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1.
Journal of Central South University(Medical Sciences) ; (12): 1-14, 2023.
Article in English | WPRIM | ID: wpr-971365

ABSTRACT

OBJECTIVES@#Ozone is widely applied to treat allergic skin diseases such as eczema, atopic dermatitis, and contact dermatitis. However, the specific mechanism remains unclear. This study aims to investigate the effects of ozonated oil on treating 2,4-dinitrochlorobenzene (DNCB)-induced allergic contact dermatitis (ACD) and the underling mechanisms.@*METHODS@#Besides the blank control (Ctrl) group, all other mice were treated with DNCB to establish an ACD-like mouse model and were randomized into following groups: a model group, a basal oil group, an ozonated oil group, a FcεRI-overexpressed plasmid (FcεRI-OE) group, and a FcεRI empty plasmid (FcεRI-NC) group. The basal oil group and the ozonated oil group were treated with basal oil and ozonated oil, respectively. The FcεRI-OE group and the FcεRI-NC group were intradermally injected 25 µg FcεRI overexpression plasmid and 25 µg FcεRI empty plasmid when treating with ozonated oil, respectively. We recorded skin lesions daily and used reflectance confocal microscope (RCM) to evaluate thickness and inflammatory changes of skin lesions. Hematoxylin-eosin (HE) staining, real-time PCR, RNA-sequencing (RNA-seq), and immunohistochemistry were performed to detct and analyze the skin lesions.@*RESULTS@#Ozonated oil significantly alleviated DNCB-induced ACD-like dermatitis and reduced the expressions of IFN-γ, IL-17A, IL-1β, TNF-α, and other related inflammatory factors (all P<0.05). RNA-seq analysis revealed that ozonated oil significantly inhibited the activation of the DNCB-induced FcεRI/Syk signaling pathway, confirmed by real-time PCR and immunohistochemistry (all P<0.05). Compared with the ozonated oil group and the FcεRI-NC group, the mRNA expression levels of IFN-γ, IL-17A, IL-1β, IL-6, TNF-α, and other inflammatory genes in the FcεRI-OE group were significantly increased (all P<0.05), and the mRNA and protein expression levels of FcεRI and Syk were significantly elevated in the FcεRI-OE group as well (all P<0.05).@*CONCLUSIONS@#Ozonated oil significantly improves ACD-like dermatitis and alleviated DNCB-induced ACD-like dermatitis via inhibiting the FcεRI/Syk signaling pathway.


Subject(s)
Animals , Mice , Dinitrochlorobenzene/metabolism , Skin/metabolism , Cytokines/metabolism , Interleukin-17/metabolism , Tumor Necrosis Factor-alpha/metabolism , Dermatitis, Allergic Contact/pathology , Dermatitis, Atopic/chemically induced , Signal Transduction , RNA, Messenger/metabolism , Mice, Inbred BALB C
2.
Chinese journal of integrative medicine ; (12): 612-619, 2022.
Article in English | WPRIM | ID: wpr-939792

ABSTRACT

OBJECTIVE@#To evaluate the therapeutic effects of acupoint autohemotherapy (A-AHT) on 1-chloro-2,4-dinitrobenzene (DNCB)-induced atopic dermatitis (AD) in mice focusing on regulating T helper 1/T helper 2 (Th1/Th2) immune responses.@*METHODS@#Thirty BALB/c mice were divided into 5 groups by a random number table, including normal control (NC), AD model (AD), A-AHT, sham A-AHT (sA-AHT), and acupoint injection of normal saline (A-NS) groups, 6 mice per group. Mice were challenged by DNCB for the establishment of experimental AD model. On the 8th day, except for the NC and AD groups, the mice in the other groups received management once every other day for a total of 28 days. For the A-AHT and sA-AHT groups, 0.05 mL of autologous whole blood (AWB) was injected into bilateral Zusanli (ST 36) and Quchi (LI 11) and sham-acupoints (5 mm lateral to ST 36 and LI 11), respectively. The A-NS group was administrated with 0.05 mL of normal saline by acupoint injection into ST 36 and LI 11. Dermatitis severity for dorsal skin of mice was determined using the Severity Scoring of Atopic Dermatitis (SCORAD) every week. The total immunoglobulin E (IgE), interleukin-4 (IL-4), and interferon-gamma (IFN-γ) cytokine levels in serum were examined by enzyme-linked immunosorbent assay (ELISA). Spleen Th1/Th2 expression were analyzed via flow cytometry and immunohistochemical assay was used to detect T-box expressed in T cell (T-bet) and GATA-binding protein 3 (GATA3) expressions in skin lesions of mice.@*RESULTS@#Compared with the AD group, both A-AHT and sA-AHT reduced the SCORAD index and serum IgE level (P<0.05 or P<0.01); A-AHT, sA-AHT and A-NS down-regulated serum IL-4 level and upregulated IFN-γ/IL-4 ratio (P<0.05 or P<0.01); A-AHT regulated the Th1/Th2 shift specifically and increased the related transcription factors such as T-bet expression and T-bet/GATA3 ratio (P<0.05).@*CONCLUSION@#A-AHT showed significant effectiveness on the AD model mice, through regulating Th1/Th2 immune responses.


