Acsl1 is essential for skin barrier function through the activation of linoleic acid and biosynthesis of ω-O-acylceramide in mice.

The long-chain acyl-CoA synthase1 (Acsl1) is a major enzyme that converts long-chain fatty acids to acyl-CoAs. The role of Acsl1 in energy metabolism has been elucidated in the adipose tissue, heart, and skeletal muscle. Here, we demonstrate that systemic deficiency of Acsl1 caused severe skin barrier defects, leading to embryonic lethality. Acsl1 mRNA and protein are expressed in the Acsl1+/+ epidermis, which are absent in Acsl1-/- mice. In Acsl1-/- mice, epidermal ceramide [EOS] (Cer[EOS]) containing ω-O-esterified linoleic acid, a lipid essential for the skin barrier, was significantly reduced. Conversely, ω-hydroxy ceramide (Cer[OS]), a precursor of Cer[EOS], was increased. Moreover, the levels of triglyceride (TG) species containing linoleic acids were lower in Acsl1-/- mice, whereas those not containing linoleic acid were comparable to Acsl1+/+ mice. As TG is considered to work as a reservoir of linoleic acid for the biosynthesis of Cer[EOS] from Cer[OS], our results suggest that Acsl1 plays an essential role in ω-O-acylceramide synthesis by providing linoleic acid for ω-O-esterification. Therefore, our findings identified a new biological role of Acsl1 as a regulator of the skin barrier.

Adrenal dysplasia in a young SD rat.

The present report describes an adrenal dysplasia in which developmental abnormality was observed in the adrenal gland of a six-week-old male Crl:CD(SD) rat. Microscopically, a localized lesion composed of mildly vacuolated adrenal fasciculata cells with a slightly disturbed cord structure and containing areas with high cell density was observed in a unilateral adrenal gland; no macroscopical changes were detected in the organ. The areas with high cell density consisted of two cell types. One type included small cells with a round nucleus and acidophilic cytoplasm, and the cells were positive for steroidogenic factor-1 (SF-1) but negative for nestin. The other type of cells had a spindle to polygonal shape, clear nucleus, and a cytoplasm with an obscure boundary; the cells were positive for nestin but negative for SF-1, neuronal nuclear antigen, and chromogranin A. These results suggested that the former type of cells were adrenal cortex cells and that the latter were immature neuronal cells. Considering that immature adrenal cortex cells and neural crest cells (future adrenal medulla) are mixed during a stage in rat adrenal gland development, we concluded that the observed lesion was caused by developmental abnormality. To the best of our knowledge, this is the first report to describe dysplasia in rat adrenal glands.

Characterization of pancreatic islet cell tumors and renal tumors induced by a combined treatment of streptozotocin and nicotinamide in male SD rats.

We herein investigated the histopathological features, including proliferative activity and immunoexpression, of pancreatic islet cell tumors (ICTs) in male SD rats induced by streptozotocin (STZ) and nicotinamide (NA), and discussed their relevance to biological behaviors and prognoses. A total of 70 and 43% of rats developed ICTs 37-45 weeks after the treatment with STZ (50 or 75mg/kg, i.v.) and NA (350mg/kg, twice, p.o.), respectively. Among the islet tumors observed in the STZ/NA-treated groups, 75% were adenomas, while 25% were carcinomas. Most STZ/NA-induced carcinomas were characterized by well-differentiated tumor cells with/without local invasion into the surrounding tissues, and weak proliferative activity. No outcome such as distance metastasis and death was noted. All of the ICTs strongly expressed insulin, part of which had hormone productivity; however there were no hypoglycemia-related clinical signs such as convulsion in these rats 36 weeks after the treatment. These results suggested that rat ICTs induced STZ/NA have small impact on biological activity or prognosis. STZ/NA treatment significantly increased of focal proliferative lesions in the kidney, liver and adrenal glands other than pancreatic islets. Of the STZ/NA-induced kidney tumors, more than 60% were renal cell adenomas, and many of them were basophilic type. The incidence of eosinophilic or clear cell type of tumors was less than 10%, respectively. Immunohistochemical analyses revealed that many of the STZ/NA-induced basophilic type of renal tumors were derived from proximal tubules, whereas the clear cell and eosinophilic types were derived from collecting tubules.

