Clinical Tooth Color Matching: In Vivo Comparisons of Digital Photocolorimetric and Spectrophotometric Analyses.

OBJECTIVE: This study aimed to compare the tooth color matching of two dental colorimetric methods: the spectrophotometric analysis (SPM) and the standardized digital photocolorimetric analysis (DPC). METHODS: The color of 60 maxillary central incisors of 30 volunteers (22.5 ± 7.6 years) was analyzed. In the DPC method, tooth photographs were standardized with the eLABor_aid protocol, processed with Adobe Photoshop Lightroom software, and the values of L*, a*, and b* were obtained with a Digital Color Meter software. For the SPM, L*, a*, and b* were measured directly with a handheld spectrophotometer. Data were submitted to paired t-test and Pearson correlation test (α=0.05). Mean color difference between the two methods was calculated with CIELAB formula. RESULTS: All color coordinates revealed different values when comparing DPC to SPM in the same tooth (p<0.0001). Mean color difference (ΔEab) between SPM and DPC was 11.5 ±3.1. A positive correlation was observed for L* (R2=0.73,p<0.0001), a* (R2=0.31, p=0.017), and b* (R2=0.83, p<0.0001). CONCLUSIONS: Even though the color coordinate values were different in both methods, they were correlated, revealing that the DPC is a viable alternative to determine the tooth color matching.

ASP2409, A Next-Generation CTLA4-Ig, Versus Belatacept in Renal Allograft Survival in Cynomolgus Monkeys.

Belatacept is the first costimulatory blockade agent approved for maintenance immunosuppression in kidney transplant recipients. Clinical results have indicated that belatacept is associated with superior renal function and improved metabolic profile; however, higher incidence of acute rejection and posttransplant lymphoproliferative disorder are the shortcomings of this agent. In this study, ASP2409, a new cytotoxic T-lymphocyte associated protein 4-immunoglobulin possessing 14-fold higher in vitro CD86 binding affinity than belatacept, was tested for renal allograft survival in cynomolgus monkeys. ASP2409 monotherapy dose-dependently prolonged renal allograft survival. Low-dose ASP2409 in combination with a subtherapeutic dose of tacrolimus showed much longer median survival time than monotherapy. Similar allograft survival results were observed in regimens based on high-dose ASP2409, belatacept, and therapeutic-dose tacrolimus. The results of renal allograft histopathology with high-dose ASP2409-based regimens were not inferior to the belatacept-based regimen. Moreover, higher frequencies of FoxP3-positive regulatory T cells in renal allografts were observed in ASP2409- and belatacept-based regimens compared with tacrolimus-based regimens. No serious side effects related to ASP2409 administration were found during the study. These data suggest that ASP2409 is a promising candidate for calcineurin inhibitor-sparing or -avoidance regimens.

Operative Results and Clinical Features of Chronic Stanford Type B Aortic Dissection: Examination of 234 Patients Over 6 Years.

OBJECTIVE/BACKGROUND: Recently, the indications for thoracic endovascular aortic repair (TEVAR) have been expanding, and the applicability of TEVAR for acute type B aortic dissection (TBAD) is proposed with regard to the high mortality of open surgery for chronic TBAD. TEVAR in the acute phase may lead to remodeling of the false lumen (FL), but it is controversial whether it completely resolves the aortic expansion in the chronic phase. In this study, operative results and the relationship between FL status and the time before surgical intervention were retrospectively analyzed. METHODS: From January 2008 to September 2013, 234 patients underwent open surgery for chronic TBAD. Most patients were on left heart bypass. By considering Japanese aortic disease treatment guidelines and the smaller physique of Japanese patients, operative indications were aneurysm >50 mm in diameter or rapid aneurysm enlargement of >5 mm in a 6 month period. RESULTS: In 180 cases, the FL was patent. The mean interval between onset of TBAD and operation was 61 ± 54 months. There was no significant difference between patients in the patent FL group and those in the thrombosed FL group (p = .44). Mean ratio of FL diameter to maximum aortic diameter (FL/AD) was 0.64 ± 0.21. There was no correlation between FL and AD before the operation (r = .12). Descending thoracic aortic replacement (DTAR) was performed in 127 cases and thoracic ascending aortic replacement (TAAR) in 107 cases (Crawford type I, n = 9; Crawford type II, n = 65; Crawford type III and IV, n = 22, respectively; Safi type V, n = 11). The overall operative mortality was 6.8%: 3.9% (5/127) for DTAR and 10.3% (11/107) for TAAR. The three year survival was 86.7, and the freedom from re-intervention rate was 97.0%. CONCLUSION: Enlargement of uncomplicated TBAD in the chronic phase was poorly related to FL status and the results of open repair have improved. However, further prospective study is necessary.

