Expression of Hedgehog signalling molecules in microcystic adnexal carcinoma.

BACKGROUND: Microcystic adnexal carcinoma (MAC) is a rare skin neoplasm that has not been characterized on a molecular basis. AIM: To assess expression profiles of Hedgehog (HH) signalling molecules in MAC and control tumours. METHODS: Immunohistochemistry was performed for Sonic Hedgehog (SHH), Indian Hedgehog (IHH), Patched 1 (PTCH1) and Smoothened (SMO) on patient MAC tissue (n = 26) and control tumour tissue, including syringoma (SyG; n = 11), trichoepithelioma (TE; n = 11) and basal cell carcinoma (BCC; n = 12) tissues. RESULTS: Patched 1 and SMO immunoreactivity was significantly higher in BCC than in SyG, TE or MAC (P < 0.001 and P < 0.03, respectively). The highest IHH expression was observed in BCC and TE compared with SyG and MAC (P < 0.04). Notably, the highest SHH protein expression was observed in SyG compared with MAC, TE and even BCC (P < 0.001). In patients with MAC, SMO immunoreactivity significantly (r = 0.51; P < 0.01) correlated with PTCH1 expression. Further correlation studies did not show significant associations between the HH expression markers assessed (P > 0.05). CONCLUSION: Our results indicate that alterations of the HH signalling are unlikely to play a major role in the pathogenesis of MAC, which is in contrast to the morphologically similar BCC and TE. Our observation provides additional information to the limited molecular pathology knowledge on this rare tumour.

Lysyl oxidase-like-2 mutations and reduced mRNA and protein expression in mid-dermal elastolysis.

BACKGROUND: Mid-dermal elastolysis (MDE) is a rare skin condition, characterized by selective loss of elastic fibres in the mid dermis. The pathogenesis of MDE is still unclear. AIM: To investigate expression of lysyl oxidase-like 2 (LOXL2) in a reasonable sample of patients with MDE and to search for mutations in LOXL2. METHODS: We investigated archived lesional tissue of 13 patients with MDE and skin tissue samples of 10 sex- and age-matched healthy controls (HCs). Gene and protein expression of LOXL2 was investigated using real-time reverse-transcription PCR and immunohistochemistry. Mutation analysis was performed using the Sanger method. RESULTS: We observed decreased LOXL2 mRNA expression in lesional skin of patients with MDE (0.48 ± 0.16) compared with healthy skin of the same patients (1.5 ± 0.51) and normal skin of HCs (1.9 ± 0.13). Compared with healthy patient skin (epidermis 2.38 ± 1.6, dermis 1.2 ± 1), LOXL2 protein expression in lesional patient skin (epidermis 1.1 ± 0.7, dermis 0.3 ± 0.45) was significantly decreased (P < 0.04 and P = 0.02, respectively). Mutation analysis of the entire LOXL2 gene could be performed for five patients, all of whom were found to have at least one mutation in the LOXL2 gene. Three of these had a mutation in the promoter region (c.967 G>C, c.1022 C>T, and c.1025 G>A, respectively), and one of them also had a mutation in the splice region of intron 11/exon 12 (IVS11-1 G>A). Of the remaining two patients, one had a mutation in exon 3 (T1391), and the other had a mutation in exon 11 (C663Y). CONCLUSIONS: Our present data suggest that decreased elastin renewal due to LOXL2 mutations and consecutive reduced LOXL2 expression contribute to the pathogenesis of MDE.

Left-sided laterality of Merkel cell carcinoma in a German population: more than just sun exposure.

