Laser-assisted photodynamic therapy for superficial basal cell carcinoma and Bowen's disease: a randomized intrapatient comparison between a continuous and a fractional ablative CO2 laser mode.

BACKGROUND: Laser-assisted photodynamic therapy is being explored as a method to enhance efficacy of photodynamic therapy (PDT). OBJECTIVE: To compare a continuous (CL) and a fractional (FL) ablative CO2 laser-assisted methyl aminolevulinate (MAL) PDT in the management of superficial basal cell carcinoma (sBCC) and Bowen's disease (BD). METHODS: Thirty treatment areas in fifteen patients with inoperable, histologically verified sBCC or BD received CL or FL after intrapatient randomization. Laser treatment was followed by MAL application and illumination occurred 3 h later. This treatment was repeated after 2 weeks. An equivalence analysis was performed on the primary endpoint efficacy, while secondary endpoints pain, side-effects and aesthetics were evaluated using paired samples tests. Patients were also asked for their preferred treatment. RESULTS: An excellent efficacy of 92.9% (sBCC, 100%; BD, 80%) was found in both CL + PDT and FL + PDT after 12 months. Equivalence could not be established. Little pain was perceived in most patients during PDT illumination. PDT treatment in FL + PDT was less painful, significantly during the second treatment (P = 0.026). Side-effects were mild to moderate with erythema being the most frequent immediate side-effect, followed by oedema, crusting and burning sensation. Pigmentary changes occurred in 21% (CL + PDT) to 29% (FL + PDT), and aesthetics were good to excellent in all patients. CL + PDT and FL + PDT did not significantly differ in side-effects (P = 0.219-1.000) or aesthetics (P = 0.157-1.000). CONCLUSIONS: Results in this pilot study support the promising role of laser-assisted PDT. Both treatment arms demonstrated the same efficacy as well as comparable side-effects and aesthetics. PDT illumination was significantly less painful in the FL + PDT group, suggesting a preference for FL + PDT. The authors recommend further investigation with a larger sample size, a subgroup analysis between sBCC and BD and comparison of different treatment protocols before one technique could be preferred to another.

Mohs micrographic surgery for basal cell carcinoma: evaluation of the indication criteria and predictive factors for extensive subclinical spread.

BACKGROUND: The incidence of basal cell carcinoma (BCC) is rising and BCC treatment has an important impact on healthcare budget. Mohs micrographic surgery (MMS) has the highest 5-year cure rate but is an expensive technique. OBJECTIVES: To study the indication criteria for MMS, using a series of 1062 patients treated for facial BCCs between 1998 and 2011. METHODS: The accuracy of the indication criteria was evaluated by comparing the characteristics of BCC requiring one vs. more than one round of MMS. Predictors for extensive subclinical spread (three or more rounds) were examined using the preoperative patient and all tumour characteristics. RESULTS: BCCs with a surface > 1 cm(2) and aggressive histology (morphoeaform and micronodular), and a patient age > 80 years are strong predictors for two or more rounds of MMS being required. Extensive subclinical spread was present in recurrent tumours, morphoeaform BCC or BCC with mixed histology. CONCLUSIONS: We found that tumour size and aggressive histology are the strongest indication criteria for MMS. Recurrence and aggressive histology are predictors for extensive subclinical spread but not for two or more rounds of MMS. Evidence-based indications for MMS are necessary to ensure cost-effective management of BCC.

The receptor for advanced glycation end products (RAGE) enhances autophagy and neutrophil extracellular traps in pancreatic cancer.

Neutrophil extracellular traps (NETs) are formed when neutrophils expel their DNA, histones and intracellular proteins into the extracellular space or circulation. NET formation is dependent on autophagy and is mediated by citrullination of histones to allow for the unwinding and subsequent expulsion of DNA. NETs have an important role in the pathogenesis of several sterile inflammatory diseases, including malignancy, therefore we investigated the role of NETs in the setting of pancreatic ductal adenocarcinoma (PDA). Neutrophils isolated from two distinct animal models of PDA had an increased propensity to form NETs following stimulation with platelet activating factor (PAF). Serum DNA, a marker of circulating NET formation, was elevated in tumor bearing animals as well as in patients with PDA. Citrullinated histone H3 expression, a marker of NET formation, was observed in pancreatic tumors obtained from murine models and patients with PDA. Inhibition of autophagy with chloroquine or genetic ablation of receptor for advanced glycation end products (RAGE) resulted in decreased propensity for NET formation, decreased serum DNA and decreased citrullinated histone H3 expression in the pancreatic tumor microenvironment. We conclude that NETs are upregulated in pancreatic cancer through RAGE-dependent/autophagy mediated pathways.

