Efficacy of a polyphenolic extract from silver fir (Abies alba) bark on psoriasis: a randomised, double-blind, placebo-controlled trial.

Silver fir (Abies alba) bark extract contains a mixture of bioactive polyphenols. We tested their effectiveness in the treatment of psoriasis in order to further investigate the potential topical anti-inflammatory activity of polyphenols by means of a randomized, double-blind, placebo-controlled add-on clinical trial, after having examined their ability to downregulate the expression of IL-1ß cytokine in monocyte/macrophage primary cell culture. 61 patients with mild psoriasis met the inclusion criteria and were willing to comply with protocol requirements, were enrolled in the study. The severity of the disease was measured by psoriasis area severity index (PASI). Treatment efficacy was evaluated by assessing erythema (E, 0 to 4-point scale), desquamation (D, 0 to 4-point scale) and induration (I, 0 to 4-point scale) of lesions before and after the treatment. All patients enrolled in the study had symmetrical psoriasis plaques on the skin. All patients received O/V ointment with 2% of silver fir bark extract and/or placebo, respectively. We compared medications by right/left intra-patient comparison, so that the control group was always contralateral of the tested one. Location of the tested or control site was randomised, using a computer-generated randomisation schedule. Silver fir extract was well-tolerated. A superiority of active treatment above placebo, based on the clinical investigational PASI score system was observed by 15 % in all volunteers and in 40% regarding the improvement of psoriasis on elbows. However, statistical analysis showed no significant differences between placebo and active treatment with the extract from silver fir bark (p < 0.05).

Inflammatory and regenerative responses in salmonids following mechanical tissue damage and natural infection.

Locale responses in muscle tissue against either a sterile tissue damage or infection were compared in salmonid fish in order to examine the inflammatory responses and regeneration of tissue. From higher vertebrates both damage and infection are known to cause inflammation since DAMPs released from injured cells as well as PAMPs from the surface of pathogens are immunogenic. To examine this in salmonid fishes, Atlantic salmon (Salmo salar) were infected with Moritella viscosus, the causative agent of winter ulcer. Muscle tissue was sampled from infected fish at 4, 7 and 14 days post infection. Samples were obtained from site of lesions and from locations without clinical signs of disease and lesions. The tissue damage was performed in rainbow trout (Oncorhynchus mykiss) by applying sterile needles to skin and muscle tissue to one side of the fish. Samples were taken 7, 14, 21, 28 and 42 days post injury from the injured side and non-injured site (internal control). From both infected and damaged fish, samples were subject to real-time RT-PCR for measuring the expression of IL-1beta, IL-8, IL-10, Hsp70, iNOS, TGF-beta, TLR-5m, TLR-9, TLR-22, TGF-beta, MMP-2, CTGF, myostatin-1alphabeta and collagen-1alpha which are coding for immunological factors and tissue regeneration. Locale, inflammatory responses were seen as strong up-regulation of IL-1beta and IL-8 in both groups of fish, but it was more pronounced in infected fish. Expression of the toll-like receptors showed induction of TLR-5m following infection, but TLR-9 and TLR-22 following damage. Further, in both studies the regenerative genes TGF-beta, MMP-2, CTGF, myostatin-1alphabeta were induced, but showed different kinetics. Collagen-1alpha was only induced in infected fish, probably due to heavier tissue damage in these.

Previously unrecognised division within Moritella viscosa isolated from fish farmed in the North Atlantic.

Previously undocumented phenotypical and genetic variation was identified amongst isolates of Moritella viscosa collected from various geographical locations and from different fish species. The studied isolates could be split into 2 major phenotypically and genetically different clusters, one of which was consistent with the species type strain (NCIMB 13548). Isolates consistent with the type strain originated exclusively from Atlantic salmon farmed in Norway, Scotland and the Faroe Isles, although a single isolate from farmed Norwegian cod clustered closely with this group. The 'variant' cluster comprised isolates originating from Norwegian farmed rainbow trout, Icelandic farmed rainbow trout and salmon, Canadian farmed (Atlantic) salmon, Icelandic lumpsucker and only exceptionally from Norwegian salmon. With the exception of the single aforementioned cod isolate, all isolates from Norwegian farmed cod belonged to the variant cluster. Phenotypically, the clusters could be absolutely separated only by elevated haemolytic activity in the variant strain, although approximately half of these isolates also produced acid from mannose, in contrast to the typical (type) strain. While 16S rRNA gene sequencing was unable to separate the 2 clusters, Western blot analyses, plasmid profile analysis, pulsed field gel electrophoresis and gyrB gene sequence analysis produced clusters consistent with the phenotypic data. Macroscopically and histologically the disease in rainbow trout caused by the variant strain was consistent with that previously described in Atlantic salmon. The results of the present study may indicate a degree of host specificity of the typical strain for Atlantic salmon.

