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1.
J Clin Aesthet Dermatol ; 17(4): 37-41, 2024 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-38638185

RESUMEN

Objective: Postinflammatory hyperpigmentation (PIH) is a common sequela of acne vulgaris. Topical treatment with hydroquinone is the standard treatment, but may be associated with complications. Cysteamine is a relatively safe depigmenting agent with an observed depigmenting effect. We designed this study to assess the efficacy of a cysteamine 5% cream in treating acne-induced PIH. Methods: Twenty-eight out of 32 participants finalized this investigator-blind, randomized, and controlled trial (registered in Iranian Registry of Clinical Trials [IRCTID: IRCT20140212016557N5]). We randomized the patients to apply either cysteamine 5% or hydroquinone 4%/ascorbic acid 3% (HC) cream. Postacne hyperpigmentation index (PAHPI) and melanin index were the assessment measures after four months of treatment. We evaluated the quality of life by the Dermatology Life Quality Index (DLQI) questionnaire. Results: Both cysteamine and HC cream significantly decreased the PAHPI score and melanin index of acne-induced PIH patients (p<0.05). The decrease in PAHPI score and melanin index were not significantly different in treatment groups after four months (p>0.05). Quality of life ameliorated significantly only with cysteamine treatment. However, no significant change in quality of life was observed between groups. Limitations: Limitations of our study include the relatively small sample size and absence of follow-up. Conclusion: Cysteamine cream is an effective treatment of post-acne PIH, with similar efficacy to the accepted treatment of PIH, i.e., hydroquinone cream.

2.
Clin Case Rep ; 12(3): e8621, 2024 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-38464583

RESUMEN

Key Clinical Message: Fox-Fordyce disease is a recognized adverse effect of depilating lasers. It should be considered among the differential diagnoses of the pruritic perifollicular papules that occur at the hair removal laser treatment site, including Alex/Diode laser. Abstract: Fox-Fordyce disease (FFD) is an uncommon skin disorder affecting the apocrine sweat glands and presents as pruritic skin-colored papules, mainly on the axillary and pubic areas. Some predisposing factors are proposed, including hormonal changes. A few cases of FFD have been reported after hair removal by light- and laser-assisted devices. Herein, we report FFD in some areas (axillary and pubic areas) treated with Alex/Diode hair removal laser.

3.
Artículo en Inglés | LILACS-Express | LILACS | ID: biblio-1521598

RESUMEN

ABSTRACT Objective: The objective of this study was to describe a case of cutaneous lichen planus (LP) that appeared following COVID-19 infection. Case description: We report a case of extensive cutaneous classic familial LP in a 4-year-old male child after an asymptomatic serologically confirmed COVID-19 infection. The patient developed intensely itchy, purple, flat-topped papules and plaques, mainly on the dorsal surface of the hands, feet, forearms, and shins. Histopathological examination of the skin biopsy showed vacuolar and apoptotic degeneration of the basal cell layer with a band-like lymphocyte infiltrate at the dermo-epidermal junction and confirmed the diagnosis of LP. Comments: LP could be considered among the differential diagnoses of pediatric post-COVID inflammatory skin lesions, either in the patients recovering from COVID-19 infection or in the suspicious asymptomatic cases in close contact with COVID-19-infected patients.


RESUMO Objetivo: Descrever um caso de líquen plano cutâneo (LP) após infecção por COVID-19. Descrição do caso: Relatamos um caso de LP familiar clássico extenso cutâneo em uma criança de quatro anos de idade após uma infecção por COVID-19 assintomática e sorologicamente confirmada. O paciente desenvolveu pápulas e placas intensamente pruriginosas, roxas e achatadas, principalmente na superfície dorsal das mãos, pés, antebraços e canelas. O exame histopatológico da biópsia de pele mostrou degeneração vacuolar e apoptótica da camada basal com infiltrado de linfócitos em faixa na junção dermoepidérmica e confirmou o diagnóstico de líquen plano. Comentários: O líquen plano pode ser considerado entre os diagnósticos diferenciais de lesões cutâneas inflamatórias pós-COVID pediátricas, tanto em pacientes em recuperação de infecção por COVID-19 quanto em casos assintomáticos suspeitos em contato próximo com pacientes infectados por COVID-19.

