Lesional CD8+ T-Cell Number Predicts Surgical Outcomes of Melanocyte-Keratinocyte Transplantation Surgery for Vitiligo.

The melanocyte-keratinocyte transplantation procedure (MKTP) treats stable and recalcitrant vitiligo. Despite careful selection of candidates based on clinical stability, the success of the procedure is unpredictable. The aim of our study was to define the immunological profile of stable vitiligo lesions undergoing MKTP and correlate them with clinical outcomes. We included 20 MKTP candidates with vitiligo and a patient with piebaldism as a control. Prior to MKTP, T-cell subsets and chemokines in the recipient skin were measured by flow cytometry and ELISA. During MKTP, melanocytes in the donor skin were quantified by flow cytometry. After MKTP, patients were followed for 12 months and repigmentation was assessed clinically and by ImageJ analysis of clinical photographs. Baseline immunologic biomarkers, duration of clinical stability, and transplanted melanocyte number were correlated to postsurgical repigmentation scores. CD8+ T cells were elevated in 43% of the clinically stable vitiligo lesions. CD8+ T-cell number negatively correlated with postsurgical repigmentation scores (r = -0.635, P = 0.002). Duration of clinical stability, skin chemokines, and transplanted melanocyte number did not influence postsurgical repigmentation. This study demonstrates that CD8+ T-cell number correlates negatively with success of postsurgical repigmentation and can be a biomarker to identify ideal surgical candidates.

Combined intralesional injection of tuberculin purified protein derivative plus cryotherapy versus each alone in the treatment of multiple common warts.

Different modalities are used for treatment of common warts, but none of them had been proved the best in achieving complete cure. We aim to compare the effect of cryotherapy, intralesional injection of tuberculin purified protein derivative (PPD) and cryotherapy combined with intralesional injection of tuberculin PPD in the treatment of multiple common warts. This study is a randomized clinical trial in which the patients were randomly divided into three groups; group (A) included 25 patients subjected to cryotherapy, group (B) included 25 patients subjected to intralesional injection of tuberculin PPD and group (C) included 25 patients subjected to cryotherapy plus intralesional injection of tuberculin PPD. All the three groups showed a significant clinical improvement (p < 0.001) with statistically significant difference between cryotherapy group (A) and intralesional injection of tuberculin PPD group (B) (p < 0.001) and between cryotherapy group (A) and cryotherapy plus intralesional injection of tuberculin PPD group (C) (p < 0.001). However, there was no statistically significant difference between both intralesional injection of tuberculin PPD group alone (B) and cryotherapy plus intralesional injection of tuberculin PPD group (C) (p = 0.213). In Conclusion the cryotherapy combined with intralesional injection of PPD and intralesional injection of PPD alone are better than cryotherapy alone in treatment of multiple common warts. However, better response could be reached in combination of both cryotherapy and intralesional PPD with less number of sessions.

Superficial cryotherapy using dimethyl ether and propane mixture versus microneedling in the treatment of alopecia areata: A prospective single-blinded randomized clinical trial.

To verify and compare the therapeutic efficacy and safety of superficial cryotherapy using dimethyl ether and propane (DMEP) mixture vs. microneedling in the treatment of mild scalp alopecia areata (AA). In a prospective randomized single-blinded clinical trial, 80 patients with clinically evident scalp mild AA were randomly assigned into two groups of 40 patients each. Group (1) was treated by superficial cryotherapy using DMEP in three freeze-thaw cycles of 5 s each. Group (2) was treated by microneedling. Both groups were treated every 2 weeks for 6 sessions and followed up for 3 months after the last session. Patients were assessed by photographic documentation, trichoscopic evaluation, severity of alopecia tool (SALT) score, and alopecia areata symptom impact scale (AASIS). An excellent response was achieved in 15 (37.5%) of group (1) compared with 14 (35%) of group (2) patients, while a good response was achieved in 23 (57.5%) of group (1) compared with 21 (52.5%) of group (1) patients, with a statistically insignificant difference. The mean SALT score change percentage was a statistically significantly higher in group (2) patients. The mean AASIS change percentage was higher in group (1) patients, but this was a statistically insignificant. In both groups, the mean numbers of trichoscopic signs of AA significantly decreased from baseline to the end of follow-up period. Both therapeutic modalities were well-tolerated, with no recurrence after the follow-up period. Both superficial cryotherapy using DMEP mixture, and microneedling are simple, effective, and safe therapeutic options for mild scalp AA, however, microneedling showed higher efficacy.

