, biopsies of the skin were taken and processed for observation in a light microscope. The study was carried out considering the recommendations of the Guide for the Care and Use of Laboratory Animals. Both groups had a closed incision at day 10 with microscopic characteristics of a proliferative phase. However, the S + U group showed a scar of shorter length and flatter appearance, whose epidermis was more organized and thinner, with differentiated basal cells. At the dermal level, wound contraction was observed, evidencing a smaller scar area. Ulmoplus®, as a coadjuvant in closing surgical wounds process using conventional suture is a good alternative, since it accelerates the healing process and allows better aesthetic results of the scar.

Skin development in 14 to 60-day-old Guinea pig fetus / Desarrollo de la piel del feto de conejillo de indias de 14 a 60 días

To contribute knowledge of an important experimental model for studies on skin embryology, a study was carried out to describe the morphological events of the skin during the intrauterine life of Cavia porcellus from the 10th to the 60th day of gestation. Embryos and fetuses were dissected, and the skin of the nasal, cranial, lumbar and anal regions was processed byoptical microscopy. At 30 days the first hairs, called lanugos, were observed in the cranial region. The morphological description showed that a few days can make a great difference in development.

Con el objetivo de contribuir al conocimiento de un importante modelo experimental para estudios sobre embriología de la piel, se llevó a cabo un estudio para describir los eventos morfológicos de la piel durante la vida intrauterina de Cavia porcellus desde el día 10 hasta el día 60 de gestación. Los embriones y los fetos se disecaron y se procesó la piel de las regiones nasal, craneal, lumbar y anal, mediante microscopía óptica. A los 30 días se observaron los primeros vellos, llamados lanugos, en la región craneal. La descripción morfológica mostró que unos pocos días pueden marcar una gran diferencia en el desarrollo.

The structure of the skin, types and distribution of mucous cell of yangtze sturgeon ( Acipenser dabryanus ) / La estructura de la piel, los tipos y distribución de las células mucosas del esturión yangtze ( Acipenser dabryanus )

The structural characteristics of the skin, types and distribution of mucous cells of Yangtze sturgeon (Acipenser dabryanus) were studied at the light microscope level, stained with Haematoxylin-eosin (HE) and Alcian blue-periodie acid Schiff (ABPAS). The skin of both was composed of epidermis and dermis. The dermis was divided into stratum spongiosum and stratum compactum. The stained color of stratum compactum was stained more deeply than that of stratum spongiosum. The skin thickness displayed differences in the fish at different body positions. The thickest of epidermis layer was on the dorsal region for Yangtze sturgeon, reversely, the thinnest was the mandibular region; Stratum spongiosum on the mandibular region was the thickest, the stratum spongiosum of the maxillary region was not obvious. In summary, keratinized spines, a kind of keratin derivative, are widely distributed in the mandibular, ventral, dorsal, and caudal peduncle skin surface for Yangtze sturgeon, and some pit organs mainly present in the skin surface of the maxillary and ventral regions. In short, the small amount of mucous cells in the skin of Yangtze sturgeon and the type of mucous cell were main Type IV, nevertheless there was a distribution of a few Type III.

Se estudiaron las características estructurales de la piel, los tipos y la distribución de las células mucosas del esturión Yangtze (Acipenser dabryanus) con microscopio de luz, teñidas con hematoxilina-eosina (HE) y azul alcián-ácido de Schiff (AB-PAS). La piel estaba compuesta por epidermis y dermis. La dermis se dividía en estrato esponjoso y estrato compacto. El grosor de la piel mostró diferencias en los peces en diferentes posiciones del cuerpo. La capa más gruesa de la epidermis se observó en la región dorsal del esturión Yangtze; a la inversa, la más delgada en la región mandibular. El estrato esponjoso en la región mandibular era el más grueso, el estrato esponjoso de la región maxilar no era visualizado. En resumen, las espinas queratinizadas, un tipo derivado de la queratina, estaban ampliamente distribuidas en la superficie de la piel del pedúnculo mandibular, ventral, dorsal y caudal en el esturión Yangtze, y algunos órganos en fosas, presentes principalmente en la superficie de la piel de las regiones mandibular y ventral. En resumen, la pequeña cantidad de células mucosas en la piel del esturión Yangtze y el tipo de célula mucosa eran células principales tipo IV, sin embargo, se observaron algunas células tipo III.

