Structure and Characterization of Eriphia verrucosa Hemocyanin.

Arthropod hemocyanins (Hcs) are a family of large extracellular oxygen-transporting proteins with high molecular mass and hexameric or multi-hexameric molecular assembly. This study reports for the first time the isolation and characterization of the structure of an arthropod hemocyanin from crab Eriphia verrucosa (EvH) living in the Black Sea. Its oligomeric quaternary structure is based on different arrangements of a basic 6 × 75 kDa hexameric unit, and four of them (EvH1, EvH2, EvH3, and EvH4) were identified using ion-exchange chromatography. Subunit 3 (EvH3) shows high similarity scores (75.0, 87.5, 91.7, and 75.0 %, respectively) by comparison of the N-terminal sequence of subunit 1 from Cancer pagurus of the North Sea (Cp1), subunits 3 and 6 of Cancer magister (Cm3 and Cm6), and subunit 2 of Carcinus aestuarii (CaSS2), respectively. Moreover, a partial cDNA sequence (1309 bp) of E. verrucosa hemocyanin encoding a protein of 435 amino acids was isolated. The deduced amino acid sequence shows a high degree of similarity with subunits 3, 4, 5, and 6 of C. magister (81-84 %). Most of the hemocyanins are glycosylated, and three putative O-linkage sites were identified in the partial amino acid sequence of EvH at positions 444-446, 478-480, and 547-549, respectively. The higher stability of native Hc in comparison to its subunit EvH4 as determined by circular dichroism (CD) could be explained with the formation of a stabilizing quaternary structure.

Multivariate analysis approach to the serum peptide profile of morbidly obese patients.

BACKGROUND: Obesity is currently epidemic in many countries worldwide and is strongly related to diabetes and cardiovascular disease. Mass spectrometry, in particular matrix-assisted laser desorption/ionization time of flight (MALDI-TOF) is currently used for detecting different pattern of expressed protein. This study investigated the differences in low molecular weight (LMW) peptide profiles between obese and normal-weight subjects in combination with multivariate statistical analysis. MATERIALS: Serum samples of 60 obese patients and 10 healthy subjects were treated by cut-off membrane (30000 Da) to remove the most abundant proteins. The filtrates containing the LMW protein/peptides were analyzed by MALDI-TOF mass spectrometry. Dataset was elaborated to align and normalize the spectra. We performed cluster analysis and principal component analysis to detect some ionic species that could characterize and classify the subject groups. RESULTS: We observed a down-expression of ionic species at m/z 655.94 and an over-expression of species at m/z 1518.78, 1536.77, 1537.78 and 1537.81 in obese patients. Furthermore we found some ionic species that can distinguish obese patients with diabetes from those with normal glucose level. CONCLUSION: Serum peptide profile of LMW associate with multivariate statistical approach was revealed as a promising tool to discriminate and characterize obese patients and it was able to stratify them in relation to comorbidity that usually are associated with this disease. Further research involving a larger sample will be required to validate these findings.

Self-reported stress reactivity and psoriasis-related stress of Nordic psoriasis sufferers.

OBJECTIVES: The purpose of the study was to investigate the perceived influence of stress on psoriasis onset and disease severity in a large sample of psoriatics and to compare stress reactors and non-reactors with respect to psoriasis-related stress, disease severity, family history of psoriasis and sociodemographic factors. PATIENTS/METHODS: A total of 5795 members of the Nordic psoriasis associations and 702 patients recruited from Nordic dermatologists or university clinics were asked whether their first outbreak of psoriasis occurred during times of worry and stress. They were also asked to rate the degree to which their psoriasis was influenced by stress and to complete the Psoriasis Life Stress Index, the Psoriasis Disability Index and a number of additional questions concerning sociodemographic factors. RESULTS: Seventy-one per cent of the members and 66% of the patients reported that their psoriasis was exacerbated by stress, and 35% in both groups reported that the onset of their psoriasis occurred during a time of worry and stress. Stress reactors, scoring above the median on stress reactivity, reported greater disease severity, psoriasis-related stress and impairment of disease-related quality of life. They also reported more frequent use of tobacco, tranquillizers and antidepressants. More women than men were stress reactors, and stress reactors were more likely to have a family history of psoriasis. CONCLUSION: Our findings confirm and extend the results of previous studies and indicate that a subgroup of psoriatics may be more psychologically reactive to their disease and its influence on everyday life. Whether this group is also physiologically more reactive to psychosocial stress remains to be investigated.

