Biallelic Mutations in PDE10A Lead to Loss of Striatal PDE10A and a Hyperkinetic Movement Disorder with Onset in Infancy.

Deficits in the basal ganglia pathways modulating cortical motor activity underlie both Parkinson disease (PD) and Huntington disease (HD). Phosphodiesterase 10A (PDE10A) is enriched in the striatum, and animal data suggest that it is a key regulator of this circuitry. Here, we report on germline PDE10A mutations in eight individuals from two families affected by a hyperkinetic movement disorder due to homozygous mutations c.320A>G (p.Tyr107Cys) and c.346G>C (p.Ala116Pro). Both mutations lead to a reduction in PDE10A levels in recombinant cellular systems, and critically, positron-emission-tomography (PET) studies with a specific PDE10A ligand confirmed that the p.Tyr107Cys variant also reduced striatal PDE10A levels in one of the affected individuals. A knock-in mouse model carrying the homologous p.Tyr97Cys variant had decreased striatal PDE10A and also displayed motor abnormalities. Striatal preparations from this animal had an impaired capacity to degrade cyclic adenosine monophosphate (cAMP) and a blunted pharmacological response to PDE10A inhibitors. These observations highlight the critical role of PDE10A in motor control across species.

Glucocorticoid receptors GRα and GRß are expressed in inflammatory dermatoses.

BACKGROUND: Glucocorticoids (GC) are the most commonly used anti-inflammatory drugs in dermatology. The actions of GCs are mediated by the glucocorticoid receptor (GR). Alternative splicing of GR mRNA gives rise to different isoforms, GRα and GRß being the most important. GRß antagonizes the activity of GRα and its up-regulation has been associated with glucocorticoid insensitivity in several non-cutaneous inflammatory diseases. METHODS: Using immunohistochemical stainings, we analyzed the expression of GRα and GRß in lesional skin samples of patients with atopic dermatitis, lichen ruber planus, eczema nummulare and lichen simplex chronicus. We also conducted a study of 13 severe atopic patients to investigate the effect of prednisolone treatment on the expression of GR isoforms using quantitative PCR, western blot and immunohistochemical analysis. RESULTS: GRα and GRß were expressed in atopic dermatitis, lichen ruber planus, eczema nummulare and lichen simplex chronicus. Our novel finding was that GRß is abundant in keratinocytes and cutaneous neutrophils. Nuclear staining of both GRα and GRß was strongest in keratinocytes of patients with lichen ruber planus, whereas the least nuclear positivity was detected in keratinocytes of patients with atopic dermatitis. In severe atopic dermatitis GRα and GRß were expressed in both peripheral blood mononuclear cells and the skin. The expression of GRα and GRß varied during prednisolone therapy but the changes were not related to treatment response or GC insensitivity. CONCLUSION: GRα and GRß are expressed in inflammatory dermatoses. In severe atopic dermatitis the increased expression of GRß mRNA is not connected to insensitivity against prednisolone treatment.

IL-17/Th17 pathway is activated in acne lesions.

The mechanisms of inflammation in acne are currently subject of intense investigation. This study focused on the activation of adaptive and innate immunity in clinically early visible inflamed acne lesions and was performed in two independent patient populations. Biopsies were collected from lesional and non-lesional skin of acne patients. Using Affymetrix Genechips, we observed significant elevation of the signature cytokines of the Th17 lineage in acne lesions compared to non-lesional skin. The increased expression of IL-17 was confirmed at the RNA and also protein level with real-time PCR (RT-PCR) and Luminex technology. Cytokines involved in Th17 lineage differentiation (IL-1ß, IL-6, TGF-ß, IL23p19) were remarkably induced at the RNA level. In addition, proinflammatory cytokines and chemokines (TNF-α, IL-8, CSF2 and CCL20), Th1 markers (IL12p40, CXCR3, T-bet, IFN-γ), T regulatory cell markers (Foxp3, IL-10, TGF-ß) and IL-17 related antimicrobial peptides (S100A7, S100A9, lipocalin, hBD2, hBD3, hCAP18) were induced. Importantly, immunohistochemistry revealed significantly increased numbers of IL-17A positive T cells and CD83 dendritic cells in the acne lesions. In summary our results demonstrate the presence of IL-17A positive T cells and the activation of Th17-related cytokines in acne lesions, indicating that the Th17 pathway is activated and may play a pivotal role in the disease process, possibly offering new targets of therapy.

Expression of cancer-related carbonic anhydrases IX and XII in normal skin and skin neoplasms.

