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.

Cartilage and subchondral bone distributions of the distal radius: a 3-dimensional analysis using cadavers.

OBJECTIVE: To quantify the spatial distributions of cartilage and subchondral bone thickness of the distal radius. DESIGN: Using 17 cadaveric wrists, three types of 3-dimensional models were created: a cartilage-bone model, obtained by laser scanning; a bone model, rescanned after dissolving the cartilage; and a subchondral bone model, obtained using computed tomography. By superimposing the bone model onto the cartilage-bone and the subchondral bone models, the cartilage and subchondral bone thickness were determined. Measurements along with the spatial distribution were made at fixed anatomic points including the scaphoid and lunate fossa, sigmoid notch and interfossal ridge, and compared at each of these four regions. RESULTS: Cartilage thickness of the interfossal ridge (0.89 ± 0.23 mm) had a larger average thickness compared to that of the scaphoid fossa (0.70 ± 0.18 mm; p = 0.004), lunate fossa (0.75 ± 0.17 mm; p = 0.044) and sigmoid notch (0.64 ± 0.13 mm; p < 0.001). Subchondral bone was found to be thickest at the scaphoid (2.18 ± 0.72 mm) and lunate fossae (1.94 ± 0.93 mm), which were both thicker than that of sigmoid notch (1.63 ± 1.06 mm: vs scaphoid fossa, p = 0.020) or interfossal ridge (1.54 ± 0.84 mm: vs scaphoid fossa, p = 0.004; vs lunate fossa, p = 0.048). In the volar-ulnar sub-regions of the scaphoid and lunate fossa, the subchondral bone thickened. CONCLUSIONS: Our data can be applied when treating distal radius fractures. Cartilage thickness was less than 1 mm across the articular surface, which may give an insight into threshold for an acceptable range of step-offs. The combined findings of subchondral bone appreciate the importance of the volar-ulnar corner of the distal radius in the volar locking plate fixation.

Cartilage wear patterns in severe osteoarthritis of the trapeziometacarpal joint: a quantitative analysis.

OBJECTIVE: The present quantitative study aimed to assess the three-dimensional (3-D) cartilage wear patterns of the first metacarpal and trapezium in the advanced stage of osteoarthritis (OA) and compare cartilage measurements with radiographic severity. DESIGN: Using 19 cadaveric trapeziometacarpal (TMC) joints, 3-D cartilage surface models of the first metacarpal and trapezium were created with a laser scanner, and 3-D bone surface model counterparts were similarly created after dissolving the cartilage. These two models were superimposed, and the interval distance on the articular surface as the cartilage thickness was measured. All measurements were obtained in categorized anatomic regions on the articular surface of the respective bone, and we analyzed the 3-D wear patterns on the entire cartilage surface. Furthermore, we compared measurements of cartilage thickness with radiographic OA severity according to the Eaton grading system using Pearson correlation coefficients (r). RESULTS: In the first metacarpal, the cartilage thickness declined volarly (the mean cartilage thickness of the volar region was 0.32 ± 0.16 mm, whereas that of the dorsal region was 0.53 ± 0.18 mm). Conversely, the cartilage evenly degenerated throughout the articular surface of the trapezium. Measurements of the categorized regions where cartilage thinning was remarkable exhibited statistical correlations with radiographic staging (r = -0.48 to -0.72). CONCLUSIONS: Our findings indicate that cartilage wear patterns differ between the first metacarpal and trapezium in the late stage of OA. There is a need for further studies on cartilage degeneration leading to symptomatic OA in the TMC joint.

Altered bone density and stress distribution patterns in long-standing cubitus varus deformity and their effect during early osteoarthritis of the elbow.