4.
Skin Health Dis ; 3(6): e299, 2023 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-38047250

RESUMEN

Background and Objectives: Granuloma annulare (GA) is a benign skin disorder with various histopathologic features that are rarely investigated in Iranian population. We performed this study to find out the clinical and histopathologic features of GA in our referral centre. Methods: One hundred-thirty-six patients with biopsy-proven GA were reviewed. Clinical data and pathological features were recorded. Results: One hundred-eight female patients and 28 male patients (Female/male ratio: 3.85) with mean age of 42.54 ± 21.2 years (range: 2-83 years) were recruited. Eighty-eight (64.7%) patients had interstitial infiltrative pattern and 48 (35.3%) patients had complete palisading granulomas. The infiltrate occupied both upper and lower dermis in most of the cases (67.7%). Significant mucin was detected more commonly in complete GA compared to interstitial GA (p = 0.019), but inflammation degree, eosinophils, plasma cells, and giant cells were not different between two subtypes of GA (p > 0.05). The significant inflammation contained more significant plasma cells (p = 0.006). The significantly more giant cells were detected in patients between 20 and 60 years of age (p = 0.015); but other factors were not different between age groups. Conclusions: In our study, the prevalence of GA in women was significantly higher than in men. Interstitial GA was the more common histological subtype and the inflammation was less severe and the infiltrate was mostly pandermal in our cases. More severe inflammation contained more plasma cells, and more dense giant cells were seen in middle aged patients.

5.
Clin Case Rep ; 11(7): e7638, 2023 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-37415589

RESUMEN

Key Clinical Message: The etiology of the Leser-Trélat sign is still unknown, it is likely that viral infections like COVID-19 can be associated with eruptive seborrheic keratosis, although the exact pathogenesis is still not clear, but this phenomenon can be due to TNF-alpha and TGF-alpha and immunosuppression condition as well as in COVID-19 infection. Abstract: Seborrheic keratosis is a typical benign skin lesion that is almost always seen in elderly populations. The sudden increase in size or an increase in the number of these lesions is called Leser-Trelat sign, this sign is suggesting as a paraneoplastic appearance of internal malignancy. But, Leser-Trelat sign is also described in some nonmalignant conditions, for example, human immunodeficiency virus infection and human papillomavirus infection. Herein, we describe a patient with Leser-Trelat sign after recovery from COVID-19 infection with no evidence of internal malignancy. This case was partially presented as a poster in the 102nd Annual Congress of British Association of Dermatologists in Glasgow, Scotland from July 5 2022 to July 7 2022. British Journal of Dermatology, 187, 2022 and 35. The patient signed written informed consent to permit the publication of the case report without identifying data and to use the photography for publication. The researchers committed to maintaining patient confidentiality. Institutional ethics committee approved the case report (ethics code: IR.sums.med.rec.1400.384).

6.
Clin Case Rep ; 10(8): e6228, 2022 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-35957778

RESUMEN

Various conditions, including infections, can cause telogen effluvium (TE). One of them is coronavirus disease 2019 (COVID-19), where hair loss usually begins between 2 and 12 weeks after the illness. TE can be acute or chronic, and the chronic type can be intermittent. Here, we present the case of a 17-year-old girl with severe and widespread hair loss following an upper respiratory infection suspected to be COVID-19, with the patient having a history of such attacks since childhood. Evidence from biopsy and dermoscopy indicated a diagnosis of TE.

