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The gastrointestinal tract is not the common infection site of Pseudomonas aeruginosa. The role of P. aeruginosa as a causative agent for diarrhea in children without preexisting disease is controversial.From 2003 to 2012, we reviewed the records of 259 diarrheal patients less than 5 years of age whose stool culture grew P. aeruginosa. Virulence phenotypes of bacterial isolates were determined in vitro, including cytotoxicity, penetration and adherence to epithelial cells.The presence of P. aeruginosa in children with diarrhea less than 5 years old is 0.91%. P. aeruginosa-associated diarrheal diseases were classified into 4 groups: Shanghai fever (enteric infection and sepsis) (5%), P. aeruginosa enterocolitis (15%), P. aeruginosa-related diarrhea (19%) and antibiotic-associated diarrhea (43%). The remaining patients had coinfection with other pathogens (18%). Shanghai fever was the most severe enteric disease with invasive infection and complications. The clinical features of P. aeruginosa enterocolitis were prolonged fever with bloody or mucoid diarrhea mimicking bacterial enterocolitis. The clinical features of P. aeruginosa-related diarrhea and antibiotic-associated diarrhea were similar to viral or toxin-mediated diarrhea. Compared with other P. aeruginosa-associated diarrheal diseases, patients with Shanghai fever were younger, usually infants, and the characteristic laboratory findings included leukopenia, thrombocytopenia, high C-reactive protein, hyponatremia and hyperglycemia. Except for Shanghai fever, antibiotic treatment is not recommended. Isolates from Shanghai fever were more cytotoxic and adherent than isolates from uncomplicated diarrheal patients.P. aeruginosa could be an enteric pathogen even in healthy children. Young age and highly virulent bacterial strains were risk factors for Shanghai fever.

Metal oxide nanoparticles are a new class of important materials used in a wide variety of biomedical applications. Bulk zinc oxide (ZnO) particles have been used for temporal or permanent luting cement because of their excellent mechanical strength and biocompatibility. ZnO nanoparticles have distinct optical and antibacterial properties and a high surface-to-volume ratio. We investigated the mechanical and antibacterial properties of luting cement with different ratios of ZnO nanospheres. We showed that luting cement with 5% and 10% ZnO nanospheres was less soluble in low-pH (pH 3) artificial saliva. Antibacterial activity was 40% higher for Streptococcus mutans and 90% higher for Porphyromonas gingivalis when >10% (w/v) of the bulk particles were replaced with ZnO nanospheres in ZnO polycarboxylate cement. ZnO nanospheres were also biocompatible with mammalian cells. Additionally, the compressive strength was 1.2 times greater and the diametral tensile strength was 1.5 times greater for cements with 10% ZnO nanospheres than for conventional ZnO polycarboxylate cement. We propose a new method for improving dental luting cement by integrating it with ZnO nanospheres. This method simultaneously adds their greater antibacterial, mechanical, and acid resistance properties and retains an outstanding degree of biocompatibility.

Purpose. An observational study was performed to investigate the carriage rate and serotypes of Streptococcus pneumoniae in the 13-valent pneumococcal conjugate vaccine (PCV13) era in Taiwan. Methodology. From March 2014 to March 2015 a total of 500 healthy children and their households (631 adults) were enrolled from two large medical centres for nasopharyngeal carriage survey. Clinical isolates were prospectively collected from June 2012 to May 2015 at Chang Gung Memorial Hospital. We applied a multiplex polymerase chain reaction in addition to culture to detect S. pneumoniae. Results. S. pneumoniae was isolated from 12.0 % of the children and 3.6 % of the households. In the children's cohort only 23.3 % of the isolates could be assigned to PCV13 serotypes; non-vaccine serotypes were predominant (76.6 %) and the most frequently detected non-vaccine serotypes were 15A/F and 15B/C (both 13.3 %), followed by 23A (6.7 %). In the household cohort, 21.7 % belonged to PCV13 serotypes, and 78.3 % to non-vaccine serotypes. Clinical analysis of culture-confirmed pneumococcal infection showed that infection caused by PCV13 serotypes decreased by 47 % from 83 % in 2012–2013 to 44 % in 2014–2015, while infection caused by non-PCV13 serotypes increased from 17 to 56 %. Among the carriage isolates a significantly higher percentage belonged to serogroup 15 compared to serogroup 19 (26.6 vs 6.66 %, 2014–2015; P=0.003). Therefore, clinical isolates belonging to serogroup 15 were more prevalent than those belonging to serogroup 19 (44.1 vs 32.3 %, 2014–2015; P=0.318). Conclusion. The isolation of non-vaccine serotypes and unknown serotypes after the introduction of PCV13 in children highlights the importance of continued surveillance for emerging serotypes.