OBJECTIVE: To quantify the bone density and stress distribution patterns in long-standing cubitus varus and clarify the effects of the deformity on bone density. DESIGN: We created three-dimensional computed tomography (CT) elbow models from 21 patients with long-standing cubitus varus deformities without advanced osteoarthritis (OA) and assessed the deformity by superimposing the affected humerus onto a mirror-image of the contralateral normal. Elbows were divided into 13 regions before measuring the bone density of each region and comparing the percentage of high-density volume (%HDV) between affected and normal sides. We constructed finite element models and quantitatively analyzed stress distribution. RESULTS: Average degrees of deformities were 20.1° of varus, 6.4° of extension, and 12.7° of internal rotation. The medial side of the affected humerus and ulna, Anteromedial trochlea (P < 0.001), Medial coronoid (P = 0.004), and Medial olecranon (P = 0.049) had significantly higher %HDVs than their normal counterparts. Conversely, %HDVs on the affected lateral side, Capitellum (P < 0.001), Anterolateral trochlea (P = 0.010), Posterolateral trochlea (P < 0.001), Lateral coronoid (P = 0.007), and Lateral olecranon (P < 0.001) were significantly lower than the normal side. The affected radial head %HDVs at Anterolateral and Posteromedial quadrants were high (P = 0.007) and low (P = 0.007), respectively. The bone density distribution coincided with stress distribution patterns revealed by finite element analysis (FEA), except in the lateral region influenced by forearm rotation. CONCLUSIONS: Repetitive stress on the medial elbow may alter bone density distribution patterns, probably presenting from early stage of OA.

Relationships between transepidermal water loss, cutaneous microcirculatory function and autonomic nervous activity.

OBJECTIVE: Several studies have shown that a deterioration of skin properties, an impaired cutaneous microcirculatory function and an imbalance of autonomic nervous activity are observed in smokers and in patients with diabetes mellitus or Raynaud's phenomenon. These observations suggest that skin properties are associated with cutaneous microcirculatory function and autonomic nervous activity in pathological conditions. However, there is no published evidence to support the concept that these two functions have any relationship with skin properties even in healthy subjects. To investigate the hypothesis that these properties are related, we conducted a survey of healthy adult subjects to investigate the relationships between cutaneous microcirculatory function and autonomic nervous activity and skin properties. METHODS: The hydration of the stratum corneum and transepidermal water loss (TEWL) were investigated as skin properties, and the responsiveness of skin blood flow (SkBF) to local warming was examined as an index of cutaneous microcirculatory function in 19 healthy adult male subjects. Electrocardiograms were monitored for 24 h and heart rate variability was analysed considering low-frequency power (LF: 0.04-0.15 Hz), high-frequency power (HF: 0.15-0.40 Hz) and a ratio of low- to high-frequency power (LF/HF) as indices of autonomic nervous activity; HF is an index of parasympathetic activity, whereas LF/HF is an index of sympathovagal balance. The relationships between those indices were then analysed. RESULTS: A moderate negative correlation was found between TEWL and the relative maximum rate of increases in the responsiveness of SkBF on local warming. A moderate positive and a moderate negative correlation were observed between TEWL and LF/HF or HF, respectively. Moreover, a moderate negative and a moderate positive correlation were shown between the responsiveness of SkBF and LF/HF or HF, respectively. The hydration of the stratum corneum showed no correlations with any indices of microcirculation or autonomic nervous activity. CONCLUSION: These results indicate that skin barrier function, cutaneous microcirculatory function and autonomic nervous activity are mutually associated in healthy adults.

A large ovarian leiomyoma discovered incidentally in a 76-year-old woman: case report.

BACKGROUND: Ovarian leiomyoma is very rare type of ovarian tumor. This benign tumor is seen in the pediatric age group to premenopausal women. CASE: A 76-year-old woman had a huge leiomyoma (19 x 11 x 10 cm) of the right ovary. The preoperative diagnosis was difficult to distinguish from a broad ligament leiomyoma or ovarian cancer. CONCLUSIONS: Although theses tumors are benign, its extreme rarity led us to report an additional and rather unusual case of ovarian leiomyoma, and to focus some attention on this type of tumor.