Classification of segmental vitiligo on the trunk.

BACKGROUND: Segmental vitiligo is characterized by a unilateral and localized distribution. A classification for segmental vitiligo on the face has been proposed, while this is still unavailable for lesions on the trunk. OBJECTIVES: To design a classification for segmental vitiligo lesions on the trunk by evaluating their distribution pattern. METHODS: In this retrospective observational study, the distribution pattern of 106 segmental vitiligo lesions on the trunk was analysed and classified into recurring subtypes. RESULTS: Segmental vitiligo was more frequently observed on the ventral side (85·8%) than the lateral side (52·8%) or the back (36·8%) of the trunk. Based on recurring similarities in distribution patterns, lesions were categorized into six distinct subtypes: types 1, 2 and 3 involved the upper part of the trunk, type 4 and 5 the middle part and type 6 the lower part of the trunk. In total, 67·9% of all segmental vitiligo lesions fitted into these subtypes. The most frequent type of lesion was subtype 3 (22·6%), which showed a characteristic V-shaped pattern on the upper trunk, followed by subtype 5 (17·9%), with a band-like pattern on the lateral side, and subtype 6 (13·2%), with a rectangular depigmentation on the lower abdomen. CONCLUSIONS: Based on our observations we suggest a new classification into six subtypes for segmental vitiligo lesions on the trunk, allowing categorization of the majority of segmental vitiligo patterns on the trunk. This may have important prognostic and diagnostic significance in early developing lesions.

A microarray analysis of temporal gene expression profiles in thermally injured human skin.

Partial-thickness burns incite a multitude of responses which eventually culminate in cutaneous wound repair. We hypothesized that these events would evoke extensive alterations in gene expression thereby orchestrating the complexity of spatial and temporal events that characterize "normal" human wound healing. In the present study, gene expression from partial-thickness areas at defined temporal periods (1-3 days, 4-6 days, and 7-18 days) after injury were compared to normal non-wounded skin. Gene alterations proved extensive (2286 genes). Statistically significant alterations were noted among increased and decreased genes expressed in the three different temporal groupings. Our foundational data (based on samples from 45 individuals) provide a comprehensive molecular gene expression portrait of the cutaneous reparative responses that are initiated during the first 17 days after injury. Our efforts also represent an initial endeavor to move beyond the historically defined "morphological phases" of wound repair toward reporting molecular clues that define the temporal sequence of healing in human subjects. Further analysis of genes that are either affected or remain not affected following injury to normal skin is expected to identify potential targets for therapeutic augmentation or silencing.

Disruption of PPARgamma signaling results in mouse prostatic intraepithelial neoplasia involving active autophagy.

Peroxisome proliferator-activated receptor-gamma (PPARgamma) regulates the interface between cellular lipid metabolism, redox status and organelle differentiation. Conditional prostatic epithelial knockout of PPARgamma in mice resulted in focal hyperplasia which developed into mouse prostatic intraepithelial neoplasia (mPIN). The grade of PIN became more severe with time. Electron microscopy (EM) showed accumulated secondary lysosomes containing cellular organelles and debris suggestive of autophagy. Consistent with this analysis the autophagy marker LC-3 was found to be upregulated in areas of PIN in PPARgamma KO tissues. We selectively knocked down PPARgamma2 isoform in wild-type mouse prostatic epithelial cells and examined the consequences of this in a tissue recombination model. Histopathologically grafted tissues resembled the conditional PPARgamma KO mouse prostates. EM studies of PPARgamma- and PPARgamma2-deficient epithelial cells in vitro were suggestive of autophagy, consistent with the prostatic tissue analysis. This was confirmed by examining expression of beclin-1 and LC-3. Gene expression profiling in PPARgamma-/gamma2-deficient cells indicated a major dysregulation of cell cycle control and metabolic signaling networks related to peroxisomal and lysosomal maturation, lipid oxidation and degradation. The putative autophagic phenotypes of PPARgamma-deficient cells could be rescued by re-expression of either gamma1 or gamma2 isoform. We conclude that disruption of PPARgamma signaling results in autophagy and oxidative stress during mPIN pathogenesis.

Electroporation-mediated delivery of a naked DNA plasmid expressing VEGF to the porcine heart enhances protein expression.

Gene therapy is an attractive method for the treatment of cardiovascular disease. However, using current strategies, induction of gene expression at therapeutic levels is often inefficient. In this study, we show a novel electroporation (EP) method to enhance the delivery of a plasmid expressing an angiogenic growth factor (vascular endothelial growth factor, VEGF), which is a molecule previously documented to stimulate revascularization in coronary artery disease. DNA expression plasmids were delivered in vivo to the porcine heart with or without coadministered EP to determine the potential effect of electrically mediated delivery. The results showed that plasmid delivery through EP significantly increased cardiac expression of VEGF compared with injection of plasmid alone. This is the first report showing successful intracardiac delivery, through in vivo EP, of a protein expressing plasmid in a large animal.

