RESUMO
OBJECTIVES: To efficiently detect somatic UBA1 variants and establish a clinical scoring system predicting patients with pathogenic variants in VEXAS (vacuoles, E1 enzyme, X-linked, autoinflammatory, somatic) syndrome. METHODS: Eighty-nine Japanese patients with clinically suspected VEXAS syndrome were recruited [81 males and 8 females; median onset age (IQR) 69.3 years (62.1-77.6)]. Peptide nucleic acid-clamping PCR (PNA-PCR), regular PCR targeting exon 3 clustering UBA1 variants, and subsequent Sanger sequencing were conducted for variant screening. Partitioning digital PCR (pdPCR) or targeted amplicon deep sequencing (TAS) was also performed to evaluate the variant allele frequency (VAF). We developed our clinical scoring system to predict UBA1 variant-positive and negative patients and assessed the diagnostic value of our system using receiver operating characteristic (ROC) curve analysis. RESULTS: Forty patients with reported pathogenic UBA1 variants (40/89, 44.9%) were identified, including a case having a variant with VAF of 1.7%, using a highly sensitive method. Our clinical scoring system considering >50 years of age, cutaneous lesions, lung involvement, chondritis, and macrocytic anaemia efficiently predicted patients with UBA1 variants (the area under the curve for the scoring total was 0.908). CONCLUSIONS: Genetic screening with the combination of regular PCR and PNA-PCR detected somatic UBA1 variants with high sensitivity and specificity. Our scoring system could efficiently predict patients with UBA1 variants.
RESUMO
In April 2015, Consumer Affairs Agency of Japan launched a new food labeling system known as "Foods with Function Claims (FFC)." Under this system, the food industry independently evaluates scientific evidence on foods and describes their functional properties. As of May 23, 2017, 1023 FFC containing 8 fresh foods have been launched. Meanwhile, to clarify the health-promoting effects of agricultural products, National Agriculture and Food Research Organization (NARO) implemented the "Research Project on Development of Agricultural Products" and demonstrated the risk reduction of osteoporosis of ß-cryptoxanthin rich Satsuma mandarins and the anti-allergic effect of the O-methylated catechin rich tea cultivar Benifuuki. These foods were subsequently released as FFC. Moreover, NARO elucidated the health-promoting effects of various functional agricultural products (ß-glucan rich barley, ß-conglycinin rich soybean, quercetin rich onion, etc.) and a healthy boxed lunch. This review focuses on new food labeling system or research examining functional aspects of agricultural products.
Assuntos
Produtos Agrícolas , Rotulagem de Alimentos/normas , Alimento Funcional/normas , Legislação sobre Alimentos , Rotulagem de Alimentos/legislação & jurisprudência , Promoção da Saúde , Humanos , JapãoRESUMO
Human immunodeficiency virus (HIV)-associated CD8+ T-cell skin infiltrative disease with severe erythroderma has rarely been reported. While HIV-positive patients are prone to develop lymphoma, which is often associated with Epstein-Barr virus, polymorphic lymphoproliferative disorder is rare, accounting for <5% of cases. We herein report a 41-year-old HIV-positive man who presented with a fever, erythroderma, and lymphadenopathy and was diagnosed with the coexistence of both diseases. His condition improved significantly with continued antiretroviral therapy. This case suggests that HIV-induced immunodeficiency is central to the pathogenesis of both entities and that improvement of the immunodeficient state is an effective treatment.