Colonization of chicken flocks by Campylobacter jejuni in multiple farms in Japan.

Campylobacter jejuni was monitored in 4 chicken farms during the period 2003 to 2006 to elucidate the mechanisms of transmission. Three farms (1 to 3), located at least 14 km from each other, belonged to an integrated poultry company, which also provided the farms with day-old chicks from several hatcheries as well as chicken feed. Another farm (4), which belonged to a different company, was located 270 m from farm 1. A total of 206 C. jejuni isolates obtained from the 4 farms were classified into 10 flaA-based RFLP types. Identical RFLP types were found in isolates obtained from chickens originating from multiple hatcheries and reared in different chicken houses on individual farms. Flocks were colonized by strains with 1 or 2 RFLP types in each production cycle, sometimes differing between cycles. Identical RFLP types were found in isolates obtained from the environment around the chicken houses. Using multilocus sequence typing, strains with different RFLP types could be distinguished from each other. Identical RFLP and multilocus sequence typing profiles were found in isolates obtained from farms 1 and 4, and from farms 1 and 2. These results suggest that C. jejuni in these farms comes from common sources external to the farms, even if the farms belong to different companies and obtain chicks from different suppliers.

An autopsy case of acute aortic dissection during postpartum period.

BACKGROUND: Aortic dissection in young women without Marfan disease is related in most instances to pregnancy. This is a potentially catastrophic occurrence. CASE: An autopsy case of acute aortic dissection type B (Stanford classification), clinically undiagnosed during late puerperium period in a young woman with no discernible risk factors (e.g. family history and signs of connective tissue diseases) is presented. Autopsy with ancillary investigations revealed that knowledge of this albeit relatively rare complication of postpartum may assist the clinician in earlier diagnosis and referral of patients for surgical treatment. CONCLUSION: This case is presented to raise awareness and review the literature for the critical care of postpartum patients.

Encephalomyelitis associated with microsporidian infection in farmed greater amberjack, Seriola dumerili (Risso).

An outbreak of a disease characterized by a peculiar spiral movement in farmed greater amberjack, Seriola dumerili (Risso), occurred in Kagoshima Prefecture, Japan, in May 2008, immediately after importing the fish from China. Although neither bacteria nor viruses were detected in routine diagnostic tests, histopathological observations of the affected fish revealed severe inflammation in the tegmentum of the brain including the medulla oblongata and the anterior part of the spinal cord. In addition, a microsporidian parasite was observed in the nerve cell bodies or axons in the inflamed tissues. We identified a microsporidian small subunit rRNA gene (SSU rDNA) from the lesion, and the sequence showed 96.1% identity with that of Spraguea lophii. Subsequent in situ hybridization using probes presumably specific to the SSU rRNA confirmed that the parasite observed in histopathology harboured the identified SSU rRNA. Apparently degenerated microsporidian cells or spores were also frequently observed in tissue sections. Thus, the disease was most probably caused by the infection of a hitherto unknown microsporidian parasite that has a genetic affinity to the genus Spraguea, in the central nervous system of the amberjack.

Peptide YY producing strumal carcinoid tumor of the ovary.

BACKGROUND: Primary ovarian carcinoid tumor is uncommon and represents less than 0.1% of ovarian malignancies. The strumal carcinoid may be complicated by carcinoid syndrome induced by peptide YY (PYY). CASE REPORT: We describe a 45-year-old woman with a bilateral ovarian tumor diagnosed through periodical gynecological examination. She presented with severe constipation. Right ovarian cyst laparoscopically resected was diagnosed as a strumal carcinoid tumor; the left one was mature cystic teratoma. No metastatic findings were seen macroscopically on the ovarian surface and pelvic peritoneum. Constipation was drastically improved by resecting the tumor. The carcinoid tumor cells were positive for tumor-producing PYY by mRNA analysis. CONCLUSION: It is important to be aware of this entity in the pathological diagnosis of ovarian tumors, in the presence of any clinical indicator of carcinoid tumor/syndrome, as it carries a markedly better prognosis and clinical outcome in comparison with most other malignant ovarian tumors.