Subject(s)
Animals , Mice , Acupuncture Points , Dermatitis, Atopic/therapy , Dinitrobenzenes , Dinitrochlorobenzene , Immunoglobulin E , Interferon-gamma , Interleukin-4 , Mice, Inbred BALB C , Saline Solution
3.
Acta cir. bras ; 36(1): e360102, 2021. tab, graf
Article in English | LILACS | ID: biblio-1152689

ABSTRACT

ABSTRACT Purpose To study the Periplaneta americana L. extract Ento-B on the treatment of chronic ulcerative colitis induced by 2,4-dinitrochlorobenzene and acetic acid in rats and to explore its primary mechanism of action. Methods Using 2,4-dinitrochlorobenzene combined with acetic acid to induce chronic ulcerative colitis (chronic UC) in rats. The sulfasalazine (400 mg/kg) and Ento-B (200 mg/kg, 100 mg/kg,50 mg/kg) were given by intragastric administration and the effect was evaluated according to the disease activity index (DAI) score, colon mucosal injury index (CMDI) score, histopathological score (HS) and the serum levels of Interleukin-4(IL-4), Interleukin-10(IL-10), Tumor necrosis factor-α(TNF-α), Malondialdehyde(MDA), Superoxide dismutase(SOD) and Inducible nitric oxide synthase(iNOS.) Results Compared with the model group, all doses of Ento-B could reduce the score of CMDI (p < 0.05), HS(p < 0.05 or p < 0.01), significantly increased the expression of IL-4, IL-10, SOD (p < 0.01) and decreased the levels of TNF-α, MDA, iNOS in serum of UC rats, significantly improving the degree of colon lesionsin UC rats. Conclusions Ento-B may play an important role in the treatment of ulcerative colitis induced byUC rats. The mechanism may be related to the increased expression of IL-4, IL-10, SOD and reduced expression of TNF-α, MDA, iNOS.


Subject(s)
Animals , Rats , Periplaneta , Colitis, Ulcerative/chemically induced , Colitis, Ulcerative/drug therapy , Plant Extracts/therapeutic use , Plant Extracts/pharmacology , Tumor Necrosis Factor-alpha , Colon , Acetic Acid , Dinitrochlorobenzene
5.
Journal of Veterinary Science ; : 307-315, 2015.
Article in English | WPRIM | ID: wpr-66455

ABSTRACT

Atopic dermatitis, one of the most important skin diseases, is characterized by both skin barrier impairment and immunological abnormalities. Although several studies have demonstrated the significant relationship between atopic dermatitis and immunological abnormalities, the role of hydroxyeicosatetraenoic acids (HETE) in atopic dermatitis remains unknown. To develop chiral methods for characterization of 12-HETE enantiomers in a 1-chloro-2,4-dinitrochlorobenzene (DNCB)-induced atopic dermatitis mouse model and evaluate the effects of 12-HETE on atopic dermatitis, BALB/c mice were treated with either DNCB or acetone/olive oil (AOO) to induce atopic dermatitis, after which 12(R)- and 12(S)-HETEs in the plasma, skin, spleen, and lymph nodes were quantified by chiral liquid chromatography-tandem mass spectrometry. 12(R)- and 12(S)-HETEs in biological samples of DNCB-induced atopic dermatitis mice increased significantly compared with the AOO group, reflecting the involvement of 12(R)- and 12(S)-HETEs in atopic dermatitis. These findings indicate that 12(R)- and 12(S)-HETEs could be a useful guide for understanding the pathogenesis of atopic dermatitis.