Discoid lupus erythematosus treated with cryotherapy.

A female is presented with multiple longstanding hypertrophic discoid lupus erythematosus lesions resistant to topical and systemic therapy for years. She suffered severe itching and pain localized to the lesions. Treatment with cryosurgery resulted in complete healing of the lesions, leaving slightly hypopigmented soft scars. There was no tendency of relapse during a follow-up period of ten years.

Treatment of psoriasis in the Nordic countries: a questionnaire survey from 5739 members of the psoriasis associations data from the Nordic Quality of Life Study.

The data from a questionnaire-based study of 5,739 members of the psoriasis associations of Denmark, Finland, Iceland, Norway, Sweden and the Faeroe Islands showed that the two most commonly used active agents were topical steroids (89.7% total use and 49.4% present use) and calcipotriol (73.1% total use and 35.8% present use), with only small variations between the countries. Marked differences between the countries were, however, found within all other types of psoriasis therapy, including the so-called alternative treatments. Significant priorities varied between the different countries. The use of dithranol in Finland was almost twice the average. While 14.2% of Danish members had received grenz-rays within the last week only 0.1% of the Finns had been given the same treatment. Psoralen plus ultraviolet A (PUVA) was being used by 13.1% of the Finnish psoriatics compared with 3.8% of Danes, while PUVA was almost non-existent on the Faeroe Islands. The use of non-PUVA phototherapy was highest in Norway and Sweden. Almost 10% of the Danes were presently on methotrexate, which was used far more than etretinate/acitretin or cyclosporine. In contrast, Finnish patients more often received etretinate than other systemic agents, and in Iceland there was a higher present use of cyclosporine than of etretinate. The popularity of alternative therapies was highest in Iceland, where 26.6% had taken such medication during the last week. The results of the study suggest that different treatment patterns should be taken into consideration when discussing the prognosis of psoriasis in different countries.

Topical calcipotriol combined with phototherapy for psoriasis. The results of two randomized trials and a review of the literature. Calcipotriol-UVB Study Group.

BACKGROUND: Calcipotrial has a well-documented effect in the treatment of psoriasis. OBJECTIVE: To confirm the beneficial effect of the combination of calcipotriol and UVB and to demonstrate that the combination is safe and well tolerated. METHODS: Data from two randomized right/left studies were analysed. Patients included in the studies had chronic stable plaque-type psoriasis with symmetrical lesions on the arms, the legs and/or the trunk. In one study, 101 patients were treated with calcipotriol on one side and calcipotriol + UVB on the other side of the body (open study). In the other study, 77 patients were treated with calcipotriol + UVB on one side and vehicle + UVB on the other side of the body (double-blind study). Calcipotriol ointment, 50 microg/g, was applied twice daily and UVB 3 times weekly for 8 weeks. UVB was increased from 0.7 MED before treatment in rapid steps up to the erythema threshold. RESULT: In both treatment series the therapeutic effect of the combination of calcipotriol and UVB was enhanced as compared to calcipotriol alone and UVB alone. In the first series there was a significant reduction of the psoriasis area and severity index (PASI) with the combination after 2 weeks as compared to calcipotriol alone. At the end of treatment significantly more sides were cleared after calcipotriol + UVB than after calcipotriol alone. In the other series there was a significantly faster onset of improvement on the sides treated with calcipotriol + UVB than on those treated with vehicle + UVB. After 2 weeks there was a significant difference in PASI in favour of calcipotriol + UVB. At the end of treatment, however, there was no difference between the treatments. There was a similar adverse event profile with either treatment. The addition of UVB to calcipotriol did not alter the tolerability or safety of topically applied calcipotriol. CONCLUSIONS: The result indicates a beneficial effect of combining calcipotriol and phototherapy. The findings are compared to other published studies.

