Presence of Phlebitis in Aseptic Nasal Septal Abscess Complicated with Ulcerative Colitis; Possible Association with Granulomatosis with Polyangiitis: A Case Report.

Although nasal septal abscesses are uncommon, their cosmetic complications can be severe. Hence, prompt diagnosis and treatment are important. Here, we report a case of aseptic nasal septal abscess in a patient with proteinase 3-antineutrophil cytoplasmic antibody (PR3-ANCA)-positive ulcerative colitis (UC), in which phlebitis was observed and granulomatosis with polyangiitis (GPA) might co-exist. A 27-year-old female suffered from intermittent abdominal pain and diarrhea for several years. She visited our hospital complaining of worsening swelling and pain in the middle forehead and fever lasting 2 weeks. Physical examination and computed tomography revealed severe swelling of the nasal septum. The patient was diagnosed with nasal septal abscess, and incision drainage and biopsy from the bilateral nasal septum were performed, which showed severe ulcerative neutrophilic mucositis with phlebitis. Simultaneously, blood examination yielded slight positivity for PR3-ANCA. Colonoscopy, including biopsy, revealed severe inflammation without vasculitis nor granuloma, which led to the diagnosis with PR3-ANCA-positive UC. Phlebitis in the nasal mucosa and elevated PR3-ANCA suggested co-existing GPA; hence, she was treated with glucocorticoids and rituximab. Following treatment, the nasal septal abscess and digestive symptoms disappeared. She was discharged on day 25 without symptom recurrence or major nasal deformity. For the prevention of nasal deformity due to persistent inflammation, prompt administration of immunosuppressive therapy should be considered with adequate evaluations for systemic diseases, including UC and GPA.

Intrinsic defect structures of polycrystalline CaKFe4As4 superconductors.

We investigated the defect structures of polycrystalline CaKFe4As4 (CaK1144) superconductors by scanning transmission electron microscopy (STEM). The STEM studies revealed the presence of a one-layer CaFe2As2 (∼1 nm size) defect along the ab-plane, as observed in single crystalline CaK1144. Step-like CaFe2As2 defects are also observed. These nanoscale defects generate fine-sized stacking faults, a lattice mismatch, and stress field defects in the matrix of CaK1144 owing to the different sizes. Correlation of the defects in polycrystalline and single crystalline samples suggests that the defects type and their density depend on the synthesis conditions. A self-field critical current density (Jc) of 15.2 kA cm-2 was obtained at 5 K, and the curves were sustained above 30 K with a considerable Jc value of 1.4 kA cm-2. We investigated the relationship between the observed intrinsic defects and the behavior of the field dependence of Jc. The intrinsically intergrown planar defects, even in polycrystalline samples, are expected to be advantageous for various high-field applications of bulk CaK1144 superconductors.

Effects of cyclophosphamide on the prognosis of Japanese patients with renal vasculitis associated with anti-neutrophil cytoplasmic antibody-positive microscopic polyangiitis.

BACKGROUND: The aim of this study was to determine the efficacy of cyclophosphamide (CY) on anti-neutrophil cytoplasmic antibody (ANCA)-positive microscopic polyangiitis (MPA) with renal involvement in Japanese patients. METHODS: Eighty-two patients with newly diagnosed ANCA-positive MPA were enrolled in this retrospective study. Patients were divided into two groups based on whether they received combination therapy with a corticosteroid (CS) plus CY (CY group) or CS alone or with other therapies (non-CY group). The primary outcome was defined as the combination of death and end-stage renal disease (ESRD). RESULTS: The CY and non-CY groups included 29 and 53 patients, respectively. In the non-CY group, 31 patients were treated with CS alone, and 22 with a combination of CS and other therapeutics. The percentage of males and mean Birmingham vasculitis activity scores were higher in the CY group than those in the non-CY group, but other factors such as age, serum creatinine, serum albumin, or CRP at baseline were equivalent in the two groups. No differences were observed in remission rates using induction therapy for the two groups. However, the survival rate 5 years after induction therapy was lower in the CY group than in the non-CY group (0.50 vs. 0.73; P = 0.041), although the hazard ratio of CY for the primary outcome adjusted for all confounding factors was 1.321 [95 % confidence interval (CI), 0.662-2.637; P = 0.171]. CONCLUSIONS: CY may not have an additive effect on induction therapy with CS for Japanese patients with renal vasculitis associated with ANCA-positive MPA.

Clinical characteristics of biopsy-proven renal sarcoidosis in Japan.