BACKGROUND: Lateral distribution of cancer has been observed previously. Most evident is this laterality in ultraviolet (UV)-induced skin cancer, based on an unequally distributed UV exposure. OBJECTIVES: The aim of this study was to explore whether patients from Germany also show asymmetrical lateral distribution of Merkel cell carcinoma (MCC). METHODS: In total, 115 patients with MCC were studied for laterality of the primary tumour. Correlation of clinical variables with lateral distribution of MCC was investigated as well. RESULTS: In 64/115 (55.7%) patients, primary tumours were present on the left side, in 37/115 (32.2%) on the right side, and in 14/115 (12.2%) in the midline (P < 0.0001). Excluding the latter localization occurrence of left-sided MCCs (64 of 101/63.4%) was significantly (P = 0.0072) more often observed (1.73-fold) when compared to right-sided tumours (37 of 101/36.6%). The excess of left-sided tumours was found on the head with a left-right ratio of 1.8, trunk of 8, arm of 1.2, and leg of 1.8. There was no significant association between laterality and gender, age, MCPyV status, and anatomic localization of primary tumours including the occurrence in sun-exposed sites. CONCLUSIONS: Occurrence of left-sided MCCs was significantly more often observed when compared to right-sided tumours. Laterality was not associated with tumour presentation at chronically ultraviolet-exposed sites. Hence, the reason for laterality in MCC remains obscure, but likely goes beyond UV exposure.

Prognostic relevance of high atonal homolog-1 expression in Merkel cell carcinoma.

BACKGROUND: It has recently been reported that atonal homolog 1 (ATOH1) gene is down-regulated in Merkel cell carcinoma (MCC) and thus may represent a tumor suppressor gene. OBJECTIVES: We aimed to test for ATOH1 gene mutations and expression levels in MCC tissues and cell lines. METHODS: Genomic DNA isolation and amplification via PCR was successfully performed in 33 MCCs on formalin-fixed paraffin-embedded tissue and three MCC cell lines, followed by Sanger sequencing of the whole ATOH1 gene to detect genomic aberrations. ATOH1 mRNA levels were determined by RT-PCR. Immunohistochemistry of ATOH1 was performed to quantify protein expression in tumor samples and cell lines. RESULTS: Neither in any of the 33 MCC tissue samples nor in the three cell lines ATOH1 mutations were present. ATOH1 was expressed in all lesions, albeit at different expression levels. Univariate analysis revealed that the total immunohistology score significantly correlated with the occurrence of tumor relapse (r = 0.57; P = 0.0008). This notion was confirmed in multivariate analysis suggesting that ATOH1 expression is a potential independent predictor for tumor relapse in MCC patients (P = 0.028). MCC-related death also correlated with ATOH1 expression (r = 0.4; P = 0.025); however, ATOH1 expression did not retain its predictive value in the regression model. CONCLUSIONS: In contrast to anecdotal reports ATOH1 expression is not lost by genetic alterations in MCC. However, protein expression of ATOH1 is increased in advanced MCC indicating that ATOH1 is involved in MCC progression.

Is MRI-based CFD able to improve clinical treatment of coarctations of aorta?

Pressure drop associated with coarctation of the aorta (CoA) can be successfully treated surgically or by stent placement. However, a decreased life expectancy associated with altered aortic hemodynamics was found in long-term studies. Image-based computational fluid dynamics (CFD) is intended to support particular diagnoses, to help in choosing between treatment options, and to improve performance of treatment procedures. This study aimed to prove the ability of CFD to improve aortic hemodynamics in CoA patients. In 13 patients (6 males, 7 females; mean age 25 ± 14 years), we compared pre- and post-treatment peak systole hemodynamics [pressure drops and wall shear stress (WSS)] vs. virtual treatment as proposed by biomedical engineers. Anatomy and flow data for CFD were based on MRI and angiography. Segmentation, geometry reconstruction and virtual treatment geometry were performed using the software ZIBAmira, whereas peak systole flow conditions were simulated with the software ANSYS(®) Fluent(®). Virtual treatment significantly reduced pressure drop compared to post-treatment values by a mean of 2.8 ± 3.15 mmHg, which significantly reduced mean WSS by 3.8 Pa. Thus, CFD has the potential to improve post-treatment hemodynamics associated with poor long-term prognosis of patients with coarctation of the aorta. MRI-based CFD has a huge potential to allow the slight reduction of post-treatment pressure drop, which causes significant improvement (reduction) of the WSS at the stenosis segment.