PUVA and cancer risk: the Swedish follow-up study.

There is concern about the long-term carcinogenic effects of psoralen and ultraviolet A radiation (PUVA) for treatment of skin disorders. Many authors have found an increased risk for cutaneous squamous cell carcinoma (SCC). Except in anecdotal reports, malignant melanoma had not been observed in patients treated with PUVA until recently. In the U.S.A., a 16-centre prospective study of 1380 patients showed for the first time that there might also be an increased risk for malignant melanoma in patients treated with high cumulative dosages of PUVA. We have therefore followed up the Swedish PUVA cohort until 1994. This cohort had previously been followed up until 1985. Information from 4799 Swedish patients (2343 men, 2456 women) who had received PUVA between 1974 and 1985 was linked to the compulsory Swedish Cancer Registry in order to identify individuals with cancer. The average follow-up period was 15.9 years for men and 16.2 for women. We did not find any increased risk for malignant melanoma in our total cohort of 4799 patients treated with PUVA or in a subcohort comprising 1867 patients followed for 15-21 years. For cutaneous SCC there was an increase in the risk: the relative risk was 5.6 (95% confidence interval, CI 4. 4-7.1) for men and 3.6 (95% CI 2.1-5.8) for women. Significant (P < 0.05) increases were also found in the incidence of respiratory cancer in men and women and of kidney cancer in women. In conclusion, we did not find any increased risk for malignant melanoma in our patients treated with high doses of PUVA and followed up for a long time. We confirm previous reports of an increase in the incidence of cutaneous SCC in patients treated with PUVA, and recommend that patients should be carefully selected for PUVA and rigorously followed up.

Hair ignition by dye laser for port-wine stain: risk factors evaluated.

Flashlamp-pumped pulsed dye laser is the preferred treatment for port-wine stain. Vascular hemoglobin and epidermal melanin are competing sites for dye laser absorption and damage. The case presented illustrates the potential hazard of ignition induced by dye laser treatment on the face of a patient receiving inhalation anesthesia. A 6-year-old girl with almost black hair was treated for a port-wine stain covering most of the right half of her face. She was treated with dye laser under general anesthesia administered by mask. A laser pulse close to the upper part of the eyebrow induced a blaze and the eyebrow was instantly destroyed by the fire. Regrowth of the eyebrow was complete after a few months. Hair specimens of various colors were exposed experimentally to dye laser irradiation in room and oxygen-saturated atmospheres. Risk factors of ignition are high laser dosage, a high oxygen level, repeated pulses and dark colored hair.

Perivulvar inflammatory linear verrucous epidermal nevus (ILVEN) treated with CO2 laser.

Inflammatory linear verrucous epidermal nevus (ILVEN) is a type of epidermal nevus with the onset usually at early childhood. It is generally rather persistent with pruritus as a prominent symptom and is resistant to various treatments. We report a case of disfiguring ILVEN of the vulvar area, which was treated successfully with low fluence CO2 laser. The patient remains improved 2 years following laser therapy.

Met-enkephalin receptors in human gliomas.

Endogenous opioid systems (opioid peptides and receptors) are involved in many functions including the regulation of cell growth. We investigated the presence of Met-enkephalin binding sites in gliomas by displacement assays. Results demonstrated that few gliomas exhibit Met-enkephalin binding sites and that the percentage of tumours which express these binding sites strongly decreases with increasing malignancy. Moreover, we observed a shift from mu Met-enkephalin binding sites in low grade gliomas to delta Met-enkephalin binding sites in high grade gliomas. These results suggest an inactivation of the Met-enkephalinergic system in gliomas which could lead to loss of the inhibitory effect exerted by Met-enkephalin on normal astrocyte growth and thus favour progression of malignancy.

VATS increases costs in patients undergoing lung biopsy for interstitial lung disease.