Purpose of the study was to evaluate the presence of hypoxia-inducible, tumour-associated carbonic anhydrases IX and XII in normal skin and a series of cutaneous tumours. Human tumour samples were taken during surgical operations performed on 245 patients and were immunohistochemically stained. A histological score value was calculated for statistical analyses which were performed using SPSS for Windows, versions 17.0 and 20.0. In normal skin, the highest expression of CA IX was detected in hair follicles, sebaceous glands, and basal parts of epidermis. CA XII was detected in all epithelial components of skin. Both CA IX and CA XII expression levels were significantly different in epidermal, appendigeal, and melanocytic tumour categories. Both CA IX and XII showed the most intense immunostaining in epidermal tumours, whereas virtually all melanocytic tumours were devoid of CA IX and XII immunostaining. In premalignant lesions, CA IX expression significantly increased when the tumours progressed to more severe dysplasia forms. Both CA IX and XII are highly expressed in different epithelial components of skin. They are also highly expressed in epidermal tumours, in which CA IX expression levels also correlate with the dysplasia grade. Interestingly, both isozymes are absent in melanocytic tumours.

A prognostic index in skin melanoma through the combination of matrix metalloproteinase-2, Ki67, and p53.

The objective of this immunohistochemical study was to explore the roles of Ki67 and p53 in conjunction with matrix metalloproteinase-2 and matrix metalloproteinase-9, tissue inhibitors of metalloproteinase-1 and tissue inhibitors of metalloproteinase-2 in a series of 157 cases of skin melanomas. Elevated Ki67 expression and positive staining for p53 correlated to the propensity to metastasize (P = .016) and to declined disease-specific survival, as well as to shortened recurrence-free survival. In patients with a high immunoreaction for Ki67, the 10-year disease-specific survival was 39% compared with 73% in patients with a low Ki67 expression (P = .03). In cases with a positive p53 expression in melanoma cells, the 10-year disease-specific survival was 59% compared with 76% in patients with a negative immunoreaction for p53 (P = .005). Overexpression of the matrix metalloproteinase 2 protein in conjunction with overexpression of Ki67 characterized melanomas with high metastatic potential and was associated with declined survival with a 10-year disease-specific survival of 33% compared with 85% in the cases with low matrix metalloproteinase-2 and low Ki-67 levels (P = .002). Similarly, in cases with overexpression of matrix metalloproteinase-2 and a positive immunoreaction for p53, the 10-year disease-specific survival was only 42% compared with 80% in patients with matrix metalloproteinase-2 less than 20% and a negative immunostaining for p53 (P < .001). The presence of all 3 adverse prognostic factors was prognostically more significant than any marker alone with a 10-year survival of only 28%. This combination of determining matrix metalloproteinase 2, Ki67, and p53 immunoreactive proteins could be beneficial in the selection of high-risk melanoma patients for future adjuvant trials.

Comparison of the prognostic value of matrix metalloproteinases 2 and 9 in cutaneous melanoma.

Previously, gelatinases matrix metalloproteinase (MMP)-2 and MMP-9 have been shown to be involved in melanoma invasion and progression. Also, overexpression of MMP-2 has been suggested to be linked to hematogenous metastasis in melanoma. This study was conducted to study the prognostic value of MMP-2 and MMP-9 in human melanoma. The expression of MMP-2 and MMP-9 immunoreactive protein was evaluated in 157 cases of primary melanomas. An immunohistochemical analysis using specific monoclonal antibodies for MMP-2 and MMP-9 on paraffin-embedded tissues was performed, and the immunostaining results were compared with the clinical course of melanoma. Overexpression of MMP-2 (>20% of malignant cells positive) was an independent prognostic marker for melanoma related death, with odds ratio of 2.6 (95% confidence interval, 1.32-5.07). The 10-year disease-specific survival rate was only 51% in patients with overexpression of MMP-2 protein compared with 79% in patients with a primary melanoma with low expression for MMP-2 (P = .001). Interestingly, male patients with a melanoma with overexpression of MMP-2 showed a 10-year disease-specific survival of only 41% compared with 77% in other male patients (P = .003). It is notable that the immunoreactive protein for MMP-9 in primary melanoma was not found to be of any prognostic importance. In the future, MMP-2 could be acknowledged as a new prognostic factor in melanoma. MMP-9, on the other hand, was not associated with the clinical course of melanoma. Based on the current data, MMP-2 could be evaluated as an inclusion factor for adjuvant studies especially in male melanoma patients.

The expression of myoglobin and ROR2 protein in Dupuytren's disease.