Establishment of a new cell line susceptible to Cyprinid herpesvirus 3 (CyHV-3) and possible latency of CyHV-3 by temperature shift in the cells.

A new cell line named CCF-K104 predominantly consisting of fibroblastic cells showed optimal growth at temperatures from 25 °C to 30 °C. Serial morphological changes in the cells induced by Cyprinid herpesvirus 3 (CyHV-3) included cytoplasmic vacuolar formation, cell rounding and detachment. Mature virions were purified from CyHV-3-infected CCF-K104 cells by sucrose gradient ultracentrifugation and had a typical herpesvirus structure on electron microscopy. Infectious CyHV-3 was produced stably in CCF-K104 cells over 30 viral passages. Our findings showed that CCF-K104 is a useful cell line for isolation and productive replication of CyHV-3. A temperature shift from 25 °C to 15 °C or 35 °C did not allow serial morphological changes as observed at 25 °C for 14 days. Under the same conditions, real-time PCR showed that CyHV-3 was present with low viral DNA loads, suggesting that CyHV-3 may establish latent infection in CCF-K104 cells. Amplification of the left and right terminal repeat sequences of the CyHV-3 genome arranged in a head-to-tail manner was detected by nested PCR following an upshift in temperature from 25 °C to 35 °C. The PCR results suggested that the circular genome may represent a latent form of CyHV-3.

Nisin A extends the shelf life of high-fat chilled dairy dessert, a milk-based pudding.

AIMS: The aims of this study were to evaluate the effectiveness of nisin A to control the growth of spore-forming bacteria, Bacillus and Paenibacillus, in chilled high-fat, milk pudding and to reduce heat treatment to improve aroma and flavour. METHODS AND RESULTS: Nisin A was added to milk pudding containing 5·0 and 7·5% fat to final concentrations of 40, 80, 120 and 240 IU ml(-1). Spores from Bacillus thuringiensis, Bacillus cereus and Paenibacillus jamilae were inoculated into samples at 10 spores ml(-1) prior to pasteurization at 130°C for 2 s. Milk pudding without inoculation was pasteurized using less heat condition (100, 110 and 120°C for 2 s) to measure the effect of adjusting the ingredients to prevent naturally occurring bacteria. The viable cells during storage at 15, 20 and 30°C showed nisin A inhibited spiked bacteria to varying degrees depending on species, sensitivities to nisin A concentration and fat content, and inhibited natural populations at 80 IU g(-1) nisin A in 5·0% fat and at 120 IU g(-1) in 7·5% fat milk pudding. An aroma compound analysis and organoleptic assessment showed processing at 110 and 120°C decreased the temperature-dependent unpleasant odours, for example, reduced dimethyl sulfide and dimethyl disulfide by 1·2-1·5 times and increased rankings in taste tests compared with 130°C treated pudding. CONCLUSIONS: Nisin A was found to be effective as a natural preservative to control spoilage bacteria in high-fat milk pudding and extend its shelf life, when using reduced heat treatments to improve the flavour and aroma without compromising food safety. SIGNIFICANCE AND IMPACT OF THE STUDY: This is the first report showing nisin A is effective in reducing spoilage bacteria in high-fat, chilled dessert, milk pudding. Therefore, nisin A can be used to improve milk puddings to satisfy both industry and consumer demand for food quality and safety.

Sensitive cardiac troponin in the diagnosis and risk stratification of acute heart failure.

