Immunohistochemical Analysis of Adhesion Molecules E-Selectin, Intercellular Adhesion Molecule-1, and Vascular Cell Adhesion Molecule-1 in Inflammatory Lesions of Atopic Dermatitis.

E-selectin, ICAM-1 (intercellular adhesion molecule-1), and VCAM-1 (vascular cell adhesion molecule-1) play a role in atopic dermatitis (AD). This study aimed to evaluate their expression in skin biopsy specimens of patients diagnosed with AD using an optimized computer program. A descriptive analysis and comparison of digitally measured surface area and cell number were performed. The number of E-selectin-positive cells did not vary between the groups. In patients with AD, decreases of 1.2-fold for ICAM-1- and 1.3-fold for VCAM-1- positive cells were observed. The E-selectin-positive epidermal surface area increased (p < 0.001), while ICAM1 and VCAM1 decreased 2.5-fold and 2-fold, respectively, compared to controls. In the AD-affected skin, the E-selectin-positive endothelial area was 3.5-fold larger (p < 0.001), and the ICAM1-positive area was almost 4-fold larger (p < 0.001). E-selectin and ICAM-1 were expressed in the control dermis moderately and weakly, respectively. A strong E-selectin signal was detected in the AD-affected skin macrophages and a strong ICAM-1 signal in the dermal vessel endothelium. In the endothelial cells of AD-affected skin, no VCAM-1 signal could be found. E-selectin, ICAM-1, and VCAM-1 expression show significant disease-specific changes between AD-affected and control skin. The combination of digital analysis and a pathologist's evaluation may present a valuable follow-up of AD activity parameters.

Contact Sensitivity in Patients with Atopic Dermatitis.

Atopic dermatitis (AD) is a common chronic and relapsing, non-contagious inflammatory skin disorder, characterized by skin barrier impairment and baseline immune irregularities. The literature on the relationship between AD and cutaneous delayed-type hypersensitivity is inconclusive. There is an ongoing debate whether contact sensibility (CS) is found more or less often among patients with AD. Aim of the study was to evaluate the incidence of contact sensitivity (positive patch test reactions) in patients with and without AD. We patch tested a total of 2143 patients (563 men and 1580 women). There were 226 patients with history of AD; 61 (27%) men and 165 (73%) women. The patient group without AD consisted of 1917 patients, 502 (26%) male and 1415 (74%) female patients, who were referred to our Department with clinical suspicion of allergic contact dermatitis (ACD). A patch test was performed with the baseline series, and readings were performed on days D2, D3, and D7. Among patients with AD, 109 (48.2%) had a positive patch test reaction to at least one allergen, whereas 1094 (57.1%) patients with no history of AD had a positive patch test reaction. The most common positive allergens in patients with AD were nickel (II) sulfate (13.3%), thimerosal (12.4%), cobalt (II) chloride (11.5%), methylisothiazolinone (MI) (8.4%), fragrance mix I (6.6%), potassium dichromate (5.3%), methyldibromo glutaronitrile (4.0%), and carba mix (4.0%). The results of our study agree with previous findings that there is no significant difference in prevalence of CS between the atopic and nonatopic populations.

Autoimmune Progesterone Dermatitis Diagnosed by Lymphocyte Transformation Test and Progesterone Provocation Test.