Cutaneous mastocytosis with a mutation in the juxtamembrane domain of c-kit in a young laboratory beagle dog.

Cutaneous mastocytosis, which resembles a subset of urticaria pigmentosa in humans, is rare in dogs. We herein report unrepresentative neoplastic proliferation of mast cells in ventral skin removed routinely from a nine-month-old female laboratory beagle dog at necropsy. A histological examination revealed diffuse extensive cellular infiltration from the superficial to deep dermis in most parts of the skin around the fourth and fifth mammary papilla without nodule formation. Tumor cells were fairly monomorphic, well-differentiated mast cells with round nuclei of small distinct nucleoli and moderate to abundant, slightly eosinophilic and granular cytoplasm. A perivascular arrangement of mast cells was noted at the margin of the lesions. Infiltration of eosinophils and degeneration of collagen were not observed in the dermis. Cutaneous mastocytosis was diagnosed based on these features. A sequence analysis of lesions revealed the deletion of Gln555 to Ile570 within the juxtamembrane domain of c-kit (exon 11).

Awa1p on the cell surface of sake yeast inhibits biofilm formation and the co-aggregation between sake yeasts and Lactobacillus plantarum ML11-11.

We examined mixed-species biofilm formation between Lactobacillus plantarum ML11-11 and both foaming and non-foaming mutant strains of Saccharomyces cerevisiae sake yeasts. Wild-type strains showed significantly lower levels of biofilm formation compared with the non-foaming mutants. Awa1p, a protein involved in foam formation during sake brewing, is a glycosylphosphatidylinositol (GPI)-anchored protein and is associated with the cell wall of sake yeasts. The AWA1 gene of the non-foaming mutant strain Kyokai no. 701 (K701) has lost the C-terminal sequence that includes the GPI anchor signal. Mixed-species biofilm formation and co-aggregation of wild-type strain Kyokai no. 7 (K7) were significantly lower than K701 UT-1 (K701 ura3/ura3 trp1/trp1), while the levels of strain K701 UT-1 carrying the AWA1 on a plasmid were comparable to those of K7. The levels of biofilm formation and co-aggregation of the strain K701 UT-1 harboring AWA1 with a deleted GPI anchor signal were similar to those of K701 UT-1. These results clearly demonstrate that Awa1p present on the surface of sake yeast strain K7 inhibits adhesion between yeast cells and L. plantarum ML11-11, consequently impeding mixed-species biofilm formation.

Screening of lactic acid bacteria that can form mixed-species biofilm with Saccharomyces cerevisiae.

The abilities of lactic acid bacteria (LAB) to form mixed-species biofilm with Saccharomyces cerevisiae in a static co-culture were investigated out of 168 LAB stock cultures, and two Lactobacillus plantarum strains (D71 and E31) and one Leuconostoc mesenteroides strain K01 were found to form mixed-species biofilm with S. cerevisiae BY4741. SEM observation showed that there was no significant difference in morphological properties among these three mixed-species biofilms and they resembled that formed by S. cerevisiae with L. plantarum ML11-11 previously isolated from a brewing sample of Fukuyama pot vinegar. The co-aggregation assays showed that L. plantarum D71 and L. plantarum E31 could co-aggregate with S. cerevisiae similarly to L. plantarum ML11-11, while L. mesenteroides K01 had no ability to co-aggregate with yeast. The above results indicate that aggregation followed by direct cell-to-cell contact is required for mixed-species biofilm formation between these L. plantarum strains and S. cerevisiae, though some different mechanism may be involved in biofilm formation between L. mesenteroides strain and S. cerevisiae.

Involvement of neutrophil gelatinase-associated lipocalin and osteopontin in renal tubular regeneration and interstitial fibrosis after cisplatin-induced renal failure.