BACKGROUND: The aim of our study was to investigate the diagnostic and prognostic value of a sensitive cardiac troponin I (s-cTnI) assay in patients with acute heart failure (AHF). METHODS: Sensitive cardiac troponin I was measured in 667 consecutive patients at presentation to the emergency department with acute dyspnoea. Three s-cTnI strata were predefined: below the limit of detection (<0.01 µg L(-1) , undetectable), detectable but still within the normal range (0.01-0.027 µg L(-1) ) and increased (≥0.028 µg L(-1) , ≥99th percentile). The final diagnosis was adjudicated by two independent cardiologists blinded to the s-cTnI levels. Median follow-up in patients with AHF was 371 days. RESULTS: Levels of s-cTnI were higher in patients with AHF (n = 377, 57%) compared to patients with noncardiac causes of acute dyspnoea (median 0.02 vs. <0.01 µg L(-1) , P < 0.001). In patients with AHF, in-hospital mortality increased with increasing s-cTnI in the three strata (2%, 5% and 14%, P < 0.001). One-year mortality also increased with increasing s-cTnI (21%, 33% and 47%, P < 0.001). s-cTnI remained an independent predictor of 1-year mortality [adjusted odds ratio 1.03 for each increase of 0.1 µg L(-1) , 95% confidence interval (CI) 1.02-1.05, P < 0.001] after adjustment for other risk factors including B-type natriuretic peptide. The net reclassification improvement was 68% (P < 0.001), and absolute integrated discrimination improvement was 0.18 (P < 0.001). The diagnostic accuracy of s-cTnI for the diagnosis of AHF as quantified by the area under the receiver operating characteristic curve was 0.78 (95% CI, 0.75-0.82). CONCLUSIONS: Sensitive cardiac troponin I is a strong predictor of short- and long-term prognosis in AHF that helps to reclassify patients in terms of mortality risk. Detectable levels of s-cTnI, even within the normal range, are independently associated with mortality.

Evaluation of a lateral flow assay for the diagnosis of Mycobacterium bovis infection in dairy cattle.

A commercially available multi-antigen lateral flow assay (LFA) for detection of antibodies to Mycobacterium bovis was evaluated by testing dairy cattle in Baja California, México. Sera and tissue samples were obtained from 268 dairy cattle at a slaughterhouse and were tested by LFA, bacteriological culture, and polymerase chain reaction (PCR). Of 107 culture positive samples, 70 (65.4%) were positive by PCR and 49 (45.8%) were positive by LFA. The PCR and LFA gave positive results with an additional 59 (36.6%) and 77 (47.8%) of the 161 culture negative samples, respectively. The false negative rate for the PCR was 34.6% and 54.2% for the LFA. Due to the high false positive rate for both PCR and LFA observed in this study, the LFA cannot be a useful test, even in combination with PCR.

Distribution of mucous cells on the body surface of Japanese flounder Paralichthys olivaceus.

The distribution of mucous cells was examined in the skin on the ocular and blind sides of Japanese flounder Paralichthys olivaceus. Observations were performed on both body sides at the following regions: cheek, lower jaw (blind side), gill cover (ocular side), dorsal side, lateral line, belly and caudal peduncle. The mucous cells observed were elliptic and positively stained for periodic acid Schiff reaction and Mayer's mucicarmine and showed a higher density and larger size on the ocular side compared to the blind side. Low densities of mucous cells were observed on the lower jaw compared with other regions of the body. The depth of the crack located between scales was deeper on the ocular side than the blind side, which might reflect total epidermis area and total number of mucous cells. Bacterial infection elucidated some information on the effect on the density and size of mucous cells, where the density and size decreased slightly after infection. Only the lower jaw, however, showed an increased number of mucous cells. The results show that the potential of skin to secrete mucus is higher on the ocular than on the blind side and bacterial infection decreases mucous secretion.

Immunogenicity of pressure inactivated Edwardsiella tarda bacterin to Anguilla japonica (Japanese eel).