Three-dimensional distribution of articular cartilage thickness in the elderly cadaveric acetabulum: a new method using three-dimensional digitizer and CT.

OBJECTIVE: The aim of this study was to evaluate the three-dimensional (3D) distribution of the acetabular articular cartilage thickness in cadaveric elderly individuals, measured using a new method with a 3D-digitizer and computed tomography (CT) and to validate this method using a thresholding technique. DESIGN: Twenty cadaveric hemipelves without fracture, previous hip surgery, or macroscopic degenerative changes were digitized by a 3D-digitizer to make 3D cartilage surface models, and scanned by 3D-CT to create 3D bone surface models. These two surface models were then merged using a surface registration method. Acetabular articular cartilage thickness was evaluated as the distance between the two surface models, and the distribution was mapped. Tests for accuracy and reproducibility were performed by comparing the cartilage thickness of five human femoral heads measured by stereomicroscopy with the distance between the cartilage and bone surface models. RESULTS: The superolateral cartilage tended to be the thickest in all acetabula. The smallest category (0-0.5 mm) of articular cartilage thickness existed at the posteroinferior lunate surface. In this new method, the mean measurement error was 0.018+/-0.044 mm for the average optimum threshold and the intraclass correlation coefficients were 0.99 in surface registration and 0.94 in data acquisition for reproducibility, indicating high accuracy and reproducibility. CONCLUSIONS: The proposed method for measuring articular cartilage using a 3D-digitizer and 3D-CT was accurate and reproducible. In the elderly individuals, acetabular articular cartilage tended to be thicker in the superolateral area and there was the thinnest category (0-0.5 mm) on the posteroinferior lunate surface of the acetabulum. The contour generated along 480 Hounsfield units (HU) was closest to the subchondral bone contour in the elderly hip.

Prevention of neural tube defects by loss of function of inducible nitric oxide synthase in fetuses of a mouse model of streptozotocin-induced diabetes.

AIMS/HYPOTHESIS: Maternal diabetes during pregnancy increases the risk of congenital malformations such as neural tube defects (NTDs). Although the mechanism of this effect is uncertain, it is known that levels of nitric oxide synthase (NOS) and nitric oxide are elevated in embryos of a mouse model of diabetes. We postulated that overproduction of nitric oxide causes diabetes-induced congenital malformations and that inhibition of inducible NOS (iNOS) might prevent diabetic embryopathy. METHODS: Mice were rendered hyperglycaemic by intraperitoneal injection of streptozotocin. The incidence of congenital malformations including NTDs was evaluated on gestational day 18.5. We assessed the involvement of iNOS in diabetes-induced malformation by administering ONO-1714, a specific inhibitor of iNOS, to pregnant mice with streptozotocin-induced diabetic mice and by screening mice with iNOS deficiency due to genetic knockout (iNos(-/-)). RESULTS: ONO-1714 markedly reduced the incidence of congenital anomalies, including NTDs, in fetuses of a mouse model of diabetes. It also prevented apoptosis in the head region of fetuses, indicating that iNOS is involved in diabetes-related congenital malformations. Indeed, no NTDs were observed in fetuses of diabetic iNos(-/-) mice and the incidence of other malformations was also markedly reduced. CONCLUSIONS/INTERPRETATION: We conclude that increased iNOS activity during organogenesis plays a crucial role in the pathogenesis of diabetes-induced malformations and suggest that inhibitors of iNOS might help prevent malformations, especially NTDs, in diabetic pregnancy.

Alpha-lipoic acid reduces congenital malformations in the offspring of diabetic mice.

