Tacrolimus (FK506) ointment combined with Nb-UVB could activate both hair follicle (HF) and dermal melanocyte precursors in vitiligo: the first histopathological and clinical study.

Topical Tacrolimus, especially when combined with Nb-UVB, has been proven clinically to be effective in the treatment of vitiligo. However, no histological study has evaluated the repigmentation mechanism of tacrolimus ointment in combination therapy with Nb-UVB. In this study, the histological findings in patients receiving Nb-UVB were compared with those receiving topical tacrolimus combined with Nb-UVB. Twenty patients were recruited and received Nb-UVB treatment. The first ten patients were selected for the combination therapy and instructed to apply tacrolimus 0.1% ointment twice daily on the specified lesion of interest. The remaining ten patients did not receive any other topical treatments. Skin biopsy was performed at baseline from the depigmented area and 2-3 months post-treatment from the repigmented area. Biopsy specimens were stained with haematoxylin-eosin-safran (HES), Fontana Masson, HMB45, Melan A, MITF, SOX10 and Nestin. Clinically, in the combination therapy group, interfollicular repigmentation in addition to the perifollicular and marginal pattern was observed. Histologically, in the combination therapy group, besides the migration of melanocytes from the bulge of the hair follicle seen in the monotherapy group, for the first time, we observed dermal melanocyte precursors located in mid- and superficial dermis.

Effect of mitomycin-C on corneal endothelial cell parameters after refractive surface ablation procedures.

Background: The effect of mitomycin-C (MMC) on the reduction of endothelial cell count in the cornea remains controversial. We aimed to evaluate the effect of MMC on corneal endothelial cell parameters after refractive surface ablation procedures, including photorefractive keratectomy (PRK) and laser epithelial keratomileusis (LASEK). Methods: In this interventional, comparative, follow-up study, 342 eyes of 171 patients were followed up for 6 months. Patients undergoing PRK or LASEK were included and were divided into two groups: group one (188 eyes of 94 patients) with an ablation depth of ≥ 65 µm and who received intraoperative 0.02% MMC for 30 s, and group two (154 eyes of 77 patients) with an ablation depth of < 65 µm and who received balanced salt solution for 30 s. Changes in endothelial cell density (ECD), central corneal thickness (CCT), coefficient of variation (CV), and hexagonality values were compared between the groups at 3 and 6 months after surgery. Results: The mean (standard deviaiton [SD]) age of the patients was 28.11 (6.56) years. The mean (SD) ECD did not change significantly in either group between the baseline and at 3 and 6 months postoperatively. The baseline mean ECD was significantly higher in group one than that in group two (P < 0.001) and remained so at 3 (P = 0.002) and 6 months (P = 0.022) postoperatively. The baseline hexagonality value was lower in group one (P = 0.173), with a gradual decrease during the postoperative follow-up as compared with that in group two (P = 0.016 and 0.001 at 3 and 6 months postoperatively, respectively). Group one had a significantly lower CCT at 3 and 6 months postoperatively (both P < 0.001) and a higher mean CV (3 months: P = 0.028; 6 months: P = 0.328). Conclusions: A single intraoperative application of MMC for 30 s as prophylaxis for corneal haze development during refractive surface ablation procedures had no significant effect on ECD up to 6 months postoperatively. Future studies with a contralateral-eye design (to neutralize factors specific to the individual patient), a larger sample size, and longer follow-up are necessary to confirm or disprove our observations.

Macrophage migration inhibitory factor polymorphism (rs755622) in alopecia areata: a possible role in disease prevention.