The kidney has a capacity to recover from ischemic or toxic insults that result in cell death, and timely tissue repair of affected renal tubules may arrest progression of injury, leading to regression of injury and paving the way for recovery. To investigate the roles of neutrophil gelatinase-associated lipocalin (NGAL/lcn2) and osteopontin (OPN/spp1) during renal regeneration, the expression patterns of NGAL and OPN in the cisplatin-induced rat renal failure model were examined. NGAL expression was increased from day 1 after injection; it was seen mainly in the completely regenerating proximal tubules of the cortico-medullary junction on days 3-35; however, the expression was not seen in abnormally dilated or atrophied renal tubules surrounded by fibrotic lesions. On the other hand, OPN expression was increased from day 5 and the increased expression developed exclusively in the abnormal renal tubules. NGAL expression level well correlated with the proliferating activity in the regenerating renal epithelial cells, whereas OPN significantly correlated with the α-smooth muscle actin-positive myofibroblast appearance, expression of transforming growth factor (TGF)-ß1, and the number of CD68-positive macrophages. Interestingly, rat renal epithelial cell line (NRK-52E) treated with TGF-ß1 decreased NGAL expression, but increased OPN expression in a dose-dependent manner. Because increases of TGF-ß1, myofibroblasts and macrophages contribute to progressive interstitial renal fibrosis, OPN may be involved in the pathogenesis of fibrosis; on the contrary, NGAL may play a role in tubular regeneration after injury. Expression analysis of NGAL and OPN would be useful to investigate the tubule damage in renal-toxicity.

Effect of basic fibroblast growth factor on angiogenesis and bone regeneration in non-critical-size bone defects in rat calvaria.

We used microcomputed tomography (micro-CT) to evaluate the effects of basic fibroblast growth factor (FGF-2) contained in absorbable collagen sponges on angiogenesis and bone regeneration in rat calvarial non-critical-size bone defects. Two symmetrical non-critical-size calvarial bone defects (diameter, 2.7 mm) were created in male Fisher rats. An absorbable collagen sponge with or without FGF-2 (0.1% or 0.3%) was implanted into each defect. Blood vessel volume and bone volume were calculated using software. On day 28, blood vessel volume and bone volume were significantly greater in the 0.3% and 0.1% FGF-2 groups than in the control group. FGF-2 concentration-dependently increased blood vessels and bone formation in non-critical-size bone defects in rat calvaria.

Effect of a chitosan sponge impregnated with platelet-derived growth factor on bone augmentation beyond the skeletal envelope in rat calvaria.

We evaluated the ability of platelet-derived growth factor (PDGF) to promote bone augmentation beyond the skeletal envelope in rat calvaria. The calvariae of 14 rats were exposed, and two plastic caps-one with 0.03% PDGF and a chitosan sponge and one with a chitosan sponge alone-were placed. Microcomputed tomography and histologic sections were used to determine the amount of bone augmentation within the plastic caps. Bone volume was calculated using measurement software. Bone volume and amount of bone augmentation were significantly greater in the PDGF group than in the control group. In conclusion, a chitosan sponge containing 0.03% PDGF enhanced bone formation beyond the skeletal envelope in rat calvaria.

FGF-2 angiogenesis in bone regeneration within critical-sized bone defects in rat calvaria.

OBJECTIVE: This study examined the effect of basic fibroblast growth factor (FGF)-2 with an absorbable collagen sponge on angiogenesis and bone regeneration in rat calvarial critical-sized bone defects using microcomputed tomography. METHODS: Forty 7-week-old male Fischer rats were used. The symmetrical critical-sized calvarial bone defects (5 mm diameter) were created. An absorbable collagen sponge with or without FGF-2 (0.1% or 0.3%) was implanted into each bone defect. RESULTS: Blood vessel and bone volumes were significantly higher in the 0.3% FGF-2 group compared with the control and 0.1% FGF-2 groups on day 28. Significantly more osteoblast- and osteoclast-like cells were seen in the 0.3% FGF-2 group. CONCLUSIONS: Thus, FGF-2 increased blood vessel and bone formation in rat calvarial critical-sized bone defects.