Background: Although some articles have described renal sarcoidosis, the incidence among biopsy cases remains unclear. Here, we defined the incidence of renal sarcoidosis among renal biopsy cases and analyzed the clinical course. Methods: We performed an epidemiological study examining renal biopsy cases treated at 5 centers between January 2000 and September 2015 and identified 16 cases (7 men, 9 women; mean (±SD) age, 59.4±18.6 years) out of a total of 14191 renal biopsy cases. Renal involvement of sarcoidosis was defined as granulomatous tubulointerstitial nephritis, tubulointerstitial nephritis without granulomatous lesions, and renal calcinosis. Fifteen of the cases were treated with steroid therapy. One case initially received steroid pulse therapy. The outcome was evaluated based on the estimated glomerular filtration rate (eGFR), CKD stage, and the change in eGFR (ΔeGFR) after treatment. A favorable response was defined as ΔeGFR ≥25%. Results: The incidence of renal sarcoidosis was 0.11%. The mean eGFR was 28.2±16.1 mL/min/1.73 m2. At the last observation, the mean eGFR was 43.7±19.7 mL/min/1.73 m2. Although a favorable response to steroid therapy was found in the majority of cases (10/15, 67%), 12 of the 15 cases (80%) had residual renal dysfunction at the last observation and 8 cases (53%) had moderate to severe renal dysfunction. Conclusion: Renal sarcoidosis is extremely rare among renal biopsy cases. Among cases with an unfavorable response to steroid therapy, pathogenetic mechanisms other than sarcoidosis and severe nephron damage were observed. (Sarcoidosis Vasc Diffuse Lung Dis 2018; 35: 252-260).

Continuous large-scale production of the cytokine CXCL8 from a novel porcine cell line.

Cytokine production from two unstimulated porcine cell lines (SL-24 and SK-L) was examined using porcine cytokine detection ELISA kits and RT-PCR. Porcine IL-1 alpha, IL-6, and CXCL8 were detected in all samples examined. In particular, the SL-24 cell line (derived from bone marrow cells of a malignant lymphoma-affected pig), produced large amounts of porcine CXCL8. Flow cytometer analysis showed the cell line to be strongly CD44 positive, and was therefore considered to be of monocyte or macrophage origin. Porcine CXCL8 production was greatest (83.86 +/- 32.33 ng/mL) at six days post-cultivation. The SK-L cell line (derived from porcine kidney) also produced CXCL8, but production was less than 1.5 ng/mL. Porcine CXCL8 from the SL-24 cell line, induced chemotactic activity in porcine neutrophils, while the production of CXCL8 from the SL-24 cell line was inhibited by dexamethasone, which suggests that the mechanism of CXCL8 production is related to an NF-kappaB binding site. The production of CXCL8 from the SL-24 cell line was enhanced by the addition of recombinant porcine IL-15, which is the first reported observation of such CXCL8 production. Cloning of the SL-24 cell line by limited dilution revealed two types of cells present in the starting population. One cell type, designated as long-form cells (LC), produced large amounts of CXCL8, while the other, designated short-form cells (SC), produced small amounts of the cytokine. The LC cells were adapted to grow in serum-free medium in which they produced large amounts of CXCL8. The large-scale production of porcine CXCL8 from the SL-24 cell line will be of value in determining the mechanism of cytokine production and as a source of naturally produced porcine CXCL8.

Abnormal retention of 99mTc-TF in a hamster model of cardiomyopathy analyzed by 99mTc-TF and 125I-BMIPP autoradiography.

OBJECTIVE: Enhanced washout of 99mTc-tetrofosmin (TF) has been reported in patients with hypertrophic cardiomyopathy (HCM). Here, using quantitative dual-autoradiography, the relationship between TF retention abnormality and metabolism depicted by 125I-BMIPP uptake was investigated quantitatively in a hamster model of cardiomyopathy. METHODS AND RESULTS: Early and delayed TF images were obtained at 5 min (7 cardiomyopathic and 5 normal hamsters) and 60 min (8 cardiomyopathic and 5 normal hamsters) after injection, respectively. BMIPP image was obtained 5 min after injection. Five cardiomyopathic and 5 normal hamsters were evaluated histologically. Percent uptake of TF and BMIPP in the heart was measured by an auto-well counter. The left ventricular wall was divided into 12 segments, and the relative regional uptake of TF and BMIPP was measured for each segment. Heterogeneity of radioactive distribution was determined by the standard deviation (SD) of radioactive counts in the left ventricular wall on autoradiogram. The uptake of early TF, delayed TF, and BMIPP in cardiomyopathic hamsters was 8.8%, 20.3%, and 25.3% lower than that in normal hamsters, p < 0.05, p < 0.01, and p < 0.001, respectively. In normal hamsters, distribution of radioactivity in all images was homogeneous, and the SD values were about 13. In cardiomyopathic hamsters, heterogeneous distribution was observed on all images, and the degree of heterogeneity was marked on delayed TF and BMIPP images. The SD was 19.7 +/- 1.2 for early TF image, 25.5 +/- 1.4 for delayed TF image, and 31.7 +/- 2.4 for BMIPP image, respectively. A weak linear correlation was observed between the relative regional uptake of the delayed TF and BMIPP in cardiomyopathic hamsters (r = 0.57). Electron microscopy demonstrated ultra-structural changes in mitochondria of cardiomyopathic hamsters. CONCLUSION: Degree of retention abnormality on delayed TF image corresponded to the metabolic abnormality, probably due to mitochondrial dysfunction, depicted on BMIPP imaging.