Subject(s)
Animals , Female , Humans , Mice , Biomarkers/blood , Chromatography, Liquid , Dermatitis, Atopic/chemically induced , Dinitrochlorobenzene/adverse effects , Hydroxyeicosatetraenoic Acids/blood , Irritants/adverse effects , Mice, Inbred BALB C , Models, Animal , Tandem Mass Spectrometry
6.
An. bras. dermatol ; 85(6): 839-842, nov.-dez. 2010. tab
Article in Portuguese | LILACS | ID: lil-573622

ABSTRACT

FUNDAMENTOS: A radiação ultravioleta B (RUVB) é o mais importante fator ambiental capaz de modificar a função imunológica da pele humana. OBJETIVO: estudar a associação entre o fenótipo de suscetibilidade ou resistência à radiação RUVB e as formas polares da hanseníase. MATERIAL E MÉTODOS: foram avaliados 38 pacientes com hanseníase virchowiana (MHV) e 87 pacientes com hanseníase tuberculoide (MHT) de acordo com a classificação de Ridley e Jopling (1966). Todos os pacientes foram submetidos ao teste para determinação do fenótipo de suscetibilidade ou resistência à RUVB por meio da aplicação de um disco de dinitroclorobenzeno (DNCB) a 2 por cento em uma área de pele previamente irradiada com duas vezes a dose eritematosa mínima (DEM). Após 21 dias, outra aplicação de um disco similar de DNCB a 0,05 por cento na região escapular (área não exposta à RUVB) foi realizada para avaliar se houve sensibilização, com leitura após 48 horas. Os pacientes que apresentaram reação positiva ao DNCB foram considerados UVB-resistentes e o oposto foi considerado para aqueles que não apresentaram resposta (UVB-suscetíveis). RESULTADOS: A frequência de UVB-suscetíveis foi de 63,2 por cento (24 pacientes) no grupo MHV e 34,4 por cento (30 pacientes) no grupo MHT (OR = 3,26; IC = 1,36-7,87; x² = 7,73; p = 0,005). CONCLUSÃO: Os resultados sugerem que a UVB-suscetibilidade é um fator de risco para o desenvolvimento da MHV.


BACKGROUNDS: Ultraviolet radiation B (UVRB) is the most important environmental factor capable of altering the immune function of human skin. OBJECTIVE: To evaluate the association of the phenotypes of susceptibility or resistance to ultraviolet radiation B (UVRB) and the polar forms of leprosy. MATERIAL AND METHODS: We evaluated 38 patients with lepromatous leprosy (LL) and 87 patients with tuberculoid (TT) leprosy, according to the classification by Ridley and Jopling (1966). All the patients were submitted to a test to determine the phenotypes of susceptibility or resistance to UVRB through the application of a 2 percent dinitrochlorobenzene (DNCB) disc to a previously irradiated area with twice the minimal erythema dose (MED). After 21 days, a similar disc soaked in 0.05 percent DNCB was applied to the scapular area (unexposed to UVRB) to check for sensitiveness, with reading of the results after 48 hours. The patients that showed a positive reaction to DNCB were considered resistant (UVB-R) and those who did not show any reaction were considered susceptible (UVB-S). RESULTS: The frequency of UVB-S individuals was 63.2 percent (24 patients) in the LL group and 34.4 percent (30 patients) in the TT group (OR=3.26; IC=1.36 - 7.87; x²=7.73; p=0.005). CONCLUSION: Our results suggest that UVB-susceptibility is a risk factor to the development of lepromatous leprosy (LL).


