Does Social Capital Benefit Older Adults' Health and Well-Being? The Mediating Role of Physical Activity.

Objectives: To assess whether social capital benefits older adults' self-rated health and well-being and whether physical activity mediates this relation. Methods: A survey study was conducted among members of a sociocultural organization (age ≥55 years), both cross-sectionally (baseline Time 1; N = 959) and longitudinally (3-year follow-up Time 2; N = 409). Results: Specific indicators of social capital were positively, though modestly, related to health and well-being at Time 1 and Time 2. Experienced connectedness with age peers emerged as the strongest predictor. Physical activity only mediated the relation with experienced safety in society. Discussion: The relative importance of older adults' experienced connectedness with their age peers underlines the importance of internalized group membership as a determinant of their health and well-being. Physical activity seems to play only a minor mediating role.

Is a single day patient friendly methyl aminolevulinate photodynamic therapy illumination scheme for superficial basal cell carcinoma feasible? A randomized multicenter pilot trial.

BACKGROUND: Topical methyl aminolevulinate photodynamic therapy (MAL-PDT) is highly effective for the treatment of superficial basal cell carcinoma (sBCC). Current European treatment protocol requires two hospital visits, which is costly and unpractical. The aim of this study was to evaluate the efficacy of fractionated MAL-PDT, using two light fractions at 3 and 4 h compared to illumination at 3 and 5 h after MAL-application. METHODS: Thirty patients were randomized into two groups. The first group received illumination at 3 and 4 h (20 + 55 J/cm2) after MAL-application (3/4 group). In the other group, two light fractions were performed at 3 and 5 h (20 + 55 J/cm2) after MAL-application (3/5 group). The lesion response was evaluated at 3 and 12 months posttreatment. RESULTS: In the 3/5 group, 70.0% showed a complete response (CR) at 3 months compared to 63.6% in the other group. At 12 months, 100% showed a CR in the 3/5 group compared to 80.0% in the other group. However, most failures/recurrences were eventually due to the presence of a more aggressive BCC subtype, mostly caused by sampling error of the primary punch biopsy. CONCLUSION: Single day protocol for MAL-PDT for sBCC is feasible and this study shows promising results.

Aortic Arch Morphology and Aortic Length in Patients with Dissection, Traumatic, and Aneurysmal Disease.

OBJECTIVES: To assess aortic arch morphology and aortic length in patients with dissection, traumatic injury, and aneurysm undergoing TEVAR, and to identify characteristics specific to different pathologies. METHOD: This was a retrospective analysis of the aortic arch morphology and aortic length of dissection, traumatic injury, and aneurysmal patients. Computed tomography imaging was evaluated of 210 patients (49 dissection, 99 traumatic injury, 62 aneurysm) enrolled in three trials that received the conformable GORE TAG thoracic endoprosthesis. The mean age of trauma patients was 43 ± 19.6 years, 57 ± 11.7 years for dissection and 72 ± 9.6 years for aneurysm patients. A standardized protocol was used to measure aortic arch diameter, length, and take-off angle and clockface orientation of branch vessels. Differences in arch anatomy and length were assessed using ANOVA and independent t tests. RESULTS: Of the 210 arches evaluated, 22% had arch vessel common trunk configurations. The aortic diameter and the distance from the left main coronary (LMC) to the left common carotid (LCC) were greater in dissection patients than in trauma or aneurysm patients (p < .001). Aortic diameter in aneurysm patients was greater compared with trauma patients (p < .05). The distances from the branch vessels to the celiac artery (CA) were greater in dissection and aneurysm patients than in trauma patients (p < .001). The take-off angle of the innominate (I), LCCA, and left subclavian (LS) were greater, between 19% and 36%, in trauma patients than in dissection and aneurysm patients (p < .001). Clockface orientation of the arch vessels varies between pathologies. CONCLUSIONS: Arch anatomy has significant morphologic differences when comparing aortic pathologies. Describing these differences in a large sample of patients is beneficial for device designs and patient selection.

The impact of image-size manipulation and sugar content on children's cereal consumption.

