Significant reduction of inflammation and sebaceous glands size in acne vulgaris lesions after intense pulsed light treatment.

Intense pulsed light (IPL) has been used for years in treatment of acne vulgaris. However, quantitative evaluation of histopathological changes after its use as a sole therapy was poorly investigated. Accordingly, this study aims to objectively evaluate inflammatory infiltrate and sebaceous glands in acne vulgaris after IPL. Twenty-four patients of acne were treated with six IPL sessions. Clinical evaluation was done at 2 weeks after last session by counting acne lesions. Patient satisfaction using Cardiff Acne Disability Index (CADI) was recorded at baseline, 2 weeks and 3 months after IPL. Using histopathological and computerized morphometric analysis, quantitative evaluation of inflammatory infiltrate and measurement of surface area of sebaceous glands were performed for skin biopsies at baseline and 2 weeks after last session. After IPL, there was significant reduction of all acne lesions especially inflammatory variety with significant decrease of CADI score at 2 weeks and 3 months after IPL (p < .05). Microscopically, there was significant decrease in density of inflammatory infiltrate and surface area of sebaceous glands (p < .05). So, IPL is fairly effective therapy in acne vulgaris especially inflammatory variety. The results suggest that IPL could improve acne lesions through targeting both inflammation and sebaceous glands.

Evaluation of apoptosis regulatory proteins in response to PUVA therapy for psoriasis.

BACKGROUND: The histopathologic changes characteristic of psoriasis might be related to suppressed apoptosis. One of the actions of psoralen ultraviolet A (PUVA) in psoriasis could be exerted through induction of apoptosis of keratinocytes and lymphocytes; however, its exact molecular mechanism is still confusing. AIM: In this study, we evaluated the expression of pro-apoptotic (P53, Fas and Bax) and anti-apoptotic (Bcl-2) proteins correlating it with apoptotic index (AI) and epidermal thickness in psoriatic skin before and after PUVA therapy. METHODS: Lesional and non-lesional skin biopsy specimens were obtained from 10 patients with generalized plaque psoriasis before and after 8 weeks of PUVA therapy. Histometric measurements of epidermal thickness as well as P53, Fas, Bax and Bcl-2 expressions were evaluated using immunoperoxidase technique and apoptotic cells were detected by terminal deoxynucleotide transferase (TdT) mediated deoxyuridine triphosphate nick end labeling (TUNEL) method. RESULTS: After PUVA therapy, the epidermal thickness of psoriatic skin was significantly decreased (P < 0.001) and keratinocytes of psoriatic skin showed significant increased expression of P53 (P < 0.001), Fas (P < 0.001) and Bcl-2 (P < 0.001) with no significant change in Bax expression (P > 0.05). Apart from significant decrease of Bcl-2 expression (P = 0.01), no significant difference in all previous markers were encountered in lymphocytes (P53, Fas and Bax; P > 0.05) after PUVA therapy. The AI was significantly increased (P = 0.008) after PUVA therapy especially in lymphocytes (P = 0.002). CONCLUSION: The present study suggests that one of the actions of PUVA therapy in psoriasis might be exerted through induction of apoptosis especially of lymphocytes by suppression of Bcl-2 expression and of keratinocytes through P53 and Fas pathways leading to healing of psoriasis.

A new morphometric technique for assessment of melanization in skin of guinea pigs.

BACKGROUND: The degree of melanization is an important finding in many skin disorders. An objective measurement of melanin density is needed to achieve an accurate evaluation. OBJECTIVES: The present work aims at translating the subjective increase of melanin particles after narrow band ultraviolet-B (NB-UVB) irradiation into objective numerical values ready for statistical analysis. MATERIALS AND METHODS: This study had involved 18 guinea pigs that were exposed to biweekly sessions of NB-UVB radiation for 4 weeks to induce skin pigmentation. Two skin biopsies were obtained from each animal; the first before treatment and the second at the end of the study, using 5 mm punch and stained with hematoxylin and eosin and Masson-Fontana (MF) stains. Surface area of both the epidermis (ESA) and the melanin particles (MPSA) were measured in µm(2) using a software supplied with Olympus light microscope. The MPSA/ESA percentage was calculated for each biopsy. The results before and after NB-UVB exposure were compared and statistically analyzed. RESULTS: In the MF-stained sections, the mean ± SD of the MPSA/ESA percentage were 0.24 µm(2) ± 0.09 and 6.21 µm(2) ± 2.45 at the start of the study and at its end, respectively, with a highly significant difference (P < 0.001). CONCLUSION: This technique offers a new methodology for an accurate numerical evaluation of epidermal melanization.

Effect of PUVA therapy on melanocytes and keratinocytes in non-segmental vitiligo: histopathological, immuno-histochemical and ultrastructural study.