Atlantic salmon bath challenged with Moritella viscosa--pathogen invasion and host response.

The Gram-negative bacterium Moritella viscosa is considered to be the main causative agent of winter ulcer, a disease that primarily affects salmonid fish in sea water during cold periods. The disease is initially characterised by localised swelling of the skin followed by development of lesions. To gain more knowledge of the role of M. viscosa in the pathogenesis of winter ulcer, 159 Atlantic salmon (80-110 g) were exposed to a bath challenge dose of 7 x 10(5) cfu ml(-1) for 1 h at 8.9 degrees C. The first mortalities were registered two days post-challenge and the mortality rate increased rapidly. Multi-organ samples were taken throughout the challenge for culture, immunohistochemistry and PCR analysis. Using real-time PCR, M. viscosa DNA was first detected in the gills of all fish examined 2, 6 and 12 h after challenge. From day 2, the bacterium was detected in the muscle/skin, head kidney, spleen and liver. This was in correlation with positive cultured samples and confirmed systemic infection. The early and consistent detection of M. viscosa DNA in gill samples, and less or not in muscle/skin or intestine, could suggest gills as a port of entry for the bacterium. Immunohistochemical analysis using a polyclonal antiserum against M. viscosa demonstrated generalised staining in the lumen of blood vessels and some positive mononuclear cells. The antigens recognised by the antiserum may have originated from extracellular bacterial products and be part of a bacterial invasion strategy. To better understand the immune response in salmon to M. viscosa infection, the expression profiles of the immune genes IL1 beta, C3, ISG15 and CD83 were studied. Increased expression of IL1 beta and C3 was not induced until day 7, which may suggest that M. viscosa might utilize escape mechanisms to evade the host's immune system by suppressing relevant immune responses.

Expression of tight-junction proteins in the inflamed and clinically uninvolved skin in patients with venous leg ulcers.

Skin around venous leg ulcers (VLUs) is often inflamed and prone to contact sensitization. Expression of tight-junction components (ZO-1, occludin, and claudins 1 and 4) was studied by immunofluorescence in inflamed and noninflamed lower leg skin (both uncovered skin and skin occluded under hydrocolloid dressings) in patients with VLUs. No major differences were found in the expression of occludin and claudin-4. ZO-1 protein had stronger and more wide-ranging expression in the inflamed epidermis. Expression of claudin-1 was lost from the basal layer of the inflamed skin and skin under the hydrocolloid dressing. The skin on the lower legs affected by VLU may have altered expression of ZO-1 and claudin-1, similar to that seen in psoriatic plaques.

Lichen planus and other cutaneous manifestations in chronic hepatitis C: pre- and post-interferon-based treatment prevalence vary in a cohort of patients from low hepatitis C virus endemic area.

BACKGROUND: Several controversies exist regarding the relationship between hepatitis C virus (HCV) infection and some cutaneous manifestations, lichen planus (LP) in particular. OBJECTIVES: To determine the prevalence of LP and other cutaneous manifestations in a cohort of patients infected with HCV from low HCV endemic area of Slovenia, to correlate findings with chosen biological variables and to assess the role of interferon (IFN)-based treatment of HCV infection in cutaneous manifestations. Methods A total of 171 consecutive HCV-seropositive patients and 171 HCV-seronegative age- and gender-matched controls were studied prospectively. Prevalence of cutaneous manifestations, comparison between study patients and controls and correlation of skin findings with demographic, biochemical, virological and liver histologic findings as well as IFN-based therapy were assessed. RESULTS: Overall presence of LP in HCV-seropositives was 2.3%; although LP was not found in controls, the difference was not statistically significant (P = 0.123). Significantly higher than in controls was the prevalence of pruritus (31.0%, P < 0.001), dry skin (16.4%, P < 0.001) and hair loss (9.9%, P < 0.001). In IFN-based treatment naïves, skin findings were more frequent compared with controls, but not significantly, with no correlation to chosen biological variables. Current IFN-based treatment was significantly connected to pruritus (P < 0.001) and dry skin (P < 0.001). Compared with treatment naïves, in post-treated patients pruritus (odds ratio, 19.13; 95% confidence interval, 6.85-53.42; P < 0.001), dry skin (odds ratio, 4.21; 95% confidence interval, 1.44-12.31; P < 0.001) and hair loss (P < 0.001) were significantly more common. CONCLUSIONS: LP was not significantly related to HCV infection. Prevalence of pruritus, dry skin and hair loss was significantly higher in post-compared with pre-treated patients. The role of IFN in post-treatment persistence of skin manifestations needs to be assessed.