Previous studies have demonstrated that portion sizes and food energy-density influence children's eating behavior. However, the potential effects of front-of-pack image-sizes of serving suggestions and sugar content have not been tested. Using a mixed experimental design among young children, this study examines the effects of image-size manipulation and sugar content on cereal and milk consumption. Children poured and consumed significantly more cereal and drank significantly more milk when exposed to a larger sized image of serving suggestion as compared to a smaller image-size. Sugar content showed no main effects. Nevertheless, cereal consumption only differed significantly between small and large image-sizes when sugar content was low. An advantage of this study was the mundane setting in which the data were collected: a school's dining room instead of an artificial lab. Future studies should include a control condition, with children eating by themselves to reflect an even more natural context.

Comparative genomic analysis of Salmonella enterica subsp. enterica serovar Weltevreden foodborne strains with other serovars.

Salmonella enterica subsp. enterica serovar Weltevreden is a dominant serovar associated with foodborne gastroenteritis in South-East Asia and emerging in Europe associated with fresh vegetables. Here we compared the genome of strain 2007-60-3289-1 linked to an alfalfa sprout outbreak in Scandinavia with a S. Weltevreden strain isolated from scallops in the USA and with other S. enterica serovars. A unique plasmid pSW82 was identified for S. Weltevreden carrying a two-component type II non-ribosomal peptide synthase/polyketide synthase. Analysis of all available complete S. enterica genomes identified differences for presence of type VI secretion systems and carbohydrate metabolic pathways. Differential transcription thereof was observed when S. Weltevreden strains were grown in vitro or on sprouts.

Factors determining social distance toward people with depression among community pharmacists.

OBJECTIVE: The aim of the present study was to measure stigma among community pharmacists regarding patients with depression, using social distance as a proxy measure. Furthermore, factors influencing stigmatization were identified. METHODS: A survey consisting of questions regarding social distance, cognitive attitude components, stereotypical beliefs and behavioural tendencies towards pharmaceutical care for patients with depression was carried out. The survey was completed by 149 community pharmacists with a response rate of 82%. Multiple linear regression analysis was applied to the results of the survey. RESULTS: Most pharmacists endorsed social distance towards patients with depression. The social distance further increased the more the situation described implied "social closeness". Age, depression care attitude, the stereotypical belief "danger to others" and the level of involvement pharmacists showed towards patients with depression were found to predict social distance. The present model explains 32.3% of the variance in social distance. CONCLUSION: The results showed that community pharmacists have a stigmatizing attitude towards patients with depression. The determinants found to influence these stigmatizing attitudes might be addressed when trying to reduce stigmatization of patients with depression.

First Report of Asian Brown Rot Caused by Monilia polystroma on Apricot in Switzerland.