Japanese eel Anguilla japonica were immunized with inactivated Edwardsiella tarda bacterin preparations (formalin-killed cells, FKC (0.4%), formalin with heat-killed cells, FHKC (0.1% and 70 degreesC for 10 min), heat-killed cells, HKC (70 degrees C for 15 min), potassium chloride-killed cells, KKC (0.6%), tannic acid-killed cells, TKC (0.9%), citric acid-killed cells, CAKC (0.9%), pressure-killed cells, PKC (600 psi for 5 min) and electric current-killed cells, ECKC (100 mA at 12 v DC for 5 sec) via intraperitoneal injection in order to develop adequate inactivating method. Immune parameters in the immunized eel were measured to compare responses to different bacterins. Generally, eel rose agglutinating antibody titer in the serum within 2 week and the maximum titer occurred at 6 weeks post immunization. Elevated and significantly higher titer was produced with the PKC of E. tarda than other bacterin preparations. An Enzyme Linked Immunosorbent Assay (ELISA), to determine specific anti-E. tarda antibody in the serum, also showed significantly higher antibody titer with PKC than the other antigen preparations. Bacteriostatic assay with serum and live E. tarda indicated significantly higher activity in the PKC-immunized fish. Immunization with PKC also showed the increased level ofphagocytosis. PKC-inactivated vaccine at an immunization dose of 10(6) cells/fish induced high protection against experimental infection. Coincident with higher immune parameters and protection in the fish immunized with the PKC bacterin strongly suggested that pressure-killing is an effective inactivating method to develop an effective vaccine against edwardsiellosis.

A qualitative study of Japanese patients' perspectives on post-treatment care for gynecological cancer.

The perspectives of cancer patients are important for designing a medically and economically effective follow-up program to help in the rapid recovery of patients. However, research focusing on the perspectives of Japanese gynecological cancer patients on follow-up programs is extremely scarce. In this study, we explored the perspectives and expectations of Japanese gynecological cancer patients with regard to post-treatment follow-up. Twenty-eight patients recruited through a gynecological cancer support group were included in focus groups 1-10 years post-treatment. Participants' accounts related to their perspectives on follow-up were coded and grouped into themes according to commonalities and differences. Seven themes emerged as follows: (1) living with uncertainty, (2) monitoring recurrence, (3) test content and frequency, (4) coping with "another illness", (5) provider communication and attitude, (6) holistic care, and (7) compromising with the reality of changed body. While these Japanese gynecological patients regarded follow-up as an opportunity for reassurance, they also wanted treatment for adverse effects and the opportunity to discuss their concerns. On the basis of the study findings, we conclude that during follow-up after cancer treatment, Japanese gynecological cancer patients not only prioritize recurrence management of cancer but also place a significant amount of importance on the management of symptoms and doctor-patient communication. However, these expectations for communication and care are often unmet. Thus, there is a need to fulfill the gap between the current follow-up programs and patients' expectations by reviewing and changing the hospital's policy that mainly focuses on the detection of recurrent diseases.

Enhancement of bone formation in an experimental bony defect using ferumoxide-labelled mesenchymal stromal cells and a magnetic targeting system.

We used interconnected porous calcium hydroxyapatite ceramic to bridge a rabbit ulnar defect. Two weeks after inducing the defect we percutaneously injected rabbit bone marrow-derived mesenchymal stromal cells labelled with ferumoxide. The contribution of an external magnetic targeting system to attract these cells into the ceramic and their effect on subsequent bone formation were evaluated. This technique significantly facilitated the infiltration of ferumoxide-labelled cells into ceramic and significantly contributed to the enhancement of bone formation even in the chronic phase. As such, it is potentially of clinical use to treat fractures, bone defects, delayed union and nonunion.

Prediction of functional outcome in stunned myocardium after myocardial infarction using BMIPP and tetrofosmin imaging.

