Hypoxic Gene Signature of Primary and Metastatic Melanoma Cell Lines: Focusing on HIF-1ß and NDRG-1

Background: Hypoxia is an important microenvironmental factor significantly affecting tumor proliferation and progression. The importance of hypoxia is, however, not well known in oncogenesis of malignant melanoma. Aims: To evaluate the difference of hypoxic gene expression signatures in primary melanoma cell lines and metastatic melanoma cell lines and to find the expression changes of hypoxia-related genes in primary melanoma cell lines at experimental hypoxic conditions. Study Design: Cell study. Methods: The mRNA expression levels of hypoxia-related genes in primary melanoma cell lines and metastatic melanoma cell lines and at experimental hypoxic conditions in primary melanoma cell lines were evaluated by using real-time polymerase chain reaction. Depending on the experimental data, we focused on two genes/proteins, the hypoxia-inducible factor-1 beta and the N-myc downstream regulated gene-1. The expression levels of the two proteins were investigated by immunohistochemistry methods in 16 primary and metastatic melanomas, 10 intradermal nevi, and a commercial tissue array comprised of 208 cores including 192 primary and metastatic malignant melanomas. Results: The real-time polymerase chain reaction study showed that hypoxic gene expression signature was different between metastatic melanoma cell lines and primary melanoma cell lines. Hypoxic experimental conditions significantly affected the hypoxic gene expression signature. In immunohistochemical study, N-myc downstream regulated gene-1 expression was found to be lower in primary cutaneous melanoma compared to in intradermal nevi (p=0.001). In contrast, the cytoplasmic expression of hypoxia-inducible factor-1 beta was higher in primary cutaneous melanoma than in intradermal nevi (p=0.001). We also detected medium/strong significant correlations between the two proteins studied in the study groups. Conclusion: Hypoxic response consists of closely related proteins in more complex pathways. These findings will shed light on hypoxic processes in melanoma and unlock a Pandora's box for development of new therapeutic strategies.

Clear cell sarcoma.

Malignant melanoma (MM) of soft tissue, also called clear cell sarcoma (CCS) of tendons and aponeuroses, derives from the neural crest. CCS is similar morphologically to MM but has no precursor skin lesion, and instead, has a characteristic chromosomal translocation. Prognosis is related to the tumor size. Early recognition and initial radical surgery is the key to a favorable outcome. The tumor has to be differentiated from other benign and malignant lesions of the soft tissues, such as fibrosarcoma. The demonstration of melanin and a positive immunohistochemical reaction for S-100 protein and HMB-45 can assist in the differential diagnosis. We report the case of a 58-year-old woman with CCS arising from the soft tissue of her little finger.

Tangier disease in a Turkish family.

In this report we describe the upper gastrointestinal tractus involvement in a rare genetic disease of lipid metabolism. A 12-year-old boy presented with sore throat and fever. On physical examination, orange-yellow tonsils and adenoid tissue were noted. Mild hepatosplenomegaly was present. Lipid profile was compatible with Tangier disease (TD). Endoscopy of the upper gastrointestinal tract showed white-yellowish fatty deposits on the gastric mucosa. Microscopically, biopsy specimens contained numerous histiocytes with a foamy cytoplasm packed in the lamina propria of the gastric mucosa and at the crypt basement of the duodenum. His sister, 8 years old, was also diagnosed with TD based on abnormal lipid profile and orange-yellow tonsils. TD is a rare familial disorder of lipid metabolism, characterized by deposition of cholesteryl esters, probably involving the entirety of the gastrointestinal tract from the mouth to the anus.

Assessment of ovarian stromal artery Doppler characteristics and serum hormone levels in patients with Behçet disease.

PURPOSE: The aim of the study was to examine serum hormone levels, ovarian volume, stromal artery Doppler parameters of patients with Behçet disease (BD) to assess whether there are vascular changes in the gonads of these patients. MATERIALS AND METHODS: Twenty patients with BD and 31 healthy controls aged between 18-45 years were examined in the early follicular phase of the menstrual cycle (day 2-3) with transvaginal ultrasound to evaluate ovarian volume and ovarian stromal artery Doppler parameters. On the same day, blood was drawn for determining serum hormone levels. RESULTS: Patients with BD and the controls were comparable with regard to age and body mass index at study inclusion. Although comparison of the ovarian stromal artery Doppler velocimetric parameters did not show significant differences, resistivity, pulsatility indexes and systolic/diastolic ratio were higher, while peak systolic and end diastolic velocities were lower, in BD patients compared to controls. The mean ovarian volume of patients with BD was smaller than the controls but this difference did not reach statistical significance. There were no statistically significant differences between serum hormone levels of either group. We did not find any correlations between hormone levels and mean ovarian stromal artery Doppler parameters of patients with BD. CONCLUSION: Ovarian stromal artery Doppler parameters of patients with BD did not show any significant differences compared to healthy controls. Therefore, we conclude that ovarian stromal artery is not involved in patients with BD as assessed by transvaginal Doppler ultrasound and serum hormone levels do not differ from the levels of healthy controls.