A survey for Monilia fructicola (G. Winter) Honey on apricots (Prunus armeniaca L.) was conducted in July and August 2009 and 2010 in Canton Wallis, Switzerland. Mummies of fruits showing brown rot were collected and isolations were conducted. Nearly 200 fungal isolates, tentatively identified as M. fructigena, were retested with a multiplex PCR (1). With the Agilent 2100 Bioanalyzer (Agilent Technologies, Basel, Switzerland) instead of 1.5% agarose gels, the 23 bp difference between the diagnostic fragments of M. fructigena and M. polystroma van Leeuwen (1) could clearly be scored. M. polystroma was diagnosed in 3 of 65 and 1 of 132 isolates collected in 2009 (13 orchards) and 2010 (10 orchards), respectively. The internal transcribed spacer (ITS) regions of four isolates (09-G4, 09-P16, 09-S5, and 10-C6) were amplified and sequenced (4). The four sequences (GenBank No. JN128835) as well as those of the Hungarian isolate UFT (AM937114 [3]) were identical and highly similar to the type sequence for M. polystroma (Y17876 [2]). The type sequence had a "T" at position 414, which was lacking in the other five sequences. The genomic region of unknown function used by Côté et al. (1) to develop their PCR diagnostic tool was sequenced for isolate 09-G4 with primers MFG.for (3) and M Poly rev 5'-CCACTTACATTTTTGGCTATTG-3'. The Swiss isolate (GenBank No. JN128836) and the Hungarian isolate UFT (AM937120) sequences were identical. The pathogenicity of isolate 09-G4 was tested on Golden delicious apples. Six apples were surface sterilized (70% ethanol), halved, and placed in sterile plastic boxes cut-side down. Further, six half apples were wounded in the center with a round scalpel with a diameter of 1 cm and inoculated with a round, potato dextrose agar (PDA) plug (1-cm diameter) of actively growing mycelium (5- to 7-day-old culture). Control apples (six halves) were treated with a PDA plug without mycelium. All fruits were incubated at 20°C with a 12-h light 12-h dark cycle. Seven days after inoculation, typical brown rot symptoms were visible on all inoculated fruits. Mock inoculated fruits remained healthy. Three inoculated halves, in addition to the brown rot symptoms, also produced sporodochia and around the inoculation point the tissue become black. With the multiplex PCR (1), M. polystroma was identified as the pathogen causing brown rot symptoms on the inoculated apples. The ellipsoid single-cell hyaline conidia of isolate 09-G4 grown on the Golden delicious apples averaged 15.2 ± 4.0 × 8.97 ± 1.1 µm and were the expected size for M. polystroma conidia (14.9 to 9.1 µm [4]). The first evidence of a new Monilia species was reported by Fulton et al. (2). They found that M. fructigena isolates from Japan were distinguishable from European isolates by five base substitutions in the ITS region (four in ITS1 and one in ITS2). Later, van Leeuwen et al. (4) found that the two groups of isolates could also be distinguished by morphological differences and described the new species as M. polystroma. According to the Centre for Agricultural Bioscience International, the impact of M. polystroma in a new area is presumed to be the same or very similar to that of M. fructigena. To our knowledge, this is the first report of M. polystroma in Swiss orchards. References: (1) M.-J. Côté et al. Plant Dis. 88:1219, 2004. (2) C. E. Fulton et al. Eur. J. Plant Pathol. 105:495, 1999. (3) M. Petróczy and L. Palkovics. Eur. J. Plant Pathol. 125:343, 2009. (4) G. C. M. van Leeuwen et al. Mycol. Res. 106:444, 2002.

Antibiosis activity of Pantoea agglomerans biocontrol strain E325 against Erwinia amylovora on apple flower stigmas.

Pantoea agglomerans E325, the active ingredient in a commercial product for fire blight control, was previously shown in vitro to produce a unique alkaline- and phosphate-sensitive antibiotic specific to Erwinia amylovora. Antibiosis was evaluated as a mode of antagonism on flower stigmas using two antibiosis-deficient mutants. On King's medium B, mutants E325ad1 and E325ad2 have stable smooth-butyrous or hypermucoid colony morphologies, respectively, and the parental strain E325 exhibits phenotypic plasticity with predominantly hypermucoid colonies accompanied by slower-growing, smooth-butyrous colonies. Mutants were tested against E. amylovora on stigmas of detached flowers of crab apple (Malus mandshurica) in growth chambers and apple (Malus domestica) in the orchard. Epiphytic fitness of the antibiosis-negative mutants was similar or greater than the parental strain as determined by relative area under the population curve (RAUPC). In laboratory and orchard trials, both mutants had significantly lower inhibitory activity against the pathogen (i.e., less reduction of E. amylovora RAUPC) compared with the parental strain. E325 and the mutants caused similar decreases in pH in a broth medium, indicating that acidification, which was previously reported as a possible mechanism of pathogen inhibition on stigmas, is not directly related to antibiosis. In this study we provide the first evidence for E325 antibiosis involved in E. amylovora growth suppression on apple flower stigmas.

First Report of the ß-Tubulin E198A Mutation Conferring Resistance to Methyl Benzimidazole Carbamates in European Isolates of Monilinia fructicola.