Evaluation of dermal collagen stained with picrosirius red and examined under polarized light microscopy

Abstract: The special picrosirius red staining highlights the natural birefringence of collagen fibers when exposed to polarized light. The results from birefringence allow to evaluate the organization of the collagen fibers in the tissues. The authors intend to elucidate all steps to obtain and capture images of histological sections stained with picrosirius red and evaluated under polarized light microscopy, as well as possible artefacts that may occur.

Histological description of the skin glands of Phyllobates bicolor (Anura: Dendrobatidae) using three staining techniques / Descripción histológica de las glándulas cutáneas de Phyllobates bicolor (Anura: Dendrobatidae) usando tres técnicas de tinción

The mechanisms to obtain and store skin toxins in frogs in of the family Dendrobatidae are not completely understood. In order to contribute to understand how toxins are stored, we provide a histological description of the cutaneous glands of the species Phyllobates bicolor. The skin of two adult frogs was examined through three histological staining techniques (hematoxilin-eosin, PAS and Masson Trichrome) using conventional optic microscopy. The skin of Phyllobates bicolor contains two types of exocrine glands: mucous and serous, which empty their products to the epidermal surface through an intra-epithelial duct that leads to a stoma. The mucous and serous glands and the intercalated ducts are surrounded by a discontinuous sheath of myoepithelial cells, which colapse the lumen of the acinus and the lumen of ducts and facilitate the secretion and release of their content. The serous glands have a polarized syncytium of tall cuboidal or columnar epithelial cells. Both glands have a mixed secretion, thus, the contents of mucous glands tend to be neutral and basophilic, while the contents of the serous glands are basophilic and acidophilic.

A la fecha no existe mayor información con respecto a los mecanismos para obtener y almacenar las toxinas cutáneas de ranas de la familia Dendrobatidae. Con el fin de contribuir y entender cómo son almacenadas estas toxinas, realizamos una descripción histológica de las glándulas cutáneas de la especie Phyllobates bicolor. La piel de dos ranas adultas se examinó mediante tres técnicas de tinción histológica (hematoxilina-eosina, PAS y tricrómico de Masson) mediante microscopía óptica convencional. La piel de P. bicolor contiene dos tipos de glándulas exocrinas: mucosas y serosas, que vierten sus productos a la superficie epidérmica a través de un conducto intra-epitelial que conduce a un estoma. Las glándulas mucosas, serosas y los conductos intercalados están rodeados por una funda discontinua de células mioepiteliales, las que colapsan el lumen de los acinos y conductos, facilitando la secreción y liberación de su contenido. Las glándulas serosas tienen un sincitio polarizado de células epiteliales columnares cúbicas. Ambas glándulas tienen una secreción mixta, por lo tanto, los contenidos de las glándulas mucosas tienden a ser neutral y basófilas, mientras que los contenidos de las glándulas serosas son basófilas y acidófilas.

Differential expression of alpha-smooth muscle actin and vascular endothelial growth factor in skin biopsy of patients with systemic sclerosis: a histological and immunohistochemical study

Vasculopathy is a hallmark of systemic sclerosis [SSc]. It contributes to many of its clinical manifestations and precedes fibrosis. The aim of this study was to investigate the expression of alpha-smooth muscle actin [alpha-SMA] in skin biopsy of patients with SSc and correlate it with other manifestations of vasculopathy, including those seen on fundus fluorescein angiography and tissue vascular endothelial growth factor [VEGF] expression. This study included 25 patients with SSc and 10 healthy individuals. Patients underwent full history taking and a clinical examination. All participants underwent fundus fluorescein angiography. Skin biopsy was examined by H and E staining, Mallory triple staining, and immunohistochemical staining for alpha-SMA and VEGF. Histological examination showed loss of dermal papillae, hypovascularity of the dermis, and subepidermal fibrosis. Immunohistochemical staining of the vessel wall in skin biopsy samples showed a statistically highly significant increase in VEGF and a highly significant decrease in alpha-SMA in patients as compared with controls. There was a highly significant positive correlation between VEGF and duration of illness, Raynaud's phenomenon, digital ulcers, disease activity score, and modified Rodnan Skin Score. As regards alpha-SMA, there was a highly significant negative correlation with Raynaud's phenomenon, disease activity score, modified Rodnan Skin Score, and VEGF, whereas there was a significant negative correlation with digital ulcers. The strongest correlation [r] for the duration of illness was found with alpha-SMA, followed by VEGF. alpha-SMA was found to be correlated to different manifestations of vasculopathy in SSc. It was found to be one of the early markers of vasculopathy among the other studied variables. Besides its diagnostic role in SSc vasculopathy, it could play a role in impaired vasculogenesis, making it a potential therapeutic target in the management of SSc