Assuntos
Linfócitos T CD8-Positivos , Infecções por Vírus Epstein-Barr , Transtornos Linfoproliferativos , Humanos , Masculino , Adulto , Linfócitos T CD8-Positivos/imunologia , Infecções por Vírus Epstein-Barr/complicações , Infecções por Vírus Epstein-Barr/imunologia , Infecções por Vírus Epstein-Barr/tratamento farmacológico , Transtornos Linfoproliferativos/tratamento farmacológico , Transtornos Linfoproliferativos/imunologia , Transtornos Linfoproliferativos/diagnóstico , Síndrome da Imunodeficiência Adquirida/tratamento farmacológico , Síndrome da Imunodeficiência Adquirida/complicações , Síndrome da Imunodeficiência Adquirida/imunologia , Herpesvirus Humano 4/imunologia , Infecções por HIV/tratamento farmacológico , Infecções por HIV/complicações , Infecções por HIV/imunologia , Resultado do Tratamento , Terapia Antirretroviral de Alta Atividade , Linfócitos B/imunologiaRESUMO
We describe a case of recalcitrant phaeohyphomycosis caused by Exophiala lecanii-corni, which was previously reported as Exophiala jeanselmei, infection. A 63-year-old Japanese woman presented with a 15-year history of multiple pruritic erythematous patches and plaques on the face. Histopathological examination and fungal culture revealed phaeohyphomycosis by E. jeanselmei. The attempted treatments included 6 g/day 5-flucytosine (5-FC), 100 mg/day itraconazole (ITCZ), and local hyperthermia. 5-FC was effective initially, but the patient deteriorated after discontinuation. Subsequently, she was referred to our hospital. Histopathological examination showed granuloma with multinucleated giant cells with infiltrating fungal hyphae in the dermis. The causative fungus was finally identified as E. lecanii-corni by ribosomal RNA gene analysis. The patient improved after receiving 200 mg/day ITCZ orally for 15 months with local hyperthermia. In the present case, we confirmed the identification of E. lecanii-corni as the causative agent by molecular methods. We also emphasize the importance of combination therapy with antimycotic agents and local hyperthermia in phaeohyphomycosis.
Assuntos
Exophiala , Feoifomicose , Exophiala/genética , Feminino , Humanos , Pessoa de Meia-Idade , Feoifomicose/diagnóstico , Feoifomicose/tratamento farmacológicoRESUMO
We developed a method to measure the rupture forces between antibody and antigen by atomic force microscopy (AFM). Previous studies have reported that in the measurement of antibody-antigen interaction using AFM, the specific intermolecular forces are often obscured by nonspecific adhesive binding forces between antibody immobilized cantilever and substrate surfaces on which antigen or nonantigen are fixed. Here, we examined whether detergent and nonreactive protein, which have been widely used to reduce nonspecific background signals in ordinary immunoassay and immunoblotting, could reduce the nonspecific forces in the AFM measurement. The results showed that, in the presence of both nonreactive protein and detergent, the rupture forces between anti-ferritin antibodies immobilized on a tip of cantilever and ferritin (antigen) on the substrate could be successfully measured, distinguishing from nonspecific adhesive forces. In addition, we found that approach/retraction velocity of the AFM cantilever was also important in the reduction of nonspecific adhesion. These insights will contribute to the detection of specific molecules at nanometer scale region and the investigation of intermolecular interaction by the use of AFM.
Assuntos
Anticorpos/imunologia , Anticorpos/metabolismo , Ferritinas/imunologia , Ferritinas/metabolismo , Imunoensaio/métodos , Microscopia de Força Atômica/métodos , Animais , Bovinos , Detergentes/farmacologia , Humanos , Ligação Proteica/efeitos dos fármacos , Especificidade por Substrato , Compostos de Sulfidrila/metabolismo , Propriedades de SuperfícieRESUMO
Trichloroacetic acid (TCA) is one of the most widely used peeling agents, and induces full necrosis of the whole epidermis, followed by reconstitution of the epidermis and the matrix of the papillary dermis. The cytotoxic effects of TCA, such as suppressing proliferation of keratinocytes and fibroblasts and protein synthesis by fibroblasts, have already been reported. However, the entire biological mechanism responsible for TCA peeling has yet to be determined. Hypothetical activation effects of TCA treatment on epidermal cells to induce production of growth factors and cytokines are examined, and are compared with its cytotoxic effects in terms of time course and applied TCA concentrations. After various periods of incubation with TCA, viability of Pam212 murine keratinocytes was investigated with MTT assay and dye exclusion assay, and production of growth factors and cytokines with reverse transcription-polymerase chain reaction (RT-PCR). Changes in platelet-derived growth factor (PDGF)-B mRNA expression and protein production in the human skin specimens after TCA application were then examined by RT-PCR and immunohistochemistry, respectively. Incubation with TCA showed cytotoxicity and induced death of Pam212 cells, depending on the incubation period and the TCA concentration. In addition, expressions of PDGF-B, tumor growth factor (TGF)-alpha, TGF- beta1 and vascular endothelial growth factor, which are the growth factors reportedly secreted from keratinocytes during wound healing, were all detected in Pam212 cells after short-term treatment with TCA. Expressions of inflammatory cytokines such as interleukin (IL)-1 and IL-10 were also induced. In TCA-treated NIH-3T3 fibroblasts, in contrast, observed was upregulation of only keratinocyte growth factor, which is reportedly secreted from fibroblasts, as well as the similar cytotoxic effect. In human skin, PDGF-B mRNA expression became significantly upregulated after TCA application, and then immediately downregulated. Immunoreactive PDGF-B in the cytoplasm of keratinocytes became detectable throughout the epidermis after TCA application, reached maximum after the peak of mRNA expression, and then declined significantly over 24 h when the epidermis became completely necrotic. The TCA-treated epidermis acts as a major source of growth factors, including PDGF-B, before undergoing full necrosis. This effect might contribute to a promotion of re-epithelialization and dermal regeneration without wound contraction and scarring.