The causal agent of brown rot on stone and pome fruits, Monilinia fructicola (G. Wint.), is a quarantine pathogen in Europe. It has been detected in Austria (later eradicated), Spain, the Czech Republic, Italy, Germany, and Switzerland (1). In the United States and other countries, M. fructicola isolates were reported to show resistance to different classes of fungicides, including methyl benzimidazole carbamates (MBC) (2). Lichou et al. (2) reported the presence of isolates resistant to the MBC carbendazim in France, but the mechanisms inducing MBC resistance in these isolates were not studied. Ma et al. (3) in California, and more recently, Zhu et al. (4) in South Carolina, demonstrated that the molecular mechanisms accounting for low and high levels of resistance to MBC fungicides in M. fructicola isolates were the mutations H6Y and E198A, respectively, in the ß-tubulin gene. Four M. fructicola isolates each from Italy, France, Spain, and Switzerland (16 isolates total), all having an unknown level of MBC resistance, were selected. In each isolate, the section of the ß-tubulin gene containing the two potentially mutant codons was PCR-amplified with the primers TubA and TubR1 (3) and the amplicons were sequenced directly. Sequence analysis revealed the amino acid histidine (H) at codon 6 in all the isolates, which would not predict MBC resistance, while alanine (A) at codon 198 (the mutation predictive of a high level of MBC resistance) was found in all isolates from Spain and Switzerland and in three isolates each from France and Italy. A representative sequence of the four identical partial ß-tubulin gene sequences from the Swiss isolates was submitted to GenBank under the Accession No. HQ709265. All isolates were tested in a potato dextrose agar (PDA) petri dish assay for resistance to the MBC fungicide thiophanate-methyl (Nippon Soda Co., Ltd., Tokyo, Japan) at the discriminatory dose of 50 µg/ml (4). All isolates with the E198A mutation were able to grow on the media, while the two isolates without the E198A mutation were not able to grow. The result indicated that most isolates had a high level of resistance to the MBC fungicide. To our knowledge, this is the first report of the presence of the E198A mutation conferring resistance to MBC fungicides in European isolates of M. fructicola. As the mutation appears to be widely distributed, we anticipate that MBC fungicides may be ineffective at controlling brown rot in countries with occurrence of M. fructicola. References: (1) M. Hilber-Bodmer et al. Plant Dis. 94:643, 2010. (2) J. Lichou et al. Phytoma 547:22, 2002. (3) Z. H. Ma et al. Appl. Environ. Microbiol. 69:7145, 2003. (4) F. X. Zhu et al. Plant Dis. 94:1511, 2010.

Itch and scratching as predictors of time to clearance of psoriasis with narrow-band ultraviolet B therapy.

BACKGROUND: Narrow-band ultraviolet (UV) B phototherapy is an effective treatment for psoriasis. However, there is considerable variability in the number of treatment sessions needed to achieve psoriasis clearance. While several clinical and treatment-related factors predict time to clearance, the effect of itching and scratching on the number of irradiation sessions is insufficiently understood. OBJECTIVE: Predictors of the time to clearance were assessed in patients with psoriasis who were referred for UVB treatment in a randomized double-blind comparison of irradiation regimens for UVB phototherapy. METHODS: After randomization to either UVB irradiation with a suberythematogenic or an erythematogenic regimen, patients were irradiated with 20% and 40% incremental doses, respectively, three times weekly. The Psoriasis Area and Severity Index (PASI) score was measured at baseline and every 4 weeks, and itching and habitual scratching were measured at baseline. RESULTS: Among the 77 patients who achieved psoriasis clearance (90% reduction of PASI), itching and scratching were correlated with the number of irradiation sessions needed to achieve clearance, with higher levels of itch and scratching predicting more sessions. These effects remained significant after controlling for the initial PASI score, irradiation schemes, minimal erythema dose (MED), skin type, cumulative dose, protocol adjustments and lifestyle factors (smoking habits and alcohol consumption). CONCLUSIONS: Patients with higher levels of itch and scratching need more irradiation sessions to achieve clearance of psoriasis with UVB phototherapy. Systematic assessment of the severity of itch and scratching, followed by short-term itch-coping programmes for patients at risk, might be a cost-effective, adjunct to UVB therapy.

Narrowband ultraviolet B therapy in psoriasis: randomized double-blind comparison of high-dose and low-dose irradiation regimens.