BACKGROUND: The mechanism of diabetes-induced congenital malformation remains to be elucidated. It has been reported that alpha-lipoic acid (LA) prevents neural tube defects (NTDs) in offsprings of rats with streptozotocin-induced diabetes. Here, we evaluate the protective effect of LA against diabetic embryopathy, including NTDs, cardiovascular malformations (CVMs), and skeletal malformations, in mice. METHODS: Female mice were rendered hyperglycemic using streptozotocin and then mated with normal male mouse. Pregnant diabetic or non-diabetic mice were treated daily with either LA (100 mg/kg body weight) or saline between gestational days 0 and 18. On day 18, fetuses were examined for congenital malformations. RESULTS: Plasma glucose levels on day 18 were not affected by LA treatment. No congenital malformations were observed either in the saline-treated or LA-treated non-diabetic group. In the saline-treated diabetic group, 39% of fetuses had external malformations and 30% had NTDs. In the LA-treated diabetic group, the corresponding proportions were 11 and 8%, respectively. LA treatment also decreased the incidence of CVMs from 30-3% and of skeletal malformations from 29-6%. CONCLUSIONS: We conclude that LA can reduce NTDs, CVMs and skeletal malformations in the offspring of diabetic mice at term delivery.

Viscosity of lunar lavas.

The viscosity of a synthetic silicate liquid with the composition of a lunar rock has been determined experimentally and found to be lower than that of any previously studied volcanic rock on earth. Course Crystallinity can be a feature of rapidly cooled surface lavas.

Thermal conductivity of lunar and terrestrial igneous rocks in their melting range.

The thermal conductivity of a synthetic lunar rock in its melting range is about half that of a terrestrial basalt. The low conductivity and increased efficiency of insulating crusts on lunar lavas will enable flows to cover great distances without being quenched by high radiant heat losses from the surface. For a given rate of heat production, the thermal gradient of the moon would be significantly steeper than that of the earth.

Exocytosis in spermatozoa in response to progesterone and zona pellucida.

Exocytosis in mammalian spermatozoa (the acrosome reaction) is a process essential for fertilization. Both progesterone and zona pellucida induce exocytosis in spermatozoa, which may encounter both during penetration of the oocyte's vestments. When mouse spermatozoa were exposed first to progesterone and then to zona pellucida, exocytosis was enhanced to a greater degree than that seen when the agonists were presented together or in the inverse order, which suggests that the steroid exerts a priming effect. Progesterone similarly primed the generation of intracellular messengers evoked by zona pellucida. The effects triggered by progesterone were mimicked by gamma-aminobutyric acid (GABA) and were blocked by bicuculline, which indicates that the steroid acts on a GABAA receptor.

Endoscopic mucosal resection for early gastric cancer: comparison of two modifications of the cap method.

BACKGROUND AND STUDY AIM: Endoscopic mucosal resection using a cap (EMR-C) is an established method for curative resection of early neoplastic lesions; prelooping of the snare may however be difficult and lead to imprecise resection. We therefore compared two modifications of the conventional technique using outer snare placement with an accessory channel in a prospective, nonrandomized study. PATIENTS AND METHODS: Between October 2004 and March 2007, 54 patients (men 37, women 17; mean age 71 years) underwent EMR. One method involved an internally retained snare (IRS) cap, with a fixed prelooped snare inside the cap; the other method used an externally guided snare (EGS) cap with the snare guided over an oblique cap. The main outcome parameters were specimen size, en bloc resection, and complications. RESULTS: There was no difference between use of the IRS and EGS cap methods in relation to specimen size (27.6 vs. 27.1 mm), or rates of en bloc resection (88.9 % vs. 83.3 %); only one perforation occurred, and this was in the EGS group. CONCLUSION: Both techniques appeared to provide similar efficacy, the inner rim of the IRS cap stabilizes aspiration of the lesion compared with the EGS cap that does not have it.

Tumorous calcification causing carpal tunnel syndrome.