BACKGROUND: Dupuytren's disease (DD) is a hand disease inherited as an autosomal dominant trait with variable penetrance, especially among populations of northern European ancestry. The etiology and pathophysiology of DD are not clear. The purpose of this study was to examine the gene expression profiles of palmar fascia of DD and healthy patients using microarray analysis to highlight the genes that might contribute to the pathogenesis of DD. MATERIALS AND METHODS: Dupuytren contracture samples were taken from excised mature cords of DD patients during aponeurectomies. Control samples were collected from healthy hand trauma patients. Microarray analysis was performed with the Affymetrix HGU133A genome array (Affymetrix, Santa Clara, CA). Expression changes of selected proteins were confirmed at the protein level with Western and dot blotting or by immunohistochemistry. RESULTS: At least an 8-fold change in gene expression was found with 127 genes, including a 90-fold down-regulation of myoglobin and a 14-fold up-regulation of tyrosine kinase-like orphan receptor 2 (= ROR2) from absent to present during the disease. The changes in myoglobin and ROR2 expression were confirmed at the protein level. CONCLUSIONS: In this study, we showed for the first time the connection of ROR2 in Dupuytren's disease. ROR2 and myoglobin may play an important role in the pathophysiology of this disease.

Intraosseous schwannoma of the middle phalanx.

Intraosseous schwannoma is an extremely rare, benign neoplasm. Only a few cases involving the bones of the hand have been reported, and none of these cases has involved middle phalanx. We present a case of intraosseous schwannoma of the middle phalanx of the right ring finger.

Biocompatibility and strength properties of nitinol shape memory alloy suture in rabbit tendon.

Nitinol (NiTi) is a promising new tendon suture material with good strength, easy handling and good super-elastic properties. NiTi sutures were implanted for biocompatibility testing into the right medial gastrocnemius tendon in 15 rabbits for 2, 6 and 12 weeks. Additional sutures were implanted in subcutaneous tissue for strength measurements in order to determine the effect of implantation on strength properties of NiTi suture material. Braided polyester sutures (Ethibond) of approximately the same diameter were used as control. Encapsulating membrane formation around the sutures was minimal in the case of both materials. The breaking load of NiTi was significantly greater compared to braided polyester. Implantation did not affect the strength properties of either material.

Bone tissue engineering: treatment of cranial bone defects in rabbits using self-reinforced poly-L,D-lactide 96/4 sheets.

This study is one of a series in which the authors evaluate various absorbable sheets to guide bone regeneration in cranial bone defects. The aim was to evaluate the use of self-reinforced poly-L,D-lactide 96/4 (SR-PLA96) sheets for cranial bone tissue engineering in experimental defects in rabbits. Square defects of 10 x 10 mm were created in the right parietal bone. SR-PLA96 implants (15 x 15 mm) were used to cover these defects in 12 New Zealand White rabbits. Similar defects were created in the left parietal bone, but no sheets were used (controls). The rabbits were killed after 6, 24, or 48 weeks. Histology and histomorphometry were used to evaluate healing of the defects. Defects covered with SR-PLA96 sheets showed more abundant bone formation than control (non-covered) defects. At 6 weeks, the defects were occupied mainly by fibrous tissue. At 24 weeks, healing with bone formation was more obvious in the covered defects. At 48 weeks, bone completely bridged defects covered with SR-PLA96 sheets, and incomplete bridging was seen in non-covered control defects. Hence, bone tissue engineering in experimental cranial bone defects in rabbits can be achieved using SR-PLA96 sheets to guide bone regeneration.

Tularemia of the middle ear.

We report the case of a 10-year-old boy with prolonged fever who was found to have tularemia of the middle ear. Otolaryngologic cases including oropharyngeal and glandular or ulceroglandular forms of the head and neck region are estimated to account for 12% of all tularemia cases, but to date we have not seen a report of tularemia in the middle ear. The possibility of tularemia may not occur to a physician because of the wide variation of clinical manifestations.

Smoking and skin: a study of the physical qualities and histology of skin in smokers and non-smokers.

Tobacco smoke is toxic to cells and could be a factor contributing to accelerated skin ageing. The aim of this study was to provide new information on the possible effects of smoking on the physical qualities of skin and the morphology of elastic fibres. The study population consisted of 98 men, including 47 current smokers and 51 never-smokers. Skin thickness and elasticity were measured from cheek, temple, abdomen, dorsal forearm and non-sun-exposed upper inner arm. Verhoeff-stained punch biopsies from the non-sun-exposed upper inner arm were assessed with a computerized image analyser in a blinded fashion to assess the amount and width of elastic fibres. The thickness of cheek skin was increased in the smokers, but skin thickness in other measured sites did not differ between the groups. The amount and width of elastic fibres in the sun-protected skin of the smokers and non-smokers did not differ significantly, nor did skin elasticity in this or any other region under evaluation, suggesting that smoking alone affects neither the amount and width of dermal elastic fibres nor the elasticity of skin in male smokers.