Intralesional injection of vitamin D3 versus zinc sulfate 2% in treatment of plantar warts: a comparative study.

BACKGROUND: Plantar warts are common skin lesions caused by the human papilloma virus. It is characterized by the presence of a horny ring of hyperkeratosis surrounding the wart, making its elimination a therapeutic challenge. Several destructive agents are available for treatment with variable success. Intralesional vitamin D3 has been reported as a successful treatment of warts. Intralesional zinc sulfate has been found to be another successful therapeutic modality for wart elimination. OBJECTIVE: To compare the efficacy and safety of intralesional vit. D3 versus zinc sulfate in treatment of plantar warts. PATIENTS AND METHODS: Forty patients were included in the study. Patients were randomly assigned to either vit. D3 group or zinc group. In vit. D3 group, patients received intralesional injection of 0.3 ml vitamin D3 (100,000 IU (2.5 mg/ml)), while zinc group patients received intralesional 2% zinc sulfate. Assessment of treatment efficacy and safety was carried out by clinical examination and comparative photographic evaluation before each session and up to 3 months after the last session. RESULTS: Eighty percent of vit. D3 treated patients and 70% of zinc sulfate patients showed complete response. CONCLUSIONS: Intralesional vit. D3 and zinc sulfate appear to be effective treatment modalities for plantar warts.

Assessment of the Effect of Metabolic Syndrome on the Autophagy Marker LC3 in Psoriasis Vulgaris Patients: A Cross-Sectional Study.

PURPOSE: Psoriasis vulgaris, one of the most prevalent chronic inflammatory skin diseases, is associated with metabolic syndrome (MetS). Autophagy, an intracellular degradation system is essential for cell survival and differentiation, and its dysfunction may contribute to metabolic diseases. A cross-sectional study was conducted on 38 psoriasis vulgaris patients and 16 healthy control subjects to 1) Assess immunohistochemical (IHC) expression of microtubule-associated protein light chain 3 (LC3); 2) Evaluate the relationship between Psoriasis Area Severity Index (PASI) score, and LC3 expression. PATIENTS AND METHODS: PASI score was evaluated for all psoriasis patients. Lipid profile, blood sugar, and CRP were done for all patients and controls. A punch biopsy was taken from lesional and perilesional skin of psoriasis patients and normal skin of the controls. Tissue sections were prepared. IHC LC3 staining was done and evaluated. RESULTS: LC3 was nearly absent, in the epidermis of the lesional skin of psoriasis while it was strong among control (p=0.001). LC3 expression in the lesional skin of psoriasis vulgaris was lower than its expression in perilesional (p=0.001). However, LC3 expression was not significantly changed with PASI or the presence/absence of MetS. CONCLUSION: A potential link between psoriasis vulgaris and autophagy as assessed by LC3 could be present. LC3 was down-regulated in psoriasis lesions than in normal skin. However, its expression did not change with PASI or MetS. An autophagy enhancer might be used as a possible therapeutic target in psoriasis vulgaris patients.

Diagnosis of onychomycosis clinically by nail dermoscopy versus microbiological diagnosis.