PURPOSE: There is little information on solitary tumorous calcification causing carpal tunnel syndrome in the literature. This study describes our experience with surgically treated cases of solitary tumorous calcification causing carpal tunnel syndrome. METHODS: Seven patients with symptomatic carpal tunnel syndrome who had tumorous calcification in the carpal tunnel confirmed by radiographical examinations and had then undergone open incisional carpal tunnel release were reviewed. The precise location and the appearance of the calcified mass were confirmed with the preoperative radiographic examinations and the operative records. The additional histology and the composition analysis of the calcified mass were also reviewed in five removed calcifications. RESULTS: Radiographs of each hand revealed a mass of calcification lying anterior to the capitate bone in the carpal tunnel. Intraoperatively, an oval calcified nodule, measuring 10 x 10 x 10 - 18 mm, was observed overlying the capitate, attached firmly to the palmar radiocarpal extrinsic ligament. A composition with an average of 60 % basic calcium phosphate was revealed by infrared absorption spectrometry. Histological sections showed a calcified deposit surrounded by fibrocartilagenous tissue in three cases. CONCLUSION: These facts suggest that the pathogenesis of tumorous calcification causing carpal tunnel syndrome is comparable with that of calcifying tendinitis of the rotator cuff in which a cell-mediated reactive process plays an important role at the tendon insertion.

Virulence of Escherichia coli Isolates Obtained from Layer Chickens with Colibacillosis Associated with Pericarditis, Perihepatitis, and Salpingitis in Experimentally Infected Chicks and Embryonated Eggs.

To evaluate the virulence of avian pathogenic Escherichia coli (APEC) isolates obtained from colibacillosis cases associated with pericarditis, perihepatitis, and salpingitis, the embryo lethality assay and experimental infection model in chicks were used in this study. According to the established criteria based on mortality in the embryo lethality assay for evaluating the virulence of E. coli isolates, 23 of the 26 APEC isolates associated with pericarditis and perihepatitis and 8 of the 20 isolates associated with salpingitis were found to be virulent. Isolate D137, which had been obtained from a case with pericarditis and perihepatitis and had an embryo mortality of 92%, and isolate D445, which had been obtained from a case with pericarditis and perihepatitis and had an embryo mortality of 17%, were used for the experimental infection. Four of the five 11-day-old chickens inoculated through the air sac with isolate D137 died 1 day postinoculation, and the challenge strain was recovered from the air sac, pericardial sac, or liver; however, colibacillosis lesions were found in only one of the five birds postmortem. All five chicks inoculated with isolate D445 survived for 7 days postinoculation and exhibited airsacculitis or pericarditis lesions at 7 days postinoculation; the challenge strain was not recovered from the lesions postmortem. The results obtained in this study suggest that the different APEC isolates tested cause illness in chickens through distinct pathogenesis.

The novel compound NO-1886 increases lipoprotein lipase activity with resulting elevation of high density lipoprotein cholesterol, and long-term administration inhibits atherogenesis in the coronary arteries of rats with experimental atherosclerosis.

We have discovered a novel compound, NO-1886, which possesses a powerful lipoprotein lipase (LPL) activity-increasing action. Administration of NO-1886 increased LPL activity in the postheparin plasma, adipose tissue, and myocardium of rats, and produced a reduction in plasma triglyceride levels with concomitant elevation of HDL cholesterol levels. Administration of NO-1886 increased LPL enzyme mass in postheparin plasma and mRNA activity in epididymal adipose tissue, and it was concluded that the mode of action of this compound is stimulation of tissue LPL synthesis. We also conducted long-term studies to assess the impact of increases in LPL activity and HDL levels on the development of atherosclerotic lesions in rats. Administration of NO-1886 for as long as 90 d significantly decreased the degree of atherosclerotic changes in the coronary arteries of vitamin D2-treated, cholesterol-fed rats. Statistical analysis indicated that increased concentration of HDL is the factor contributing mostly to the prevention of coronary artery sclerosis. In summary, the results of our study indicate that compound NO-1886 increases LPL activity, causing an elevation in HDL levels, and that long-term administration of NO-1886 to rats with experimental atherosclerosis provides significant protection against the development of coronary artery lesions.