Análise ultraestrutural e implicação do RNA de interferência na fibrilogênese do colágeno V em pacientes com esclerodermia / Ultrastructural analysis and implication of RNA interference in collagen V fibrillogenesis in patients with scleroderma

Introdução: Recentemente muitas são as funções atribuídas ao colágeno V em condições fisiológicas normais e em algumas doenças, como na esclerodermia. Esta proteína apresenta características estruturais singulares de regulação do diâmetro das fibrilas heterotípicas, e imunogenicidade, sendo capaz de desencadear uma resposta imune independente. Em descobertas recentes, desenvolvidas por nosso grupo, ficou evidenciado que o colágeno V apresenta histoarquitetura anômala, aumentando a sua imunoexpressão na pele e pulmão em estágios iniciais da doença, assim como, aumento do RNAm de suas cadeias, principalmente alfa2(V). Por esta razão e com o intuito de entender sobre os processos moleculares e ultraestruturais que correlacionam o COL V à fibrose na ES, a proposta do presente estudo foi realizar análise morfológica e ultraestrutural das cadeias, alfa1(V) e alfa2(V), da pele de pacientes com ES, assim como avaliar a influência do gene COL5A2 na fibrilogênese. Pacientes e Métodos: As análises foram desenvolvidas utilizando fragmento de pele obtidas de biópsia, tanto de pacientes como controles, sob aprovação do comitê de ética (CAPPesq 0331/10) e de acordo com Declaração de Helsinque. Participaram do estudo 7 pacientes (homens=3, mulheres=4) diagnosticados com ES há dois anos ou menos, e indivíduos voluntários saudáveis (n=7) utilizados como grupo controle. A biópsia das peles foi dividida para dois grupos de análises: estudos histológicos e cultura de fibroblastos; sob solução de formol a 10%, foram realizadas análises histomorfométricas [coloração de Hematoxilina e Eosina (H e E), Tricrômico de Masson, imunofluorescência (IM) para cadeias alfa1(V) e alfa2(V), e reconstrução tridimensional (Zeiss LSM 510 META/UV)], e em solução de glutaraldeído 2% para análise ultraestrutural (microscopia eletrônica de transmissão com imunomarcação com ouro coloidal). A quantificação das cadeias foi realizada por histomorfometria, através de análise de imagem utilizando o software...

Introduction: Recently many functions are attributed to type V collagen in normal physiological conditions and in some diseases, such as scleroderma. This protein presents unique structural features for regulating the diameter of fibrils heterotypic and immunogenicity being capable of eliciting an immune response independent. In recent discoveries, developed by our group, it was evident that collagen V presents anomalous histoarchitecture, increasing the immunoexpression in the skin and lung in the early stages of the disease, as well as increased mRNA of his chains, mainly alpha2(V). For this reason and in order to understand the molecular and ultrastructural processes that correlate COL V fibrosis in SSc, the purpose of this study was to perform morphological and ultrastructural analysis of the chains alpha1(V) and alpha2(V) of the patient´s skin with ES, as well as to assess the influence of the COL5A2 gene in fibrillogenesis. Patients and Methods: The analyses were developed using skin fragment obtained from biopsy, both of patients and controls, under the approval of the ethics committee (CAPPesq 0331/10) and in accordance with the Declaration of Helsinki. The study included 7 patients (male = 3, female = 4) diagnosed with ES for two years or less, and healthy volunteers (n = 7) used as a control group. The biopsy of the skin was divided in two groups of analyzes: histological studies and cultured fibroblasts; Under formaldehyde solution 10%, histomorphometric analysis [staining with hematoxylin and eosin (H e E), Masson's trichrome, immunofluorescence (IM) to alpha1(V) and alpha2(V) chains, and three-dimensional reconstruction (Zeiss LSM 510 META were performed/UV), and in a solution of 2% glutaraldehyde for ultrastructural analysis (transmission electron microscopy with immunostaining with colloidal gold). Quantitation was performed by the chain histomorphometry by image analysis using Image Pro-Plus 6.0 software. Already a fibroblast culture...