Diagnosis of onychomycosis requires microbiological studies, which are time-consuming. Dermoscopy is non invasive, easy and coastless method. To evaluate the diagnostic role of dermoscopy in onychomycosis and comparing its findings with microbiological results. Eighty patients with onychomycosis and 40 controls were studied for nail dermoscopic finding, and microbiological examinations in the form of microscopic examination by 20% KOH, Sabouraud dextrose agar (SDA), and HiCrome Candida Differential Agar. 72.5% of the patients were females. Most of the patient were presented with one finger (35%) and two fingers (35%). 85% of the patient were presented clinically with distal lateral subungual onychomycosis followed by total dystrophic onychomycosis (12.5%) and lastly with superficial white onychomycosis (2.5%). 52.5% and 75% of the patients were positive by direct microscopic examination with 20%KOH and SDA, respectively. Dermatophytes isolated from 7.5% of the patient, non-dermatophytes (Aspergillus) was isolated from 2.5%, and 65% had Candida by SDA. C. albicans was the commonest species (75%), followed by C. tropicalis (17.3%), and lastly C. krusei (7.7%). Dermoscopic examinations of patients showed nail spikes, longitudinal striations, and color changes in 75%, 82.5%, and 95%, respectively, with statistically significant P value (P < 0.001). There was significant difference regarding long striations and yellow coloration dermoscopic finding with positive KOH patients. All patients with positive culture showed nail spikes on dermoscopic examination. Dermoscopy is a rapid tool for diagnosis of onychomycosis. Longitudinal striations is the best diagnostic dermoscopic finding. Microbiological test are still needed for accurate and reliable diagnosis.

Fractional Erbium-YAG Laser and Platelet-Rich Plasma as Single or Combined Treatment for Atrophic Acne Scars: A Randomized Clinical Trial.

INTRODUCTION: Acne scarring is a common undesirable complication of acne vulgaris. Fractional erbium-yttrium aluminum garnet (YAG) 2940 nm laser and platelet-rich plasma have been used in treating acne scars with variable outcomes. The objective of this study is to assess the efficacy of fractional erbium-YAG 2940 nm laser and platelet-rich plasma as a single line of treatment in comparison with combined treatment in atrophic postacne scars. METHODS: Seventy-five patients were included in this trial and randomized into three equal groups (25 each). Group A was subjected to six sessions of erbium-YAG laser for 6 months, group B was treated with 12 sessions of platelet-rich plasma over the same period, and group C was subjected to six sessions of erbium-YAG laser plus 12 sessions of platelet-rich plasma over the same period. Each subject was evaluated by acne scar grading, photography, and subjective evaluation. RESULTS: Both treatment modalities showed improvement of acne scars, but the improvement with combined treatment was better than that with erbium-YAG laser or platelet-rich plasma alone regarding scar grade improvement (P = 0.007 and 0.001), clinical improvement (P = 0.001 and 0.001), and patient satisfaction (P = 0.005 and 0.001), respectively. CONCLUSIONS: The combination of platelet-rich plasma plus erbium-YAG laser is superior to either treatment alone for acne scars, with trivial side effects for all treatment modalities. TRIAL REGISTRATION: ClinicalTrials.gov identifier; NCT03933033.

Epidemiologic Trends of Viral Skin Infections in Egypt: A Cross-Sectional Hospital-Based Study.

Viral skin infections (VSIs) were ranked among the top 50 prevalent diseases in 2010. The objective of this study was to determine the epidemiologic features of VSIs in patients attending a dermatology clinic in Egypt from June 2010 to May 2011. Patient's residence, occupation, housing data, and family history of similar conditions were recorded. Categorical data were recorded as frequencies and percentages and were compared by Chi square test. P value < 0.05 was significant. Diagnosis of VSIs was made in 1000/20322 (4.9%) patients. Out of the 1000 patients with VSIs, 580 (58.0%) were residents of rural areas and 420 (42.0%) were residents of urban areas (p = 0.02). Out of the 1000 patients, 489 (48.9%) were females and 511 (51.1%) were males (p = 0.25). The breakdown of 1000 patients with VSIs indicated diagnosis of viral warts in 673 (67.3%), chickenpox (CP) in 200 (20.0%), herpes simplex (HS) facialis in 50 (5.0%), herpes zoster (HZ) in 42 (4.2%), molluscum contagiosum (MC) in 27 (2.7%.0), and anogenital warts in 8 (0.8%) cases. Overcrowding (sharing a bedroom by more than 3 persons) was recorded in 652/1000 (65.2%) of the patients with VSIs [165/200 (82.5.3%) in CP, 36/50 (72%) in HS facials, 427/673 (63.4%) in viral warts, 14/27 (51.9%) in MC, and 10/42 (23.8%) in HZ]. Family history of a similar condition was positive in 329/1000 (32.9%) of the patients with VSIs [142/200 (71.0%) in CP, 177/673 (26.3%) in viral warts, 5/27 (18.5%) in MC, and 4/50 (8%) in HS facialis]. In conclusion, viral warts and CP were the commonest VSIs diagnosed in patients who attended a dermatology clinic in Egypt. Viral skin infections were more prevalent among patients who lived in rural areas and under crowded conditions. These data may have important public health implications particularly in developing countries.