[Basal cell carcinoma and rare form variants]. / Das Basalzellkarzinom und seine seltenen Formvarianten.

Basal cell carcinomas are the most common primary cutaneous malignant neoplasms. The diagnosis of basal cell carcinoma represents a common and routine task for pathologists and dermatopathologists. The aim of this review is the clinical and histopathological presentation of the most common subtypes of basal cell carcinoma. Furthermore, the rare variants of basal cell carcinoma and their differential diagnoses are also discussed.

[Cutaneous infection due to Fusarium oxysporum in a female diabetic: molecular biological detection of the mold from formalin-fixed paraffin embedded tissue using sequencing of the ITS region of the rDNA]. / Kutane Infektion durch Fusarium oxysporum bei einer Diabetikerin: Molekularbiologischer Schimmelpilznachweis aus Formalin-fixiertem, Paraffin-eingebettetem Gewebe mittels Sequenzierung der ITS-Region der rDNA.

BACKGROUND: Skin lesions due to Fusarium spp. occur either secondarily following hematological spread in systemic infection or represent primary cutaneous infections following traumatic inoculation. CASE REPORT: A 34-year-old woman with insulin-dependent diabetes mellitus presented with a most likely posttraumatic leg ulcer present for 4 weeks. The ulcer showed superficial necrosis with cellular debris, neutrophils, and leukocytoclasia. Septate hyphae were detected both in the necrotic area and between the collagen fibers on initial H & E stained sections. Using PAS and Grocott-Gomori silver staining, the dichotomous branching hyphae were clearly visible. Unfortunately, cultural detection of the fungi was impossible. After extraction and purification of the fungal DNA from formalin-fixed and paraffin embedded (FFPE) tissue sections, the amplification of the ITS region of rDNA was done. Using sequencing and comparison with reference sequences of a gene bank, Fusarium oxysporum was identified. THERAPY: Therapy was performed by surgical excision of the entire ulcer followed by topical antiseptic treatment and wound conditioning. No systemic antifungal treatment was given. The lesion healed without any problems. DISCUSSION: Cutaneous fusarium infections are rare but emerging opportunistic infections. Histological examination represents the quickest diagnostic method for detection of the fungal infection. An alternative approach represents the species identification based on molecular techniques.

The impact of MRI-based inflow for the hemodynamic evaluation of aortic coarctation.

Aortic coarctation (CoA) accounting for 3-11% of congenital heart disease can be successfully treated. Long-term results, however, have revealed decreased life expectancy associated with abnormal hemodynamics. Accordingly, an assessment of hemodynamics is the key factor in treatment decisions and successful long-term results. In this study, 3D angiography whole heart (3DWH) and 4D phase-contrast magnetic resonance imaging (MRI) data were acquired. Geometries of the thoracic aorta with CoAs were reconstructed using ZIB-Amira software. X-ray angiograms were used to evaluate the post-treatment geometry. Computational fluid dynamics models in three patients were created to simulate pre- and post-treatment situations using the FLUENT program. The aim of the study was to investigate the impact of the inlet velocity profile (plug vs. MRI-based) with a focus on the peak systole pressure gradient and wall shear stress (WSS). Results show that helical flow at the aorta inlet can significantly affect the assessment of pressure drop and WSS. Simplified plug inlet velocity profiles significantly (p < 0.05) overestimate the pressure drop in pre- and post-treatment geometries and significantly (p < 0.05) underestimate surface-averaged WSS. We conclude that the use of the physiologically correct but time-expensive 4D MRI-based in vivo velocity profile in CFD studies may be an important step towards a patient-specific analysis of CoA hemodynamics.