Community-based system, reports, and substantiated cases of elder abuse: disparities between municipalities and relating factors in Japan.

This study examines (1) the staffing and financial characteristics of systems for elder abuse detection and intervention in the municipal governments of Japan and (2) the relationship among the development of detection and intervention systems, the reporting rates of suspected elder abuse cases, and substantiated abuse rates in 927 municipalities across Japan. Progressive systems for the detection and intervention of elder abuse were significantly associated with a larger number of public officers than in non-progressive systems. Furthermore, greater rates of both suspected and substantiated cases of abuse were associated with progressive systems for elder abuse detection and intervention. Per capita annual expenditures on the comprehensive support project and the community general support center's catchment under the Long-Term Care Insurance (LTCI) program showed no significant association with the development of systems, the rate of suspected cases, or the number of substantiated cases. National social policy makers should examine strategies that would help municipalities assign sufficient staff to elder abuse detection and intervention programs.

Cardiac-specific over-expression of epidermal growth factor receptor 2 (ErbB2) induces pro-survival pathways and hypertrophic cardiomyopathy in mice.

BACKGROUND: Emerging evidence shows that ErbB2 signaling has a critical role in cardiomyocyte physiology, based mainly on findings that blocking ErbB2 for cancer therapy is toxic to cardiac cells. However, consequences of high levels of ErbB2 activity in the heart have not been previously explored. METHODOLOGY/PRINCIPAL FINDINGS: We investigated consequences of cardiac-restricted over-expression of ErbB2 in two novel lines of transgenic mice. Both lines develop striking concentric cardiac hypertrophy, without heart failure or decreased life span. ErbB2 transgenic mice display electrocardiographic characteristics similar to those found in patients with Hypertrophic Cardiomyopathy, with susceptibility to adrenergic-induced arrhythmias. The hypertrophic hearts, which are 2-3 times larger than those of control littermates, express increased atrial natriuretic peptide and ß-myosin heavy chain mRNA, consistent with a hypertrophic phenotype. Cardiomyocytes in these hearts are significantly larger than wild type cardiomyocytes, with enlarged nuclei and distinctive myocardial disarray. Interestingly, the over-expression of ErbB2 induces a concurrent up-regulation of multiple proteins associated with this signaling pathway, including EGFR, ErbB3, ErbB4, PI3K subunits p110 and p85, bcl-2 and multiple protective heat shock proteins. Additionally, ErbB2 up-regulation leads to an anti-apoptotic shift in the ratio of bcl-xS/xL in the heart. Finally, ErbB2 over-expression results in increased activation of the translation machinery involving S6, 4E-BP1 and eIF4E. The dependence of this hypertrophic phenotype on ErbB family signaling is confirmed by reduction in heart mass and cardiomyocyte size, and inactivation of pro-hypertrophic signaling in transgenic animals treated with the ErbB1/2 inhibitor, lapatinib. CONCLUSIONS/SIGNIFICANCE: These studies are the first to demonstrate that increased ErbB2 over-expression in the heart can activate protective signaling pathways and induce a phenotype consistent with Hypertrophic Cardiomyopathy. Furthermore, our work suggests that in the situation where ErbB2 signaling contributes to cardiac hypertrophy, inhibition of this pathway may reverse this process.

Contraction behaviors of Vorticella sp. stalk investigated using high-speed video camera. I: Nucleation and growth model.

The contraction process of living Vorticella sp. has been investigated by image processing using a high-speed video camera. In order to express the temporal change in the stalk length resulting from the contraction, a damped spring model and a nucleation and growth model are applied. A double exponential is deduced from a conventional damped spring model, while a stretched exponential is newly proposed from a nucleation and growth model. The stretched exponential function is more suitable for the curve fitting and suggests a more particular contraction mechanism in which the contraction of the stalk begins near the cell body and spreads downwards along the stalk. The index value of the stretched exponential is evaluated in the range from 1 to 2 in accordance with the model in which the contraction undergoes through nucleation and growth in a one-dimensional space.