Efficacy of microneedling with topical vitamin C in the treatment of melasma.

BACKGROUND: Despite the wide therapeutic options available for the treatment of melasma, including many active topical medications, technologies with lights and peelings, clinical control of this disorder is extremely challenging. OBJECTIVES: To evaluate the effect of microneedling with topical vitamin C in the treatment of melasma. METHODS: Thirty female patients with melasma received six sessions of microneedling with addition of topical vitamin C every two weeks. At each session, photos were taken and Melasma Area and Severity Index (MASI) score was calculated to assess the clinical improvement. RESULTS: Mean age of the eligible patients was 33.2 ± 5.77 years. About 50% of cases were of Fitzpatrick skin type III. All patients showed improvement at the end of the sessions. Mean MASI score in the first session was 8.61 ± 4.45 and there was a gradual decline in its value till it reached a mean of 5.75 ± 4.16 in the last session (P < 0.0001). CONCLUSION: Microneedling with topical vitamin C is an effective and safe treatment option for epidermal melasma especially in Fitzpatrick skin phototypes I-III.

Lesional and circulating levels of interleukin-17 and 25-hydroxycholecalciferol in active acne vulgaris: Correlation to disease severity.

OBJECTIVES: The immunological aspects of inflammatory acne are still incompletely understood, so this study aimed to investigate the possible role of IL-17 and 25 hydroxycholecalciferol (25(OH)D3) in the disease pathogenesis and progression. MATERIALS AND METHODS: Across-sectional study has been conducted on 135 patients with active acne vulgaris of various severities and 150 matched controls. ELISA assays of serum and tissue levels of IL-17 and 25(OH)D3, also immunohistochemical and Western blotting demonstration of the expression patterns of lesional IL-17 in comparison with control group, were performed. RESULTS: The mean serum levels of IL-17 were 544.2 pg/mL ± 477.4 SD and 42.2 pg/mL ± 8.1 SD for acne patients and controls, respectively, with significantly higher levels among the patient group (P < 0.05). Higher IL-17 expression levels in active acne lesions when compared with its level in healthy skin of the controls. The mean serum levels of 25(OH)D3 among patients and controls were 33.3 ng/mL ± 9.7 SD and 51.7 ng/mL ± 2.7 SD, respectively, with significantly lower levels among the patient group (P < 0.05). There were significantly negative correlations between IL-17 and 25(OH)D3 levels (P < 0.001 for both). CONCLUSIONS: Deficiency of vitamin D3 accompanied with higher IL-17 in an inverse pattern may have a possible role in active acne vulgaris.

Possible role of interleukin-17 and macrophage migration inhibitory factor in cutaneous warts.

BACKGROUND/OBJECTIVES: Cutaneous warts (CW), or verrucae, are benign proliferation of skin that result from infection with human papilloma viruses. Cellular immune reactivity plays a significant role in wart regression. The aim of this study was to elucidate the cellular immune status of patients with CW through measurements of their serum levels of interleukin-17 (IL-17) and macrophage migration inhibitory factor (MIF,) and, identify the possible role of IL-17 and MIF in CW. We assessed serum IL-17 and MIF levels in patients with different forms of CW and compare the results with controls. PATIENT AND METHODS: Serum levels of IL-17 and MIF were measured using commercially available ELISA assay kits in 60 patients with CW and 20 healthy controls. RESULTS: Serum levels of IL-17 and MIF were significantly lower in patients with CW when compared with the controls (P-value <.01, <.05, respectively). There was nonsignificant correlation between IL-17 and MIF. CONCLUSION: Low IL-17 and MIF levels may have a contributory role in occurrence, maintenance, severity, and recurrence of different types of CW which depend mainly on the defect of cell-mediated immunity. This may shed new light on nontraditional strategies for the future medical treatments of CW through regulation of IL-17 and MIF.