Subject(s)
Adolescent , Adult , Female , Humans , Male , Middle Aged , Young Adult , Leprosy, Lepromatous/etiology , Leprosy, Tuberculoid/etiology , Ultraviolet Rays/adverse effects , Disease Susceptibility , Dinitrochlorobenzene , Indicators and Reagents , Phenotype , Risk Factors , Radiation Tolerance/physiology
7.
Laboratory Animal Research ; : 95-102, 2010.
Article in English | WPRIM | ID: wpr-153255

ABSTRACT

The various murine models have contributed to the study of human atopic dermatitis (AD). However limitations of the models involve low reproducibility and long time to develop AD. In an attempt to overcome these limitations and establish an atopic dermatitis murine model, we repeated the application of 2, 4-dinitrochlorobenzene (DNCB) patch in NC/Nga and BALB/c mice, which has advantages in reproduction and cost. For the sensitization, a 1 cm2 gauze-attached patch, where 1% or 0.2% DNCB was periodically attached on the back of NC/Nga and BALB/c mice. To estimate how homologous our model was with human atopic dermatitis, clinical, histological and immunological alterations were evaluated. Both strains showed severe atopic dermatitis, increase in subiliac lymph node weight, mast cells, epidermal hyperplasia and serum IgE levels. Though both exhibited a high IL-4/IFN-gamma and IL-4/TNF-beta ratio in the expression of mRNA, the shifting of DNCB-treated BALB/c mice was increased to more than double that of NC/Nga mice. These results suggest that our DNCB patched model using BALB/c mice were more suitable than NC/Nga mice in demonstrating the immune response. We anticipate that our novel model may be successfully used for pathogenesis of atopic dermatitis and assessment of therapeutic approaches.


Subject(s)
Animals , Humans , Mice , Dermatitis, Atopic , Dinitrochlorobenzene , Hyperplasia , Immunoglobulin E , Lymph Nodes , Mast Cells , Reproduction , RNA, Messenger
8.
Annals of Dermatology ; : 269-278, 2010.
Article in English | WPRIM | ID: wpr-60738

ABSTRACT

BACKGROUND: Although reactive oxygen species (ROS) have been produced in both mouse bone marrow-derived dendritic cells (DCs) and XS-106 DCs by contact sensitizers and irritants in previous studies, the generation of ROS in human monocyte-derived DCs (MoDCs) and their role in contact hypersensitivity (CHS) has yet to be elucidated. OBJECTIVE: The purpose of this study was to determine whether contact allergens and irritants induce ROS in MoDCs and, if so, to evaluate the role of contact allergen and irritant induced-ROS in MoDCs in CHS. METHODS: Production of ROS was measured by 5-(and-6)-chloromethyl-2',7'-dichlorodihydrofluorescein diacetate (CM-H2DCFDA) assay. Surface CD86 and HLA-DR molecules were detected by flow cytometry. Protein carbonylation was detected by Western blotting. RESULTS: ROS were produced by contact allergens such as dinitrochlorobenzene (DNCB) and thimerosal and the irritant benzalkonium chloride (BKC). DNCB-induced, but not BKC-induced, ROS increased surface CD86 and HLA-DR molecules on MoDCs and induced protein carbonylation. These changes were reduced in the presence of antioxidant N-acetyl cysteine. CONCLUSION: Our results suggest that DNCB-induced ROS may be different from those induced by irritant BKC. The DNCB-induced ROS may be associated with the CHS response, because they activate surface molecules on DCs that are important for generating immune reactions.


Subject(s)
Animals , Humans , Mice , Allergens , Benzalkonium Compounds , Blotting, Western , Cysteine , Dendritic Cells , Dermatitis, Contact , Dinitrochlorobenzene , Flow Cytometry , HLA-DR Antigens , Irritants , Protein Carbonylation , Reactive Oxygen Species , Thimerosal
9.
Korean Journal of Dermatology ; : 1105-1112, 2009.
Article in Korean | WPRIM | ID: wpr-220716

ABSTRACT

BACKGROUND: Atopic dermatitis is a chronic itchy, inflammatory skin disease that usually relapses. Although the etiology of atopic dermatitis remains unclear, it has been shown that both Th1 and Th2 cytokines play pathogenic roles in the generation of atopic dermatitis. DA-9102 is a fraction from the Actinidia species and DA-9102 displays immune modulating activity for allergy related disease. OBJECTIVE: We have developed the atopic dermatitis model of NC/Nga mice using DNCB and we examined whether DA-9102 suppresses the development of atopic dermatitis-like skin lesions on NC/Nga mice. METHODS: NC/Nga mice were challenged with DNCB during 5 weeks to develop atopic dermatitis-like skin lesions. Daily DA-9102 or cyclosporine A or HPMC (control) were then given orally. The efficacy of DA-9102 in NC/Nga mice was judged by measurement of the skin lesion severity (a modified SCORAD score), the serum IgE and IgG2a levels and the cytokine levels (IFN-gamma and IL-4) from spleen cells cultured with ConA. RESULTS: Atopic dermatitis-like lesions were developed on the NC/Nga mice by using topical DNCB. Oral administration of 100 mg/kg DA-9102 significantly suppressed the development of dermatitis, as was analyzed by a modified SCORAD score (p<0.01). The serum IgE level increased gradually with age, but treatment with DA-9102 suppressed the increment of the serum IgE level (p<0.01). The mean values of IFN-gamma in the NC/Nga mice of the DA-9102 group were lower than those of the control mice group (p<0.05). The mean values of IL-4 were undetectable in all the experimental groups. The serum IgG2a level were not significantly different among all the experimental groups. CONCLUSION: We successfully developed an atopic dermatitis model in NC/Nga mice. Based on our in in vitro data, we suggest that DA-9102 can be useful for the treatment of atopic dermatitis.