Assuntos
Cáusticos/farmacologia , Epiderme/efeitos dos fármacos , Fibroblastos/efeitos dos fármacos , Queratinócitos/efeitos dos fármacos , Proteínas Proto-Oncogênicas c-sis/metabolismo , Ácido Tricloroacético/farmacologia , Administração Cutânea , Adulto , Animais , Cáusticos/administração & dosagem , Cáusticos/toxicidade , Sobrevivência Celular/efeitos dos fármacos , Relação Dose-Resposta a Droga , Epiderme/metabolismo , Epiderme/patologia , Feminino , Fator 7 de Crescimento de Fibroblastos/metabolismo , Fatores de Crescimento de Fibroblastos/metabolismo , Fibroblastos/metabolismo , Fibroblastos/patologia , Humanos , Imuno-Histoquímica , Interferon gama/metabolismo , Interleucinas/metabolismo , Queratinócitos/metabolismo , Queratinócitos/patologia , Camundongos , Células NIH 3T3 , Necrose , Proteínas Proto-Oncogênicas c-sis/genética , RNA Mensageiro/metabolismo , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Fatores de Tempo , Fator de Crescimento Transformador alfa/metabolismo , Fator de Crescimento Transformador beta1/metabolismo , Ácido Tricloroacético/administração & dosagem , Ácido Tricloroacético/toxicidade , Regulação para Cima , Fator A de Crescimento do Endotélio Vascular/metabolismoRESUMO
It has been reported that gamma-linolenic acid contained in borage oil is effective against atopic dermatitis. The clinical effects of undershirts coated with borage oil rich in gamma-linolenic acid on atopic dermatitis were evaluated. Thirty-two children, aged 1-10 years, were involved in the clinical control study. Sixteen had worn undershirts coated with borage oil everyday for 2 weeks, and 16 had worn non-coated undershirts as a placebo. Their symptoms were assessed on a 4-point scale. Those children who had worn undershirts coated with borage oil for 2 weeks showed improvements in their erythema and itch, which were statistically significant. Transepidermal water loss from the back was decreased. In the placebo group, there were no statistically significant differences. The undershirts coated with borage oil were found to be statistically effective, and had no side-effects on children with mild atopic dermatitis.
Assuntos
Antirreumáticos/administração & dosagem , Vestuário , Dermatite Atópica/tratamento farmacológico , Óleos de Plantas/administração & dosagem , Ácido gama-Linolênico/administração & dosagem , Administração Cutânea , Criança , Pré-Escolar , Dermatite Atópica/patologia , Método Duplo-Cego , Feminino , Humanos , Lactente , MasculinoRESUMO
Cyclophosphamide (CPA) is one of the therapeutic agents for systemic inflammatory disorders. In murine dermal endothelial cells (F-2), 4-hydroxycyclophosphamide (4-HC), which is active metabolite of CPA, enhanced TNF-alpha-induced DNA fragmentation. In addition, 4-HC was shown to elevate TNF-alpha-induced caspase-3 activation. Caspase-8 activation was identified by the treatment of TNF-alpha, whereas 4-HC was no effect. In contrast, only when treated with 4-HC, caspase-9 activation and the increase in the intracellular expression of Bax were detected. These results suggest that CPA may sensitize endothelial cells to TNF-alpha-induced apoptosis through a mitochondria-dependent pathway and clinically may contribute to the limitation of inflammatory process.