Contraction behaviors of Vorticella sp. stalk investigated using high-speed video camera. II: Viscosity effect of several types of polymer additives.

The contraction process of living Vorticella sp. in polymer solutions with various viscosities has been investigated by image processing using a high-speed video camera. The viscosity of the external fluid ranges from 1 to 5mPa·s for different polymer additives such as hydroxypropyl cellulose, polyethylene oxide, and Ficoll. The temporal change in the contraction length of Vorticella sp. in various macromolecular solutions is fitted well by a stretched exponential function based on the nucleation and growth model. The maximum speed of the contractile process monotonically decreases with an increase in the external viscosity, in accordance with power law behavior. The index values approximate to 0.5 and this suggests that the viscous energy dissipated by the contraction of Vorticella sp. is constant in a macromolecular environment.

[Pulmonary mycobacteriosis caused by Mycobacterium peregrinum in a young, healthy man].

A 24-year-old male never-smoker with no contributory medical history was referred to our division because of chronic productive cough and fever. His chest CT showed pulmonary consolidation including nodular shadows around the bronchovascular bundle in both upper lobes and right middle lobe. Sputum smears were negative but cultures were positive for Mycobacterium peregrinum, twice. He was given a diagnosis of pulmonary mycobacteriosis caused by M. peregrinum and treated with clarithromycin, levofloxacin, and ethambutol. His symptoms and the infiltrates improved, and further sputum cultures were negative for mycobacteria. M. peregrinum is a species included in the M. fortuitum group, of rapidly-growing non-tuberculous mycobacteria. To the best of our knowledge, only 2 cases with lung infection caused by M. peregrinum have been previously reported, both of which occurred in patients in their 60s with underlying diseases, including low immunity due to immunosuppressants for polymyositis, or bronchiectasia. We describe a rare case of pulmonary mycobacteriosis due to M. peregrinum in a healthy man without any underlying diseases.

The effect of emphysema on lung function and survival in patients with idiopathic pulmonary fibrosis.

BACKGROUND AND OBJECTIVE: In this study the prevalence, lung function and prognosis of IPF combined with emphysema were evaluated. METHODS: Consecutive patients with usual interstitial pneumonia (UIP) on high-resolution computed tomography (HRCT), with or without emphysema, were assessed retrospectively. The area of fibrosis in the base of the lungs was assessed by HRCT as minimal (<2 cm from the subpleura), moderate (>or=2 cm from the subpleura, <1/3 of the area of the base of the lungs) or severe (>or=1/3 of the area of the base of the lungs). RESULTS: Among 660 patients with UIP on HRCT, 221 showed upper-lobe emphysema. Pulmonary function results for patients with UIP and UIP/emphysema, respectively, were: FVC, 71.8% and 87.1%; FEV1%, 86.7% and 87.9%; and DL(CO), 74.3% and 65.2% of predicted. The relationship between FVC, the extent of fibrosis and survival was investigated in 362 patients with records of pulmonary function tests and no lung cancer at the time of entry into the study. Although the extent of fibrosis was similar between the groups, 71.3% of UIP patients met the lung volume criteria for IPF (FVC <80% of predicted), whereas only 26.5% of UIP/emphysema patients met the lung volume criteria for IPF. Median survival was 7.5 years in the UIP group and 8.5 years in the UIP/emphysema group. CONCLUSIONS: Emphysema was a common finding in patients with UIP. Patients with UIP and emphysema had greater lung volumes and better survival compared with those with UIP alone.

Evaluation of the usefulness of urinary biomarkers for nephrotoxicity in rats.