Scanning electron microscopy of a blister roof in dystrophic epidermolysis bullosa / Microscopia eletronica de varredura do teto de uma bolha de epidermolise bolhosa distrofica

In dystrophic epidermolysis bullosa the genetic defect of anchoring fibrils leads to cleavage beneath the basement membrane, with its consequent loss. We performed scanning electron microscopy of an inverted blister roof of a case of dystrophic epidermolysis bullosa, confirmed by immunomapping and gene sequencing. With a magnification of 2000 times a net attached to the blister roof could be easily identified. This net was composed of intertwined flat fibers. With higher magnifications, different fiber sizes could be observed, some thin fibers measuring around 80 nm and thicker ones measuring between 200 and 300 nm.

Na epidermólise bolhosa distrófica, o defeito genético das fibrilas ancorantes leva à clivagem abaixo da membrana basal, com sua consequente perda. Realizamos microscopia eletrônica de varredura do teto invertido de uma bolha de um caso de epidermólise bolhosa distrófica, cujo diagnóstico foi confirmado com imunomapeamento e com sequenciamento gênico. Com uma ampliação de 2.000 vezes, pôde ser facilmente identificada uma rede ligada ao teto da bolha. Essa rede era composta por fibras achatadas e entrelaçadas. Com grandes aumentos, fibras de diferentes tamanhos puderam ser observadas: algumas finas, medindo cerca de 80 nm, e outras mais largas, medindo entre 200 nm e 300 nm.

Effect of subcutaneous phosphatidylcholine injection used in the treatment of localization obesity on the histological structures of human skin and subcutaneous tissue: a pilot study

Multiple clinical trials have supported the idea that subcutaneously injected phosphatidylcholine [PPC] leads to a reduction in localized fat collection. However, only a few histological studies that explain the mechanism of action of PPC have been published. This study aimed to evaluate the clinical and detailed histological changes in the skin and subcutaneous tissue after PPC injection. Ten female patients with local fat deposits [upper outer thigh] were assessed after a single session of subcutaneous injection with PPC on the basis of thigh circumference measurement, and histological examination of skin biopsy specimens was carried out before and 1 and 2 months after treatment. Histological sections were stained with H and E, Masson's trichrome [for collagen fibers], and by aldehyde fuchsin [for elastic fibers], followed by morphometric study and statistical analysis. Two months after injection, a statistically significant reduction in thigh circumference was found [P=0.045], with leathery tight skin texture at the injected area. Histological examination revealed dermal inflammatory responses 1 month after injection, with destruction of fat cells. These observations were reduced after 2 months with evidence of regenerating fat cells. Statistical analysis showed a significant increase in collagen area% [P=0.025] and statistically nonsignificant increase in elastic fiber area% at the end of the study. A single session of subcutaneous PPC injection had an evident lipolytic effect, with noticeable contouring and skin tightening due to regenerative effect on skin connective tissue, particularly dermal collagen. However, lipolytic effect was partially temporary because of regeneration of fat cells

Morphological abnormalities in mitochondria of the skin of patients with sporadic amyotrophic lateral sclerosis / Alteraciones morfológicas en las mitocondrias en la piel de enfermos con esclerosis lateral amiotrófica esporádica

OBJECTIVES: Mitochondrial dysfunction has been reported in the central nervous system, hepatocytes and peripheral blood lymphocytes from patients with sporadic amyotrophic lateral sclerosis (SALS). However, the status of skin mitochondria has not been reported, in spite of the fact that SALS patients present skin abnormalities. The objective of the present study was to compare mitochondrial ultrastructural parameters in keratinocytes from patients with SALS and healthy controls. METHODS: Our study was based on the analysis of 112 skin mitochondria from 5 SALS patients and 99 organelles from 4 control subjects by electron microscopy. RESULTS: Computerized image analysis showed that mitochondrial major axis length, area and perimeter of the organelle were significantly smaller in SALS respect of healthy control subjects. Morphologically, SALS mitochondria presented cristolysis and breakage of the outer membrane. CONCLUSIONS: Mitochondrial dysfunction in the skin may possibly reflect changes occurring in mitochondria of the central nervous system. The analysis of mitochondrial morphology in this tissue may be of value to follow disease progression and, eventually, the effectiveness of current therapies for SALS.