A case of brucellosis presenting with widespread maculopapular rash.

Human brucellosis is a multisystemic infectious disease with a broad spectrum of clinical manifestations. Severe complications involving musculoskeletal, nervous, genitourinary, and cardiovascular systems may be encountered during the course of the disease; however, cutaneous complications have been reported rarely. We report a patient with brucellosis in whom the main presenting clinical feature was a maculopapular rash and fever. He was initially diagnosed as brucellosis based on the standard tube agglutination test and blood culture positivity. Histopathologic examination of these maculopapular lesions showed perivascular and periadnexal inflammation with loose granuloma formation including giant cells. We emphasize that brucellosis is an infectious disease that should always be kept in mind in the differential diagnosis of a patient with rash and fever, especially in endemic areas.

Diffuse plane xanthomatosis in a patient with Budd-Chiari syndrome and monoclonal gammopathy.

Diffuse plane xanthomas are characterized by the presence of yellowish plaques on the eyelids, neck, upper trunk, buttocks, and flexural folds. Histology shows foamy histiocytes in the dermis. Approximately half of the cases are associated with lymphoproliferative disorders. Budd-Chiari syndrome is an uncommon condition induced by thrombotic or nonthrombotic obstruction of hepatic venous outflow. We present a case of diffuse plane xanthoma in a 62-year-old man who developed normolipemic plane xanthomas coinciding with Budd-Chiari syndrome and monoclonal gammopathy. We review the English-language literature regarding the rare association of xanthomas and Budd-Chiari syndrome.

Absence of estrogen and progesterone receptors around the affected vessels of angioma serpiginosum: case report.

Angioma serpiginosum is a disorder of dilated superficial dermal capillaries. The disease is considered by some authors to be a consequence of increasing levels of estrogens. We present a case of angioma serpiginosum in a 46-year-old White woman. The lesions consisting of red-purple puncta were grouped as vaguely annular and angular patches with serpiginous borders, located on the right side of the neck, the right side of the abdomen, and on the left arm. Histopathologic examination revealed clusters of dilated capillaries in the dermal papillae, without endothelial proliferation. Periodic acid-Schiff (PAS) stain-positive deposits were noted around the affected blood vessels. However, immunohistochemical analysis revealed the absence of estrogen and progesterone receptors within the involved blood vessels. Hormonal assays were also normal. Our observations suggest that a hormonal stimulus probably plays no role in the pathogenesis of angioma serpiginosum. The significance of PAS-positive deposits as a diagnostic marker for angioma serpiginosum needs further confirmation.

Bilateral upper eyelid edema in Melkersson-Rosenthal syndrome.

Melkersson-Rosenthal syndrome is an uncommon disorder characterized by a triad of facial nerve palsy, orofacial edema, and fissured tongue. A 42-year-old woman with Melkersson-Rosenthal syndrome presented with painless, nonpitting, bilateral asymmetric upper eyelid edema. The left eyelid was a bit larger than the right eyelid. CT and MRI demonstrated periorbital soft tissue thickening compatible with the microscopic findings of infiltration of lymphocytes, edema, and cystic dilatation of lymphatic vessels. After treatment by systemic doxycycline and corticosteroid, she showed some improvement of the eyelid edema. Isolated bilateral eyelids swelling may be observed in Melkersson-Rosenthal syndrome. In the case of unexplained nonpitting eyelid edema, biopsy should be performed.

Focal acral hyperkeratosis: a rare cutaneous disorder within the spectrum of Costa acrokeratoelastoidosis.

Acrokeratoelastoidosis and focal acral hyperkeratosis share similar clinical features and identical histologic epidermal alterations. These disorders are distinguished solely on the basis of the absence of elastorrhexis in the latter. We present a case of focal acral hyperkeratosis in a 9-year-old girl. The lesions consisted of translucent polygonal papules clustered on the thenar regions of the palms and over the metacarpophalangeal and interphalangeal joints. Histopathologic examination revealed orthohyperkeratosis within focal clavus-like depressions of the epidermis and prominent hypergranulosis. There was no evidence of alterations in elastic tissue. The clinicopathologic distinction between focal acral hyperkeratosis and acrokeratoelastoidosis is blurred. There is enough evidence to consider the former as a histologic variant of Costa acrokeratoelastoidosis syndrome, and a better nomenclature for this disorder would be "acrokeratoelastoidosis without elastorrhexis."