BACKGROUND: Ultraviolet (UV) B phototherapy is an established treatment option for psoriasis. The optimum dosage regimen still has to be determined. Within-subject comparisons do not take into account the systemic effects of UVB phototherapy. The area of the body treated with low-dose UVB may benefit from the systemic effects of the site treated with a higher UVB dose. OBJECTIVES: To study the time to clearance in patients with psoriasis in a randomized controlled trial, in which patients were treated with narrowband UVB in either a high-dose or a low-dose regimen. METHODS: One hundred and nine patients were randomized to a high-dose regimen (group 1) or to a low-dose regimen (group 2). Patients of group 1 and 2 were irradiated with 40% and with 20% incremental doses, respectively, three times weekly. Psoriasis Area and Severity Index (PASI) was measured at baseline and at every 4-week control visit. Treatment was stopped in cases of clearance (90% reduction of baseline PASI). RESULTS: No significant differences were found in the number of patients achieving clearance. The high-dosage scheme resulted in four fewer treatments with no significant differences in cumulative UV dose, although more protocol adjustments were required in the beginning of the study because of erythema. After 3 months a significantly better clinical outcome was seen after high-dose UVB therapy. CONCLUSIONS: High-dose UVB therapy results in fewer treatments with better long-term efficacy, with cost-effective benefits for hospital and patients. Therefore UVB phototherapy in a high-dose regimen for psoriasis is recommended. However, a protocol adjustment in the second week with a high-dose regimen is desirable to prevent erythema.

Induction of protoporphyrin IX by aminolaevulinic acid in actinic keratosis, psoriasis and normal skin: preferential porphyrin enrichment in differentiated cells.

BACKGROUND: In photodynamic therapy the endogenous photosensitizer protoporphyrin IX (PpIX) is synthesized following topical application of aminolaevulinic acid (ALA). However, different tissues have distinct PpIX-accumulating properties, due to differences in penetration of ALA through the stratum corneum and/or alterations in the haem cycle. Preferential PpIX accumulation has been reported in terminally differentiated cell cultures but ex vivo data are lacking. OBJECTIVES: To study the intrinsic PpIX-accumulating capabilities of skin explants from lesional and nonlesional skin in psoriasis and actinic keratosis, with normal skin serving as a control, and to study PpIX accumulation in relation to differentiation status in normal skin. METHODS: Skin explants from patients with psoriasis, actinic keratosis and normal skin were incubated with ALA and PpIX was measured spectrofluorometrically. PpIX was measured in basal (beta1-integrin-positive) and suprabasal (beta1-integrin-negative) keratinocytes in normal skin. In addition, PpIX accumulation was studied in cell cultures at different levels of confluence and after induction of terminal differentiation. RESULTS: No significant differences in PpIX content were found between the different tissues. However, increased PpIX was found in beta1-integrin-negative compared with beta1-integrin-positive cells. In addition, in subconfluent cell cultures less PpIX was found compared with confluent cell cultures. Induction of terminal differentiation in vitro, however, resulted in less PpIX, which was likely to be related to cell volume. CONCLUSIONS: As no significant differences in PpIX synthesis could be found between the different tissue types, these data emphasize the importance of the penetration route of ALA through the stratum corneum. Preferential PpIX accumulation observed in suprabasal epidermal keratinocytes and confluent cell cultures points towards a terminal differentiation-specific effect.

Pretreatment to enhance protoporphyrin IX accumulation in photodynamic therapy.

The response rates of photodynamic therapy (PDT) vary widely. Limited uptake of topically applied 5-aminolaevulinic acid (ALA), or its methyl ester (MAL), and suboptimal production of protoporphyrin IX (PpIX) may account for these differences. Recently, we demonstrated that hyperkeratosis is an important negative factor in ALA uptake. This review has its focus on pretreatment of the skin in order to improve the clinical outcome of ALA/MAL PDT. Pretreatment of hyperkeratosis can be achieved with keratolytics, curettage/debulking, tape stripping, microdermabrasion or laser ablation. Penetration enhancers may alter the composition or organization of the intercellular lipids of the stratum corneum. Several studies have been performed on the use of dimethyl sulfoxide, azone, glycolic acid, oleic acid and iontophoresis to increase the penetration of ALA. As PpIX production is also dominated by temperature-dependent processes, elevating skin temperature during ALA application may also improve treatment results. Another approach is the use of additives that interact with the heme biosynthetic pathway, e.g. by removing ferrous iron with iron-chelating substances such as: ethylenediaminetetraacetic acid; 3-hydroxypyridin-4-ones; 1,2-diethyl-3-hydroxypyridin-4-one-hydrochloride; and desferrioxamine. In conclusion, simple pretreatments or additions to the regular practice of PDT, aimed to optimize intralesional PpIX content, improve the clinical outcome.

