Avaliação in vitro do potencial reparador de matriz de Quitosana associada a Madecassoside: aplicações no tratamento de lesões cutâneas / In vitro evaluation of the matrix repairing potential of Chitosan associated with Madecassoside: applications in the treatment of skin lesions

A cicatrização de úlceras cutâneas depende de fatores-chave que incluem a interação adequada dos diferentes constituintes celulares da epiderme, derme e tecido subcutâneo. A restauração da barreira epidérmica é altamente eficiente durante o período embrionário, com um relativo decréscimo na vida adulta. Distúrbios sistêmicos, como diabetes e hanseníase, podem comprometer a capacidade de reparação da pele, gerando ulceras crônicas, que são consideradas como relevantes problemas de saúde pública. O presente estudo se propôs a estabelecer parâmetros de eficiência de membranas bioativas, preparadas com o biopolímero quitosana (QT), em associação ao extrato vegetal, madecassoside (MA). Nas preparações obtidas, foram avaliadas características físico-químicas e propriedades antimicrobianas. A biocompatibilidade das preparações, e sua capacidade de promover migração celular, foi testada in vitro em fibroblastos da linhagem NIH/3T3. As membranas foram divididas em grupos: QT 2%; QT/MA 0,10% (QTMA010); QT/MA 0,25% (QTMA025); QT/MA 0,50% (QTMA050). Os grupos foram avaliados em diferentes intervalos de tempo, de 0 a 96 horas (T0, T24, T48, T72, T96). Nossos dados indicam que membranas bioativas, preparadas com quitosana (QT 2%) e madecassoside (MA 0,10%, 0,25%, 0,50%), são biocompatíveis e possuem propriedades físico-químicas adequadas. As preparações contendo associação de ambos os compostos se mostraram superiores à QT. A capacidade de promover migração de fibroblastos, in vitro, foi estatisticamente superior em todos os grupos acrescidos de MA, indicando um papel relevante desse composto em preparações de utilização tópica para cicatrização úlceras cutâneas.

Papain immobilization on heterofunctional membrane bacterial cellulose as a potential strategy for the debridement of skin wounds.

We combined the chemical and physical methods of papain immobilization through the aldehyde groups available on oxidized bacterial cellulose (OxBC) to provide high proteolytic activity for future applications as bioactive dressing. Bacterial cellulose (BC) was obtained by the fermentation of Komagataeibacter hansenii in Hestrin-Schramm medium for 5 days, followed by purification and oxidation using NaIO4. Surface response methodology was used to optimize papain immobilization (2%, w/v) for 24 h. The independent variables: pH (3-7) and temperature (5 to 45 °C) were investigated. The mathematically validated optimal conditions of 45 °C and pH 7 had a statistical effect on the immobilization yield (IY) of papain in OxBC (52.9%). These ideal conditions were also used for papain immobilization in BC (unoxidized). The IY of 9.1% was lower than that of OxBC. OxBC-Papain and BC-Papain were investigated using thermal analysis, confocal microscopy, and diffusion testing. The OxBC support exhibited a more interactive chemical structure than the BC support, and was capable of immobilizing papain by covalent bonds (-C-NHR) and adsorption (ion exchange), with 93.3% recovered activity, 49.4% immobilization efficiency, and better thermal stability. Papain immobilized to OxBC by adsorption displayed 53% widespread papain activity. The results indicate the potential of prolonged bioactivity in debrided chronic wounds.

Fatores de risco para a ocorrência das úlceras plantares decorrente da hanseníase de acordo com a árvore de decisão / Risk factors for plantar ulcers from leprosy according to the decision tree

Objective:To identify the risk factors for plantar ulcers in patients with leprosy.Methods:This is an epidemiological, observational, cross-sectional and analytical study. The population was composed of leprosy cases reported from 2005 to 2016. Pearson's Chi-square test or Fisher's exact test andMann-Whitney test were used for the univariate analysis, with a statistical significance of 5% (p < 0.05). In the multivariate analysis, a decision tree was elaborated using the CHAID algorithm. Results:Clinical form, degree of physical incapacity at discharge, affected nerve and the lack of insoles or adapted footwear before appearing to ulcer are risk factors for plantar ulcer. Conclusion:the need for an early diagnosis of leprosy was highlighted, as well as the efficient association of non-drug interventions with disability prevention techniques and the use of accommodating insoles and/or special shoes.