Assuntos
Antirreumáticos/farmacologia , Apoptose/efeitos dos fármacos , Ciclofosfamida/análogos & derivados , Endotélio Vascular/efeitos dos fármacos , Fator de Necrose Tumoral alfa/farmacologia , Animais , Inibidores de Caspase , Caspases/metabolismo , Células Cultivadas , Ciclofosfamida/farmacologia , Cisteína Endopeptidases/farmacologia , Fragmentação do DNA/efeitos dos fármacos , Endotélio Vascular/enzimologia , Ativação Enzimática , Camundongos , Poli(ADP-Ribose) Polimerases/metabolismo , Regulação para Cima , Proteína X Associada a bcl-2/metabolismoRESUMO
A 54-year-old man with hepatitis C fell into shock with symptoms similar to enterocolitis after ingesting an undercooked barbecued mackerel. Most of his eruptions developed into annular erythema with small vesicles. He had taken high dose corticosteroids with intravenous cefotiam. His eruptions improved, but his shock state was exacerbated on Day 2. Treatment for endotoxin shock was initiated using piperacillin, intravenous immunoglobulin (IVIg), and hemoperfusion with Polymyxin B immobilized fiber (PMX-F), which resulted in shock reversal. The serum IL-6 value was 118,000 pg/mL on admission, and decreased to 2040 pg/mL on Day 3. On Day 6, the results from the culture of skin biopsy specimens showed the diagnosis as Vibrio vulnificus septic shock. Debridement was not needed, which is thought to be essential to Vibrio vulnificus sepsis. The changes in the serum IL-6 levels demonstrated that hemoperfusion with PMX-F and IVIg therapy was practical for Vibrio vulnificus septic shock.
Assuntos
Choque Séptico/etiologia , Vibrioses/complicações , Vibrio vulnificus , Humanos , Masculino , Pessoa de Meia-IdadeRESUMO
Dystrophic epidermolysis bullosa (DEB) is caused by mutations in the COL7A1 gene encoding collagen, the major component of anchoring fibrils. Premature termination codon (PTC) mutations in both alleles usually lead to the Hallopeau-Siemens variant that shows the most severe phenotype. We experienced a case of the non-Hallopeau-Siemens variant (nHS-RDEB), which had a mild clinical severity although it has PTC mutations in both alleles. Our patient was a compound heterozygote for a nonsense mutation (R669X) in exon 15 and a nonsense mutation (E2857X) in exon 116. But we confirmed the existence of some anchoring fibrils on electron micrograph. This suggested that a PTC close to the 3' end of COL7A1 does not completely abolish the collagen VII mRNA. We hypothesized that the truncated procollagen VII from the mutant allele with a nonsense mutation (E2857X) in exon 116 included two out of eight cysteines needed for disulfide bond formation, and hence a few functional anchoring fibrils could be formed.
Assuntos
Códon de Terminação , Colágeno/genética , Epidermólise Bolhosa Distrófica/diagnóstico , Epidermólise Bolhosa Distrófica/genética , Predisposição Genética para Doença , Adulto , Alelos , Diagnóstico Diferencial , Epidermólise Bolhosa Distrófica/patologia , Feminino , Genes Recessivos , HumanosRESUMO
Alpha-hydroxy acid (AHA) agents, such as glycolic acid and lactic acid, have been used as therapeutic agents for more than a quarter of a century. Recently, they have been used as agents to rejuvenate photo-aged skin. It is believed that these AHA agents induce the epidermis to remodel and accelerate desquamation, thus exerting their therapeutic effects. In this study, we investigated the histological differences in skin treated with glycolic, lactic, citric and acetic acids once daily for 6 weeks. The melanin pigments in the basal layer were less prominent in the glycolic and lactic acid-treated skin than in the citric and acetic acid-treated skin. The melanin deposits in the horny layers were equal for all AHA. However, the melanin deposits in the squamous layers were less prominent in the glycolic and lactic acid-treated skins than in the citric and acetic acid-treated skins; this was analogous to observations of the basal layers. Collagen I and procollagen I were increased after treatment with glycolic, lactic and citric acid in the upper dermis, but were not increased with acetic acid treatment. However, the staining of the epidermis and dermis for matrix metalloproteinase-1 (MMP-1) after treatment was not significantly different among the agents. Our data suggest that longer treatment intervals with glycolic and lactic acid can cause improvements in both the epidermal and dermal components and support the usefulness of AHA for rejuvenating photo-damaged skin.