Prevalence of acne vulgaris and its impact of the quality of life among secondary school-aged adolescents in Sohag Province, Upper Egypt.

BACKGROUND: Acne vulgaris is the most common dermatological condition encountered in adolescents. AIM: It was to determine the prevalence of acne vulgaris and its impact of the quality of life among adolescents attending secondary schools in Sohag Province, Upper Egypt. PATIENTS AND METHODS: This was a cross-sectional study conducted in randomly selected governmental and technical secondary schools in Sohag Province, Upper Egypt. Approval was taken from the scientific research committee of Sohag Faculty of Medicine and also from Ministry of Education. Every student with acne was subjected to full medical history and local examination of head and neck to assess the severity of acne vulgaris. Assessment of the impact of acne vulgaris on their quality of life was carried out using self-reported validated specific questionnaire, the Cardiff Acne Disability Index (CADI). RESULTS: This study included 994 teenage secondary school students. The overall prevalence of acne vulgaris was 333 (33.5%). The mean age of the students with acne was 16.84±0.87. Acne vulgaris was more common among females than among males (200, 60% vs 133, 40%). The most common form of acne vulgaris was mild 178, 53%, followed by moderate form 135, 41%, and severe form 20, 6%. CADI score was significantly related to the disease grade and it was maximum among those with severe grade, followed by moderate and lastly mild disease grade. CONCLUSION: Acne vulgaris is a common skin disease and has a valuable impact on quality of life among adolescents attending secondary schools in Sohag Province, Upper Egypt.

Platelets rich plasma versus minoxidil 5% in treatment of alopecia areata: A trichoscopic evaluation.

Alopecia areata is a common cause of nonscarring alopecia that occurs in a patchy, confluent, or diffuse pattern. Dermoscopy is a noninvasive technique for the clinical diagnosis of many skin diseases. Topical minoxidil solution 5% and platelet rich plasma are important modalities used in treatment of alopecia areata. We aimed to evaluate the efficacy of PRP versus topical minoxidil 5% in the treatment of AA by clinical evaluation and trichoscopic examination. Ninety patients were allocated into three groups; the first was treated with topical minoxidil 5% solution, the second with platelets rich plasma injections, and the third with placebo. Diagnosis and follow up were done by serial digital camera photography of lesions and dermoscopic scan before and every 1 month after treatment for 3 months. Patients treated with minoxidil 5% and platelets rich plasma both have significant hair growth than placebo (p < .05). Patients treated with platelets rich plasma had an earlier response in the form of hair regrowth, reduction in short vellus hair and dystrophic hair unlike patients treated with minoxidil and control (p < .05). In conclusion, platelets rich plasma is more effective in the treatment of alopecia areata than topical minoxidil 5% as evaluated by clinical and trichoscopic examination.

Relation Between Erectile Dysfunction and Silent Myocardial Ischemia in Diabetic Patients: A Multidetector Computed Tomographic Coronary Angiographic Study.

INTRODUCTION: Erectile dysfunction (ED) can precede coronary artery disease. In addition, silent myocardial ischemia (SMI) is more common in diabetic patients and is a strong predictor of cardiac events and death. AIM: To evaluate the presence of SMI in patients with diabetes and ED using multidetector computed tomographic coronary angiography (MDCT-CA). METHODS: This study evaluated patients with diabetes and ED without any history of cardiac symptoms or signs. Erectile function was evaluated with the Sexual Health Inventory for Men score, erection hardness score (EHS), and maximal penile circumferential change by an erectometer. MDCT-CA was used for the detection of coronary artery stenosis. MAIN OUTCOME MEASURES: Sexual Health Inventory for Men score, EHS, maximal penile circumferential change, and coronary artery stenosis by MDCT-CA. RESULTS: Of 20 patients (mean age = 61.45 ± 10.7 years), MDCT-CA showed coronary artery stenosis in 13 (65%) in the form of one-vessel disease (n = 6, 30%), two-vessel disease (n = 2, 10%), and three-vessel disease (n = 5, 25%). Fifty percent of patients showed at least 50% vessel lumen obstruction of the left anterior descending coronary artery, which was the most commonly affected vessel (55%). Fifteen percent (3 of 20) of patients had greater than 90% stenosis, and two of them underwent an immediate coronary angioplasty with stenting to prevent myocardial infarction. Maximum coronary artery stenosis was positively correlated with age (P = 0.016, r = 0.529) and negatively correlated with EHS (P = .046, r = -0.449). Multivariate regression analysis using age and EHS showed that age was the only independent predictor of SMI (P = .04). CONCLUSION: MDCT-CA can be a useful tool to identify SMI in diabetic patients with ED, especially in those of advanced age and/or with severe ED.