Objetivo:Identificar os fatores de risco para a ocorrência das úlceras plantares em pacientes com hanseníase. Métodos:Trata-se de um estudo epidemiológico, do tipo observacional, transversal e analítico. A população foi composta pelos casos de hanseníase notificados no período de 2005 a 2016. Para a análise univariada foram utilizados os testes Qui-quadrado de Pearson ou teste exato de Fisher e teste de Mann-Whitney, com significância estatística de 5% (p < 0,05). Na análise multivariada, foi elaborada árvore de decisão utilizando o algoritmo CHAID. Resultados: A forma clínica, grau de incapacidade física na alta, nervo acometido e o não uso de palmilhas ou calçado adaptado antes de surgir a úlcera são fatores de risco para a ocorrência de úlcera plantar. Conclusão:evidenciou a necessidade do diagnóstico precoce da hanseníase, como também da eficiente associação das intervenções medicamentosas e não medicamentosas por meio das técnicas de prevenção de incapacidade e uso de palmilhas acomodativas e/ou calçados especiais

Differential expression of MicroRNAs in leprosy skin lesions

Leprosy, a chronic infectious disease caused by Mycobacterium leprae, is a major public health problem in poor and developing countries of the Americas, Africa, and Asia. MicroRNAs (miRNAs), which are small non-coding RNAs (18-24 nucleotides), play an important role in regulating cell and tissue homeostasis through translational downregulation of messenger RNAs (mRNAs). Deregulation of miRNA expression is important for the pathogenesis of various neoplastic and non-neoplastic diseases and has been the focus of many publications; however, studies on the expression of miRNAs in leprosy are rare. Herein, an extensive evaluation of differentially expressed miRNAs was performed on leprosy skin lesions using microarrays. Leprosy patients, classified according to Ridley and Jopling's classification or reactional states (R1 and R2), and healthy controls (HCs) were included. Punch biopsies were collected from the borders of leprosy lesions (10 tuberculoid, 10 borderline tuberculoid, 10 borderline borderline, 10 borderline lepromatous, 4 lepromatous, 14 R1, and 9 R2) and from 9 HCs. miRNA expression profiles were obtained using the Agilent Microarray platform with miRBase, which consists of 1,368 Homo sapiens (hsa)-miRNA candidates. TaqMan quantitative real-time reverse transcription polymerase chain reaction (RT-PCR) was used to validate differentially expressed miRNAs. Sixty-four differentially expressed miRNAs, including 50 upregulated and 14 downregulated (fold change ≥2.0, p-value ≤ 0.05) were identified after comparing samples from patients to those of controls. Twenty differentially expressed miRNAs were identified exclusively in the reactional samples (14 type 1 and 6 type 2). Eight miRNAs were validated by RT-PCR, including seven upregulated (hsa-miR-142-3p, hsa-miR-142-5p, hsa-miR-146b-5p, hsa-miR-342-3p, hsa-miR-361-3p, hsa-miR-3653, and hsa-miR-484) and one downregulated (hsa-miR-1290). These miRNAs were differentially expressed in leprosy and several other diseases, especially those related to the immune response. Moreover, the integration of analysis of validated mi/mRNAs obtained from the same samples allowed target pairs opposite expression pattern of hsa-miRNA-142-3p and AKR1B10, hsa-miRNA-342-3p and FAM180b, and hsa-miRNA-484 and FASN. This study identified several miRNAs that might play an important role in the molecular pathogenesis of the disease. Moreover, these deregulated miRNAs and their respective signaling pathways might be useful as therapeutic markers, therapeutic targets, which could help in the development of drugs to treat leprosy

Autochthonous lepromatous leprosy in a Spanish woman with burns on both feet and skin lesions.

The incidence of leprosy is decreasing worldwide and it is considered a rare disease in developed countries. In Spain, leprosy is mainly an imported disease with only few autochthonous cases seen. The diagnosis is difficult because of a low index of suspicion and the absence of visualisation of fast-acid bacilli in the lesions. Here, we report an autochthonous case of leprosy diagnosed after 4 years of evolution of skin lesions. Mechanical rupture of the biopsy helped finally to make the correct diagnosis of the disease.