Alopecia areata (AA) is an organ-specific autoimmune disease that targets the bulb of the hair follicles and results in non-scarring hair loss that can range from patchy lesions to involvement of the entire scalp. AA develops when the hair follicles lose their physiologic state of immune privilege. One of the key factors that help in maintaining this immune privilege by suppressing natural killer cells is macrophage migration inhibitory factor (MIF). Surprisingly, MIF is also known to provoke autoimmunity by upregulating cytokines. To address this dilemma and understand the exact nature of the involvement of MIF in disease pathogenesis we investigated the association of MIF gene polymorphisms (- 173 G > C, rs755622) with AA by conducting a case-control study of 274 subjects. We observed that the frequency of the C allele in the patients was significantly lower than the control group (0.15, 0.23, respectively, p = 0.01) and the combined frequencies of the CC and GC genotypes (dominant Mendelian pattern) had the most prevalent difference between the two groups (odds ratio 0.60, 95% confidence interval 0.36-0.99; p = 0.048).Since the C allele is associated with higher MIF transcription levels, this could infer that MIF is more likely to attribute to the preservation of the immune privilege rather than acting as a proinflammatory factor.

Is the +405 G/C single nucleotide polymorphism of the vascular endothelial growth factor (VEGF) gene associated with late-onset vitiligo?

The increasing body of evidence for the relationship between the vascular endothelial growth factor (VEGF) polymorphism and autoimmune disorders combined with the enhanced expression of this angiogenic factor in vitiligo makes VEGF a very interesting candidate gene to be investigated in vitiligo. The aim of this study was to evaluate the possible associations between the +405 G/C single nucleotide polymorphisms (SNP) of the VEGF gene (rs2010963) and vitiligo. The independent case-control population sample of 152 patients with vitiligo and 152 matched controls was evaluated in this study. A questionnaire was completed for each vitiligo patient to document the demographic and clinical characteristics of the patients. All enrolled individuals had a venous blood sample collected. Genotype frequencies for +405 G/C VEGF gene polymorphism were determined using polymerase chain reaction (PCR) amplification and restriction fragment length polymorphism (RFLP) analysis. There were no significant differences in genotype or allele distributions for this SNP between cases and controls. However, we observed a significant association between GG genotype and higher age at onset of vitiligo (p = 0.04). Moreover, patients stratification revealed a significant increase in the frequency of GG genotype compared to CC + CG genotypes in patients with the late onset (≥20 years) vitiligo (p = 0.05). Although these results are not conclusive, they could potentially lead to considering the angiogenic factors as a potential target for therapy in late-onset vitiligo.

The Association between Genetic Variation in Wnt Transcription Factor TCF7L2 (TCF4) and Alopecia Areata.

Background: Alopecia areata (AA) is a non-scarring hair loss with a polymorphous presentation ranging from patchy lesions to involvement of the entire scalp. The disease is the consequence of an autoimmune attack on hair bulbs that results in a premature transition of hair follicles to catagen and telogen. Thus the Wnt/ß-catenin signaling pathway that regulates the hair cycling might be involved in the pathogenesis of AA. Genetic variations in the components of Wnt/ß-catenin could greatly alter their adaptive mechanisms against an immunologic attack. Objectives: Our aim was to investigate the association between AA and genetic polymorphisms in the TCF7L2 gene, one of the most important components of the Wnt/ß-catenin pathway. Methods: This is a case-control study of 145 patients with AA and 152 healthy controls. Genotyping of the TCF7L2 gene (rs7903146) was performed via the ARMS-PCR method (amplification refractory mutation system- polymerase chain reaction). The allele and genotype distribution was compared between the two groups. Results: The frequency of the T allele (0.38 vs. 0.28, odds ratio = 1.56, 95% CI = 1.09-2.17, p = 0.013) and TT + CT genotypes (0.68 vs. 0.53, odds ratio = 1.88, 95% CI = 1.17-3.02, p = 0.008) were significantly higher in AA patients. Conclusions: This study indicates that the TCF7L2 gene variant is associated with AA. Its contribution to disease pathogenesis could either be through a hair cycling defect or dendritic cell dysregulation.

CDH1 and DDR1 common variants confer risk to vitiligo and autoimmune comorbidities.