In vivo antitumor activity of novel water-soluble taxoids.

The antitumor activity of newly developed taxoids possessing a sugar moiety (glucose, galactose or mannose) to improve water solubility was evaluated. Galactose-bound taxoid (10-alpha-GAG-DT) proved superior to all other taxoids in both water solubility and antitumor activity. Therapeutic efficacy of 10-alpha-GAG-DT in terms of in vivo antitumor activity was found to be approximately equivalent to that of docetaxel, which is known to be superior to that of paclitaxel. The observation that the sugar moiety was gradually hydrolyzed in serum to release docetaxel indicates that 10-alpha-GAG-DT acts as a prodrug.

Effect of N-terminal mutation of human lysozyme on enzymatic activity.

Using mutant signal peptide introduced Pro at the C-terminal region, 5 kinds of N-terminal mutants of human lysozyme (HLY) were produced. The genes coding chicken lysozyme signal peptide - human lysozyme (HLY) hybrid preproteins were altered as follows: -2Leu to Pro and -lGly to either Ala, Val, Leu, Asn or Lys, and were expressed in yeast cells. The N-terminal sequence data of HLYs secreted from yeast cells, showed that each signal peptide was cleaved only after -2Pro and either Ala, Val, Leu, Asn or Lys was added at their N-terminals of HLYs. This result suggested that the cleavage site of signal peptide can be shifted by the introduction of turn-promoting residue like Pro and N-terminal sequence of protein can be altered. The differences of additional residues were reflected in lytic activities of HLYs. Especially, Lys-HLY exhibited significantly wide optimal pH range and resistance to higher ionic strength conditions than other HLYs including native HLY. This result indicates that the importance of positive charge on the HLY surface to interact with substrate.

Effect of tranilast in early-stage diabetic nephropathy.

BACKGROUND: Tranilast is an antifibrotic drug known to suppress collagen synthesis by fibroblasts by interfering with the effects of TGF-beta. We recently reported that it slowed the progression rate of advanced diabetic nephropathy (DN) by reducing the accumulation of collagens in renal tissue. The present study was undertaken to examine the effect of tranilast on early-stage DN. METHODS: Among out-patients with diabetes mellitus, we selected patients with (i) urinary albumin excretion of 30-1000 mg/g creatinine (/gCr) in the first morning urine, (ii) serum creatinine (SCr) < or =1.2 mg/dl and no haematuria and (iii) currently taking an angiotensin-converting enzyme inhibitor or angiotensin receptor blocker. Twenty patients fulfilled the criteria, of whom 10 were selected at random and commenced on tranilast [100 mg, 3 times daily; T(+) group]. The remaining 10 patients comprised the T(-) group. Excretion of both urinary type IV collagen (U-IV) and albumin (U-A) in the first morning urine was measured every 3 months. The follow-up period was 1 year. RESULTS: At baseline, no significant differences were observed in SCr, HbA(1c), blood pressure and U-A excretion between the T(+) and T(-) groups, but U-IV excretion in the T(+) group was higher than in the T(-) group (6.4 +/- 0.66 vs 3.7 +/- 0.36 microg/gCr, mean +/- SEM, P < 0.01). At 1 year, SCr was not different from the baseline in either group. In the T(+) group, however, excretion rates of both U-IV and U-A tended to decrease with time, and after 1 year, were significantly decreased compared with excretion at baseline (U-A: 279 +/- 78 to 191 +/- 62 mg/gCr; P = 0.049, U-IV: 6.4 +/- 0.66 to 4.4 +/- 0.99 microg/gCr; P = 0.02). In contrast, in the T(-) group, excretion of both U-A and U-IV tended to increase with time. The changes of both U-A and U-IV excretions in the two groups took statistically different trends through tranilast treatment (P = 0.01 and P = 0.04, respectively). CONCLUSIONS: Our results suggest that tranilast could be therapeutically beneficial in early-stage DN.