Effect of the triterpene 3ß, 6ß, 16ß-trihydroxylup-20(29)-ene isolated from the leaves of Combretum leprosum Mart. on cutaneous wounds in mice.

ETHNOPHARMACOLOGICAL RELEVANCE: Combretum leprosum Mart. is a native species of the Caatinga, an important biome in the Brazilian semi-arid region. This species is commonly used in Brazil as a healing agent, as well as for the treatment of skin diseases. AIM OF THE STUDY: This study investigated the healing potential of the ethanolic extract (EECL) and the bioactive triterpene 3ß, 6ß, 16ß-trihydroxylup-20 (29)-ene (CLF-1) isolated from the leaves of C. leprosum. MATERIALS AND METHODS: Skin wounds (1cm(2)) were created in the dorsal zone of mice with a scalpel blade number 15. The treatment consisted in a daily topical application of 100 µl of 150 mM NaCl, EECL and CLF-1 (at 10 µg/100 µl) for 12 days. The lesions were then macro and microscopically evaluated. RESULTS: On postoperative day (POD) 2, the lesions treated with EECL and CLF-1 showed a moderate presence of vessels of the granulation tissue progressing in the dermis. The same effect was not observed in the control group. The treatment with EECL and CLF-1 stimulated angiogenesis, resulting in a rapid deposition of extracellular matrix (ECM). Moreover, the animals treated with EECL and CLF-1 showed smaller lesions on POD 7, primarily due to the contraction in the reticular dermis induced by organization of myofibroblasts, which was not observed in the group treated with NaCl. In addition, the lesions treated with EECL and CLF-1 showed ECM restructuration and presence of epithelium coating, which was not observed in the group treated with NaCl, in which the lesions showed no epithelial lining, suggesting delayed healing. CONCLUSION: CLF-1 isolated from the leaves of C. leprosum may be considered to be an important molecule for the treatment of skin lesions. However, further investigations are necessary to establish its role in chronic lesions and to elucidate the mechanism of action involved in the cutaneous healing process. To the best of our knowledge, this is the first report describing the pro-healing activity of the ethanolic extract and the triterpene 3ß, 6ß, 16ß-trihydroxylup-20(29)-ene isolated from leaves of C. leprosum.

Genome-wide screening of mRNA expression in leprosy patients

Leprosy, an infectious disease caused by Mycobacterium leprae, affects millions of people worldwide. However, little is known regarding its molecular pathophysiological mechanisms. In this study, a comprehensive assessment of human mRNA was performed on leprosy skin lesions by using DNA chip microarrays, which included the entire spectrum of the disease along with its reactional states. Sixty-six samples from leprotic lesions (10TT, 10BT, 10BB, 10BL, 4LL, 14R1, and 10R2) and nine skin biopsies from healthy individuals were used as controls (CC) (ages ranged from 06 to 83 years, 48 were male and 29 female). The evaluation identified 1580 differentially expressed mRNAs [Fold Change (FC) ≥ 2.0, p ≤ 0.05] in diseased lesions vs. healthy controls. Some of these genes were observed in all forms of the disease (CD2, CD27, chit1, FA2H, FAM26F, GZMB, MMP9, SLAMF7, UBD) and others were exclusive to reactional forms (Type "1" reaction: GPNMB, IL1B, MICAL2, FOXQ1; Type "2" reaction: AKR1B10, FAM180B, FOXQ1, NNMT, NR1D1, PTX3, TNFRSF25). In literature, these mRNAs have been associated with numerous pathophysiological processes and signaling pathways and are present in a large number of diseases. The role of these mRNAs maybe studied in the context of developing new diagnostic markers and therapeutic targets for leprosy.

Diagnostic challenges of single plaque-like lesion paucibacillary leprosy.