Proliferation in the use of video-assisted thoracic surgery (VATS) has occurred without data demonstrating benefit. Few studies have critically compared VATS with limited thoracotomy (LT) in homogeneous patient populations undergoing standardized procedures. We retrospectively reviewed the hospital records of 37 consecutive patients referred for elective lung biopsy as part of an ongoing interstitial lung disease protocol to determine whether VATS improved outcome or reduced costs. Sixteen patients underwent VATS, and 21 patients underwent LT lung biopsy over a 31 month period. The two groups were homogeneous in regard to clinical symptoms, radiologic findings, age, sex, and preoperative pulmonary function. The operative mortality was not different between the two groups (VATS, 0/16, and LT, 1/21). The postoperative stay was 4.8 +/- 1.0 days for VATS and 5.0 +/- 0.5 days for LT (p = not significant). Operating time, number of specimens obtained, chest tube output, and day of chest tube removal did not differ. There was no difference in the amount of analgesics required during the postoperative period. Operating room cost for VATS was significantly greater than that for LT ($2,663 +/- $384 versus $1,801 +/- $94; p = 0.04) despite the use of nondisposable equipment. Anesthesia-related costs were also greater for VATS ($309 +/- $11 versus $244 +/- $15; p = 0.002). In conclusion, lung biopsy in patients with interstitial lung disease can be performed safely and efficiently with either VATS or LT, but VATS results in higher procedure-related costs.

Immunodetection of endogenous opioid peptides in human brain tumors and associated cyst fluids.

The antitumorigenic effects of endogenous opioid peptides and their presence in extracerebral tumors are well documented. In this study, methionine-enkephaline (met-enkephalin) was measured by radioimmunoassay in 108 glial and nonglial brain tumors and in 44 associated cyst fluids. By immunohistochemistry, the distribution of the peptide and its precursor, preproenkephalin A, was also analyzed. Met-enkephalin and preproenkephalin were detected in the cytoplasm and cell processes of all tumors. Moreover, for neuroectodermal tumors (i.e., gliomas, gangliogliomas, and dysembryoplastic neuroepithelial tumors), a strong inverse correlation (P < 0.0001) was observed between the met-enkephalin levels and the degree of malignancy (242.9, 148.3, 55.3, and 30.3 pg/mg protein for grade 1, 2, 3, and 4, respectively). When compared to normal tissue, this differential expression mainly results from a decrease in the opioid peptide content in high-grade neuroectodermal tumors. Meningiomas and cerebral metastases displayed low met-enkephalin levels, similar to those of grade 4 neuroectodermal tumors. Large amounts of met-enkephalin were found in all cyst fluids. These data suggest that the endogenous opioid system is an integral component of brain tumors and that met-enkephalin may represent a useful malignancy marker in neuroectodermal tumors.

Glutaric acidemia type II. Heterogeneity in beta-oxidation flux, polypeptide synthesis, and complementary DNA mutations in the alpha subunit of electron transfer flavoprotein in eight patients.

We studied metabolic, polypeptide and genetic variation in eight glutaric acidemia type II (GA II) patients with electron transfer flavoprotein (ETF) deficiency. As measured by 3H-fatty acid oxidations in fibroblasts, beta-oxidation pathway flux correlated well with clinical phenotypes. In six patients with severe neonatal onset GA II, oxidation of [9,10(n)-3H]-palmitate ranged from 2% to 22% of control and of [9,10(n)-3H]myristate, from 2% to 26% of control. Of two patients with late onset GA II, one had intermediate residual activities with these substrates and the other normal activities. Radiolabeling and immunoprecipitation studies revealed that three of the six neonatal onset GA II patients had greatly diminished or absent alpha- and beta-ETF subunits, consistent with a failure to assemble a stable heterodimer. Another neonatal onset patient showed normal synthesis of beta-ETF but decreased synthesis of alpha-ETF. Two neonatal onset and two late onset GA II patients showed normal synthesis of both subunits. Analysis of the pre-alpha-ETF coding sequence revealed seven different mutations in the six patients with neonatal onset GA II. The most common mutation was a methionine for threonine substitution at codon 266 found in four unrelated patients, while all the other mutations were seen in single patients. No mutations were detected in the two patients with late onset GA II.