Heterogeneity of fluorescence in psoriasis after application of 5-aminolaevulinic acid: an immunohistochemical study.

BACKGROUND: Psoriasis has been shown to highly accumulate protoporphyrin IX (PpIX), but a variable distribution within plaques after fluorescence diagnosis is seen. It is unknown what causes this heterogeneity of fluorescence in psoriatic skin, despite adequate keratolytic treatment. Variations in fluorescence might explain the variable and the mostly partial clinical response of psoriasis seen after photodynamic therapy (PDT). OBJECTIVES: This study examines morphological and immunohistochemical differences in inhomogeneous PpIX-induced fluorescence in stable plaque psoriasis. MATERIALS AND METHODS: Fourteen patients with stable plaque psoriasis were included in this study. In each patient one psoriatic plaque was incubated with 20% 5-aminolaevulinic acid (ALA) ointment for 3 h after keratolytic treatment. Fluorescence diagnosis with ALA-induced porphyrins (FDAP) was performed and subsequently high- and low-fluorescent psoriatic skin samples were biopsied. Biopsies were investigated with respect to histological hyperkeratosis (thickness of stratum corneum), proliferation (Ki-67 antigen), keratinization (K10, filaggrin) and inflammation (CD3). Digital images acquired with FDAP were analysed using image analysis software. RESULTS: Inhomogeneous fluorescence was seen in 12 of the 14 plaques. A significantly thicker stratum corneum was found in low-fluorescent psoriatic skin compared with highly fluorescent skin. Fluorescence intensity and thickness of the stratum corneum proved to be negatively correlated. The variable-fluorescent parts of the lesional psoriatic skin showed no differences in epidermal proliferation, keratinization or inflammation. CONCLUSIONS: Heterogeneous ALA-induced fluorescence in psoriasis plaques related to inhomogeneous distribution of PpIX in the epidermis may result from differences in penetration of ALA and/or light within a plaque caused by differences in stratum corneum thickness. The variable clinical response seen after PDT in psoriasis could be explained by this. These findings are consistent with the general assumption that optimal desquamation prior to FDAP or PDT is required for the most favourable results.

A placebo-controlled randomized study on the clinical effectiveness, immunohistochemical changes and protoporphyrin IX accumulation in fractionated 5-aminolaevulinic acid-photodynamic therapy in patients with psoriasis.

BACKGROUND: Topical 5-aminolaevulinic acid (ALA)-photodynamic therapy (PDT) for the treatment of psoriasis has been evaluated in a few studies. In these studies different treatment parameters were used, there was a variable clinical response, and a nonhomogeneous fluorescence was seen after irradiation with Wood's light. OBJECTIVES: To study the clinical effectiveness, immunohistochemical changes and protoporphyrin IX accumulation in ALA-PDT in patients with psoriasis. Eight patients with stable plaque psoriasis with symmetrical involvement were included in the study. Two symmetrical plaques were randomly allocated to PDT either with 10% ALA or with placebo. Irradiation consisted of 2 and 8 J cm(-2) with a dark interval of 2 h (Waldmann PDT 1200 L, 600-750 nm, 40 mW cm(-2)) once weekly for 4 weeks. Before, during and after irradiation, fluorescence diagnosis was performed. Biopsies were taken at baseline, week 1 and week 6 for immunohistochemical assessment. Psoriatic plaques were clinically assessed using the plaque severity (sum) score. Fluorescence diagnosis was performed and expression of immunohistochemical markers for proliferation, differentiation and T-cell infiltration [Ki67, keratin 10 (K10), CD4, CD8 and CD45RO] was assessed. RESULTS: From week 1 up to week 6, ALA-PDT gave a significant reduction in the number of Ki67+ nuclei, while the K10 expression increased. After 6 weeks significant improvement was observed for CD8 and CD45RO. These changes were absent in the placebo-treated lesions. The sum scores were also significantly lower in the ALA-treated plaques. Heterogeneity of macroscopic fluorescence was observed during treatment despite keratolytic treatment. CONCLUSIONS: The present study shows that clinical improvement during fractionated ALA-PDT in psoriasis parallels histological improvement as seen in normalization of epidermal proliferation, differentiation and infiltration of relevant T-cell subsets. Optimizing the current treatment protocol may increase clinical efficacy further.