Assuntos
Hidroxiácidos/farmacologia , Ceratolíticos/farmacologia , Pele/efeitos dos fármacos , Ácido Acético/administração & dosagem , Ácido Acético/farmacologia , Administração Cutânea , Adulto , Abrasão Química , Ácido Cítrico/administração & dosagem , Ácido Cítrico/farmacologia , Esquema de Medicação , Feminino , Glicolatos/administração & dosagem , Glicolatos/farmacologia , Humanos , Hidroxiácidos/administração & dosagem , Ceratolíticos/administração & dosagem , Ácido Láctico/administração & dosagem , Ácido Láctico/farmacologia , Masculino , Metaloproteinase 1 da Matriz/efeitos dos fármacos , Pessoa de Meia-Idade , Envelhecimento da PeleRESUMO
Anti SS-A/Ro antibodies are involved in the pathogenesis of cutaneous lupus erythematosus (CLE) in a part through antibody dependent cellular cytotoxicity (ADCC). However, it is still obscure which and how anti 60 and anti 52 kDa Ro antibodies are involved. To address the issue, we examined both types of anti Ro antibodies of sera from patients with systemic LE (SLE) or discoid LE (LE). The titer of anti 60 kDa antibody in SLE was significantly much higher than that of DLE or control. The positive ratio of DLE showed more higher tendency than control, but it was not statistically significant. The similar tendency was observed in the titer of anti 52 kDa antibody. An association between the anti 60 kDa antibody and the anti 52 kDa antibody was statistically significant in SLE patients. Although the relative index (RI) was statistically significant in DLE, they included many negative sera, which biased the statistics. Both anti 60 kDa antibody and anti 52 kDa antibody could induce significant ADCC of ultraviolet B (UVB) light-irradiated keratinocytes from neonatal foreskins and/or normal adults, in which anti 60 kDa antibody showed higher cytotoxicity than anti 52 kDa antibody. The autologous combination studies (keratinocytes and monospecific anti 60 or anti 52 kDa antibody from patients) suggested anti 60 kDa antibody was more potent to induce ADCC activity than anti 52 kDa antibody. Taken together, it is likely that anti 60 kDa antibody dependent keratinocyte damage plays more significant role in the pathogenesis of SLE skin lesions than anti 52 kDa dependent damage. However, both type antibodies seemed to have little contribution to the pathogenesis of DLE skin lesion.
Assuntos
Anticorpos Antinucleares/análise , Lúpus Eritematoso Discoide/imunologia , Adulto , Anticorpos Antinucleares/imunologia , Citotoxicidade Celular Dependente de Anticorpos/imunologia , Feminino , Humanos , Japão , Queratinócitos/imunologia , Queratinócitos/patologia , Lúpus Eritematoso Discoide/etiologia , Lúpus Eritematoso Discoide/patologia , Lúpus Eritematoso Sistêmico/etiologia , Lúpus Eritematoso Sistêmico/imunologia , Lúpus Eritematoso Sistêmico/patologia , Masculino , Pessoa de Meia-Idade , Peso MolecularRESUMO
Sairei-to, one of the Japanese-Chinese herbal medicines has been used for the treatment of various diseases, especially collagen disease and edema in nephrotic syndrome. However, the mechanism of the therapeutic effects remains uncertain. Therefore, we investigated the immunological changes of skin, kidney, spleen cells and serum in autoimmune-prone MRL/lpr, MRL/n and C57BL/6J mice treated with Sairei-to. In MRL/lpr mice treated with Sairei-to, the improvement of proteinuria, reduction in the number of hematoxylin bodies in kidney, and reduced serum levels of blood urea nitrogen were observed. These results indicate that Sairei-to can improve or inhibit the progression of lupus nephritis. The proportion of CD19 and the serum levels of IgG1, which is one of the pathogenesis of lupus dermatoses and lupus nephritis, were significantly reduced in Sairei-to-treated MRL/lpr mice. Therefore, it is suspected that the B cell function was suppressed by Sairei-to. In addition, CD4/8 ratio in spleen cells and the degree of lymphoproliferation in MRL/lpr mice also decreased. Interestingly, IL-4 producing spleen cells were increased significantly by ELISPOT assay, and IFN-gamma mRNA expressions were reduced in Sairei-to-treated MRL/lpr mice. Regarding the Th balance, an imbalance towards Th1 predominance may play a significant role in MRL/lpr mice, and the Th1 axis was suppressed and the Th2 axis became predominant in Sairei-to-treated MRL/lpr mice. On the other hand, Th2 cell type immunoglobulins (IgG1) were suppressed. These results suggested that Sairei-to is potential for impairing shifted Th1/Th2 balance and hypergammaglobulinemia resulting in therapeutic effects.