OBJETIVOS: Existen alteraciones en la función mitocondrial en el sistema nervioso central, en hepatocitos y en linfocitos de sangre periférica en SALS. Aunque, no se ha estudiado si existen cambios estructurales en las mitocondrias de la piel. Nuestro objetivo fue comparar la ultraestructura de mitocondrias en queratinocitos de enfermos con SALS con la de controles sanos. MÉTODO: Fueron analizadas en el microscopio electrónico 112 mitocondrias dérmicas de 5 pacientes y 99 provenientes de 4 controles. RESULTADOS: EL análisis computarizado mostró que el eje mayor mitocondrial, el área y el perímetro de las organelas fueran significativamente menor que en controles. Morfológicamente, las mitocondrias de SALS presentaron cristólisis y ruptura de la membrana externa. CONCLUSIÓN: La alteración mitocondrial en la piel posiblemente refleje cambios que también ocurran en las mitocondrias neuronales. Este análisis morfológico de las mitocondrias podría tener valor en el seguimiento de la enfermedad y eventualmente en la evaluación de la efectividad de futuras terapias.

Classical Ehlers-Danlos syndrome: clinical, Histological and ultrastructural aspects / Síndrome de Ehlers-Danlos clássica: aspectos clínicos, histológicos e ultraestruturais

A 12-year-old boy with difficulty in wound healing and scars of unusual appearance was examined. Thin, shiny and protruding skin, characterizing pseudotumoral lesions, was observed on his knees and elbows. Loose joints and increased skin elasticity were also observed. Light microscopy showed rare collagen bundles and loosely dispersed collagen fibers. Irregularly distributed elastic tissue was observed, and transmission electron microscopy revealed disorganized collagen fibers. In cross sections under high magnifications, fibers of varying diameters could be seen, as well as an irregular fiber outline. Scanning electron microscopy of the dermis showed disorganized collagen fibers, which were rarely arranged in bundles. Isolated and crossed-over fibers were also found.

Paciente masculino de 12 anos relatou dificuldade em cicatrizar, seguido por cicatrizes de aspecto infrequente . A pele dos joelhos e cotovelos é fina, brilhante e protrusa, caracterizando pseudo- tumores. Além disso apresenta hiperelasticidade cutânea e articular. A microscopia óptica demonstrou raros feixes colágenos, as fibras colágenas encontram-se dispersas. O tecido elástico está presente e secundariamente irregular. A microscopia eletrônica de transmissão também observou fibras colágenas desorganizadas e com cortes transversais em grande aumento evidenciou diâmetros variados e contorno irregular das mesmas. A microscopia eletrônica de varredura da derme demonstrou raros feixes colágenos e fibras colágenas isoladas e entrecruzadas.

Uso de la microscopia confocal de reflectancia en dermatología / Use of confocal reflectante microscopy in dermatology

La microscopia confocal de reflectancia (MCR) es una herramienta de imágenes no invasiva que permite la visualización in vivo y en tiempo real de estructuras microscópicas de la epidermis, unión dermo-epidérmica y dermis superficial con una resolución cuasi histológica. Ha sido utilizada en la evaluación de lesiones benignas y malignas, demostrando un gran potencial para aplicaciones en investigación básica y en dermatología clínica. En el presente trabajo se revisan conceptos acerca del desarrollo y principios básicos del funcionamiento de la MCR, así como de sus aplicaciones en la práctica clínica.

Reflectance confocal microscopy (RCM) is a noninvasive imaging tool that allows in-vivo and realtimevisualization of microscopic structures of the epidermis, dermo-epidermal junction and superficialdermis with a quasi-histological resolution. It has been used in the evaluation of benignand malignant lesions, showing a great potential for applications in basic research and clinicaldermatology. This paper reviews concepts about the development and basic principles of the RCMand its applications in clinical practice.

Light and electron microscopic studies of the effect of combined topical calcipotriol and PUVA therapy on patients with vitiligo

