Non-invasive transcriptomic analysis using mRNAs in skin surface lipids obtained from children with mild-to-moderate atopic dermatitis.

BACKGROUND: Specimens for analysing the molecular pathology of skin disease are generally obtained through invasive methods, such as biopsy. However, less burdensome methods are desirable for paediatric patients. We recently established a method that comprehensively analyses RNA present in sebum (skin surface lipid-RNAs: SSL-RNAs) using a next-generation sequencer. Using this method, biological information can be obtained from the skin in a completely non-invasive manner. OBJECTIVES: To verify the applicability of the SSL-RNA method for analysis of paediatric skin and analyse the molecular pathology of mild-to-moderate atopic dermatitis (AD) in children. METHODS: We collected sebum specimens from the whole faces of 23 healthy children and 16 children with mild-to-moderate AD (eczema area and severity index (EASI) score: 5.9 ± 2.6) ranging in age from 6 months to 5 years, using an oil-blotting film. We then extracted SSL-RNAs from the samples and performed an AmpliSeq transcriptomic analysis. RESULTS: The expressions of genes related to keratinization (LCE, PSORS1C2, IVL and KRT17), triglyceride synthesis and storage (PLIN2, DGAT2 and CIDEA), wax synthesis (FAR2), ceramide synthesis (GBA2, SMPD3 and SPTLC3), antimicrobial peptides (DEFB1) and intercellular adhesion (CDSN), all of which are related to the skin barrier, are lower in children with AD than in healthy children. The children with AD also have higher expression of CCL17, a Th2-cytokine and an increased Th2-immune response as demonstrated by a gene set variation analysis. Moreover, KRT17 and CCL17 expression levels are significantly correlated with the EASI score. CONCLUSIONS: Molecular changes associated with abnormal immune responses and the epidermal barrier in children with mild-to-moderate AD can be determined using the SSL-RNA method. This non-invasive method could therefore be a useful means for understanding the molecular pathology of paediatric AD.

Upsizing the femoral component increases patellofemoral contact force in total knee replacement.

In posterior stabilised total knee replacement (TKR) a larger femoral component is sometimes selected to manage the increased flexion gap caused by resection of the posterior cruciate ligament. However, concerns remain regarding the adverse effect of the increased anteroposterior dimensions of the femoral component on the patellofemoral (PF) joint. Meanwhile, the gender-specific femoral component has a narrower and thinner anterior flange and is expected to reduce the PF contact force. PF contact forces were measured at 90°, 120°, 130° and 140° of flexion using the NexGen Legacy Posterior Stabilized (LPS)-Flex Fixed Bearing Knee system using Standard, Upsized and Gender femoral components during TKR. Increasing the size of the femoral component significantly increased mean PF forces at 120°, 130° and 140° of flexion (p = 0.005, p < 0.001 and p < 0.001, respectively). No difference was found in contact force between the Gender and the Standard components. Among the patients who had overhang of the Standard component, mean contact forces with the Gender component were slightly lower than those of the Standard component, but no statistical difference was found at 90°, 120°, 130° or 140° of flexion (p = 0.689, 0.615, 0.253 and 0.248, respectively). Upsized femoral components would increase PF forces in deep knee flexion. Gender-specific implants would not reduce PF forces.

Enlarged post-operative posterior condyle tightens extension gap in total knee arthroplasty.

We investigated whether the extension gap in total knee replacement (TKR) would be changed when the femoral component was inserted. The extension gap was measured with and without the femoral component in place in 80 patients with varus osteoarthritis undergoing posterior-stabilised TKR. The effect of a post-operative increase in the size of the femoral posterior condyles was also evaluated. The results showed that placement of the femoral component significantly reduced the medial and lateral extension gaps by means of 1.0 mm and 0.9 mm, respectively (p < 0.0001). The extension gap was reduced when a larger femoral component was selected relative to the thickness of the resected posterior condyle. When the post-operative posterior lateral condyle was larger than that pre-operatively, 17 of 41 knees (41%) showed a decrease in the extension gap of > 2.0 mm. When a specially made femoral trial component with a posterior condyle enlarged by 4 mm was tested, the medial and lateral extension gaps decreased further by means of 2.1 mm and 2.8 mm, respectively. If the thickness of the posterior condyle is expected to be larger than that pre-operatively, it should be recognised that the extension gap is likely to be altered. This should be taken into consideration when preparing the extension gap.

Mevastatin reduces cartilage degradation in rabbit experimental osteoarthritis through inhibition of synovial inflammation.

OBJECTIVE: To examine the therapeutic efficacy of an HMG-CoA reductase inhibitor (statin) in rabbit osteoarthritis (OA) in vitro and in vivo. METHODS: In the presence or absence of mevastatin, rabbit chondrocytes and synoviocytes were incubated with Interleukin-1beta (IL-1beta), and analyzed by biochemical methods. Thirty-two mature rabbits that underwent bilateral anterior cruciate ligament transaction (ACLT) received six consecutive weekly intra-articular injections of mevastatin at three different concentrations or a control solution. All animals were sacrificed 6 weeks after ACLT, and the knee joints were assessed by morphological, histological, immunohistochemical, and biochemical methods. RESULTS: Mevastatin inhibited IL-1beta stimulation of gene expression of monocyte chemoattractant protein-1 (MCP-1) and matrix-metalloproteinases 3 (MMP-3), in synoviocytes but not chondrocytes. The levels of MCP-1 and MMP-3 productions in synoviocytes were significantly reduced by statin-treatment. In rabbit with OA, intra-articular injection of mevastatin significantly reduced cartilage degradation, as assessed by morphological and histological examinations. Synovial tissues of knees treated with mevastatin showed less severe inflammatory responses with reduced thickness of synovial cell lining and less infiltration of subsynovial CD68+monocyte lineage cells compared to untreated control knees. Relative mRNA expressions of MCP-1, IL-1beta, MMP-3, and MMP-13 were reduced in synovial tissues, but not articular cartilage, of knees treated with mevastatin compared with untreated control knees. CONCLUSION: During the development of experimental OA, intra-articular administration of HMG-CoA reductase inhibitor (statin) reduces inflammatory cell infiltration and matrix-degrading enzyme expression, thus limiting cartilage degradation.