Subject(s)
Animals , Mice , Actinidia , Administration, Oral , Cyclosporine , Cytokines , Dermatitis , Dermatitis, Atopic , Dinitrochlorobenzene , Hypersensitivity , Immunoglobulin E , Immunoglobulin G , Interleukin-4 , Recurrence , Skin , Skin Diseases , Spleen
11.
Korean Journal of Dermatology ; : 1470-1477, 2008.
Article in Korean | WPRIM | ID: wpr-204242

ABSTRACT

BACKGROUND: Various allergens and irritants induced the production of reactive oxygen species (ROS) in the well-established mouse dendritic cell (DC) line XS106 and this production of ROS was inhibited by antioxidants. OBJECTIVE: To investigate the production and functions of ROS in mouse bone marrow-derived DCs (BM-DCs) by various haptens and irritants, we examined the production of ROS, the expression of surface molecules, and the production of interleukin-12 (IL-12) in mouse BM-DCs. METHODS: Six to eight-week-old female C57/BL6 mice were used in this study. Mouse BM-DCs were co-cultured with DNFB, DNCB, TNBS, hydroquinone, NiSO4, CoCl2, MnCl2, thimerosal, SDS, and BKC. The production of ROS and the expression of surface molecules (CD40, CD80, CD86, and MHC-II) were measured by flow cytometry in chemical-treated mouse BM-DCs. In addition, the cells were pretreated with antioxidants to determine whether the production of ROS can be inhibited. The production of IL-12 was also measured in DNCB and SDS-treated mouse BM-DCs using ELISA. Results: The production of ROS in mouse BM-DCs was induced by various allergens, including DNFB, DNCB, TNBS, hydroquinone, MnCl2 and irritants like SDS, BKC. The expression of surface molecules was induced by various chemicals and NiSO4 was the most potent inducer of surface molecules in mouse BM-DCs. The production of ROS in DNCB and SDS-treated mouse BM-DCs was partially inhibited by diphenylene iodonium, but not by rotenone, vitamin E, allopurinol, glutathione. The production of IL-12 was not detected in DNCB and SDS-treated mouse BM-DCs. CONCLUSION: The production of ROS was induced in mouse BM-DCs by various allergens and irritants. The expression of surface molecules was also induced by various chemicals. The production of ROS was partially inhibited by DPI. The production of IL-12 was not detected.


Subject(s)
Animals , Female , Humans , Mice , Allergens , Allopurinol , Antioxidants , Chlorides , Dendritic Cells , Dinitrochlorobenzene , Dinitrofluorobenzene , Flow Cytometry , Glutathione , Haptens , Hydroquinones , Interleukin-12 , Irritants , Manganese Compounds , Onium Compounds , Reactive Oxygen Species , Rotenone , Thimerosal , Vitamin E , Vitamins
12.
Chinese Journal of Industrial Hygiene and Occupational Diseases ; (12): 147-150, 2008.
Article in Chinese | WPRIM | ID: wpr-303988