Autoimmune progesterone dermatitis (APD) is rare autoimmune response to endogenous progesterone or to earlier exposure to exogenous progesterone (1). Skin lesions typically occur due to increases in progesterone during the luteal phase of the menstrual cycle (2). A-31-year-old mother of two children presented to our Department with a 5-year history of pruritic and painful erythematosus macules, papules, and patches on her neck, pectoral region, and face, which appeared 2-3 days before the onset of menses and gradually resolved 7-10 days later (Figure 1). The lesions first appeared 10 months after her second pregnancy and a few months after she had started using oral contraceptive pills (OCP) containing gestodene combined with ethinyloestradiol. A few months before presenting to us, the lesions had started spreading on her forearms, elbows, and pretibial areas. Since one year prior to our visit she had complained of occasional urticaria with angioedema one week prior to menses, which resolved after menses. The lesions were accompanied by malaise, headache, and fatigue. The patient was asymptomatic between the outbreaks. She reported that she had been using various local corticosteroids, peroral antihistamines, and prednisone for the treatment of her skin lesions, but this treatment had not improved her symptoms. She suffered from mild seasonal rhinoconjunctivitis. We performed multiple laboratory tests that were unremarkable. Histopathological examination of a biopsy taken from a lesion on the neck showed epidermal hyperplasia and nonspecific mild dermal inflammation. Since progesterone was not available in aqueous solution in our country, we did not perform an intradermal test, but we performed a lymphocyte transformation test (LTT) to medroxyprogesterone and estradiol. The patient's lymphocytes showed markedly enhanced proliferation to medroxyprogesterone in vitro, while being negative to estradiol. We had performed control LTT in 10 healthy controls and 10 patients with atopy, and such hyperactivity was not observed in any of them. We performed an oral provocation test with OCP containing gestodene combined with ethinyloestradiol. Two days after commencing treatment, the patient developed widespread dermatitis (Figure 2) with nausea, malaise, and angioedema. The patient was informed about treatment options and possible side-effects. She started with OCP with the lowest amount of progesterone, containing ethinyloestradiol and dropirenone for treatment of APD, but terminated treatment after the second cycle due to a worsening of the skin lesions and urticaria accompanied with angioedema. At the time of writing, our patient continues to have premenstrual flares. The typical symptoms of APD are skin lesions such as eczema, erythema multiforme, prurigo, stomatitis, papulopustular lesions, folliculitis, urticaria, angioedema, and rarely anaphylaxis (2) that develop 3-10 days before and subside 1-2 days after menses, with recurrent cyclic aggravation (1,3,4). Frequently, patients have a history of exogenous progesterone intake (1,5,6), as in our patient, which could have resulted in antibody formation. The diagnosis of APD is established by an appropriate clinical history (premenstrual flare of skin lesions), a progesterone intradermal test, an intramuscular (7), oral (8), or intravaginal (1, 6) progesterone challenge test, and circulating antibodies to progesterone. Progesterone testing has not been standardized. Most of the sex hormones are not suitable for testing since they contain an oily component that can produce an irritant test reaction. Gestodene, which was used for the oral provocation test in our patient, is a potent progesterone (9). The LTT shows reactions to circulating lymphocytes and reflects immune reactions within the body. The goal of treatment is suppression of ovulation. Currently, the first-line choice of therapy is a combination oral contraceptive (3). We believe that OCP have a limited effect because all of them contain a progesterone component. If this treatment is ineffective, patients have been treated with danazol, gonadotropin releasing hormone analogs (3,4,6), conjugated estrogens (7), tamoxifen, oophorectomy (8), and progestogen desensitization (10) with varying success.

[Prevention of leg ulcer]. / Prevencija potkoljenicnog vrijeda.

Lower leg ulcers is the most common form of ulceration of the lower extremities. The prevalence of leg ulcer varies among studies from 0.1% to 0.6%. In the majority of studies, 1% of the population develop leg ulcer at least once in lifetime. The prevalence is higher in elderly people. There are several hypotheses used to explain the pathophysiological steps leading from the popliteal venous hypertension in value. Currently, the treatment of leg ulcer relies on due knowledge of ulcer pathophysiology and making an accurate diagnosis. Venous disease has a significant impact on quality of life and work productivity. In addition, costs associated with the prevention and treatment of lower leg ulcers are significant.

Patch test reactions to metal salts in patients with different types of dermatitis.

Metal allergies can be a clinical problem, especially in atopic individuals. This study is unique and contributes with new knowledge in everyday life skin care of irritant and atopic dermatitis patients. The aim of the study was to determine the frequency of positive patch test reactions to metal contact allergens (potassium dichromate, cobalt chloride, nickel sulfate, white mercury precipitate) in patients diagnosed with allergic contact dermatitis, irritant contact dermatitis, and atopic dermatitis. Between 2007 and 2011, patch testing was performed in 2185 patients according to the International Contact Dermatitis Research Group technique. Study results showed statistically significant differences in patch test responses to 2 allergens, nickel sulfate (χ(2)=24.22; p<0.001) and cobalt chloride (χ(2)=22.72; p<0.001). Nickel sulfate was the most common allergen in allergic contact dermatitis and atopic dermatitis, while for irritant contact dermatitis the most common allergen was cobalt chloride. Among the 4 tested metal allergens, the most common and relevant was nickel sulfate (χ(2)=17.25; p<0.004), found in almost all study subjects. In conclusion, the increased awareness of allergens and their potential sources may help limit the use of these chemicals in consumer product manufacturing.

5-azacytidine enhances proliferation in transplanted rat fetal epiglottis.