Real-time PCR detection of Moritella viscosa, the likely causal agent of winter-ulcer in Atlantic salmon Salmo salar and rainbow trout Oncorhynchus mykiss.

We report the development of a real-time PCR protocol for specific detection of Moritella viscosa. This bacterium is considered to be the main aetiological agent in development of winter-ulcer, a disease severely affecting salmonid aquaculture in Norway. From a newly elaborated draft version of the genome of M. viscosa, the tonB gene sequence was selected as a suitable basis for the development of the real-time PCR assay. The real-time PCR demonstrated the presence of M. viscosa DNA sequences in 88.1% of samples collected from 35 outbreaks of winter-ulcer in Norwegian fish farms. In contrast, standard culturing on blood agar identified M. viscosa in only 39.7% of fish. While the culturing method revealed a similar prevalence (26 to 27%) of M. viscosa in kidney and ulcer samples, substantially more ulcer (81.5%) than kidney (49.7%) samples were shown positive by real-time PCR.

Plasma vitamin E and blood selenium concentrations in Norwegian dairy cows: regional differences and relations to feeding and health.

Plasma alpha-tocopherol (vit E) and blood selenium (Se) concentrations in February were determined in samples from 314 dairy cows in Norway, selected to provide a representative subset of the Norwegian dairy cow population. Each sample was followed by a questionnaire with information about feeding of the cow at the time of sampling. The results were correlated to herd data and to calving and health data for each cow from the Norwegian Dairy Herd Recording System and the Norwegian Cattle Health Recording System. The mean concentrations were 6.9 microg vit E per ml plasma and 0.16 microg Se per g blood. Both levels were highest in mid lactation. Plasma vit E varied with the amount of silage fed to the cow, while blood Se varied with the amount of concentrates and mineral supplements, and with geographical region. No differences in vit E or Se levels were found between cows with recorded treatments for mastitis, parturient paresis or reproductive disorders in the lactation during or immediately prior to sampling, and those without such treatments. For ketosis, a small difference in blood Se was found between the groups with or without recorded treatments. It is concluded that winter-fed lactating cows in Norway had an adequate plasma level of vit E and a marginal-to-adequate level of Se.

Grouping by plasmid profiles of atypical Aeromonas salmonicida isolated from fish, with special reference to salmonid fish.

Plasmid profile analyses were performed for 113 strains of atypical Aeromonas salmonicida and the reference strain A. salmonicida subsp. salmonicida ATCC 14174. The atypical A. salmonicida strains comprised 98 strains obtained from fish originating from 54 farms and 2 lakes in Norway, 10 strains from Canada (2), Denmark (2), Finland (1), Iceland (1) and Sweden (4), the reference strains NCMB 1109 and ATCC 15711 (Haemophilus piscium) of A. salmonicida subsp. achromogenes, and the type cultures A. salmonicida subsp. achromogenes NCMB 1110, A. salmonicida subsp. masoucida ATCC 27013 and A. salmonicida subsp. smithia CCM 4103. A total of 95 strains of atypical A. salmonicida were separated into 7 groups (I to VII) based on the plasmid profiles. Eighteen strains of atypical A. salmonicida had no common plasmid profile. The type strain NCMB 1110 and the reference strain NCMB 1109 were included in group IV, and the type strain ATCC 27013 in group V, but the other reference and type strains had plasmid profiles different from all the other strains. An epidemiological link was documented between strains collected from different farms/localities in each of groups I, III, V and VII. Physiological and biochemical characterizations were performed for 93 of the strains to investigate phenotypic differences between the plasmid groups. Group VII strains and 3 strains with no common plasmid profile differed from the other groups in being catalase-negative. Differences in phenotypic characteristics were shown between the plasmid groups. However, significant variations in reactions for several phenotypic characteristics also occurred within each of the groups I to VII. The present study indicates that plasmid profiling may give useful epidemiological information during outbreaks of atypical A. salmonicida infections in fish. Additional comprehensive phenotypic characterisation is of limited value since the phenotypic characteristics in each plasmid group are not uniform.