The differential effect of broad band vs narrow band UVB with respect to photodamage and cutaneous inflammation.

Since Fischer reported on the superiority of 313 nm UVB compared with broad band UVB in the treatment of psoriasis, narrow band UVB has become the main phototherapeutical modality in several countries. There is some discussion about the safety and photobiological effects of narrow band UVB. In the present study, narrow and broad band UVB have been compared with respect to parameters for photodamage and inflammation. Fourteen healthy volunteers were randomized in two groups. Both groups were irradiated with three minimal erythema doses (MED) of narrow or broad brand UVB, respectively. Before and 4, 24 and 48 h after irradiation, 6 mm biopsies were taken from immunohistochemical analysis of p53, apoptosis and p16 (photodamage parameters) and T-cells, polymorphonuclear leukocytes (PMN) and Langerhans' cells (inflammatory cells). Mean MED for narrow band UVB was 8.125 times higher than broad band UVB. Significant changes in expression were seen for all parameters except for p16. P53, apoptosis, T-cells and PMN increased, while Langerhans' cell count decreased significantly. No significant differences were seen between the narrow band UVB. In conclusion, following irradiation of three MED narrow band UVB and broad band UVB safety parameters for carcinogenesis and inflammation were induced to the same extent. As narrow band UVB is more effective than broad band UVB, the present study suggests superiority of narrow band UVB as a treatment with a better benefit risk ration.

Gene cloning and characterization of multiple alkane hydroxylase systems in Rhodococcus strains Q15 and NRRL B-16531.

The alkane hydroxylase systems of two Rhodococcus strains (NRRL B-16531 and Q15, isolated from different geographical locations) were characterized. Both organisms contained at least four alkane monooxygenase gene homologs (alkB1, alkB2, alkB3, and alkB4). In both strains, the alkB1 and alkB2 homologs were part of alk gene clusters, each encoding two rubredoxins (rubA1 and rubA2; rubA3 and rubA4), a putative TetR transcriptional regulatory protein (alkU1; alkU2), and, in the alkB1 cluster, a rubredoxin reductase (rubB). The alkB3 and alkB4 homologs were found as separate genes which were not part of alk gene clusters. Functional heterologous expression of some of the rhodococcal alk genes (alkB2, rubA2, and rubA4 [NRRL B-16531]; alkB2 and rubB [Q15]) was achieved in Escherichia coli and Pseudomonas expression systems. Pseudomonas recombinants containing rhodococcal alkB2 were able to mineralize and grow on C(12) to C(16) n-alkanes. All rhodococcal alkane monooxygenases possessed the highly conserved eight-histidine motif, including two apparent alkane monooxygenase signature motifs (LQRH[S/A]DHH and NYXEHYG[L/M]), and the six hydrophobic membrane-spanning regions found in all alkane monooxygenases related to the Pseudomonas putida GPo1 alkane monooxygenase. The presence of multiple alkane hydroxylases in the two rhodococcal strains is reminiscent of other multiple-degradative-enzyme systems reported in Rhodococcus.

New alkane-responsive expression vectors for Escherichia coli and pseudomonas.

We have developed Escherichia coli and Pseudomonas expression vectors based on the alkane-responsive Pseudomonas putida (oleovorans) GPo1 promoter PalkB. The expression vectors were tested in several E. coli strains, P. putida GPo12 and P. fluorescens KOB2Delta1 with catechol-2,3-dioxygenase (XylE). Induction factors ranged between 100 and 2700 for pKKPalk in E. coli and pCom8 in Pseudomonas strains, but were clearly lower for pCom8, pCom9, and pCom10 in E. coli. XylE expression levels of more than 10% of total cell protein were obtained for E. coli as well as for Pseudomonas strains.