BACKGROUND AND AIMS: Psoralen ultraviolet A (PUVA) is an important modality in treating vitiligo. Its effect on melanocytes and keratinocytes is not sufficiently studied. In this work, we investigated 30 cases of non-segmental vitiligo regarding the changes of melanocytes and keratinocytes in both vitiliginous and nearby areas before and after PUVA therapy. METHODS: Three skin biopsies were obtained from each patient from the vitiliginous, marginal and perilesional areas before and after 12 months of PUVA. Biopsies were examined histologically using haematoxylin and eosin, Masson-Fontana stains and 3,4-dihydroxyphenylalanine (DOPA) reaction and histochemically using human melanoma black-45 (HMB-45) antibody while ultrastructural examination was performed on six patients. Control biopsies were taken from five healthy volunteers. RESULTS: In 10% of pretreated biopsies from the centre of vitiligo lesions, scanty melanocytes were detected histologically and ultrastructurally, while they did not stain with DOPA or HMB-45 antibody suggesting that these melanocytes were inactive. Moreover, degenerative changes were detected by electron microscopy in both melanocytes and keratinocytes in all areas. After PUVA therapy, obvious improvement of the histopathological changes occurred with significant increase in active melanocytes. The degeneration of melanocytes and keratinocytes was also reduced at the ultrastructural level. CONCLUSION: Vitiligo affects both melanocytes and keratinocytes causing degenerative changes. These changes were present in both the leucodermic and the apparently normal perilesional skin. PUVA increases the number of active epidermal melanocytes in the three tested areas and recovers the melanocyte and keratinocyte degeneration.

The expression and distribution of deoxyribonucleic acid repair and apoptosis markers in testicular germ cells of infertile varicocele patients resembles that of old fertile men.

OBJECTIVE: To explore the expression and distribution of DNA damage repair and apoptosis marker proteins in human testicular germ cells of infertile varicocele patients; and to compare the expression and distribution with that of young and old fertile men. DESIGN: Retrospective case-control study. SETTING: Academic institutions. PATIENT(S): Testicular specimens were obtained from 8 infertile varicocele patients aged 20-30 years and from 16 fertile volunteers aged 20-82 years. INTERVENTION(S): Testicular germ cell DNA repair markers were assessed using immunohistochemical staining for the cell proliferation marker (proliferating cell nuclear antigen), DNA repair markers [poly(ADP-ribose) polymerase-1 (PARP-1), poly(ADP-ribose), X-ray repair cross-complementing 1, and apurinic/apyrimidinic endonuclease 1], and apoptosis markers (caspase 9, active caspase 3, and cleaved PARP-1). MAIN OUTCOME MEASURE(S): The prevalence and cellular localization of the above markers in testicular tissues of varicocele patients and fertile men of varying ages. RESULT(S): Statistically significant differences in DNA damage repair-associated proteins and apoptosis markers were observed in infertile men with varicocele compared with fertile young men. Old fertile men showed similar expression of the same markers when compared with infertile varicocele patients. CONCLUSION(S): The study demonstrates that there is an increase in human testicular germ cell DNA repair and apoptosis in infertile varicocele patients and that their profile resembles that of premature aging.

Deoxyribonucleic acid repair and apoptosis in testicular germ cells of aging fertile men: the role of the poly(adenosine diphosphate-ribosyl)ation pathway.

OBJECTIVE: To explore the relationship between men's age and DNA damage repair proteins related to apoptosis in human testicular germ cells. DESIGN: Retrospective case-control study. SETTING: Academic institutions. PATIENT(S): Testicular specimens were obtained from 22 fertile volunteers aged 20-82 years. INTERVENTION(S): Deoxyribonucleic acid repair markers were assessed using immunohistochemical staining for the cell proliferation marker [proliferating cell nuclear antigen (PCNA)]; DNA repair markers [poly(adenosine diphosphate-ribose) polymerase-1 (PARP-1), poly(adenosine diphosphate-ribose) (PAR), X-ray repair cross-complementing1(XRCC1), and apurinic/apyrimidinic endonuclease 1 (APE1)]; and apoptosis-associated markers (caspase 9, active caspase 3, and cleaved PARP-1). MAIN OUTCOME MEASURE(S): The prevalence and cellular localization of the above markers in testicular tissues of young, middle aged, and old men. RESULT(S): Statistically significant differences in DNA damage repair-associated proteins (PARP-1, PAR, XRCC1, and APE1), and apoptosis markers (caspase 9, active caspase 3, and cleaved PARP-1) were observed in testicular samples from older men. These differences were most marked in spermatocytes. CONCLUSION(S): The study demonstrates that there is an age-related increase in human testicular germ cell DNA break repair and apoptosis with age.