Assuntos
Autoimunidade/efeitos dos fármacos , Autoimunidade/fisiologia , Citocinas/metabolismo , Medicamentos de Ervas Chinesas/farmacologia , Células Th1/metabolismo , Células Th2/metabolismo , Animais , Nitrogênio da Ureia Sanguínea , Relação CD4-CD8 , Citocinas/genética , Imunoglobulinas/sangue , Imuno-Histoquímica , Rim/citologia , Rim/efeitos dos fármacos , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Endogâmicos MRL lpr , Tamanho do Órgão/efeitos dos fármacos , Fenótipo , Proteinúria/urina , RNA Mensageiro/metabolismo , Valores de Referência , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Baço/citologia , Baço/efeitos dos fármacos , Baço/fisiologiaRESUMO
A new method was developed to allow direct visualization of damaged sites on individual DNA molecules. Fluorescence in situ hybridization on extended DNA molecules was modified to detect a single abasic site. Abasic sites were specifically labeled with a biotinylated aldehyde-reactive probe and fluorochrome-conjugated streptavidin. The light emitted by a single fluorochrome-DNA complex was calibrated. The number of abasic sites on the DNA molecule was estimated by counting each fluorochrome-DNA complex. The present study directly visualized and characterized the abasic sites of single DNA molecules.
Assuntos
Dano ao DNA , DNA/química , Microscopia de Fluorescência , FotoquímicaRESUMO
Lambda DNA was stretched and patterned on mica surface using soft lithography. A highly diluted solution of amino propyl trimethoxy silane in hexane was deposited on a line patterned polydimethylsiloxane (PDMS) stamp. The functionalized stamp was then used to pick up DNA by molecular combing while the line patterns are parallel to the liquid surface. The stamp was then microcontact printed on freshly cleaved mica. We successfully obtained stretched DNA pattern on mica surface. DNA was found to be stretched in patterns perpendicular to those carved on the stamp. The stretched DNA population was large enough to be used for molecular biology mapping studies. Furthermore, the possibility of locating stretched DNA molecules in the desired position by stamping makes this method a good candidate for assembling non-semiconductor molecular devices.
Assuntos
DNA/química , Microscopia de Força Atômica/métodos , Microscopia de Fluorescência/métodos , Conformação de Ácido Nucleico , Silicatos de Alumínio , DNA/ultraestrutura , Dimetilpolisiloxanos/química , Oligonucleotídeos/química , Silicones/químicaRESUMO
We applied atomic force microscopy (AFM) to investigate the surface structure of barley chromosome in combination with a chemical treatment method. As a result, we have obtained high-resolution topographic images of granular structures with a diameter of ca. 50 nm on the surface of critical-point dried metaphase chromosomes. Treatment with 2M NaCl significantly modified the chromosome surface structure: surface roughness was increased and chromosome thickness was decreased. The NaCl treatment extracted two major proteins with molecular weights of 4000 and 20,000 Da. These proteins might be belonging to non-histone protein families that do not contain any aromatic amino acid. The results demonstrate the advantage of the combined method of high-resolution AFM imaging and chemical treatments for understanding nano-scale surface structures of the chromosome.