Presentation of dermatological images on the Internet.

In this paper, we focused on selected problems of integrating and presenting medical images organised in a World Wide Web (WWW) database. To solve these problems we developed a prototype of a bilingual (Slovenian and English) WWW database of medical images for the field of dermatology. This dermatology database includes a graphic interface with four modes of access: (1) browsing, (2) searching, (3) comparison of images, and (4) self-testing. The quantity and quality of requests to this WWW database was estimated with log file analysis. There was a steady increase in the number of users and volume of data transferred from the dermatology WWW database.

Organization and dissemination of multimedia medical databases on the WWW.

In the paper, we focus on the problem of building and disseminating multimedia medical databases on the World Wide Web (WWW). The current results of the ongoing project of building a prototype dermatology images database and its WWW presentation are presented. The dermatology database is part of an ambitious plan concerning an organization of a network of medical institutions building distributed and federated multimedia databases of a much wider scale.

Increase in contact allergy to fragrances: patch-test results 1989-1998.

We report the results of patch tests with fragrance-mix as a part of the standard series carried out over the last 10 years (1989-1998) during routine testing of 6129 patients in our department. 5.9% of the total number of patients who were patch tested were positive to fragrance mix. The sex ratio was 2.3:1 with a female predominance. In 1989-1993, the frequency of contact sensitivity to fragrance mix was 3.9% (4.9% for females and 2.1% for males). This rate rose both in female and male patients during the observed period of time and attained 8.9% (females) and 4.1% (males) in 1994-1998; the overall frequency in 1994-98 was 7.5%. This rising trend, which was statistically significant, might be the consequence of an increased use of cosmetics and toiletries containing fragrances in our population.

Phenotypic and genotypic characterization of Vibrio viscosus sp. nov. and Vibrio wodanis sp. nov. isolated from Atlantic salmon (Salmo salar) with 'winter ulcer'.

Two groups of Vibrio strains isolated from Atlantic salmon with 'winter ulcer' were characterized phenotypically and genotypically. The data obtained indicated that each of the two groups represented a new species in the genus Vibrio. The names Vibrio viscosus sp. nov. [type strain NVI 88/478T (= NCIMB 13584T)] and Vibrio wodanis sp. nov. [type strain NVI 88/441T (= NCIMB 13582T)] are proposed for the new species. V. viscosus strains exhibited a similar total DNA RFLP pattern and a similar plasmid DNA profile. DNA relatedness (hydroxyapatite method) of the V. viscosus type strain to nine other V. viscosus strains was 81-93% at 60 degrees C. Divergence within related sequences was 0.0-1.5% and relatedness at 75 degrees C was 74-100%. V. wodanis strains exhibited marked heterogeneity on the basis of RFLP analysis and plasmid profiles. DNA relatedness of the V. wodanis type strain to 10 other V. wodanis strains was 66-94% at 60 degrees C. Divergence within related sequences was 0.0-1.5% and relatedness at 75 degrees C was 55-97%. Relatedness between V. viscosus and V. wodanis type strains was approximately 20%. Among other Vibrio species, the closest relative of V. viscosus was Vibrio marinus (ATCC 15381T) (43% relatedness at 60 degrees C) and that of V. wodanis was Vibrio logei (ATCC 15382) (57% relatedness at 60 degrees C). These same pairs were the closest phenotypic relatives. DNA sequence analysis of the 16S rRNA gene of V. viscosus indicated an intimate relationship to V. marinus. A total evaluation of the results, however, supports V. viscosus to be a separate species in the genus Vibrio. The analysis of the sequence of the 16S rRNA gene of V. wodanis supports that V. logei (ATCC 15382) was the most related species. Ability to degrade casein, oxidative production of acid from trehalose and production of lysine decarboxylase are important biochemical tests that will differentiate between V. viscosus, V. wodanis, V. marinus (ATCC 15381T) and V. logei (ATCC 15382).