The weaker expression of the two main proteins adhering melanocytes to the epidermis basal layer, Epithelial Cadherin (E-cadherin) and Discoidin Domain Receptor Tyrosine kinase 1 (DDR1), has been implicated as one of the aggravating factors in the loss of melanocytes in vitiligo. The present study was designed to assess the association between single nucleotide polymorphisms (SNP) in the genes encoding these proteins, CDH1 and DDR1, and the risk of developing vitiligo. The independent case-control study was conducted on the sample including152 patients with vitiligo and 152 matched controls. A questionnaire was completed for recording demographic and clinical characteristics of vitiligo patients. Venous blood samples were taken from all the subjects. Genotype frequencies were determined for CDHI C/T (rs 10431924) and DDRI A/C (rs 2267641) genes polymorphisms using polymerase chain reaction (PCR) amplification method and Restriction Fragment Length Polymorphism (RFLP) analysis. The CDH1 CC genotype was found to be significantly associated with the risk of developing vitiligo. The results of stratified analysis revealed a correlation between CDH1 CC genotype and late age of onset, clinical type of vitiligo, the absence of autoimmune comorbidities and family history of autoimmune disorders. However, the expression level of CDH1 TT genotype increased significantly in patients with autoimmune comorbidities. There was also a significant relationship between the DDR1 CC genotype and the risk of developing vitiligo. The results of stratified analysis revealed a correlation between DDR1 CC genotype and early age of onset, clinical type of vitiligo and absence of family history of autoimmune disorders. The findings of the study confirm the conjecture previously made in the literature regarding the melanocytes' adhesion deficit as an initial step for pigment loss in vitiligo and emphasize the substantial role of friction and koebner phenomenon in the pathogenesis of vitiligo. Moreover, a probable association can be proposed between the adhesion deficit involved in vitiligo and autoimmune disorders.

Considerable variation among Iranian dermatologists in the knowledge and attitudes regarding the use of biologic agents to manage psoriasis.

BACKGROUND: Many international guidelines have been introduced with the aim of helping clinicians by providing evidence-based advice for the prescription of biologic therapies in psoriasis. Because no local or regional guideline is available, the treatment of psoriasis with biologics is mainly based on clinical experiences regarding the international guideline among Iranian dermatologists. OBJECTIVE: To assess the knowledge and attitudes among Iranian specialists regarding the use of biologic agents to manage psoriasis. METHODS: Data were collected using an electronic questionnaire specifically designed for this study based on a review of the literature. The designed Google form consisted of 53 multiple choice questions divided into five sections. RESULTS: A total of 111 dermatologists agreed to participate in this study. There was considerable variation among the responding dermatologists in terms of their knowledge and attitudes toward biologics. There were some significant associations between knowledge and attitudes of dermatologists toward biologics and their personal and professional characteristics. CONCLUSIONS: More comprehensive educational approaches, both in the dermatology residency and postgraduation periods, could be very beneficial to promote the knowledge and attitude of the dermatologist in treating psoriasis with biologic agents. This study could be one of the first steps to develop a country-based or a region-based plan to improve the knowledge and attitude among dermatologists regarding the use of biologic drugs in psoriasis and possibly to reach a better status to prescribe these agents in the management of psoriasis.

The Risk of Skin Necrosis Following Hyaluronic Acid Filler Injection in Patients With a History of Cosmetic Rhinoplasty.

BACKGROUND: As the number of patients using dermal filler for face augmentation increases, the number of adverse events associated with injection may increase. Unpredictable repositioning of blood vessels and a more tenuous blood supply in the operated nose may increase the risk of ischemia, necrosis, and vascular embolism following the filler injection. OBJECTIVES: To highlight the importance of the patient's history of previous cosmetic procedures including rhinoplasty in the emergence of vascular complications. METHODS: Our medical records over a two-year period were reviewed retrospectively to identify all patients who were treated at our center for vascular complications associated with facial hyaluronic acid filler injections. In each case, the subject's demographic data (gender and age), habitual status, past medical and surgical history, the symptoms and clinical presentation at the first visit, the time interval between the injection and the onset of symptoms, injected filler material and brand, injection sites, the introduced treatment, and photographs were reviewed carefully. RESULTS: A total of seven patients were identified, each developing skin necrosis following injection of the hyaluronic acid filler. All patients reported a cosmetic rhinoplasty more than three years ago. CONCLUSIONS: Our finding confirms the conjecture previously made in the literature and suggests that the distinctive vascularity of the nose and the surrounding area may cause filler augmentation induced vascular complications in patients whose vascular circulation has already been compromised by a previous nose surgery.