Vitiligo is an acquired depigmenting disorder that is characterized by loss of melanocytes from the epidermis. A large variety of therapeutic agents are being tried for the treatment of vitiligo, but an optimal treatment is not yet available. This study was conducted to estimate the efficacy of combination of topical calcipotriol and PUVA therapy in patients with vitiligo. Twenty-one patients with vitiligo were enrolled in this study, and were classified into three groups: group 1 [control group], nonlesional [normal] skin biopsies from all included patients in the study; group 2 [pretreated group], biopsies from lesional [vitiligo] skin from all included patients in the study; and group 3 [post-treated group], all included patients in the study, randomly divided into three subgroups: group 3a, included seven patients treated with topical calcipotriol cream [0.005%] twice daily for six months; group 3b, included seven patients treated with PUVA for six months; and group 3c, seven included patients treated with combined topical calcipotriol cream and PUVA twice daily for 6 months. Skin biopsies were taken, fixed, processed, and stained to assess histological changes [H and E], melanin pigment distribution [Masson-Fontana stain], and intercellular adhesion molecule 1 [ICAM-1] expression in the dermis and epidermis [immunohistochemical techniques]. Moreover, for statistical analysis, the mean area percentage of melanin pigment distribution in the epidermis, optical density of ICAM-1 expression in the epidermis and dermis of the immunostained sections, and number of melanosomes [using electron microscope] in the basal and spinous keratinocytes were measured and counted, respectively, in each of the studied groups. Vitiligo biopsies treated with combination of topical calcipotriol and PUVA showed disappearance of mononuclear cell infiltration, highly significant increase in the amount of melanin pigment concentration, highly significant decrease in ICAM-1 expression in the epidermis and dermis, and significant increase in the number of melanosomes when compared with the vitiligo group. Moreover, reappearance of melanocytes and regeneration of keratinocytes ultrastructure were observed. Combined topical calcipotriol and PUVA therapy showed complete repigmentation in vitiligo skin associated with regeneration of keratinocytes and reappearance of melanocytes in a shorter duration when compared with topical calcipotriol or PUVA therapy alone

Testes físico-mecânicos e físico-químicos do couro da tilápia vermelha / Physical-mechanical and physical-chemical tests of red tilapia leather

Realizaram-se testes físico-mecânicos e físico-químicos em couro de tilápia vermelha (Oreochromis spp.) a fim de testar a sua resistência. As amostras foram distribuídas em delineamento inteiramente ao acaso com dois tratamentos: no T1, procedeu-se à retirada do corpo-de-prova no sentido longitudinal e, no T2, à retirada do corpo-de-prova no sentido transversal. Para os testes de determinação da resistência à tração, alongamento e rasgamento progressivo, foi utilizado o dinamômetro EMIC, com velocidade de afastamento entre as cargas de 100 ± 20mm/min, em ambiente climatizado (± 23ºC e UR do ar de 50 por cento), por 24 horas. A espessura do couro variou de 0,61 a 0,75mm, mas não houve diferença entre os sentidos analisados. O couro apresentou maior resistência à tração no sentido transversal, 25,89N/mm², (P<0,01), comparado ao sentido longitudinal, 14,20 N/mm². O alongamento foi significativamente (P<0,05) maior no sentido longitudinal, 80,8 por cento, em relação ao transversal, 62,6 por cento. Não houve diferença para o rasgamento progressivo entre os tratamentos. O couro apresentou teor de óxido de cromo de 3,8 por cento, graxa de 15,1 por cento e pH e cifra diferencial de 3,5 e 0,5, respectivamente. Os valores nos testes de resistência e físico-químicos apresentados pelo couro indicam que ele pode ser utilizado para a confecção de vestuário e artefatos de couro em geral.

Physical-mechanical and physical-chemical tests were carried out on red tilapia leather. They were distributed in a completely randomized design with two treatments: T1 = longitudinal body of proof; T2 = transversal body of proof. It was used the EMIC dynamometer for the tests of resistance to traction and elongation and the progressive tearing, with the speed of 100±20mm/min away between the charges, in an acclimatized room (±23ºC and relative humidity of 50 percent) during 24 hours. The thickness of the analyzed leathers ranged from 0.61 to 0.75mm, without differing between the analyzed ways. The leather demonstrated a higher resistance to traction in the transversal direction (25.89N/mm²) (P<0.01), when compared to the longitudinal one (14.20N/mm²). However, the elongation was significantly higher (P<0.05) in the longitudinal direction (80.8 percent) when compared to the transversal (62.6 percent). There was no significant difference for the progressive tearing between the treatments (longitudinal = 18.56N/mm; transversal = 21.90N/mm). The leather demonstrated a content of 3.8 percent of chromium oxide, 15.1 percent of grease, and pH and difference value of 3.5 and 0.5, respectively. The values in the resistance and physical-chemical tests shown by the leather indicate that it may be used for clothing and leather artifacts in general.