Reproducibility of image-based analysis of cerebral aneurysm geometry and hemodynamics: an in-vitro study of magnetic resonance imaging, computed tomography, and three-dimensional rotational angiography.

BACKGROUND AND STUDY AIMS: Image-based computational fluid dynamics (CFD) provides a means for analysis of biofluid mechanical parameters of cerebral aneurysms. This may enable patient-specific rupture risk analysis and facilitate treatment decisions. Application of different imaging methods may, however, alter the geometrical basis of these studies. The present study compares geometry and hemodynamics of an aneurysm phantom model acquired by means of magnetic resonance imaging (MRI), computed tomography (CT), and rotational angiography (3DRA). MATERIALS AND METHODS: The phantom model of a basilaris artery aneurysm was fabricated based on data generated by CT angiography. This model underwent imaging by means of CT, MRI, and 3DRA. We compared the geometrical reconstructions using the original dataset with those obtained from CT, MRI, and 3DRA. Similarly, CFD analyses were performed using the four reconstructions (3DRA, MRI, CT, and original dataset). RESULTS: MRI and the 3DRA-based reconstructions yield mean reconstruction errors of 0.097 mm and 0.1 mm, which are by a factor of 2.5 better than the CT reconstruction. The maximal error for the aneurysm radius (7.11 mm) measurement was found in the 3DRA reconstruction and was 3.8% (0.28 mm). A comparison of calculated time-averaged wall shear stress (WSS) shows good correlations for the entire surface and, separately, for the surface of the aneurysmal sack. The maximal error of 8% of the mean WSS calculation of the whole surface was found for the CT reconstruction. The calculations of the aneurysmal sack mean WSS from the MRI reconstruction were estimated to have a maximal error of 7%. CONCLUSION: All three imaging techniques (CT, MRI, 3DRA) adequately reproduce aneurysm geometry and allow meaningful CFD analyses.

Effects of low-dosed imidacloprid pulses on the functional role of the caged amphipod Gammarus roeseli in stream mesocosms.

Effects of two series of imidacloprid pulses on caged amphipods (Gammarus roeseli) and their shredder efficiency for litter decomposition were studied for 70 days as part of a comprehensive stream mesocosm experiment. The duration of each imidacloprid pulse of 12µgL(-1) was 12h. About 250mL cages with an initial stock of 10 adult gammarids together with different conditioned litter substrates were used. Beside alder leaves (Alnus glutinosa), straw (× Triticosecale) was also used in different trials and tested for its suitability to serve as litter substrate. Results from tracer and microprobe measurements approved the suitability of the test system under low-flow condition of 10cms(-1) in the surrounding stream water. Population development followed a logistic growth function with a carrying capacity of 200 Ind cage(-1) for alder and 161 for straw. In the course of the study, the F1 generation reached sexual maturity and F2 offspring appeared. Increased nitrogen contents of gammarid-free trials compared to stocked ones after 70 days indicated that biofilm on both substrates was an important food source for G. roeseli. However, increased shredding activity by gammarids was only detected for alder during the second pulse series. During the remaining time and also for straw, losses of coarse particular organic matter were quite constant and slow indicating the dominance of transport limited decomposition processes on the litter surfaces. No effect of imidacloprid pulses on population levels and litter decomposition could be detected. However, the number of brood carrying females was reduced in the treatments compared to the control groups in the last 3 weeks of the study. In conclusion, repeated low-level and short-term exposition may have adverse long-term effects on G. roeseli in the field with regard to both the population size and the functional role as key shredder.

Closer to reality--the influence of toxicity test modifications on the sensitivity of Gammarus roeseli to the insecticide imidacloprid.