Clinical markers and driving mechanisms in melanoma progression: VEGF-C, RhoC, c-Ski/SnoN and EGFR.

This research project aimed at evaluating the clinical and prognostic value of different molecules involved in signalling transduction pathways involved in melanoma progression. Vascular endothelial growth factor-C or VEGF-C induces lymphangiogenesis. This study showed high VEGF-C expression to be associated with the presence of a positive sentinel lymph node. The presence of VEGF-C expression in melanoma cells was associated with reduced disease free and overall survival. RhoC is important in the organization of the actin filamental system. We observed RhoC mRNA and protein expression to be upregulated in a highly metastatic melanoma cell line (DX3aza), whereas only low expression levels were found in a melanoma cell line with low proliferative and invasive capacity (MeWo). RhoC immunoreactivity in melanoma tissue was associated with high Breslow tumour thickness and the presence of ulceration. C-Ski and SnoN have been identified as negative regulators in the TGF-beta pathway. We found a significant association between the presence of nuclear c-Ski and thicker, ulcerated melanomas. SnoN expression was associated with the presence of ulceration and a positive sentinel lymph node. Epidermal Growth Factor Receptor (EGFR) expression has been associated with tumour progression and poor outcome in a variety of solid tumours, EGFR immunoreactivity was more frequently present in patients with a positive sentinel lymph node. EGFR gene amplification was not observed; however, the presence of polysomy was associated with higher Breslow tumour thickness. Treating BLM melanoma cells with different concentrations of cetuximab reduced the invasive capacity of the melanoma cells, without impact on cell viability and growth.

Proximal row carpectomy versus four-corner arthrodesis as a treatment for SLAC (scapholunate advanced collapse) wrist.

Four-corner arthrodesis and proximal row carpectomy are motion-preserving salvage solutions for the scapholunate advanced collapse wrist. We compared both procedures in a non-randomised, retrospective study of 30 cases with a mean follow-up of 3(1/2) years. Pain relief and functional gain were equal in both treatment groups. However, the complication rate was higher in the four-corner arthrodesis group, partly because of the need for internal fixation. The higher incidence of carpal tunnel syndrome following four-corner arthrodesis is intriguing. We also found significant differences in the duration of hospital stay and of postoperative sick leave in favour of proximal row carpectomy. If the cartilage on the capitate head is well preserved (Stages I-II), we prefer proximal row carpectomy because of the socio-economic benefits, the lower complication rate and the ease of the procedure. Degenerative changes following proximal row carpectomy may be a concern in young manual labourers, but have not been observed.

Effect of soil depth and texture on fecal bacteria removal from septic effluents.

This study evaluated the effectiveness of soils with different texture and depth to treat fecal bacteria eluted from a house-hold septic effluent. The assessments were accomplished by leaching undisturbed soil monoliths of 30, 45, and 60cm thickness and 25cm in diameter, representing the four different textural groups and hydraulic loadings recommended by the Kentucky Health Department, with domestic wastewater effluent collected regularly from a house-hold septic system. Eluent concentrations were monitored daily over a 15 day period for fecal coliform and fecal streptococci concentrations. The results of the study indicate an alarming frequency of failure to comply with United States Environmental Protection Agency (USEPA) criteria for depth to groundwater, when using a 30 cm vertical separation distance between the bottom of the drain-field and a limiting soil interface. The treatment performance was especially poor in coarse-textured soils. Although biomat development over time is expected to improve treatment, the high influent levels of fecal bacteria pose great concerns for surface and groundwater contamination. Fine-textured soils generally provided better treatment efficiency and more consistent compliance with EPA standards. Treatment efficiency and compliance usually improved with increasing soil depth, with the 60cm thickness providing the most consistent performance and compliance with maximum discharge limit (MDL) requirements. The findings of this study document a general inadequacy of the 30cm vertical separation distance to provide effective treatment of septic effluents in Kentucky soils, particularly in coarse-textured soils. Considering that increasing the soil depth thickness may be impractical in many marginal soils, complementary or alternative treatment technologies should be adopted to improve treatment efficiency and prevent further deterioration of the quality of water resources.

Nutrient removal from septic effluents as affected by soil thickness and texture.