Effects of isoflavones on the skin of postmenopausal women: a pilot study

OBJECTIVE: The aim of this study was to evaluate the effects of isoflavones on the skin of postmenopausal women. DESIGN: A prospective study was performed with 30 postmenopausal women before and immediately after the end of treatment with 100 mg/day of an isoflavones-rich, concentrated soy extract for six months. A skin punch was performed in the gluteal region for sample collection before and immediately after the treatment program. Morphometric determination of epidermal thickness, the papillary index (wrinkling), and the amount of dermal elastic and collagen fibers was assessed. In addition, the number of blood vessels in the sample was also evaluated. The paired Student¡¯s t-test was used for statistical analysis (P ¡Ü 0.05). RESULTS: Isoflavone treatment resulted in a 9.46% increase in the thickness of the epidermis in 23 patients. In addition, the papillary index was reduced in 21 women. The papillary index was inversely proportional to skin wrinkling, i.e., there were a large number of papillae after treatment. The amount of collagen in the dermis was increased in 25 women (86.2%). In 22 women (75.8%) we observed that the number of elastic fibers increased. The number of dermal blood vessels was significantly increased in 21 women.CONCLUSION: Our data show that the use of a concentrated, isoflavone-rich soy extract during six consecutive months caused significant increases in epithelial thickness, the number of elastic and collagen fibers, as well as the blood vessels.

Stydy of clinical, biochemical, electroencephalographic, radiological and electron microscopic features in patients with batten disease

Neuronal ceroid lipofuscinoses "NCLs" are a group of common hereditary neurodegenerative metabolic disease. Batten disease is its juvenile type that usually occurs between the age of 4-10 years. The aim of this work is to study the clinical picture, biochemical, enzymatic changes, electroencephalographic abnormalities, neuroradiological imaging and ultrastructural alterations in skin biopsies using transmission electron microscope. Methods: The study was carried out on five patients suspected having neuronal ceroid lipofucinoses. All patients subjected to complete history taking, with special concern to detailed family history. Full physical, neurological, ophthalmologic examination was done with developmental assessment. Routine lab investigations as well as a batteries of metabolic screen and MRI were performed. Electroencephalogram examination was done under standerd conditions with different provocations. Skin biopsies were taken for histological and ultrastructural examination using transmission electron microscope. The current study included 5 males aged between 6 to 13 years with a mean of 8.5 +/- 1.2 years, two of them had positive family history. The main presentations were; epilepsy in all cases [100%]; generalized tonic clonic convulsion in 3 cases [60%] and myoclonic epilepsy in 2 cases [40%]. Mental retardation and rapidly progressing visual loss were present in all cases [100%]. Motor deficits in the form of spastic quadriparesis were found in 4 cases [80%], extra pyramidal manifestations in one case [20%] and cerebellar ataxia manifested in one patient [20%]. MRI showed atrophic brain and cerebellar hemispheres in all cases. Light microscopic study of skin biopsies displayed unremarkable changes, while ultrastructural examination revealed presence of cytoplasmic membrane bound osmophilic granular, curvilinear and finger print inclusion bodies

Comparative study on the effect of retinoic acid, glycolic acid and salicylic acid on the thin skin of adult female guinea pig. light and electron microscopic study

Many products had been used as chemical peeling agents to renew thin skin. Retinoic, Glycolic and Salicylic acids were used in many dermatological diseases with hypdrkeratinization and/or hyperpigmentation. To evaluate and compare the effects of these acids on the thin skin of adult female guinea pig. Twenty animals were used and were divided into four equal groups. Group I served as the control group, whereas the other treated groups were topically applicated daily on shaved area of back skin with Retinoic [0.05%], Glycolic [12%] and Salicylic [10%] acids in Groups II, III and IV, respectively. Thin skin specimens were processed for light and transmission electron microscopic studies. Morphometric and statistical studies were done. Groups II, III and IV showed high significant increase in mean thickness of epidermal nucleated keratinocytes as compared to Control group. Group II showed proliferation hyperkeratosis and acanthosis and shrinkage of the sebaceous glands with decreased sebum production. Group III showed skin peeling by removing superficial layers of epidermis, cytoplasmic and nuclear degeneration with disruption of intercellular junctions and degeneration in melanocytes with marked decrease of melanin. Both Retinoic and Glycolic acids apparently increased the production of collagen and elastic fibers as compared to control. Group IV showed skin peeling mainly by direct action on intercellular cement substance and partially by inducing proliferation hyperkertosis. Both Retinoic and Glycolic acids showed complementary actions in treatment of hyperpigmentation and as chemical peeling agents. Salicylic acid also is a peeling agent, but its effects could be covered by either Retinoic or Glycolic acids. It is recommended to use combination of both Retinoic and Glycolic acids to give better effects on various skin disorders with hyperkeratinization and hyperpigmentation