Background Shanghai fever, a community-acquired enteric illness associated with sepsis caused by Pseudomonas aeruginosa, was first described in 1918. The understanding of Shanghai fever is incomplete. Objective To delineate the clinical features and to examine the host and microbial factors associated with Shanghai fever. Methods We prospectively enrolled 27 consecutive previously healthy children with community-acquired P aeruginosa enteritis and sepsis between July 2003 and June 2012. An immunological investigation, including measurement of serum immunoglobulin levels and lymphocyte subpopulations, was performed. The clonal relationship of bacterial isolates was determined by multilocus sequence typing (MLST) and the virulence of isolates was measured using cellular and animal models. Results The median age of the patients was 7 months; 24 (89%) were aged

With shrinking device size in deep submicron process, many non-ideal effects impact circuit performances critically. Since those effects are often not considered in traditional analog synthesis tools, several sizing-layout iterations are still required to reach the desired performance and design yield. In this paper, an integrated analog synthesis tool is presented to consider the process variation, layout effects, and final layout generation simultaneously, with a user-friendly GUI to help users complete the design flow efficiently. With the consideration of those non-ideal effects in early design stages, blind design margins and time-consuming re-design cycles can be avoided in the proposed tool, which significantly reduces the design overhead. As shown in the experimental results, this analog synthesis tool is able to generate the required circuits in seconds and effectively guarantees the post-layout performance and design yield with less hardware cost.

Summary Gelsolin has important cellular functions, including cell motility, proliferation and apoptosis. Altered gelsolin expression has been reported in several types of human malignancies, but has not been evaluated in oral carcinogenesis. In this study, all normal oral mucosa ( n = 12) had high gelsolin expression, whereas only 7.7% of oral precancerous lesions ( n = 26) had positive gelsolin expression. A significant increased positive staining was found in primary ( n = 51; 37.3%) and metastatic ( n = 26; 30.8%) oral squamous cell carcinoma lesions. Tumors with high gelsolin expression were associated with greater tumour size ( P = 0.007), invasive growth ( P = 0.02), and younger age ( P = 0.006). High gelsolin expression conferred a poor clinical outcome in patients with metastatic disease ( P = 0.005). In conclusion, a biphasic profile in gelsolin expression was observed during the progression of oral carcinogenesis. This may be due to a balance of its multiple cellular functions in tumour invasion and cell growth. The underlying mechanisms, however, remain to be elucidated.

The effect of nordihydroguaiaretic acid (NDGA) on Ca(2+) signaling in C6 glioma cells has been investigated. NDGA (5-100 microM) increased [Ca(2+)]i concentration-dependently. The [Ca(2+)]i increase comprised an initial rise and an elevated phase over a time period of 4 min. Removal of extracellular Ca(2+) reduced NDGA-induced [Ca(2+)]i signals by 52+/-2%. After incubation of cells with NDGA in Ca(2+)-free medium for 4 min, addition of 3 mM CaCl2 induced a concentration-dependent increase in [Ca(2+)]i. NDGA (100 microM)-induced [Ca(2+)]i increases in Ca(2+)-containing medium was not changed by pretreatment with 10 microM nifedipine or verapamil. In Ca(2+)-free medium, pretreatment with the endoplasmic reticulum Ca(2+) pump inhibitor thapsigargin (1 microM) abolished 100 microM NDGA-induced [Ca(2+)]i increases. Inhibition of phospholipase C with 2 microM U73122 had little effect on 100 microM NDGA-induced Ca(2+) release. Several other lipoxygenase inhibitors had no effect on basal [Ca(2+)]i. Collectively, the results suggest that NDGA increased [Ca(2+)]i in glioma cells in a lipoxygenase-independent manner, by releasing Ca(2+) from the endoplasmic reticulum in a manner independent of phospholipase C activity and by causing Ca(2+) influx.