Comparison of artificial synthesis methods of gene.

The comparative study of the methods for constructing the artificial genes have done. Equine interferon alpha-1 gene and porcine interferon alpha-1 gene were synthesized respectively by several methods from short DNA oligomers and each gene was evaluated about the accuracy of the gene sequence. The gene made by the SPR method was the most accurate.

High-energy phase-contrast X-ray imaging using a two-crystal X-ray interferometer.

To broaden the scope of phase-contrast X-ray imaging in biomedical applications, the energy of X-rays in a large-area imaging system using a two-crystal X-ray interferometer has been increased from 17.7 keV to 35 keV. The use of higher-energy X-rays makes it possible to observe larger samples having greater variation in density within shorter measurement periods, at higher spatial resolution, and at lower X-ray doses. After optimizing the imaging system for 35 keV X-rays, a 25 mm x 30 mm interference pattern with 50% visibility was generated at the Photon Factory. The system's capabilities are demonstrated by this pattern and a high-quality three-dimensional image of a rat heart.

Coherence-contrast x-ray imaging based on x-ray interferometry.

Coherence-contrast x-ray imaging--which detects changes in the degree of coherence caused by the placement of a sample in an x-ray interferometer--was developed for biomedical applications. Because the technique's sensitivity depends on the density gradient in the sample, it is particularly suitable for observing biomedical samples with large density differences, such as samples that include both biological soft tissue and bone. A measurement principle and method of this technique are described, and a fine coherence-contrast image of a mouse leg is given as an example result.

Gene design of signal sequence for the effective secretion of recombinant protein using insect cell.

In order to increase the proteinic secretion quantity of production in insect cell, the chicken lysozyme signal peptide (CLSP) was altered, and the effect of mutations on secretion of human lysozyme (HLY), the bovine interferon beta-3, and the equine interferon alpha-1 were studied. A hybrid gene composed of CLSP gene and HLY gene was used for the production of recombinant HLY in insect cell (BmN4) using vacuole virus expression system. In contrast with the Leu rich signal peptide in previous reports, Val rich type, or Ile rich type decreased the secretion of HLY, and Phe rich type prevented it completely, in spite of the increase of hydrophobicity in the central core region of each signal peptide. Besides the hydrophobicity, the tertiary structure in this region also might influence the secretion of HLY in insect cells. Then, Leu-rich signal peptide was used for the secretion of the bovine interferon beta-3, or the equine interferon alpha-1, resulted in the remarkable increase of the amount of the secretion in insect cells.

Structural requirements of signal peptide in insect cell.

In order to increase the proteinic secretion quantity of production in insect cell, the N-terminal and the hydrophobic core region of the chicken lysozyme signal peptide (CLSP) was altered. A hybrid gene composed of CLSP gene and human lysozyme (HLY) gene from short DNA oligomers was used for the production of recombinant HLY in insect cell using vacuole virus expression system. In contrast with the results in yeast, the introduction of basic residues in the N-terminal region did not make the amount of secretion increase in insect cell. On the other hand, when the sequence of CLSP was changed into Leu-rich type, the amount of HLY secretion increased sharply. Although they are eukaryotes, the structural requirements of signal peptides may have some differences in yeast and insect cell.

Gene design of signal sequence for effective secretion of protein.

To improve the secretion level of recombinant protein, the gene sequence coded the chicken lysozyme signal peptide (CLSP) connected with human lysozyme (HLY) was altered to substitute the amino acid sequence, and the effect of mutations on secretion of HLY in yeast was elucidated. The effect of mutations on secretion of recombinant HLY was elucidated in yeast. As the result of experiments, it has been shown that the positive charge in the N-terminal region of signal peptide plays an important role in the function of the eukaryotic signal peptide as well as that of prokaryote. This result enable us to design the ideal sequence for effective secretion of recombinant protein. Using such signal sequence containing additional Arginine residues, secretion levels of HLY in yeast were notably increased.

[Phase-contrast x-ray imaging with x-ray interferometer for medical applications].

Phase-contrast x-ray imaging with x-ray interferometer can depict the minute difference within the biological object, and its sensitivity is about 1000 times higher than that of absorption-contrast method. For biomedical use of this technique, a large monolithic x-ray interferometer and 2 crystal interferometer having a field of view with 25 mm x 25 mm is being developed. Phase-contrast x-ray CT could reveal detail structures within tumor and surrounded tissue, and the vessel imaging of rat liver is also possible using physiological saline at 17.7 keV x-ray energy. Recently, human breast tissues were imaged at 35 keV and the contrast of image was much better than usual absorption contrast x-ray image obtained at 17.7 keV energy.