The diagnosis of single-lesion paucibacillary leprosy remains a challenge. Reviews by expert dermatopathologists and quantitative polymerase chain reaction (qPCR) results obtained from 66 single-plaque biopsy samples were compared. Histological findings were graded as high (HP), medium (MP) or low (LP) probability of leprosy or other dermatopathy (OD). Mycobacterium leprae-specific genes were detected using qPCR. The biopsies of 47 out of 57 clinically diagnosed patients who received multidrug therapy were classified as HP/MP, eight of which were qPCR negative. In the LP/OD (n = 19), two out of eight untreated patients showed positive qPCR results. In the absence of typical histopathological features, qPCR may be utilised to aid in final patient diagnosis, thus reducing overtreatment and delay in diagnosis.

Diagnostic challenges of single plaque-like lesion paucibacillary leprosy

The diagnosis of single-lesion paucibacillary leprosy remains a challenge. Reviews by expert dermatopathologists and quantitative polymerase chain reaction (qPCR) results obtained from 66 single-plaque biopsy samples were compared. Histological findings were graded as high (HP), medium (MP) or low (LP) probability of leprosy or other dermatopathy (OD). Mycobacterium leprae-specific genes were detected using qPCR. The biopsies of 47 out of 57 clinically diagnosed patients who received multidrug therapy were classified as HP/MP, eight of which were qPCR negative. In the LP/OD (n = 19), two out of eight untreated patients showed positive qPCR results. In the absence of typical histopathological features, qPCR may be utilised to aid in final patient diagnosis, thus reducing overtreatment and delay in diagnosis.

Pathophysiology of psoriasis.

Psoriasis is a chronic inflammatory papulosquamous disease characterized by multiple remissions and relapses. For long, it was believed to be primarily a disorder of keratinization. However, the successful use of traditional immunosupressants and newer immunomodulatory agents in the treatment of psoriasis led to the belief that psoriasis is primarily a disease of Th1 cell immune dysregulation. Recent developments have brought up several new findings such as the role of Th17 cells and evidence of skin barrier dysfunction in psoriasis, akin to atopic dermatitis. The present review aims to focus on these new developments and explain the pathogenesis of psoriasis on the basis of currently available information.

Leprosy patients: neurotrophic factors and axonal markers in skin lesions / Pacientes com hanseníase: fatores neurotróficos e marcadores axonais em lesões de pele

Neurotrophins are growth factors with crucial roles in neural pathophysiology. These mediators functionally modulate nociceptive fibers, and changes in neurotrophins expression have been correlated with early loss of nociception in leprosy. This study investigated the expression of NGF, BDNF, and NT3 in dermal nerves of leprosy patients. Characterization of Remak bundles was achieved by p75NTR, and axonal markers NF-L and PGP 9.5 immunostaining. Clinical parameters of neural impairment have been evaluated by Semmes-Wenstein monofilaments. Our findings demonstrated decrease of NGF in borderline leprosy, when compared to control specimens. Similar results were observed in PGP 9.5 expression (borderline: p<0.001 and lepromatous: p<0.05) and NF-L (lepromatous: p<0.05), suggesting advanced Remak bundles degeneration in multibacillary leprosy. It has also been observed positive correlation between p75NTR and PGP 9.5, indicating association between Schwann cells and axons in Remak bundles. Present data indicate that neurotrophins imbalance may participate in the establishment of peripheral nerve damage.

Neurotrofinas são fatores de crescimento com papel fundamental na fisiopatologia neural. Esses mediadores modulam funcionalmente fibras nociceptivas. Mudanças em sua expressão têm sido relacionadas à perda precoce da nocicepção na hanseníase. Este estudo investigou a expressão de NGF, BDNF e NT3 em nervos dérmicos de pacientes hansenianos. A caracterização de fibras nervosas não mielinizadas foi feita por p75NTR e marcadores axonais NF-L e PGP 9.5. Os parâmetros clínicos de dano neural foram avaliados por monofilamentos Semmes-Wenstein. Nossos achados demonstram diminuição de NGF nos pacientes dimorfos em comparação aos controles. Resultados similares foram observados para PGP 9.5 (dimorfos: p<0,001; virchowianos: p<0,05) e NF-L (virchowianos: p<0.05), sugerindo degeneração avançada das terminações nervosas na hanseníase multibacilar. Foi observada correlação positiva entre p75NTR e PGP 9.5, indicando associação entre células de Schwann e axônios em fibras nervosas não mielinizadas. Os resultados indicam que o desequilíbrio na expressão das neurotrofinas pode participar do dano neural periférico.