This study evaluated the effectiveness of soils with different textures and thickness to treat BOD, N and P eluted from household septic effluent. The assessments were accomplished by leaching undisturbed soil monoliths of 30, 45 and 60 cm thickness and 25 cm in diameter, representing the four different textural groups and hydraulic loadings recommended by the Kentucky Health Department, with domestic wastewater effluent collected regularly from a household septic system. Effluent concentrations were monitored daily over a 15d period for biochemical oxygen demand (BOD), total-N, NH4-N, NO3-N and total-P concentrations. The results of the study indicate an alarming frequency of failure to comply with EPA criteria for BOD, total-N and NH4-N concentrations when using a 30 cm vertical separation distance between the bottom of the drain field and a limiting soil interface. The treatment performance was particularly poor in coarse-textured soils, apparently due to insufficient reactive surface area. Although biomat development over time is expected to improve the treatment for some of these parameters, the high influent levels of BOD pose great concerns for surface and groundwater contamination during the early stages of operation. Fine-textured soils generally provided better treatment efficiency and more consistent compliance with EPA standards for BOD, total-N, NH4-N and total-P, as well as greater nitrification/denitrification potential. Treatment efficiency and compliance usually improved with increasing soil depth, with the 60 cm thickness providing the most consistent performance and compliance with MDL requirements. Considering that increasing soil thickness requirements may be impractical in many marginal soils, complementary or alternative treatment technologies should be adopted to improve treatment efficiency and prevent further deterioration of the quality of water resources.

Nam1p, a protein involved in RNA processing and translation, is coupled to transcription through an interaction with yeast mitochondrial RNA polymerase.

Alignment of three fungal mtRNA polymerases revealed conserved amino acid sequences in an amino-terminal region of the Saccharomyces cerevisiae enzyme implicated previously as harboring an important functional domain. Phenotypic analysis of deletion and point mutations, in conjunction with a yeast two-hybrid assay, revealed that Nam1p, a protein involved in RNA processing and translation in mitochondria, binds specifically to this domain. The significance of this interaction in vivo was demonstrated by the fact that the temperature-sensitive phenotype of a deletion mutation (rpo41Delta2), which impinges on this amino-terminal domain, is suppressed by overproducing Nam1p. In addition, mutations in the amino-terminal domain result specifically in decreased steady-state levels of mature mitochondrial CYTB and COXI transcripts, which is a primary defect observed in NAM1 null mutant yeast strains. Finally, one point mutation (R129D) did not abolish Nam1p binding, yet displayed an obvious COX1/CYTB transcript defect. This mutation exhibited the most severe mitochondrial phenotype, suggesting that mutations in the amino-terminal domain can perturb other critical interactions, in addition to Nam1p binding, that contribute to the observed phenotypes. These results implicate the amino-terminal domain of mtRNA polymerases in coupling additional factors and activities involved in mitochondrial gene expression directly to the transcription machinery.

Up-regulation of nuclear and mitochondrial genes in the skeletal muscle of mice lacking the heart/muscle isoform of the adenine nucleotide translocator.

Mice deficient in the heart/muscle specific isoform of the adenine nucleotide translocator (ANT1) exhibit many of the hallmarks of human oxidative phosphorylation (OXPHOS) disease, including a dramatic proliferation of skeletal muscle mitochondria. Because many of the genes necessary for mitochondrial biosynthesis, OXPHOS function, and response to OXPHOS disease might be expected to be up-regulated in the Ant1(-/-) mouse, we used differential display reverse transcription-polymerase chain reaction techniques in an effort to identify these genes. 17 genes were identified as up-regulated in Ant1-deficient mice, and they fall into four categories: 1) nuclear and mitochondrial genes encoding OXPHOS components, 2) mitochondrial tRNA and rRNA genes, 3) genes involved in intermediary metabolism, and 4) an eclectic group of other genes. Among the latter genes, we identified the gene encoding anti-apoptotic Mcl-1, the Skd3 gene, and the WS-3 gene, which were previously unknown to be related to mitochondrial function. These results indicate that identification of genes up-regulated in the skeletal muscle of the Ant1-deficient mouse provides a novel method for identifying mammalian genes required for mitochondrial biogenesis.

Delivery of di-2-ethylhexyl phthalate to patients during hemodialysis.

It has been determined that the plasticizer di-2-ethylhexylphthalate (DEHP) can be leached from plastic bags used to store blood. During hemodialysis patient's blood is in contact with various plastic tubings; thus, studies were undertaken to determine how much, if any, DEHP was delivered into patients undergoing hemodialysis. Serial arterial and venous blood samples were drawn into glass syringes; serum was separated in acid-washed glass test tubes and stored at -80 degrees C. until analyzed for DEHP by gas-liquid chromatography. DEHP was delivered to each of nine patients studied. The amount delivered increased with the length of dialysis, up to 150 mg. during a 5-hour hemodialysis. The metabolic fate and toxicity, if any, of infused DEHP remains to be determined.