Fetal rat epiglottis and its developmental potential in ectopic transplants under the influence of the epigenetic drug was investigated. Epiglottises from 17-days-old rat embryos were transplanted under kidney capsules of adult rats for 14 days. 5-azacytidine (5 mg/kg) was injected intraperitoneally during first three days and controls were sham treated. TEM, immunohistochemical detection and quantitative stereological analysis of the Proliferating Cell Nuclear Antigen (PCNA) expression (numerical density N(v)) were performed. Typical chondroblasts with long surface processes and sparse lipid droplets were found in fetal epiglottis and chondrocytes with shorter processes, numerous lipid droplets and elastic fibers in adult. PCNA was expressed in almost all cells of the fetal epiglottis while in the adult it was expressed in minority of cells. In transplants, differentiation progressed towards the differentiation found in the adult. Application of 5-azacytidine increased the capacity for proliferation (N(v PCNA)) in comparison to controls but no difference in differentiation was observed. Data about the developmental potential and induction of proliferation in mammalian epiglottis by epigenetic modulation is of importance for regenerative medicine.

Expression of e-selectin in the skin of patients with atopic dermatitis: morphometric study.

Adhesion molecules may play an important role in the homing of T-cell subsets into allergen-exposed skin of atopic individuals. The aim of this study was to examine the expression of adhesion molecules in atopic dermatitis skin lesions. Biopsies were obtained from lesions in 30 adult patients with atopic dermatitis and 10 healthy adults as controls. Biopsy specimens were studied by immunohistochemistry for the expression of E-selectin in epidermis and dermis cells. Results showed significant changes in the epithelial cell expression of E-selectin, which were especially pronounced in vascular endothelium of the dermis of atopic dermatitis patients.

[What should we know about venous and arterial ulcers?]. / Sto treba znati o venskim i arterijskim ulkusima?

Chronic venous insufficiency starts by promoting the case of events. This results in pathologic venous and tissue changes consequential to chronic venous insufficiency. In this article, chronic venous insufficiency is defined as an abnormal function of the venous system due to venous valvular incompetence with or without associated venous outflow obstruction, which may involve the superficial venous system, deep venous system, or both. Long-term venous hypertension may present by a variety of clinical manifestations in terms of the severity and duration of venous circulation damage. Venous leg ulcer occurs in distal part of the shin, mostly around medial malleolus. The base of the ulcer is often covered by a yellowish fibrin layer. The surrounding skin is hyperpigmented, usually irritated by ulcer secretion, and inflamed. Arterial leg ulcer is caused by arterial insufficiency. It is round-shaped with dry basis and involves deeper structures, up to tendons. The skin is dry, atrophic, cool, pale, smooth, and hairless. Therapy of ulcers is demanding and consists of numerous general measures, topical and/or systemic therapy. Topical therapy consists of cleansing, mechanical debridement, disinfection and stimulation of granulations and epithelialization with prevention of secondary infection. Therapy with currently available bio-occlusive wound dressings is most effective. These dressings are hydrocolloids, hydropolymers, transparent films, alginate, and ionic silver. Vascular protectors, venetonics and antiaggregation medications can be used in systemic therapy.

Development of epithelia in experimental teratomas derived from rodent embryos.

Investigation of developmental potential of various embryonal tissues is important for design of new approaches to regenerative medicine aimed at supplementing tissues damaged by trauma or disease. Rodent embryos have been extensively used in experiments designed for investigation of developmental potential to give rise to various types of epithelia such as superficial epithelia, neuroepithelium and sometimes even malignantly transformed epithelium in teratoma-like structures. These experiments have been done in vitro, in transplants in vivo and by combined in vitro-in vivo methods.

Macular amyloidosis: a case report.

Macular amyloidosis is a major cause of skin pigmentation and a rare form of localized primary cutaneous amyloidosis in western countries, with a higher prevalence in Asia and South America. The etiopathogenesis of macular amyloidosis has not yet been fully clarified; a number of risk factors are involved, among them chronic friction in particular. A 54-year-old patient with macular amyloidosis is presented. The diagnosis of macular amyloidosis was based on history data on long-term persistence of the disorder, localized pruritus and constant scratching urge, grayish-brown pigmentation over the scapula, and detection of amyloid in histologic slides. Three-month tretinoin therapy resulted in pruritus alleviation, with no change in the appearance of hyperpigmentation. The exact incidence of macular amyloidosis in Croatia is not known, however, the issue appears to be underestimated or neglected in dermatology routine.

Most common inhalant allergens in atopic dermatitis, atopic dermatitis/allergic rhinitis, and atopic dermatitis/bronchial asthma patients: a five-year retrospective study.

Atopic dermatitis (AD) is a chronic relapsing inflammatory skin disease with a distinctive clinical appearance and distribution. Around 85% of patients have positive immediate skin reaction or specific IgE to different airborne allergens that are in association with respiratory allergy. The aim of this retrospective, open and uncontrolled study was to identify the most common inhalant allergens in AD patients, AD/allergic rhinitis patients, and AD/bronchial asthma patients by skin prick test per year during the 2001-2005 period.