NMDA receptors interact with the retrieval memory enhancing effect of pioglitazone in mice.

PURPOSE: The present study has been undertaken to investigate the possible involvement of the glutamatergic pathway in the beneficial effects of pioglitazone on consolidation and retrieval phases of memory. MATERIALS AND METHODS: The Y-maze task was used to assess short-term spatial recognition memory in animals. Scopolamine (1mg/kg, i.p.) or MK-801 (dizocilpine) (0.03, 0.1 and 0.3mg/kg, i.p.) were administered immediately after the training session to impair memory consolidation or 30min before the retention trial to impair memory retrieval. Pioglitazone (10, 20, 40 and 80mg/kg, p.o.) was administered 2h before the retention session in memory retrieval experiments or immediately after the training session in consolidation experiments. And finally NMDA (N-methyl-d-aspartate) (75mg/kg, i.p) was administered 15min before the administration of pioglitazone. RESULTS: 1) MK-801 (0.3mg/kg) impaired the retrieval of spatial recognition memory. 2) Pioglitazone failed to improve MK-801 induced impairment of retrieval of spatial recognition memory. 3) The 20mg/kg dose of pioglitazone significantly improved memory in mice with scopolamine induced impairment of memory retrieval. 4) Sub-effective dose of MK-801 (0.1mg/kg) was capable of reversing the beneficial effect of pioglitazone on retrieval of memory in scopolamine-treated mice, 5) Administration of NMDA (75mg/kg) and a sub-effective dose of pioglitazone (10mg/kg) reversed the effect of scopolamine and promoted memory retrieval. 6) MK-801 did not affect the consolidation phase of spatial recognition memory. 7) Pioglitazone did not affect scopolamine-induced impairment of memory consolidation. CONCLUSIONS: Sub-effective dose of MK-801 is capable of reversing the protective action of pioglitazone on scopolamine-induced impairment of memory retrieval. Additionally, co-administration of 75mg/kg NMDA and a sub-effective dose of pioglitazone potentiated the effect of pioglitazone on memory retrieval impaired by scopolamine. These results support the idea that pioglitazone plays its memory retrieval enhancement role through the glutamatergic pathway.

Short-time mitomycin-C application during photorefractive keratectomy in patients with low myopia.

PURPOSE: To evaluate the safety and efficacy of 5 seconds of mitomycin-C (MMC) application during photorefractive keratectomy (PRK) for low myopia. SETTING: Basir Eye Clinic, Tehran, Iran. DESIGN: Prospective randomized sham-controlled double-masked clinical trial. METHODS: Patients with low myopia and an ablation depth of less than 65 µm were recruited. One eye of each patient was included in the study and was randomly assigned to receive intraoperative topical MMC 0.02% for 5 seconds or a balanced salt solution in the same manner (control group). Corneal haze development during the 6 months after the PRK was the main outcome measure. RESULTS: Of the 184 eligible patients (MMC = 93; control = 91), 152 (78 and 74, respectively) completed the follow-up. The postoperative haze grade was significantly lower in the MMC group (P=.01). The mean endothelial cell density was not significantly different between the MMC group and the control group preoperatively (2879.97 cells/mm(2) ± 298.04 [SD] versus 2819.69 ± 303.89 cells/mm(2); P=.22) or 6 months postoperatively (2878.79 ± 283.04 cells/mm(2) versus 2878.79 ± 283.04 cells/mm(2); P=.25). No notable ocular complication occurred throughout the study. CONCLUSION: Short-time application of MMC 0.02% was safe and effective in preventing haze formation in eyes having PRK with an ablation depth of less than 65 µm compared with matched controls up to 6 months. FINANCIAL DISCLOSURE: No author has a financial or proprietary interest in any material or method mentioned.