Since nephrotoxicity affects the development of drug candidates, it is important to detect their toxicity at an early stage of drug development. In this study, we measured twelve urinary nephrotoxic biomarkers [total protein, albumin, kidney injury molecule-1 (KIM-1), clusterin, beta2-microglobulin, cystatin-c, alpha-glutathione S-transferase, mu-glutathione S-transferase, N-acetyl-beta-d-glucosaminidase, lactate dehydrogenase (LDH), aspartate aminotransferase and neutrophil gelatinase-associated lipocalin (NGAL)] and two conventional blood nephrotoxic biomarkers (creatinine and blood urea nitrogen) in rat models treated intravenously with puromycin aminonucleoside (PAN) or cisplatin (CDDP), which are known to induce glomerular injury or proximal tubular injury, respectively, and evaluated their usefulness by receiver operating characteristic analysis. In the PAN-treated rats, urinary albumin and (NGAL) were dramatically increased, which were thought to be caused by the dysfunction of proximal tubule in addition to glomerular injury. Conversely, based on its early and time-dependent increase, its large magnitude of alteration and its high accuracy and sensitivity of detection, (KIM-1) in urine appeared to be the best biomarker for detection of CDDP-induced proximal tubular injury. Moreover, (LDH) was considered useful for broad detection of damaged nephrons, because of its broad distribution along the nephron. Therefore, combinatorial measurement of these biomarkers may be a powerful tool for highly effective screening of nephrotoxicity.

[Clinical features of pneumothorax in patients with active nontuberculous mycobacterial lung disease].

UNLABELLED: This study was retrospectively conducted to clarify the clinical features of pneumothorax in patients with active nontuberculous mycobacterial (NTM) lung disease. The patients were 9 men and 7 women, with a median age of 70. Pathogenic mycobacteria were 12 Mycobacterium avium complex, 2 Mycobacterium kansasii, and 2 Mycobacterium fortuitum. More than one lung field was affected by NTM disease in 11 patients, whereas 4 patients were given a diagnosis of mild NTM disease only at the onset of pneumothorax. Five patients recovered with rest only, 4 with thoracic drainage, 4 needed surgery, and 2 developed chronic pneumothorax. Five patients experienced more than one recurrence of pneumothorax during treatment for NTM disease. The rate of the complication of pneumothorax in patients with NTM lung disease was estimated at around 2.3%. CONCLUSION: Pneumothorax associated with NTM lung disease is not rare and is sometimes difficult to control.

Spontaneous Erythroid Leukemia in a 7-Week-Old Crl:CD (SD) Rat.

A young male Crl:CD (SD) rat with erythroid leukemia that presented with emaciation, abdominal distension and a pale visible mucosal membrane was euthanized at 7 weeks of age. At necropsy, enlargement of liver, spleen and pancreatic lymph node was noted. Analysis of blood smear samples revealed many mono- or binucleated erythroblasts that had PAS-positive vacuoles in the cytoplasm. Histopathologically, neoplastic proliferation of atypical cells was observed in the hepatic sinusoids, splenic red pulp, bone marrow, pancreatic lymph node, kidney and lung. Neoplastic cells showed a round to spindle shape, and some neoplastic cells had deeply stained small nuclei and small cytoplasms and resembled erythroblasts. Immunohistochemically, many neoplastic cells were positive for hemoglobin. To our knowledge, this is the first report of erythroid leukemia in a rat of this age. The observed features were similar to those of pure erythroid leukemia in humans.

[A case of Ehlers-Danlos syndrome suspected from pulmonary hematoma due to disruption of the lung].

A 20-year-old man was referred to our hospital due to hemoptysis. Chest CT showed a ground-glass opacity, suggesting pulmonary bleeding; however, a diagnosis was not obtained. At a follow-up examination after 2 months, Chest CT showed improvement of the ground-glass opacity, however a cavitary nodule had newly appeared. Four months later, another new nodule was found on chest X-ray film. Video-assisted thoracoscopic lung biopsy was performed for pathological diagnosis; disruption of the pleural, lung and blood vessels, and pulmonary hematoma were found. We thought of the fragility of the pleuro-pulmonary connective tissue and also thought of the probability of Ehlers-Danlos syndrome (EDS). A biochemical analysis of cultured dermal fibroblasts and molecular biological examination revealed decreased production of type III collagen in fibroblasts and COL3A1 mutation. We diagnosed this case as vascular EDS. EDS is one of the differential diagnoses in patients presenting hemoptysis and pulmonary hematoma due to disruption of the lung.