Milker's nodule--case report.

Milker's nodule (noduli mulgentium) is a benign viral skin disease caused by parapoxvirus, a poxvirus that is endemic in cattle. The virus is usually transmitted to cattle handlers from infected cows. We present a case of a 25-year-old housewife who milked cows on her family farm and developed itchy, purplish red nodules on her fingers and hands, and lymphangiitis of the right arm. Two weeks before, several cows on her family farm were treated by a veterinarian under the diagnosis of pseudocowpox. She was treated with orally administered amoxicillin with clavulanic acid 2 g for 10 days, with complete resolution of lymphangiitis. The nodules resolved in several weeks.

Moisturizers.

Moisturizers combine occlusives and humectants to enhance the water-holding capacity of the skin. Further diversity in moisturizer formulation is created through the addition of special ingredients, designed to enhance the functions of the skin. These agents mimic natural ingredients. Application of moisturizers can serve as important adjunctive therapy for patients with various dermatologic disorders.

Contact allergy and sociodemographic characteristics.

The aim of the study was to determine the frequency of positive patch test reaction to different contact allergens according to patients age, sex, occupation and clinical features. Between 1999 and 2003, patch testing was performed in 3,293 patients with respective clinical diagnoses. Patch testing was done by the standard technique proposed by the International Contact Dermatitis Research Group (ICDRG). Study results showed statistically significant differences in patch test response according to sex and age for three allergens (cobalt chloride, nickel sulphate and thiomersal); according to occupation for nine allergens (cobalt chloride, nickel sulphate, balsam of Peru, fragrance mix, thiuram mix, wood tars, neomycin sulphate, thiomersal and detergents), and clinical diagnosis for two allergens (nickel sulphate, and wood tars). The most common and relevant allergens were: nickel sulphate, cobalt chloride and carba mix. They were found in all examinees regardless of age, sex, occupation and diagnoses. The increased awareness of allergens and their potential sources may help to limit the usage of these chemicals in manufacture of consumer products.

Retrospective survey of patch testing at department of dermatology and venerology, zagreb university hospital center in zagreb, croatia.

Epicutaneous (patch) test is a standard test to confirm contact allergy. During a six-year period (1998-2003), contact allergic dermatitis was diagnosed in 4132 (65%) of 6341 patients using standard patch test with a Croatian series of allergens according to the International Contact Dermatitis Research Group (ICDRG). The most common clinical diagnoses related to contact allergy (male; female) were contact allergic dermatitis (65.4%; 72.2%); atopic dermatitis (15.9%; 11.7%); contact irritant dermatitis (7.0%; 6.6%) and nummular eczematoid dermatitis (4.2%; 2.4%). According to occupation, positive patch test reaction was most commonly observed in workers, whereas 65% of patients showed positive reactions to one or more substances. There was an increased frequency of positive patch test reactions to potassium dichromate, nickel sulfate, thimerosal and neomycine sulfate, along with a significant decrease in the frequency of positive patch test reactions to cobalt chloride, carba mix, wood tars, detergents, parabens mixture, urushiol and rubber mixture. We strongly recommend keeping up-to-date with the epidemiology of contact allergy in Croatia.

Fungal infections of urogenital system.

We analyzed the frequency of isolation of individual fungal species in the samples of urine, vaginal and cervical swabs from 107 patients (72% women and 28% men) aged 16-82 years, who were treated in primary care for cystitis, vulvovaginitis, and cervicitis. The samples were analyzed at the Microbiological Laboratory of Zagreb Institute of Public Health, Croatia, between September 1, 2002 and June 31, 2003. Eight species of yeast were isolated from the samples. Candida (C.) albicans, C. glabrata, and C. krusei were the most common isolates, with the frequency of 61.7%, 10.3% and 8.4%, respectively. Other species (C. guilliermondii, C. famata, C. tropicalis, C. parapsilosis, and C. kefyr) were less frequently isolated, between 0.9% and 7.5%. In women, the frequency of isolation of C. albicans species from urine samples ranged from 83.3% to 30.8%, and from uterine cervical swabs from 85.7% to 50%, showing a decreasing trend with patients' age. The frequency of C. albicans isolates from vaginal swabs was equal in younger (<30 years) and older women (l51 years), but twice less frequent in middle-aged women. In men, the frequency of C. albicans species isolated from urine samples decreased with age from 100% to 52.4%. In the 107 analyzed samples with positive fungal culture, yeast isolations were significantly more frequent than pure culture (93.5%) and had a larger number of colony counts (57%). This suggested that these yeast species might have a pathogenic role in the causation of urogenital system infections.