BCL-2-related apoptosis markers in cutaneous human papillomavirus-associated lesions.

BACKGROUND: Human papilloma virus (HPV) is an etiological agent in benign and malignant epithelial tumors. Resistance to apoptotic stimuli by viral strategies represents an immunologic escape mechanism during virus-induced tumor development and is critical for efficient replication of the virus. OBJECTIVE: The aim of the present study was to investigate a role of bcl-family proteins in the anti-apoptotic pathways modulated by low-risk HPVs in the development of benign HPV-associated cutaneous tumors. METHODS: Forty lesional biopsy specimens from HPV-associated cutaneous lesions and 11 non-lesional control skin biopsies were studied by immunohistochemical analysis for the differential expressions of HPV antigens, the pro-apoptotic bax protein, and the anti-apoptotic bcl-2 and bcl-x proteins. RESULTS: Compared with the normal epidermis, bcl-2 and bcl-x expression were significantly reduced in the lesional epidermis. Bax was expressed in HPV-associated cutaneous lesions, although the expression did not reveal a significant deviation from that in normal skin. CONCLUSION: These findings indicate a discordant expression of bcl-2/ bcl-x and bax proteins in HPV-associated skin lesions and suggest that low-risk HPVs mediate other pathways that bypass the action of anti-apoptotic bcl-2 and bcl-x proteins. The presence of bax expression with a prominent decrease in bcl-2/ bax ratio and the lack of massive apoptosis in HPV-associated benign epithelial lesions may imply that interference with the pro-apoptotic proteins of bcl-family may constitute one of the several mechanisms mediated by HPV oncoproteins for the suppression of apoptotic process.

Examination of Bcl-2, Bcl-X and bax protein expression in psoriasis.

BACKGROUND: Psoriasis is an inflammatory skin disease characterized by epidermal hyperplasia and greatly accelerated epidermal turnover. The blockage of normal apoptotic process in the epidermis is one of the factors implicated in the pathogenesis of psoriasis. OBJECTIVE: The purpose of the present study was to elucidate whether bcl-family proteins are significantly involved in the hypothetical antiapoptotic cascade in lesional psoriatic epidermis. METHODS: Twenty-six lesional biopsy samples of 26 patients with psoriasis and five control specimens from normal skin were studied by immunohistochemical method for the differential expression of pro-apoptotic bax and antiapoptotic bcl-2 and bcl-x proteins. RESULTS: Compared with the normal epidermis, bcl-2 expression was significantly reduced, whereas bax and bcl-x were significantly overexpressed in the psoriatic epidermis. The localization of bcl-2/bax/bcl-x proteins in the psoriatic epidermis did not show a significant deviation from that in the normal epidermis. CONCLUSION: These findings indicate a discordant expression of bcl-2 and bax/bcl-x in psoriatic epidermis. Increased bcl-x expression might contribute to the antiapoptotic response in psoriatic keratinocytes. The presence of strong bax expression with a concomitant decrease in bcl-2 expression suggests either a functional defect in bax protein or an inherent/acquired resistance to bax-mediated apoptosis in psoriatic keratinocytes.

Juvenile subcorneal pustular dermatosis: a case report.

Subcorneal pustular dermatosis is a chronic, relapsing, pustular dermatosis that usually develops in elderly women. It is rarely seen in childhood and adolescence. The hallmark of the disease is formation of a subcorneal pustule detected on histopathologic examination. Herein we present a 13-year-old girl diagnosed with subcorneal pustular dermatosis based on the typical clinical and histologic findings. Direct and indirect immunofluorescence and serum protein electrophoresis had negative results. The patient partially recovered after 5 weeks of treatment consisting of oral dapsone and a topical steroid ointment.

Permethrin 5% cream versus metronidazole 0.75% gel for the treatment of papulopustular rosacea. A randomized double-blind placebo-controlled study.