Three-dimensional characteristics of the interface epithelium-connective tissue surface of fingeres lamina propria of Cebus apella monkey: scanning electron microscopy study / Características tri-dimensionales superficiales de la lámina propia de los dedos de monos Cebus apella: interfase del tejido epitelial-conectivo del estudio con microscopía electrónica de barrido

In the present paper were analysed the three-dimensional characteristics of the interface epithelium-connective tissue surface of finger prints of Cebus apella monkey employing the scanning electron microscopic methods. The connective tissue core (CTC) and epithelial papillae were examined verifying the three-dimensional configuration of the tissue projections. The samples were fixed in Bouin solsution for histologic preparations and in modified Karnovsky for examine to observe in scanning electron microscopy. After treatment in the 10 percent NaOH solution during 3 to 5 day s, the surface of finger prints revealed a distribution of CTC of lamina propria in situ showing original three-dimensional SEM images. The linear and circular dispositions CTC, and the furrows were clearly identified. Each pointed papilla presented a large base and longitudinal disposition of thick collagen fiber bundles and in some areas with a complex reticular formations. The longitudinal furrows between the pointed papillae exhibited a dense layer of connective tissue and showed only low CTC or laminar in shape. The presence of numerous foramina of sweat gland were noted in three-dimensional SEM images.

Se analizaron tridimensionalmente las características de la interfase del tejido epitelio-conectivo en la superficie de huellas dactilares de mono Cebus apella, empleando el microscopio electrónico de barrido. El núcleo del tejido conectivo (NTC) y las papilas epiteliales se examinaron en las tres dimensiones. Las muestras fueron fijadas en solución histológica Bouin para los preparativos y en Karnovsky modificada para examinar y observar en el microscopio electrónico de barrido. Después del tratamiento en solución de NaOH al 10 por ciento, durante 3 a 5 días, la superficie de las huellas dactilares revelaron una distribución de NTC de lámina propia in situ mostrando imágenes en tres dimensiones originales con microscopio electrónico de barrido. Las disposiciones lineal y circular del NTC están claramente identificadas. Cada papila señalada presenta una gran base y la disposición longitudinal de gruesos haces de fibras de colágeno y, en algunas zonas con formaciones de un complejo reticular. Los surcos longitudinales entre las papilas señaladas mostraron una densa capa de tejido conectivo con NTC bajos o de forma laminar. La presencia de numerosos forámenes de las glándulas sudoríparas se observaron en tres dimensiones.

Histological and immunohistochemical study on the effect of tretinoin on the thin skin of adult male albino rat

Vitamin A is important for epithelial cell proliferation and differentiation. Synthetic vitamin A derivatives, known as retinoids, have been extensively used in the last few years to treat a variety of clinical skin conditions. The aim of this study was to evaluate the histological changes in rat thin skin that might result from long term exposure to a retinoid derivative [tretinoin] and the possibility of recovery after its withdrawal. Thirty adult male albino rats were divided into three equal groups; control group, tretinointreated group, and the third group was topically treated with tretinoin cream for four weeks, then the animals were left without treatment for another four weeks. Skin specimens were processed and stained with haematoxylin and eosin and with Mallory's trichrome stain while ultrathin sections were contrasted with uranyl acetate and lead citrate. The epidermal thickness was measured and the results were statistically analysed. The effect of tretinoin on cell proliferation was further evaluated immunohistochemically using the proliferating cell nuclear antigen [PCNA]. Specimens from tretinoin-treated animals showed focal vacuolation of the keratinocytes with widening of intercellular spaces, an increase in keratohyalin granules of stratum granulosum and disorganized cells of stratum corneum. There was also a statistically highly significant increase in the epidermal thickness. The dermis showed increased collagen content and cellular infiltration mainly formed of fibroblasts. Immunohistochemical study revealed a highly significant increase in PCNA-positive nuclei in the epidermal cells. The recovery group showed mild affection of skin structure. Long-term exposure to tretinoin may induce structural changes in rat skin, being partially reversible after withdrawal of the drug. Therefore, the use of tretinoin should be restricted to the advice of dermatologists