Cicatrização de feridas decorrentes da hanseníase utilizando laser de baixa intensidade / Wound healing leprosy induced by low intensity laser

A hanseníase é causada pelo bacilo Mycobacterium leprae(M. leprae), apresentando grande capacidade de infectar vários indivíduos com contágio pelas vias aéreas superiores. A hanseníase considerada um problema de saúde pública, principalmente nos países subdesenvolvidos e nos em desenvolvimento, devido à presença de incapacidades e estigma social...

Leprosy is caused by the bacillus Mycobacterium leprae(M. leprae), presenting the capacity to infect multiple individuals with infection of the upper airways. Hansen considered a public health problem, especially in under developed countries and developing countries, due to the presence of disabilities and social stigma...

Skin manifestations of child abuse.

Child abuse is a major public health problem all over the world. There are four major types of abuse: physical abuse, sexual abuse, emotional abuse and neglect. The most common manifestations of child abuse are cutaneous and their recognition; and differential diagnosis is of great importance. Clinicians, especially dermatologists, should be alert about the skin lesions of child abuse. In the diagnosis and management of child abuse, a multidisciplinary approach with ethical and legal procedures is necessary. In this manuscript, cutaneous manifestations of physical, sexual, emotional abuse and neglect are reviewed and discussed.

Manifestaciones neurológicas de la lepra del rey Balduino IV de Jerusalén / Neurological evaluation of the leper king Balwing IV of Jerusalem

Introducción. Durante la Edad Media, los médicos fueron más conscientes de las manifestaciones de la lepra. El caso de Balduino, el rey leproso de Jerusalén (1161-1185), probablemente contribuyó a que en el mundo cristiano se incrementase el interés hacia esta enfermedad y la tolerancia hacia quienes la padecían. Revisamos las descripciones históricas de las manifestaciones neurológicas del cuadro clínico de este conocido rey. Desarrollo. Guillermo de Tiro proporciona una descripción de los primeros síntomas que presenta a los 9 años de edad, consistentes en hipoestesia en la mitad de su mano y brazo derechos, sin otras alteraciones cutáneas o nerviosas. Su cuadro clínico va empeorando y, al comienzo de la tercera década de la vida, la debilidad le impide caminar. Desarrolla una ceguera, en probable relación con una queratopatía secundariaa la afectación del VII par. Ataques repetidos de fiebre provocan un progresivo empeoramiento de su estado, y muere en Jerusalén,a la edad de 24 años, como consecuencia de la infección de sus úlceras. El primer signo de la enfermedad de Balduino es la anestesia; aunque no se describen lesiones cutáneas, es probable que sufriera una forma tuberculoide de lepra. La evolución posterior a una forma lepromatosa con grandes lesiones cutáneas hace pensar en un inicio con una forma borderline, inmunológicamente inestable. Conclusión. La biografía del rey leproso de Jerusalén proporciona interesantes descripciones de las manifestaciones neurológicas de su enfermedad. Además, da pistas acerca de los conocimientos que la medicina del siglo XII tenía de los síntomas de la lepra (AU)

Introduction. In the medieval period, physicians became more aware of leprosy symptoms and differentiated it from other similar diseases. Baldwin, the leper king of Jerusalem (1161-1185), probably contributed to an increasing interest and tolerance to this disease in medieval Christian states. We review historical descriptions of the neurological manifestations he developed. Development. William of Tyre gives us a description of first symptoms experienced by the prince when aged nine. He notices that half of his right arm and hand were partially numb. No skin or nervous lesions are described. By his early twenties, muscle weakness makes him unable to walk. He gets blinded, probably due to keratopathy related to facial nerves involvement. Repeated attacks of fever lead to progressive worsening of his disease. He finally dies in Jerusalem, aged twentyfive, probably due to a septicaemia from infected sores. The earliest sign of Baldwin’s disease is anaesthesia. Though skin lesions are not described, it is likely that at this point he had a tuberculoid form of leprosy. As his disease finally takes alepromatous form, we suspect that it began as a borderline, immunologically unstable form. Conclusion. Leper king Baldwin biography gives us interesting descriptions of neurological clinical features of leprosy. Besides, it helps us to discover twelfth century medicine knowledge about this disease (AU)