7.
Dermatol Ther ; 35(9): e15726, 2022 09.
Artículo en Inglés | MEDLINE | ID: mdl-35864578

RESUMEN

Cutaneous warts are treated by different treatments with various efficacy. Intralesional injection of 5-fluorouracil (IL 5-FU) has been compared with placebo and a few other wart treatments. Our objective was to compare the efficacy of IL 5-FU injection with cryotherapy to treat common and palmoplantar warts. Forty-five patients with common and palmoplantar warts were treated with either IL 5-FU admixed with lidocaine and epinephrine (5-FU + LE) or cryotherapy, once weekly for six sessions. The patients were assessed for clinical response score, the thickness of warts, pain score, and quality of life in treatment sessions, 2 and 8 weeks after the last session. Nine patients (64.3%) in the 5-FU + LE group, and five patients (35.7%) in cryotherapy group experienced complete response (p = 0.285). Both treatments caused improvement of clinical response score in the patients during the treatment time (p < 0.001), but the efficacy of treatments was not significantly different (p = 0.793). Changes in wart thickness were significantly more marked in the 5-FU + LE group compared to the cryotherapy group (p = 0.021). The patients in the cryotherapy group experienced more severe pain compared to the 5-FU + LE group (p = 0.011). The quality of life better improved in the 5-FU + LE group compared to the cryotherapy group (p = 0.02). To conclude, most of the patients that experienced complete clinical response had received 5-FU + LE injection, though the difference was not significant. However, 5-FU + LE injection accompanied less pain and caused more reduction in wart thickness and more improvement in quality of life compared to the cryotherapy.


Asunto(s)
Lidocaína , Verrugas , Crioterapia , Epinefrina/uso terapéutico , Fluorouracilo , Humanos , Inyecciones Intralesiones , Lidocaína/uso terapéutico , Dolor/tratamiento farmacológico , Dolor/etiología , Calidad de Vida , Resultado del Tratamiento , Verrugas/diagnóstico , Verrugas/terapia
8.
J Cosmet Dermatol ; 21(7): 2871-2878, 2022 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-35510765

RESUMEN

BACKGROUND: Few safe and effective treatments are available for melasma. Cysteamine, a non-melanocytotoxic molecule is a safer alternative to hydroquinone and usable for long-term use. AIM: To evaluate the effect of cysteamine 5% cream in the treatment of melasma. METHODS: Sixty-five of 80 patients completed this single-blind, randomized, controlled trial. The patients received cysteamine 5% or hydroquinone 4%/ascorbic acid 3% (HC) cream. The therapeutic response was evaluated by modified MASI (mMASI) and melanin index (SkinColorCatch) after 2 and 4 months of treatment. The effect of treatment on the quality of life was also assessed. RESULTS: The decrease in mMASI score was from 6.69 ± 2.96 to 4.47 ± 2.16 in the cysteamine group and from 6.26 ± 3.25 to 3.87 ± 2.00 in the HC group after 4 months (p values < 0.001). The melanin index decreased from 37.72 ± 10.17 to 31.47 ± 11.90 in the cysteamine group and from 36.37 ± 10.80 to 23.16 ± 8.83 in the HC group after 4 months (p-value = 0.003 and <0.001, respectively). The difference between mMASI score at baseline and month 4 was not significant between both groups (p-value > 0.05). The difference between the melanin index at baseline and month 4 was significantly more pronounced in the HC group (p-value = 0.002). Quality of life improved in both groups (p-value < 0.05), but was not significantly different between groups (p-value > 0.05). CONCLUSION: Cysteamine was confirmed to be an effective treatment for melasma, with equivalent results to HC in reducing mMASI score and improving quality of life, despite lesser melanin index reduction observed. Cysteamine and HC efficacy was confirmed in patients recalcitrant to previous treatments, by a significant reduction of mMASI and melanin index.


Asunto(s)
Hidroquinonas , Melanosis , Ácido Ascórbico/efectos adversos , Cisteamina/efectos adversos , Emolientes/uso terapéutico , Humanos , Hidroquinonas/efectos adversos , Melaninas , Melanosis/diagnóstico , Melanosis/tratamiento farmacológico , Calidad de Vida , Método Simple Ciego , Resultado del Tratamiento
9.
Clin Case Rep ; 10(2): e05490, 2022 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-35228886

RESUMEN

Radiation Recall Dermatitis (RRD) is an inflammatory process in the site of irradiation, induced by physical and medical agents. Few cases of RRD in the skin and lung have been reported after COVID-19 vaccination. Here, we report radiation recall dermatitis after both doses of inactivated SARS-CoV-2 vaccine (Sinopharm, China).