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the use of dendritic cells derived from mice bone marrow to evaluate the cutaneous allergic reaction induced by chemical sensitizers.</p><p><b>METHODS</b>Dendritic cells derived from mice bone marrow were cultured and administrated with 2, 4-dinitrochlorobenzene (DNCB), nickel sulfate (NiSO4), sodium dodecyl sulfate (SDS) and hexyl cinnamic aldehyde (HCA), respectively. Cell membrane molecule CD86 and extracellular IL-1 beta, IL-6 and IL-12 were detected after 0, 1, 6, 12, 24, 36, 48 hour's administration, respectively.</p><p><b>RESULTS</b>CD86 expression reached the highest level after exposure to DNCB for 48 h, and increased by about 279% compared with the control (P < 0.05), while it was lower than that of control after administrated with NiSO4 and HCA for 1 h and 6 h, and SDS for 36 h, respectively (P < 0.05). Extracellular IL-1 beta increased greatly after exposure to NiSO4 just for 1 h, with the maximum at 48 h (298 pg/ml, P < 0.05), and after exposure to HCA for 6 h, with maximum at 48 h (84 pg/ml, P < 0.05). However, it didn't fluctuate significantly after administrated with DNCB and SDS respectively, compared with the control. Extracellular IL-6 increased significantly after exposure to NiSO4 for 1 h, with the maximum at 24 h (2152 pg/ml, P < 0.05). After exposure to HCA, extracellular IL-6 reached the maximum at 1 h (1403 pg/ml), and then it was decreased quickly, but still higher than the control (P < 0.05), while it didn't change significantly after treatment with DNCB and SDS, compared with the control (P > 0.05). Extracellular IL-12 was not detected out among all the groups.</p><p><b>CONCLUSION</b>Chemical sensitizer DNCB could induce the high expression of CD86 on DC membrane, and NiSO4 and HCA could induce DC to release IL-1 beta and IL-6. However, the irritant SDS had no such effect.</p>


Subject(s)
Animals , Mice , B7-2 Antigen , Metabolism , Cells, Cultured , Dendritic Cells , Allergy and Immunology , Metabolism , Dinitrochlorobenzene , Pharmacology , Interleukin-12 , Metabolism , Interleukin-1beta , Metabolism , Interleukin-6 , Metabolism , Mice, Inbred C57BL , Nickel , Pharmacology , Sodium Dodecyl Sulfate , Pharmacology
13.
Acta Pharmaceutica Sinica ; (12): 108-112, 2008.
Article in Chinese | WPRIM | ID: wpr-268162

ABSTRACT

To identify the inhibitor of glutathione S-transferase (GST), a high-throughput screening method was established in a 384-well microplate with total 35 microL volume, and the absorbance at 340 nm is detected. The concentrations of substrates, CDNB and GST were determined by chromatometry. The optimal enzyme kinetics reaction time and temperature are 2 h and 30 degrees C , respectively. The established model was evaluated by NaOCl, a known GST inhibitor, and the parameter Z' was 0.77, which showed a high feasibility and stability of the assay. A total of 31,098 compounds were screened, of which 4 compounds were shown to inhibit GST activity, high inhibiting activity for their IC50 of GST inhibition was 3.94, 4.05, 74.85, and 77.41 mg x L(-1), separately. The results indicated that the colorimetric method by using CDNB and GSH as substrate is stable, sensitive, reproducible and also suitable for high throughput screening.


Subject(s)
Dinitrochlorobenzene , Chemistry , Drug Evaluation, Preclinical , Methods , Enzyme Inhibitors , Glutathione , Chemistry , Glutathione Transferase , Substrate Specificity
14.
Yeungnam University Journal of Medicine ; : 232-239, 2006.
Article in Korean | WPRIM | ID: wpr-158973

ABSTRACT

Treatments for alopecia areata include topical corticosteroid treatment, corticosteroid intralesional injection, systemic corticosteroid treatment, PUVA(psoralen-UVA) and topical immunotherapy. The therapeutic effects are variable. Alopecia totalis is hard to treat completely. Topical immunotherapy with dinitrochlorobenzene (DNCB), squaric acid dibutyl ester (SADBE) or diphenylcyclopropenone (diphencyprone, DPCP) represents the most accepted therapeutic modality for the treatment of extensive alopecia areata. We report two cases of alopecia totalis treated with DPCP. After DPCP treatment, total scalp hair was completely recovered.