The predictive value of combined (123)iodine-labelled 15-(p-iodophenyl)-3R,S-methyl pentadecanoic acid imaging ((123)I-BMIPP) and early technetium-99m ((99m)Tc)-tetrofosmin imaging was compared with combined (123)I-BMIPP and delayed (99m)Tc-tetrofosmin or (123)I-BMIPP and thallium-201 ((201)Tl) imaging for functional outcome of stunned myocardium after acute myocardial infarction (AMI) in 37 patients with AMI. All patients underwent successful percutaneous coronary intervention with/without stenting within 24 h of symptoms. Resting (201)Tl, (99m)Tc-tetrofosmin and (123)I-BMIPP imaging were performed within 10 days of hospital admission; (99m)Tc-tetrofosmin imaging was also performed 6 months later. Segments were mismatched when the (123)I-BMIPP score was greater than the (99m)Tc-tetrofosmin or (201)Tl scores, and were matched when all scores were the same. Left ventricular function was estimated using wall motion score. Sensitivity and regional wall motion were significantly better in mismatching (99m)Tc-tetrofosmin-early/(123)I-BMIPP segments than mismatching (201)Tl/(123)I-BMIPP or (99m)Tc-tetrofosmin-delayed/(123)I-BMIPP segments. It is concluded that mismatching of (123)I-BMIPP and early (99m)Tc-tetrofosmin uptake can predict improvement in wall motion of stunned myocardium better than the other two imaging combinations.

Recoil effect of photoelectrons in the Fermi edge of simple metals.

High energy resolution photoelectron spectroscopy of conduction electrons in the vicinity of the Fermi edge in Al and Au at excitation energies of 880 and 7940 eV was carried out using synchrotron radiation. For the excitation energy of 7940 eV, the observed Fermi energy of Al shows a remarkable shift to higher binding energy as compared with that of Au, with accompanying broadening. This is due to the recoil effect of the emitted photoelectrons. The observed spectra are well reproduced by a simple model of Bloch electrons based on the isotropic Debye model.

Hyperkalemia in familial mitochondrial cytopathy.

AIM: To contribute to understanding the pathogenesis of hyperkalemia that often occurs in patients with diabetes. MATERIALS AND METHODS: We describe 3 familial cases of mitochondrial diabetes mellitus. The mitochondrial A3243G point mutation was confirmed in a mother and her 2 children. We examined their clinical features and pathological findings, and assessed heteroplasmy of mutant mitochondria DNA (mtDNA) by molecular analysis. RESULTS: The second son had spontaneous hyperkalemia and hyporeninemic hypoaldosteronism. Histopathological examination revealed severe tubulointerstitial and vascular changes around the juxtaglomerular apparatus. The mother only showed intermittent hyperkalemia concurrently with the aggravation of heart failure, and the pathological changes in her kidneys were mild. Heteroplasmy was more severe in the second son than in the mother. CONCLUSION: Heteroplasmy of mitochondrial cytopathy combined with diabetes mellitus led to abnormalities resembling those seen in Type IV renal tubular acidosis.

Single-operator method for double-balloon endoscopy: a pilot study.

Double-balloon endoscopy (DBE) is a new technique that allows diagnosis and treatment throughout the entire small intestine. We have recently developed a method that allows a single operator to perform DBE, by employing "grip and pinch" and "hold (or 'keep') and slide" techniques. This one-person method obviates the need for two operators who must cooperate closely.

Characterization of cleavage sites and protease activity in the polyprotein precursor of Japanese marine aquabirnavirus and expression analysis of generated proteins by a VP4 protease activity in four distinct cell lines.

A polyprotein precursor NH(2)-pVP2-VP4-VP3-COOH is encoded in genomic segment A of members of the family Birnaviridae. By N-terminal sequencing analysis, primary cleavage sites of a marine birnavirus (MABV) polyprotein were identified as Ala(508) downward arrow Ser(509) and Ala(734) downward arrow Ser(735), where the cleavage motif was the same as that of infectious pancreatic necrosis virus (IPNV). However, further VP4 and VP3 cleavages occurred at novel sites. Ser(633) and Lys(674) mutations affected the cleavage activity by site-directed mutagenesis. Additional catalytic residues including Ile(543) and Val(686) were MABV-specific. As shown by electron microscopy, pVP2 and further cleaved VP3s (fcVP3s) could not form virus-like particles (VLPs). This suggests that VP3 is necessary for VLP formation. By Western blot analysis of the VP3 expression, fcVP3s were found in RSBK-2 cells and FHM cells, while VP3 was cleaved less in EPC cells, suggesting that fcVP3s might merely be a degraded form. Alternatively, if fcVP3s play functional roles other than in viral assembly, the further VP3 cleavage is, at least, not restricted in FHM cells. Strangely, VP3 was not completely further cleaved in CHSE-214 cells despite the fact that this cell line has a potential proteolytic factor, implying that complicated factors are associated with the further VP3 cleavage.