10.
Clin Rheumatol ; 41(1): 95-104, 2022 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-34471968

RESUMEN

INTRODUCTION: Systemic sclerosis (SSc) is a systemic multi-organ disease. Raynaud's phenomenon (RP) and digital ulcers (DUs) in SSc patients can be resistant to usual treatments. We studied the clinical benefits, capillaroscopy changes, and cost-effectiveness of local injection of botulinum toxin-A (BTX-A) and intravenous prostaglandin analogs (iloprost/alprostadil) in patients with SSc with resistant DUs. METHOD: In a clinical trial study, we evaluated 26 patients fulfilling the ACR/EULAR SSc criteria with resistant DUs. Visual analog scale of pain and RP, skin color and type of ulcers, and capillaroscopy were assessed before and 1 month after treatment. In the first group, 20 units of BTX-A was injected at the base of each involved fingers by a dermatologist. In the second group, 20 µg iloprost or 60 µg alprostadil was infused daily. The cost of these treatments was compared. RESULT: In 26 patients (43 fingers), there were 16 patients (22 fingers) in the BTX-A and 10 patients (21 fingers) in the prostaglandin group. In 95.5% of the BTX-A and 90.5% of the prostaglandin group, the ulcers were healed. In both groups, a significant decrease in pain was seen (p < 0.0001). Capillaroscopy patterns in both groups were not changed although the microhemorrhages disappeared significantly (p value: BTX-A: 0.03 and prostaglandin: 0.002). The cost was significantly lower in the BTX-A injection group (p < 0.0001). CONCLUSION: Both BTX-A and prostaglandins helped in the healing and pain control of DUs. In capillaroscopy, microhemorrhages were significantly decreased in both groups. In the BTX-A group, the cost was significantly lower as an outpatient treatment and was more time-saving. KEY MESSAGES: • BTX-A and prostaglandin analogs both contributed to the healing of digital tip ulcers and improving the pain • In capillaroscopy, microhemorrhages were significantly decreased or disappeared after both treatments • There was no significant side effect in both groups • Comparing both groups, in the BTX-A group, the cost was significantly lower when performed on an outpatient treatment and more time-saving.


Asunto(s)
Toxinas Botulínicas Tipo A , Enfermedad de Raynaud , Esclerodermia Sistémica , Úlcera Cutánea , Toxinas Botulínicas Tipo A/uso terapéutico , Análisis Costo-Beneficio , Dedos/diagnóstico por imagen , Humanos , Iloprost , Angioscopía Microscópica , Prostaglandinas , Enfermedad de Raynaud/tratamiento farmacológico , Esclerodermia Sistémica/complicaciones , Esclerodermia Sistémica/diagnóstico por imagen , Esclerodermia Sistémica/tratamiento farmacológico , Úlcera Cutánea/tratamiento farmacológico , Úlcera Cutánea/etiología , Úlcera
11.
Caspian J Intern Med ; 12(3): 283-289, 2021 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-34221277

RESUMEN

BACKGROUND: Cutaneous pseudolymphoma can histologically and clinically simulate various types of cutaneous lymphoma. We conducted the current study to evaluate the clinicopathological and immunohistochemical (IHC) aspects of cutaneous pseudolymphoma (PSL). METHODS: 30 cases of cutaneous PSL were selected from the archives of 2013-2017 in Shahid Faghihi Hospital pathology lab, Shiraz University of Medical Sciences. Available clinical data, histopathological features and IHC findings were statistically analyzed. RESULTS: The female: male ratio was 2:1 and the median age was 47±14.9 years. The lesions were located on the head and neck 26 (86.7%), trunk 2 (6.7%) and extremities 2 (6.7%). 23 (76.7%) cases had nodular pattern, while 7 (23.3%) showed diffuse pattern. The grenz zone was seen in 24 (80%) cases. Sixteen (53.3%) cases showed top heavy infiltration, 11 (36.7%) showed the same distribution of infiltration at the superficial and deep dermis, often involving the subcutaneous fat and 3(10%) showed bottom heavy infiltration. In IHC, 11(36.6%) cases showed the B cell type, 10 (33.3%) T cell type and 9 (30%) mixed type (B and T cells). CONCLUSION: None of the cases was suspicious for cutaneous lymphoma, applying IHC staining. Gender distribution, and the site of cutaneous lesions were similar to previous studies. The most common histological subtype was nodular, while the B-cell cutaneous pseudolymphoma was slightly more common compared to the T-cell type.