Subject(s)
Alopecia Areata , Alopecia , Dinitrochlorobenzene , Hair , Immunotherapy , Injections, Intralesional , Scalp
15.
Asian Pac J Allergy Immunol ; 2004 Dec; 22(4): 219-28
Article in English | IMSEAR | ID: sea-36790

ABSTRACT

An adult stage Opisthorchis viverrini cDNA library was constructed and screened for abundant transcripts. One of the isolated cDNAs was found by sequence comparison to encode a glutathione S-transferase (GST) and was further analyzed for RNA expression, encoded protein function, tissue distribution and cross-reactivity of the encoded protein with other trematode protein counterparts. The cDNA has a size of 893 bp and encodes a GST of 213 amino acids length (OV28GST). The most closely-related GST of OV28GST among those published for trematodes is a 28 kDa GST of Clonorchis sinensis as shown by multiple sequence alignment and phylogenetic analysis. Northern analysis of total RNA with a gene-specific probe revealed a 900 nucleotide OV28GST transcriptional product in the adult parasite. Through RNA in situ hybridization OV28GST RNA was detected in the parenchymal cells of adult parasites. This result was confirmed by immunolocalization of OV28GST with an antiserum generated in a mouse against bacterially-produced recombinant OV28GST. Both, purified recombinant and purified native OV28GST were resolved as 28 kDa proteins by SDS-PAGE. Using the anti-recOV28GST antiserum, no or only weak cross-reactivity was observed in an immunoblot of crude worm extracts against the GSTs of Schistosoma mansoni, S. japonicum, S. mekongi, Eurytrema spp. and Fasciola gigantica. The enzyme activity of the purified recombinant OV28GST was verified by a standard 1-chloro-2, 4-dinitrobenzene (CDNB) based activity assay. The present results of our molecular analysis of OV28GST should be helpful in the ongoing development of diagnostic applications for opisthorchiasis viverrini.


Subject(s)
Amino Acid Sequence , Animals , Blotting, Northern , Blotting, Southern , Cloning, Molecular , DNA, Complementary/analysis , Dinitrochlorobenzene/diagnosis , Gene Library , Glutathione Transferase/genetics , Indicators and Reagents , Opisthorchis/enzymology
16.
Korean Journal of Dermatology ; : 452-455, 2004.
Article in Korean | WPRIM | ID: wpr-99987

ABSTRACT

We report a case of vitiligo developed on the sites of DNCB application for the treatment of alopecia areata in a 53-year-old woman. Histopathologic studies of the depigmented patches showed absence of melanin pigmentation at the basal layer of the epidermis and a negative reaction for the Fontana-Masson stain. The current theories explaining vitiligo development after topical sensitizer application are that topical sensitizers induce the vitiligo by a direct cytotoxic effect on the melanocytes, or by unmasking latent vitiligo as a result of the Koebner phenomenon due to contact dermatitis, but the exact mechanism is still unclear.


Subject(s)
Female , Humans , Middle Aged , Alopecia Areata , Alopecia , Dermatitis, Contact , Dinitrochlorobenzene , Epidermis , Melanins , Melanocytes , Pigmentation , Vitiligo
17.
Arq. neuropsiquiatr ; 61(3B): 731-737, Sept. 2003. tab, graf
Article in English | LILACS | ID: lil-348650

ABSTRACT

STUDY OBJECTIVE: The purpose of this study was to assess the extent of immune dysfunction in a well-defined group of epileptic patients: children with diagnosis of West syndrome (WS) or with transitions to another age-related EEG patterns, the multifocal independent spikes (MIS), and the slow spike-wave complexes (Lennox-Gastaut syndrome - LGS). Thus, WS was studied at different points of the natural evolutive history of the disease. METHOD: A group of 50 patients (33 with WS, 10 with LGS and 7 with MIS) and 20 age-matched healthy controls were submitted to enumeration of T lymphocyte subsets: CD1, CD3, CD4, CD8, CD4/CD8 ratio and lymphocyte proliferation assay to phytohaemagglutinin (PHA), in the presence of autologous and AB, homologous plasma. Dinitrochlorobenzene (DNCB) skin test sensitization was performed only in patients. Determinations of IgG, IgA, and IgM serum levels were compared to standard values for Brazilian population in different age ranges. RESULTS: Sensitization to DNCB showed absent or low skin reactions in 76 percent of the patients. High levels of IgG (45.7 percent) and IgM (61.4 percent), and lower levels of IgA (23.9 percent) were detected in the serum of the patients. Enumeration of lymphocyte subsets in peripheral blood showed: low CD3+ (p<0.05), low CD4+ (p<0.05), high CD8+ (p<0.01) and low CD4+ / CD8+ ratio (p<0.001). The proportion of CD1+ cells in the control group was less than 3 percent, while ranged between 6 and 11 percent in 18 percent of the patients. The in vitro PHA-induced T cell proliferation showed significantly low blastogenic indices only when patients, cells were cultured in presence of their own plasma. No differences in blastogenic indices were observed when the cells of patients and controls were cultured with human AB plasma. CONCLUSION: The immunodeficiency in WS was mainly characterized by anergy, impaired cell-mediated immunity, altered levels of immunoglobulins, presence of immature thymocytes in peripheral blood and functional impairment of T lymphocytes induced by plasma inhibitory factors