Oxidative stress and epididymal sperm transport, motility and morphological defects.

OBJECTIVES: Radical oxidative species (ROS) have an important effect on sperm quality and quantity. Oxidative stress (OS) occurs when production of potentially destructive reactive oxygen species (ROS) exceeds the body's own natural antioxidant defenses, resulting in cellular damage. OS is a common pathology seen in approximately half of all infertile men. Increased ROS generation and reduced antioxidant capacity is negatively correlated with sperm concentration and motility in infertile men. For the first time, we used a more stable and reliable sensitive carbonyl protein (CP) detection method to determine ROS in seminal plasma than measuring ROS directly to clarify the effect of OS on spermatozoa in terms of protein dysfunction. This is the first report to measure CP in seminal plasma as an indicator of OS. Furthermore, for the first time we correlated the results of CP measurement with light microscopy in combination with ultrastructural analysis by electron microscopy. MATERIAL AND METHODS: 20 patients with idiopathic oligoasthenoteratozoospermia (iOAT) and 10 fertile controls were enrolled in this study. CP values were measured by enzyme-linked immuno sorbent assay (ELISA) to detect the level of OS. Transmission electron microscope (TEM) was used to detect axonemal anomalies. RESULTS: Compared to fertile controls, statistically highly significant higher degrees of abnormal sperm parameters (P<0.001) could be found in iOAT patients. CP values were highly significantly elevated in iOAT patients than in normal controls (P<0.001). A statistically highly significant difference in different axonemal anomalies were found between iOAT patients and normal controls (P<0.001). CP values have been found to be positively correlated with different axonemal anomalies (absence of axoneme (r(2)=0.841), missing of central singlet tubules (r(2)=0.702) and missing of outer doublet tubules (r(2)=0.869). A statistically negative correlation were found between different axonemal anomalies (absent axoneme (r(2)=-0.780), missing of central singlet tubules (r(2)=-0.611), and missing of outer doublet tubules (r(2)=-0.738) and forward progressive sperm motility. CONCLUSION: High levels of CP can be measured in iOAT patients, indicating that OS could underlie the aetipopathogenesis of the syndrome. OS negatively affects flagellar axonemal structure with subsequent impairment of forward progressive sperm motility. This can put an attention for antioxidants as a therapy for iOAT syndrome and further research to find how to decrease ROS production.

Phenotypical characteristics of the immune cells in allergic contact dermatitis, atopic dermatitis and pityriasis rosea.

Allergic contact dermatitis (ACD) is a cell-mediated, delayed type IV immunologic reaction. Atopic dermatitis (AD) is a chronic inflammatory skin disease that results from a complex interaction between immunologic, genetic, and environmental factors. Pityriasis rosea (PR) is a self-limited eruption of unknown etiology. Immune cell infiltrate is a constant feature in the inflammatory skin diseases. Here, we performed phenotypical characterization of the immune cells in ACD, AD and PR (ten cases each). We performed immunohistochemical stains for B cells (CD20), T cells (CD3), histiocytes (CD68) and T cells with cytotoxic activity (granzyme-B). The data were compared with findings in 20 specimens of normal skin. The results were scored as mean values of positively stained immune cells. Immunohistochemistry showed significantly high counts of immune cells in lesional skin (ACD, AD and PR) compared to the normal one (p < 0.05). In the lesional skin, the immune cells were composed predominantly of CD3(+) T lymphocytes and CD68(+) cells (histiocytes). Some of the CD3(+) cells were granzyme B(+). The counts of some immune cells (CD3(+) and CD68(+)) were high in ACD compared to AD and PR. The counts of CD20(+) and granzyme B(+) cells were high in PR compared to ACD and AD. However, these differences did not reach the level of statistical significance. The present data describe the profile of the immune cell infiltrate in AD, ACD and PR. The cell-mediated immunity seems to have critical role in the development of these lesions.