12.
Iran J Pathol ; 15(4): 299-305, 2020.
Artículo en Inglés | MEDLINE | ID: mdl-32944042

RESUMEN

BACKGROUND & OBJECTIVE: Thiopurine drugs are considered as a treatment modality in various autoimmune disorders including pemphigus vulgaris (PV). These drugs are metabolized by an enzyme "Thiopurine S-methyl transferase" (TPMT). Various variants of this enzyme may have decreased activity leading to serious drug side effects. To investigate the phenotype and genotype of TPMT in PV patients receiving thiopurine drugs. METHODS: A total of 50 patients (29 women and 21 men) with pemphigus vulgaris treating with standard dose of Thiopurine drugs were selected. Sex, age, result of liver function test and complete blood count were recorded. Genotyping of two common non-functional allele (TPMT*2 and TPMT*3C) by Allele-specific and RFLP-PCR was performed. TPMT enzymatic level was determined by an ELISA based method. RESULTS: Of patients, 36 (72%) were found to have normal TPMT level; and 12, (24%) had higher level of enzyme and 2, 4% had low TPMT enzyme, but none of the patients showed mutant TPMT*2 and TPMT*3C alleles. None of the patients showed hepatotoxicity and bone marrow suppression. CONCLUSION: The phenotypic assay based on ELISA method may have false positive and misleading results but genotyping using PCR-RFLP and allele specific PCR is accurate, simple and cost-effective and can be used in patients decided to undergo thiopurine treatment.

13.
Int J Trichology ; 12(2): 62-67, 2020.
Artículo en Inglés | MEDLINE | ID: mdl-32684677

RESUMEN

BACKGROUND: Differentiating scarring alopecia secondary to lichen planopilaris (LPP) and discoid lupus erythematosus (DLE) has always been a challenge clinically and pathologically. Plasmacytoid dendritic cells (PDCs) have been reported in the cutaneous lupus erythematosus by CD123 immunostaining. This study assesses CD123 marker positivity and patterns of elastic fiber loss in scalp biopsy to differentiate DLE from LLP. PATIENTS AND METHODS: Forty-three cases with clinical and pathological diagnosis of LPP and DLE were selected, and CD123 immunohistochemistry staining and elastic staining were performed on them. The presence of CD123-positive cells, clustering and distribution of cells, and patterns of elastic fibers loss in the dermis were evaluated. To analyze the data, the Chi-square test was used; moreover, the sensitivity and specificity of CD123 were calculated based on a diagnostic test for 2-by-2 tables. RESULTS: Infiltration of PDC was seen in 90% of DLE cases. The presence of more than 10% and 20% PDC cells in inflammatory cells had 90% and 85% sensitivity and 34.7% and 91.3% specificity, respectively. PDC clusters more than 20 cells had 100% specificity for DLE. Location and patterns of PDC infiltration were not statistically significant (P = 0.378). The wedge-shaped loss of elastic fibers and the diffuse loss were the dominant patterns in LPP and DLE, respectively (P = 0.006). CONCLUSION: Our results suggested that CD123 along with elastic staining and histological features might be useful to diagnose challenging cases of lymphocytic scarring alopecia with clinical differential diagnosis of LPP and DLE.