BACKGROUND: Permethrin 5% cream used against human ectoparasites suggests that it may be effective in papulopustular rosacea. METHODS: This study included 63 patients diagnosed as having papulopustular rosacea based on the clinical and histological findings. Patients were randomly assigned into permethrin (n = 23), metronidazole (n = 20) and placebo (n = 20) groups. Scores of erythema, telangiectasia, edema and rhinophyma and the numbers of papules, pustules, inflammatory nodules and Demodex folliculorum were determined. Twenty-three patients were given permethrin 5% cream (Zalvor 5% skin cream, 20 patients metronidazole 0.75% gel (Roza gel and 20 patients placebo cream (Basis cream, in packages looking identical to those of metronidazole and permethrin creams, and were recommended to apply them to their faces twice a day. All patients were also given SPF 20 cream for protection against sunlight. Two months of treatment were planned, and the patients were invited to the clinic for fortnightly controls. Scores of erythema, telangiectasia, edema and rhinophyma and the numbers of papules, pustules, inflammatory nodules and D. folliculorum were recorded at each visit. The mean scores of erythema and the mean numbers of papules, pustules and D. folliculorum were determined at baseline and on days 15, 30, 45 and 60. Side effects were also detected. RESULTS: The effect of permethrin 5% cream on D. folliculorum was superior to that of metronidazole 0.75% gel. The effect of permethrin 5% cream on erythema and papules was found to be more effective than placebo and as effective as metronidazole 0.75% gel. However, it had no effect on telangiectasia, rhinophyma and pustules. CONCLUSION: It can be concluded that the application of permethrin 5% cream twice daily for 2 months can be as effective and reliable as metronidazole in the treatment of rosacea and a greater benefit can be gained when it is combined with other systemic and/or topical treatments.

Detection of human herpesvirus 7 in pityriasis rosea by nested PCR.

BACKGROUND: Clinical presentation, immunologic, light microscopic, and electron microscopic studies suggest a viral etiology for pityriasis rosea (PR). OBJECTIVE: To evaluate whether human herpesvirus 7 (HHV-7) is an etiologic factor for PR. PATIENTS AND METHODS: Twenty-one PR patients (12 female, nine male) aged between 12 and 52 years, whose diagnoses were confirmed clinically and histopathologically, were included in the study. The duration of the disease was questioned. Tissue samples of 5-mm punch biopsy material were collected from the patients and from six healthy volunteers (three female, three male) as the controls. Nested polymerase chain reaction (PCR) with specific primers for HHV-7 DNA sequences (OPERON technologies Inc., HV-7S/HV-8A external sences and HV-10S/HV11A internal sences) was performed on each tissue sample. Polymerase chain reaction products were analyzed by electrophoresis on 2% agarose gels. After molecular weight markers (Haphi174) had been placed and visualized on an ultraviolet transilluminator, the gels were immersed and photographs were taken. RESULTS: The mean age was 29.86 +/- 11.77 for the PR patients and 25.33 +/- 11.69 for the controls. The mean duration of the disease was 16.28 +/- 15.74 days. Human herpesvirus 7 DNA sequences were detected in six of the PR patients (28.57%). The mean duration of the disease was calculated as 11.67 +/- 9.85 for the HHV-7-positive patients (patient nos. 3, 4, 5, 7, 8, 9) and 18.13 +/- 17.05 for the HHV-7-negative patients, and there was no statistically significant differences in either of the groups (U = 29.5, W = 50.5, P = 0.2241, using the Mann-Whitney U and Wilcoxon's rank sum W-tests). Nested PCR was negative for HHV-7 in all of the specimens from the controls. There was no statistically significant difference for the presence of HHV-7 DNA sequence between the PR patients and the controls (P = 0.2843, Fisher's exact two-tail analysis test). CONCLUSION: Our results failed to support a possible role for HHV-7 in the pathogenesis of PR.

Bcl-2-related proteins, alpha-smooth muscle actin and amyloid deposits in aggressive and non-aggressive basal cell carcinomas.

Aberrant expression of bcl-gene products has been implicated in the development of non-melanoma skin cancers. Recently, altered expression of alpha-smooth muscle actin has been proposed as predictive of tumour behaviour in basal cell carcinomas. The purpose of this study was to compare the aggressive and non-aggressive basal cell carcinomas in terms of bcl-gene products and alpha-smooth muscle actin expression. Fifty excisional biopsy samples were studied by immunohistochemical technique for the differential expressions of bcl-2, bax, bcl-x and alpha-smooth muscle actin. Bcl-2, bcl-x and bax were expressed in 34 (68%), 38 (76%) and 41(82%) specimens, respectively. Immunoreactivity for alpha-smooth muscle actin was noted both in tumour nests (64%) and within the stroma (54%). There was a significant difference between aggressive and non-aggressive basal cell carcinomas in terms of bcl-2 and stromal alpha-smooth muscle actin immunoreactivity. Non-aggressive basal cell carcinomas display a concordant expression of bcl-family proteins, whereas aggressive tumours reveal a discordant pattern. An increased expression of stromal alpha-smooth muscle actin with a concomitant decrease or loss of bcl-2 expression may be highly suggestive of aggressiveness in basal cell carcinoma.