Expression of nerve growth factor and its high-affinity receptor, tyrosine kinase A proteins, in the human scalp skin.

BACKGROUND: Nerve growth factor (NGF) and its high-affinity receptor, tyrosine kinase A (TrkA), are members of the neurotrophin family. NGF-TrkA are involved in murine hair morphogenesis and cycling. To date, their expression in human hair follicle (HF) is unknown. In this investigation, we hypothesize that NGF-TrkA proteins are expressed in the human scalp skin. Moreover, NGF-TrkA expression in HF changes with the transitions from anagen-->>catagen-->>telogen stages. MATERIALS AND METHODS: To test our hypothesis and to fill this existing gap in literature, the immunostaining values (semiquantitative evaluation of protein expression: SI, staining intensity; PP, percentage of positive cells; and IR score, immunoreactivity score) of NGF and TrkA proteins were examined in human scalp skin by immunofluorescent and immunoperoxidase staining methods. Fifty normal human scalp skin biopsy specimens were examined (healthy females, 53-57 years). In each case, 50 HFs were analyzed (35, 10, and five follicles in anagen, catagen, and telogen, respectively). RESULTS: The IR scores were statistically significantly higher (p < 0.001) in anagen as compared with either catagen or telogen HF (9.61 +/- 0.12 vs. 1.4 +/- 0.10 vs. 0.6 +/- 0.10 for NGF and 3.31 +/- 0.02 vs. 0.5 +/- 0.10 vs. 0.2 +/- 0.10 for TrkA). In the anagen HF, high expression values were seen in the distal region, followed by upper distal, lower distal, and bulb regions for both NGF (10.6 +/- 0.21 vs. 10.3 +/- 0.21 vs. 9.2 +/- 0.40 vs. 8.1 +/- 0.30) and TrkA (3.54 +/- 0.07 vs. 3.45 +/- 0.07 vs. 3.31 +/- 0.06 vs. 3.13 +/- 0.04). Both NGF and TrkA proteins showed prominent expression in the melanocytes (7.6 +/- 0.15 vs. 2.50 +/- 0.07), keratinocytes (10.2 +/- 0.40 vs. 2.71 +/- 0.06), sebaceous glands (10.2 +/- 0.40 vs. 2.72 +/- 0.06), and sweat glands (10.4 +/- 0.40 vs. 2.84 +/- 0.05). CONCLUSIONS: Our findings report, for the first time, the expression pattern of NGF and TrkA proteins in human scalp skin and HF. The differential expression of these proteins during HF cycling suggests their possible roles in human HF biology. The clinical ramifications of these observations mandate further investigations.

Phenotypic characterization of the immune and mast cell infiltrates in the human testis shows normal and abnormal spermatogenesis.