Involvement of nitric oxide in pioglitazone memory improvement in morphine-induced memory impaired mice.

INTRODUCTION: Pioglitazone, a PPAR-γ agonist, which is clinically used in treating diabetic patients, has been recently reported to have crucial roles in improving cognition and memory performance. Since the mechanisms involved in the neuroprotective effect of pioglitazone are not entirely understood, the current study was designed to investigate the possible interaction of pioglitazone with morphine in memory-impaired mice and the probable role of nitric oxide (NO) in this effect. MATERIALS AND METHODS: All the experiments were performed in passive avoidance and Y-maze paradigms. To induce memory impairment, mice were administered morphine (1, 3 and 10mg/kg, s.c.) immediately before the training trial. Pioglitazone (20, 40 and 80mg/kg, p.o.) was gavaged 2h prior to the training trial. Further, an NO synthase inhibitor, L-NAME (10mg/kg, i.p.), or an inducible NO synthase inhibitor, aminoguanidine (100mg/kg, i.p.) was administered 30 min before the training trial to determine the possible involvement of NO in the restorative effect of pioglitazone. RESULTS: 1) Morphine dose dependently impaired the acquisition of spatial memory and passive avoidance task. 2) Treatment with pioglitazone significantly improved the memory performance in morphine-treated mice in both tests. 3) In the passive avoidance task, L-NAME, but not aminoguanidine, altered the effect of pioglitazone on morphine-induced memory impairment. 4) In Y-maze discrimination, the memory improving effect of pioglitazone was reversed by both NO synthase inhibitors, L-NAME and aminoguanidine. DISCUSSION: Our results demonstrate that the pioglitazone improving effect on the morphine-induced impairment of memory acquisition is at least in part through the NO pathway. It is suggested that in short term spatial recognition memory, both inducible and constitutive NO synthases are involved, but in the long term fear memory, only the constitutive NO synthases indicated a prominent role in the anti-amnestic effect of pioglitazone on morphine-induced memory impairment.

Involvement of NMDA receptors in the beneficial effects of pioglitazone on scopolamine-induced memory impairment in mice.

INTRODUCTION: Pioglitazone, a peroxisome proliferator activated receptor γ (PPARγ) agonist, is widely used in clinical medicine as a treatment for type 2 diabetes and is recently proved to have beneficial effects on improving cognition in early stages of Alzheimer's disease (AD). Moreover, it has been shown that pioglitazone reduces N-methyl-D-aspartate (NMDA, a glutamate agonist) mediated calcium currents and transients. Since enhanced calcium transients are present in AD models, we tested the hypothesis whether pioglitazone manifests its acquisition memory enhancement role through glutamatergic pathway. MATERIAL AND METHODS: Memory performance was evaluated in a two-trial recognition Y-maze test and passive avoidance in mice. Pioglitazone (20 or 40 mg/kg, p.o.) was administered 2h before each trial, NMDA (75 mg/kg i.p.), 15 min before pioglitazone, and scopolamine, an M1 (muscarinic) receptor antagonist (0.3 or 1.0 mg/kg i.p.) and MK-801 (dizocilpine) (0.01, 0.03 or 0.1 mg/kg, i.p.), the highly selective, non-competitive NMDA antagonist--30 min beforehand. RESULTS: (1) We induced the memory impairment by scopolamine or MK-801 before trials. (2) Pioglitazone did not improve the memory impairment induced by MK-801. (3) Pioglitazone significantly improved the memory impairment induced by scopolamine. (4) Subeffective dose of MK-801 nullified the beneficial effects of pioglitazone in scopolamine induced memory impaired mice. (5) NMDA promoted the effects of subeffective dose of pioglitazone on memory impaired by scopolamine. DISCUSSION: In conclusion, the present study suggests that glutamatergic pathway is involved in the pioglitazone induced memory performance.