FTY720 suppresses CD4+CD44highCD62L- effector memory T cell-mediated colitis.

FTY720, a sphingosine-derived immunomodulator, causes immunosuppression via enhancement of lymphocyte sequestration into secondary lymphoid organs, thereby preventing their antigen-activated T cell egress to sites of inflammation. FTY720 is highly effective in inhibiting autoimmunity in various animal models. However, there is little known about how FTY720 controls the migration property of memory T cells. Here, we demonstrated that FTY720 prevents the development of colitis induced by the adoptive transfer of lamina propria (LP) colitogenic effector memory CD4+ T cells (TEM cells; CD45RB(low)CD44(high)CD62L-) into severe combined immunodeficiency (SCID) mice and suppresses interferon-gamma, interleukin-2, and tumor necrosis factor-alpha production by LP CD4+ T cells. The numbers of spleen, peripheral blood, mesenteric lymph node, and LP CD4+ T cells in FTY720-treated mice were significantly reduced compared with those in control mice. Notably, LP CD4+ TEM cells as well as splenic CD4+CD45RBhigh T cells expressed several spingosine-1-phosphate receptors that are targets for FTY720. Furthermore, FTY720 also prevented the development of colitis induced by the adoptive transfer of splenic CD4+CD45RBhigh T cells into SCID mice. Collectively, the present data indicate that FTY720 treatment may offer the potential not only to prevent the onset of disease but also to treat memory T cell-mediated autoimmune diseases including inflammatory bowel diseases.

Toxicity and roles of reactive oxygen species.

The history of studies in biology regarding reactive oxygen species (ROS) is approximately 40 years. During the initial 30 years, it appeared that these studies were mainly focused on the toxicity or microbicidal-related agents of ROS. However, recent studies have identified another action regarding oxidative signaling, other than toxicity of ROS. Basically, it is suggested that ROS are reactive, and degenerate to biomacromolecules such as DNA and proteins, leading to deterioration of cellular functions as an oxidative stress. On the other hand, recent studies have shown that ROS act as oxidative signaling in cells, resulting in various gene expressions. For example, NADPH oxidase, a major source of superoxide radicals (O(2)(-)), expresses in various tissues such as leukocytes and cardiovascular systems, and ROS derived from the enzyme play important roles in cell proliferation, differentiation, and cell death. In this review, we have focused on and described the basic properties, toxicity, and roles of ROS.

Safety analysis after tacrolimus immunosuppression in renal transplant recipients in Japan: 5-year results in >1500 patients.

Tacrolimus was approved in Japan in April 1996 for the prevention of allograft rejection in patients receiving kidney transplants. There has been a concern that immunosuppressive therapy may be associated with cardiovascular and metabolic complications, including hyperlipidemia, hypertension, and posttransplant diabetes mellitus. A multicenter (59 institutions) study was conducted in Japan in patients who underwent renal transplantation and received tacrolimus immunosuppression. Patients were followed for >5 years, from April 1996 to December 2002. Of the 1569 patients enrolled, 1542 were evaluated. In this analysis, graft survival rate and medication usage patterns of antihyperlipidemics, antihypertensives, insulin, and oral hypoglycemics were observed for >5 years in patients receiving tacrolimus immunosuppression. The graft survival rates of patients requiring antihyperlipidemic therapy and experiencing acute rejection were significantly lower compared with all other patients (P < .05). The risk of graft rejection was significantly greater in patients with cardiovascular complications requiring antihyperlipidemics or antihypertensives. Graft survival was significantly lower in patients with acute rejection and antihyperlipidemic therapy than in other patients.