OBJECTIVE: To assess the types, distributions, and numbers of immune cell infiltrates in the testes of men with azoospermia. DESIGN: Prospective study. SETTING: University research and clinical institutes. PATIENT(S): Thirty-one men with azoospermia showing normal spermatogenesis (n = 10), germ cell maturation arrest (GA, n = 12), and Sertoli cell-only syndrome (SCO, n = 9). INTERVENTION(S): Testicular tissue biopsies. Sections were stained with routine (hematoxyline and eosin), special (Masson Trichrome, Gordon, Periodic Acid Schiff, Aldehyde Fuchsin, and Orcein stains), and immunoperoxidase stains (using monoclonal antibodies for B and T cells and CD68 macrophages). MAIN OUTCOME MEASURE(S): Serum levels of T, FSH, and LH in addition to histopathological analysis. RESULT(S): Hormonal profiles were unremarkable in all patients. Marked deposition of the reticular and collagen fibers was seen more in abnormal than normal spermatogenesis. The immune (B and T lymphocytes and CD68 macrophages) and mast cells were found in the interstitium, tubular walls, and lumens of all the testes analyzed. The differential counts of these cells (B and T lymphocytes, CD68 macrophages, and mast cells, respectively) were higher in SCO (1.66 +/- 0.46, 9.14 +/- 1.30, 2.26 +/- 1.68, 3.35 +/- 0.23) and GA (2.03 +/- 0.48, 4.70 +/- 1.00, 2.61 +/- 0.70, and 4.18 +/- 0.13) when compared with those in normal spermatogenesis (1.22 +/- 0.19, 5.41 +/- 0.58, 1.55 +/- 0.33, and 2.26 +/- 0.13). Increased cellular counts were not statistically significant for T and B cells and macrophages. However, these differences were statistically significant for mast cells. CONCLUSION(S): Abnormal spermatogenesis is associated with increased numbers of the immune and mast cells. Our findings may reflect an exaggerated immune response in these cases.

Negative effects of increased sperm DNA damage in relation to seminal oxidative stress in men with idiopathic and male factor infertility.

OBJECTIVE: To examine the effects of increased sperm DNA damage in relation to seminal oxidative stress in men with idiopathic and male factor infertility. Prospective study. SETTINGS: Infertility clinic at a tertiary care academic institution. PATIENT(S): Ninety-two infertile men with normal female partners. Sixteen fertile donors served as the control group. INTERVENTION(S): Standard semen analysis and assessment of levels of seminal oxidative stress. Assisted reproductive techniques in 33 of the 92 patients (IUI [n = 19], IVF [n = 10], and intracytoplasmic sperm injection [n = 4]). MAIN OUTCOME MEASURE(S): Sperm DNA damage by sperm chromatin structure assay. Results were expressed as DNA fragmentation index. RESULT(S): Patients were classified as having either idiopathic (n = 23) or male factor infertility (n = 69). Patients with idiopathic and male factor infertility had significantly higher DNA fragmentation index and oxidative stress compared with the case of fertile donors. A clinical pregnancy was achieved in 9 (27%) of 33 patients with assisted reproductive techniques. Significantly higher DNA fragmentation index and oxidative stress were found in men who failed to initiate a pregnancy after assisted reproductive techniques (n = 24), compared with the cases of those who succeeded and of the fertile donors. DNA fragmentation index was correlated positively with oxidative stress (r = 0.27), and negatively with fertilization (r = -0.70) and embryo quality (r = -0.70). CONCLUSION(S): Sperm DNA damage is significantly increased in men with idiopathic and male factor infertility and in men who failed to initiate a pregnancy after assisted reproductive techniques. Such an increase may be related to high levels of seminal oxidative stress.

Leukocytospermia is associated with increased reactive oxygen species production by human spermatozoa.

To investigate the role of increased seminal leukocytes in enhancing reactive oxygen species (ROS) production by human spermatozoa.A prospective study. Male infertility clinic.Forty-eight infertile men. Standard semen analysis. Assessment of sperm nuclear DNA damage by sperm chromatin structure assay. Incubation of spermatozoa from nonleukocytospermic samples with blood neutrophils. Spontaneous and phorbol 12-myristate 13-acetate (PMA)-induced ROS production in pure-sperm suspensions (after removal of leukocytes) as measured by a chemiluminescence assay. Levels of spontaneous and PMA-induced ROS production in pure-sperm suspensions from the infertile men with a diagnosis of leukocytospermia (n = 16) were significantly higher compared with the case of infertile men without leukocytospermia (n = 32) and with the case of a control group of healthy volunteers (n = 13). A similar pattern of increased ROS was observed when spermatozoa were incubated with blood neutrophils. Leukocytospermia was associated with a significant decrease in sperm motility and increase in DNA damage. Increased seminal leukocytes may play a role in stimulating ROS production by human spermatozoa. Such stimulation may be mediated via direct cell-cell contact or by soluble products released by leukocytes. Poor sperm quality in leukocytospermic samples may be due to leukocyte-mediated oxidative stress.