14.
Cureus ; 12(4): e7606, 2020 Apr 09.
Artículo en Inglés | MEDLINE | ID: mdl-32399340

RESUMEN

Background Although macular amyloidosis is a relatively rare disease, it is a common cutaneous disease in Asia and the Middle East. On hematoxylin and eosin (H&E) stained slides, early lesions could easily be missed without the use of special stains and/or immunohistochemistry. Methods We enrolled 42 patients with the clinical impression of macular amyloidosis who had undergone two 4-mm punch biopsies from 2015 to 2016 at a dermatology clinic affiliated to Shiraz University. Besides, 14 cases with a clinical diagnosis other than macular amyloidosis were selected as the negative control group. Congo red, crystal violet, and immunohistochemical (IHC) staining of CK5 and high molecular weight keratin (HMWK) were performed for each specimen. Results H&E slides showed globular depositions in 15 (35.7%) out of 42 patients. None of the patients showed apple-green birefringence with Congo red stain. Evaluation of crystal violet stained sections revealed purplish violet amyloid deposits in 15 (35.7%) patients. IHC study showed expression of CK5 in 52.4% and HMWK in 50% of the patients, which was not a significant difference (p = 0.715). The findings of both IHC markers had a significant difference with H&E stains (p = 0.039) and crystal violet (p = 0.008). Additionally, we found that two punch biopsies from two sites in the involved area did not have a significant preference over one punch biopsy. All of the cases in the control group were negative for amyloid deposition in H&E, special stains, and IHC stained slides as expected. Conclusions IHC evaluation using CK5 and HMWK might be a useful tool for diagnosing macular amyloidosis.

15.
Cureus ; 12(4): e7517, 2020 Apr 02.
Artículo en Inglés | MEDLINE | ID: mdl-32377465

RESUMEN

Hirudotherapy (leech therapy) is one of the oldest practices in medical history, and nowadays it is used for several purposes in medicine. Salvage of flaps, wound healing, pain management, and treatment of varicose veins are among the common therapeutic applications of leeches. Complications associated with leech therapy include infections, bleeding, anemia, and allergic reaction. Cutaneous pseudolymphoma (benign proliferation of lymphoid cells in the skin) follows several underlying conditions. Although persistent arthropod bite reaction is one of the conditions associated with cutaneous pseudolymphoma, it has been rarely reported after medicinal leech therapy. Here we describe the case of a patient who presented with cutaneous pseudolymphoma after leech therapy as a rare cutaneous complication of hirudotherapy.

17.
Dermatol Online J ; 25(6)2019 Jun 15.
Artículo en Inglés | MEDLINE | ID: mdl-31329390

RESUMEN

Amyloidosis cutis dyschromica (ACD) is a rare pigmentary disorder with about 50 cases having been reported in the English literature. Only one case of ACD has been reported from Iran. We present three patients who presented with generalized hyper- and hypopigmented patches, sparing face, hands, and feet in all three cases. The presence of amorphous eosinophilic deposits in the papillary dermis confirmed the diagnosis of ACD; the deposits were stained by crystal violet in the histopathological examination of the lesions. In all three cases, similar lesions were present in some of the family members. ACD should be considered in the differential diagnosis of diffuse hyperpigmentation studded with hypopigmentation, especially when beginning in childhood.


Asunto(s)
Amiloidosis Familiar/patología , Hiperpigmentación/patología , Hipopigmentación/patología , Adulto , Femenino , Humanos , Irán , Adulto Joven
18.
Rev Soc Bras Med Trop ; 52: e20180278, 2019 Apr 11.
Artículo en Inglés | MEDLINE | ID: mdl-30994790

RESUMEN

INTRODUCTION: The promising non-clinical antileishmanial effects of gentian violet (GV) encouraged us to evaluate the additive effect of GV on cryotherapy. METHODS: For 8 weeks, 59/161 cutaneous leishmaniasis patients/lesions underwent cryotherapy alone (group 1) or cryotherapy accompanied by 1% GV application (group 2). The primary endpoint was clinical response. RESULTS: Ultimately, 54.7% and 45.3% of the significantly cured lesions belonged to groups 1 and 2, respectively, which was not statistically significant. The clinical response was significantly different between the two groups at the end of the fourth week. CONCLUSIONS: Although the clinical response of the two groups was significantly different at the end of the fourth week, application of GV did not increase the efficacy of cryotherapy.