Laboratory toxicity test designs are far from reality and therefore extrapolations to field situations may be more difficult. In laboratory experiments with the amphipod Gammarus roeseli exposed to the insecticide imidacloprid it was investigated if test conditions closer to reality influences its sensitivity and if it is possible to extrapolate results from these laboratory tests to results from a stream mesocosm study. Experiments were run by varying medium, temperature, size, and seasonal origin of gammarids. Age and seasonal aspects had strongest effects with juveniles and animals taken from a spring population being most sensitive with an EC50 (96 h) of 14.2 µg L⁻¹ imidacloprid. The test designs closest to the conditions in the stream mesocosms reflected best the results in mesocosms study on basis of LOEC values. However, the EC(x) extrapolation failed to predict the effects of short term imidacloprid pulses in the field.

Statistical wall shear stress maps of ruptured and unruptured middle cerebral artery aneurysms.

Haemodynamics and morphology play an important role in the genesis, growth and rupture of cerebral aneurysms. The goal of this study was to generate and analyse statistical wall shear stress (WSS) distributions and shapes in middle cerebral artery (MCA) saccular aneurysms. Unsteady flow was simulated in seven ruptured and 15 unruptured MCA aneurysms. In order to compare these results, all geometries must be brought in a uniform coordinate system. For this, aneurysms with corresponding WSS data were transformed into a uniform spherical shape; then, all geometries were uniformly aligned in three-dimensional space. Subsequently, we compared statistical WSS maps and surfaces of ruptured and unruptured aneurysms. No significant (p > 0.05) differences exist between ruptured and unruptured aneurysms regarding radius and mean WSS. In unruptured aneurysms, statistical WSS map relates regions with high (greater than 3 Pa) WSS to the neck region. In ruptured aneurysms, additional areas with high WSS contiguous to regions of low (less than 1 Pa) WSS are found in the dome region. In ruptured aneurysms, we found significantly lower WSS. The averaged aneurysm surface of unruptured aneurysms is round shaped, whereas the averaged surface of ruptured cases is multi-lobular. Our results confirm the hypothesis of low WSS and irregular shape as the essential rupture risk parameters.

Silicon supply modifies C:N:P stoichiometry and growth of Phragmites australis.

Silicon is a non-essential element for plant growth. Nevertheless, it affects plant stress resistance and in some plants, such as grasses, it may substitute carbon (C) compounds in cell walls, thereby influencing C allocation patterns and biomass production. How variation in silicon supply over a narrow range affects nitrogen (N) and phosphorus (P) uptake by plants has also been investigated in some detail. However, little is known about effects on the stoichiometric relationships between C, N and P when silicon supply varies over a broader range. Here, we assessed the effect of silicon on aboveground biomass production and C:N:P stoichiometry of common reed, Phragmites australis, in a pot experiment in which three widely differing levels of silicon were supplied. Scanning electron microscopy (SEM) showed that elevated silicon supply promoted silica deposition in the epidermis of Phragmites leaves. This resulted in altered N:P ratios, whereas C:N ratios changed only slightly. Plant growth was slightly (but not significantly) enhanced at intermediate silicon supply levels but significantly decreased at high levels. These findings point to the potential of silicon to impact plant growth and elemental stoichiometry and, by extension, to affect biogeochemical cycles in ecosystems dominated by Phragmites and other grasses and sedges.

[Cutaneous malignant peripheral nerve sheath tumor in neurofibromatosis type 1]. / Kutaner maligner peripherer Nervenscheidentumor bei Neurofibromatose Typ 1.

Patients with neurofibromatosis have an increased risk of developing malignant tumors in comparison to the general population. We describe a woman who developed a malignant peripheral nerve sheath tumor in a pre-existing neurofibroma.

[Juvenile hyaline fibromatosis]. / Juvenile hyaline Fibromatose.

Juvenile hyaline fibromatosis is a rare autosomal recessive disease of the connective tissue. We present the case of a 6-year-old normal mental developed boy with confluent pearly papules behind the ears and in the paranasal folds, firm nodules of the scalp, the back and metaphalangs, and severe gingival hypertrophy.