Subject(s)
Humans , Male , Female , Infant , Child, Preschool , Child , Antigens, CD , Epilepsy , Case-Control Studies , Dinitrochlorobenzene , Epilepsy , Immunoglobulin A , Immunoglobulin G , Immunoglobulin M , Lymphocyte Count , Phytohemagglutinins , Plasma , Skin Tests
18.
Indian J Exp Biol ; 2002 Jan; 40(1): 111-4
Article in English | IMSEAR | ID: sea-62188

ABSTRACT

Ciprofloxacin (10 mg/kg body weight, iv, twice daily for 4 days) failed to alter specific antibody titres, total immunoglobulin concentration, total serum protein concentration, total leukocyte count, lymphocyte percentage, phagocytic index and skin thickness in DNCB skin sensitivity test against Brucella plain killed antigen in New Zealand White rabbits. It can be concluded that ciprofloxacin at the dose and duration employed did not adversely affect specific immune response in normal rabbits.


Subject(s)
Animals , Anti-Infective Agents/pharmacology , Brucella Vaccine/immunology , Ciprofloxacin/pharmacology , Dinitrochlorobenzene/immunology , Female , Hemolytic Plaque Technique , Immune System/drug effects , Immunity, Cellular , Immunoglobulin G/immunology , Injections, Intramuscular , Leukocyte Count , Leukocytes/immunology , Lymphocyte Activation/drug effects , Male , Phagocytosis/drug effects , Rabbits , Skin Tests
19.
Rev. chil. dermatol ; 17(2): 105-8, 2001.
Article in Spanish | LILACS | ID: lil-296063

ABSTRACT

La alopécia areata (AA) es una patología realtivamente frecuente del folículo piloso. Su etiología, si bien es desconocida, probablemente corresponda a un fenómeno autoinmune, aunque factores genéticos y ambientales también estarían involucrados. La inmunoterapia tópica es la modalidad terapéutica más efectiva y aceptada en el tratamiento de la AA crónica severa. La inmunoterapia tópica se define como la inducción y posterior provocación periódica de una dermatitis de contacto alérgica, a través de la aplicación de un potente alergeno de contacto en una zona cutánea determinada. Han sido utilizados tres alergenos de contacto en el tratamiento de la alopecia areata: dinitroclorobenceno (DNCB), dibutil éster del ácido escuárico (SADBE) y difenciprona (DPC). En el presente trabajo se describen las principales características de estos agentes tópicos


Subject(s)
Humans , Alopecia Areata/drug therapy , Desensitization, Immunologic , Hair Follicle , Administration, Topical , Adjuvants, Immunologic/pharmacology , Cyclobutanes/administration & dosage , Cyclobutanes/pharmacology , Cyclopropanes/administration & dosage , Cyclopropanes/adverse effects , Cyclopropanes/pharmacology , Dinitrochlorobenzene/administration & dosage , Dinitrochlorobenzene/pharmacology
20.
Indian J Pathol Microbiol ; 1999 Oct; 42(4): 471-4
Article in English | IMSEAR | ID: sea-74819

ABSTRACT

In the present study delayed cutaneous hypersensitivity response (DNCB test) and humoral response (by uantification of immunoglobulins) ware carried out in 20 cases of leukaemias. None of the cases was found to be anergic or immunodeficient. In remission also patients showed the normal response.


Subject(s)
Adult , Dinitrochlorobenzene/administration & dosage , Female , Humans , Hypersensitivity, Delayed/immunology , Immunoglobulins/blood , Leukemia, Myelogenous, Chronic, BCR-ABL Positive/immunology , Lymphocyte Count , Male
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