Asunto(s)
Antiinfecciosos Locales/administración & dosificación , Crioterapia/métodos , Violeta de Genciana/administración & dosificación , Leishmaniasis Cutánea/tratamiento farmacológico , Adulto , Femenino , Estudios de Seguimiento , Humanos , Masculino , Proyectos Piloto , Método Simple Ciego , Resultado del Tratamiento
19.
Heliyon ; 5(12): e03073, 2019 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-31890980

RESUMEN

BACKGROUND: This study aims to evaluate the use of fluorescent dye Dil and super vital dye acridine orange (AO) in vitro tracking of labeled L. major in the fibroblast cells. METHODS: Dil crystal and AO were used to stain L. major in a co-culture of the fibroblasts with the parasite. AO staining solution was added to 1 × 106 parasites. After 10 min, the stained parasites were centrifuged and washed seven times with phosphate buffered saline (PBS). The stained promastigote was incubated with fibroblasts for 6-8 h. The presence of stained parasites with AO in the fibroblast was assessed using a fluorescence microscope. 1 × 106/mL promastigote of L. major was gently suspended and mixed by Dil staining solution with an ultimate concentration of 0.002 µg/mL and it was kept for 20 min at the room temperature. Subsequently, after washing it in PBS for seven times, it was centrifuged at 3000 rpm for 10 min. The supernatant was removed and the precipitate containing stained promastigote was suspended in fresh DMEM F12 with fibroblasts at 37 °C for 6 h. The presence of stained parasites with Dil in fibroblast was assessed using a fluorescence microscope. Fibroblast characterization was undertaken by a real-time polymerase chain reaction (PCR). RESULTS: Acridine orange staining assisted in detection of the live parasite in the fibroblast cells. Free promastigote looked green before entering into the fibroblasts after 12 h culture. The parasite entered the cytoplasm of fibroblasts at the beginning of the exposure and gradually entered the nucleus of the fibroblast. The fibroblast nucleus was entirely stained green by AO. The L. major appeared green under the fluorescent microscope. Dil staining revealed that the internalized parasites with red/orange color were localized within the cytoplasm after 6-hours and the nucleus of the fibroblasts after 72-hours following culture. Human fibroblasts were positive at the expression of CD10, CD26, matrix metalloproteinase-1 (MMP-1) and matrix metalloproteinase-3 (MMP-3) and negative for CD106 and integrin alpha 11. CONCLUSION: The fluorescent dye Dil staining is a safe, easy to use, inexpensive and fast method for labeling of the Leishmania parasite in the fibroblast cells. Acridine orange staining could be useful for tracing the parasites in the fibroblasts too. In this study, both Dil and AO were compared and considered as suitable vital dyes for identifying labeled Leishmania in the fibroblast in vitro, but Dil was superior to AO with its feature does not transfer from the labeled to unlabeled cells.

20.
Rev. Soc. Bras. Med. Trop ; 52: e20180278, 2019. tab, graf
Artículo en Inglés | LILACS | ID: biblio-1041586

RESUMEN

Abstract INTRODUCTION: The promising non-clinical antileishmanial effects of gentian violet (GV) encouraged us to evaluate the additive effect of GV on cryotherapy. METHODS: For 8 weeks, 59/161 cutaneous leishmaniasis patients/lesions underwent cryotherapy alone (group 1) or cryotherapy accompanied by 1% GV application (group 2). The primary endpoint was clinical response. RESULTS: Ultimately, 54.7% and 45.3% of the significantly cured lesions belonged to groups 1 and 2, respectively, which was not statistically significant. The clinical response was significantly different between the two groups at the end of the fourth week. CONCLUSIONS: Although the clinical response of the two groups was significantly different at the end of the fourth week, application of GV did not increase the efficacy of cryotherapy.


Asunto(s)
Humanos , Masculino , Femenino , Adulto , Leishmaniasis Cutánea/tratamiento farmacológico , Crioterapia/métodos , Violeta de Genciana/administración & dosificación , Antiinfecciosos Locales/administración & dosificación , Método Simple Ciego , Proyectos Piloto , Estudios de Seguimiento , Resultado del Tratamiento
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