Hypertrophy of the posterior longitudinal ligament in the thoracic spine.

STUDY DESIGN: This is a case report of a patient with hypertrophy of the posterior longitudinal ligament (HPLL) in the thoracic spine. This patient was followed for 10 years after surgery. OBJECTIVES: The purpose of this study was to report the long-term outcome of HPLL in the thoracic spine. SETTING: Department of orthopedic surgery, Hiroshima Red Cross and Atomic-bomb Survivors Hospital, Hiroshima, Japan. METHODS: A 58-year-old-woman with thoracic HPLL was reported. Magnetic resonance image (MRI) and computed tomography (CT) showed the expanded spinal cord compression from Th4 to Th12 due to HPLL. Anterior decompression and fusion (Th10-12) was performed. Histological findings of the surgical specimens showed thickening of the posterior longitudinal ligament with proliferation of chondroid tissue. The clinical outcome and the radiological findings (CT and MRI) were evaluated 10 years after surgery. RESULTS: The patient was asymptomatic postoperatively. However, the subsequent CT examination revealed ossification of the previously hypertrophied posterior longitudinal ligament. CONCLUSIONS: HPLL in the thoracic spine is a rare pathological condition causing myelopathy. The results of this study support the hypothesis that HPLL is one of the prodromal conditions of HPLL.

Engagement of human monocyte-derived dendritic cells into interleukin (IL)-12 producers by IL-1beta + interferon (IFN)-gamma.

Dendritic cells (DCs) are potent antigen-presenting cells and can induce tumour- or pathogen-specific T cell responses. For adoptive immunotherapy purposes, immature DCs can be generated from adherent monocytes using granulocyte macrophage colony stimulating factor (GM-CSF) and interleukin (IL)-4, and further maturation is usually achieved by incubation with tumour necrosis factor (TNF)-alpha. However, TNF-alpha-stimulated DCs produce low levels of IL-12. In this study, we compared the effects of TNF-alpha, interferon (IFN)-gamma, IL-1beta or IFN-gamma + IL-1beta on the phenotypic and functional maturation of DCs. Our results show that IFN-gamma, but not IL-1beta, augmented the surface expression of CD80, CD83 and CD86 molecules without inducing IL-12 production from DCs. However, IL-1beta, but not IFN-gamma, induced IL-12 p40 production by DCs without enhancing phenotypic maturation. When combined, IFN-gamma + IL-1beta treatment profoundly up-regulated the expression of CD80, CD83, CD86 and major histocompatibility complex (MHC) class II antigens. Furthermore, IFN-gamma + IL-1beta-treated DCs produced larger amounts of IL-12 and induced stronger T cell proliferation and IFN-gamma secretion in primary allogeneic mixed lymphocyte reaction (MLR) than did TNF-alpha-treated DCs. Our results show that IFN-gamma + IL-1beta induced human monocyte-derived DCs to differentiate into Th1-prone mature DCs.

[Mating activity of male SHR rats infected with sialodacryoadenitis virus].

The male SHR rats infected with sialodacryoadenitis (SDA) virus did not show any abnormal mating activity. However, high mortality was seen in embryos of diseased dams after cage-mating with infected males on Day 4 to Day 11 postinoculation.

Reproductive disorders in female rats infected with sialodacryoadenitis virus.

Effects of sialodacryoadenitis virus infection on the reproduction of Wistar and Sprague-Dawley (SD) rats were studied. Estrous cycle was considerably out of order in about 40% of infected rats of both strains, starting on days 4 to 20 postinfection and persisting for 2 to 12 days. Of Wistar and SD dams infected on day 0 of gestation about 30% and 10% of the fetuses were found dead, while only 4 and 3% of non-infected dams. In severely diseased Wistar and SD dams infected on day 15 of gestation, the offspring showed death rates of 57% and 13%, respectively, because of inadequate nursing.

[2 cases of granular cell tumor of the breast].

Two cases of granular cell tumor of the breast are reported. This disease is uncommon; only 54 cases have been reported so far. Clinically, it resembled breast cancer. Its palpation is solid. It sometimes has a dimpling sign, and, on occasion, the X-ray diagnosis shows a spicule. Pathologically, the tumor has a tendency to infiltration. The tumor cells contain granules of different sizes. Straited muscles, mesenchymal cells and Schwann's cells have been pointed out as possible roigins of the tumor. It has also been explained as histiocytic granuloma. A decisive theory about its organic development, however, has yet to be established.

[Current status and problems of gastric cancer in the aged].

Based on our analysis of 1) gastric cancer patients surgically treated between 1972-1980 in Niigata Prefecture, 2) mortality statistics, and 3) 381 gastric cancer patients older than 70 years operated in our hospital, we concluded that 1) The number of geriatric surgical patients with gastric cancer, especially early gastric cancer, is on the increase in Niigata Prefecture. 2) The mortality from gastric cancer has decreased markedly in 70-74 year-old patients, has declined slightly in patient ranging from 75-79 years and has increased in patients over 80 years. 3) In patients between 75-79 years and 70-74 years, surgery as almost equally safe and their long-term survival rate was similar. Our results suggest that gastric cancer patients ranging between 75-79 years should be operated as radically as younger patients.