Assuntos
Estruturas Cromossômicas/ultraestrutura , Hordeum/ultraestrutura , Microscopia de Força Atômica/métodos , Estruturas Cromossômicas/química , Cromossomos de Plantas/química , Cromossomos de Plantas/ultraestrutura , Eletroforese em Gel de Poliacrilamida , Desenho de Equipamento , Hordeum/genética , Microscopia de Força Atômica/instrumentação , Peso Molecular , Proteínas de Plantas/isolamento & purificação , Cloreto de Sódio/farmacologia , Propriedades de Superfície/efeitos dos fármacosRESUMO
Nano-scale structures of the YOYO-1-stained barley chromosomes and lambda-phage DNA were investigated by scanning near-field optical/atomic force microscopy (SNOM/AFM). This technique enabled precise analysis of fluorescence structural images in relation to the morphology of the biomaterials. The results suggested that the fluorescence intensity does not always correspond to topographic height of the chromosomes, but roughly reflects the local amount and/or density of DNA. Various sizes of the bright fluorescence spots were clearly observed in fluorescence banding-treated chromosomes. Furthermore, fluorescence-stained lambda-phage DNA analysis by SNOM/AFM demonstrated the possibility of nanometer-scale imaging for a novel technique termed nano-fluorescence in situ hybridization (nano-FISH). Thus, SNOM/AFM is a powerful tool for analyzing the structure and the function of biomaterials with higher resolution than conventional optical microscopes.
Assuntos
Bacteriófago lambda/genética , Cromossomos de Plantas/genética , DNA de Plantas/ultraestrutura , DNA Viral/ultraestrutura , Hordeum/genética , Microscopia de Força Atômica/métodos , Benzoxazóis/metabolismo , DNA de Plantas/genética , DNA Viral/genética , Corantes Fluorescentes/metabolismo , Nanotecnologia , Compostos de Quinolínio/metabolismoRESUMO
Tacrolimus is a prototype of a class of topical immunosuppressive agents with great potential for the treatment of inflammatory skin diseases. Topical tacrolimus therapy was applied to facial skin lesions in 11 cases of cutaneous lupus erythematosus (LE) and dermatomyositis. Of the 11 patients, 6 (3 systemic LE, one discoid LE and 2 dermatomyositis) showed a marked regression of their skin lesions after tacrolimus therapy, but 4 patients (3 discoid LE and one dermatomyositis) were resistant to the therapy. A good response was observed for facial erythematous lesions with edematous or telangiectatic changes in systemic LE and dermatomyositis. In discoid LE with typical discoid lesions, tacrolimus brought no improvement. Topical tacrolimus will become a new tool for managing the skin lesions of collagen diseases.
Assuntos
Dermatomiosite/tratamento farmacológico , Dermatoses Faciais/tratamento farmacológico , Imunossupressores/uso terapêutico , Lúpus Eritematoso Cutâneo/tratamento farmacológico , Tacrolimo/uso terapêutico , Administração Tópica , Adolescente , Adulto , Ensaios Clínicos como Assunto , Dermatoses Faciais/etiologia , Feminino , Humanos , Lúpus Eritematoso Cutâneo/complicações , Masculino , Pessoa de Meia-IdadeRESUMO
In the present case study, the patient was a 65-year-old man who suddenly developed purpuric and necrotic lesions with severe pain in his fingers and toes. Laboratory investigations revealed marked eosinophilia (77.9%), but there was no evidence to support a diagnosis of parasitic infections, allergic disease, neoplasm or connective tissue disorder. The histopathological findings did not show any distinct vasculitis, but there were obliterative changes of the arterioles. The digital gangrene gradually progressed and was unresponsive to corticosteroid therapy. The patient eventually underwent amputation of the distal phalanges. We applied allogeneic cultured dermal substitute (CDS) to the skin defect. The allogeneic CDS was prepared by culturing fibroblasts on a two-layered sponge of hyaluronic acid and atelo-collagen. This CDS is able to release a number of cytokines including VEGF. The present case had a good clinical result.
Assuntos
Dedos/patologia , Gangrena/diagnóstico , Síndrome Hipereosinofílica/diagnóstico , Dedos do Pé/patologia , Idoso , Amputação Cirúrgica , Diagnóstico Diferencial , Dedos/cirurgia , Gangrena/complicações , Gangrena/patologia , Gangrena/terapia , Humanos , Síndrome Hipereosinofílica/complicações , Síndrome Hipereosinofílica/patologia , Síndrome Hipereosinofílica/terapia , Masculino , Pele Artificial , Dedos do Pé/cirurgiaRESUMO
Pseudoxanthoma elasticum (PXE) is a heritable disease characterized by dermal, ocular, and vascular lesions that result from degeneration of the elastic fibers. Recently, the ATP-binding cassette subfamily C member 6 (ABCC6) gene has been demonstrated to be responsible for PXE, and 43 mutations have been identified to date. However, it is still unknown now mutations in the ABCCC6 gene can lead to manifestations of PXE.