The effect of the estrogen diethylstilbestrol (DES) on cytosolic free Ca(2+) levels ([Ca(2+)](i)) in MG63 human osteoblasts was explored by using fura-2 as a Ca(2+) indicator. DES at concentrations between 5--20 microM induced an immediate increase in [Ca(2+)](i) in a concentration-dependent manner with an EC(50) of 10 microM. Removing extracellular Ca(2+) reduced the Ca(2+) signal by 70%. Pretreatment with 50 microM La(3+) or 10 microM of nifedipine, verapamil and diltiazem did not change 20 microM DES-induced [Ca(2+)](i) increases. Addition of 3 mM Ca(2+) increased [Ca(2+)](i) in cells pretreated with 20 microM DES in Ca(2+)-free medium. Pretreatment with 1 microM thapsigargin (an endoplasmic reticulum Ca(2+) pump inhibitor) to deplete the endoplasmic reticulum Ca(2+) store partly inhibited 20 microM DES-induced Ca(2+) release, but addition of carbonylcyanide m-chlorophenylhydrazone (CCCP; a mitochondrial uncoupler) and thapsigargin together abolished DES-induced Ca(2+) release. Conversely, pretreatment with 20 microM DES abrogated CCCP- and thapsigargin-induced Ca(2+) release. Inhibition of phospholipase C activity with 2 microM U73122 did not alter 20 microM DES-induced Ca2+ release. Another estrogen 17beta-estradiol also increased [Ca(2+)](i) in a concentration-dependent manner with an EC50 of 7 microM. Together, the data indicate that in human osteoblasts, DES increased [Ca(2+)](i) via causing Ca(2+) release from both mitochondria and the endoplasmic reticulum in a phospholipase C-independent manner, and by causing Ca(2+) influx.

Background: The effect of bradykinin on intracellular free Ca2+ levels ([Ca2+]i) in MG63 human osteosarcoma cells was explored using fura-2 as a Ca2+ dye. Methods/Results: Bradykinin (0.1 nM–1 µM) increased [Ca2+]i in a concentration-dependent manner with an EC50 value of 0.5 nM. The [Ca2+]i signal comprised an initial peak and a fast decay which returned to baseline in 2 min. Extracellular Ca2+ removal inhibited the peak [Ca2+]i signals by 35 ± 3%. Bradykinin (1 nM) failed to increase [Ca2+]i in the absence of extracellular Ca2+ after cells were pretreated with thapsigargin (an endoplasmic reticulum Ca2+ pump inhibitor; 1 µM). Bradykinin (1 nM)-induced intracellular Ca2+ release was nearly abolished by inhibiting phospholipase C with 2 µM 1-(6-((17β-3-methoxyestra-1,3,5(10)-trien-17-yl)amino)hexyl)-1H-pyrrole-2,5-dione (U73122). The [Ca2+]i increase induced by 1 nM bradykinin in Ca2+- free medium was abolished by 1 nM HOE 140 (a B2 bradykinin receptor antagonist) but was not altered by 100 nM Des-Arg-HOE 140 (a B1 bradykinin receptor antagonist). Pretreatment with 1 pM pertussis toxin for 5 h in Ca2+ medium inhibited 30 ± 3% of 1 nM bradykinin-induced peak [Ca2+]i increase. Conclusions: Together, this study shows that bradykinin induced [Ca2+]i increases in a concentration-dependent manner, by stimulating B2 bradykinin receptors leading to mobilization of Ca2+ from the thapsigargin-sensitive stores in a manner dependent on inositol-1,4,5-trisphosphate, and also by inducing extracellular Ca2+ influx. The bradykinin response was partly coupled to a pertussis toxin-sensitive G protein pathway.

Automatically retargeting analog designs to new technology is an efficient solution for reusing analog IPs. However, most of previous approaches focus on layout retargeting only. How to obtain the new device sizes for another technology is often not discussed. Simply scaling the device sizes may not reach the desired performance due to the non-ideal effects. Therefore, a layout-aware automatic sizing flow for retargeting analog circuits is proposed in this paper. Based on the layout template extracted from the original design, the layout-induced parasitic effects in new technology are also considered in the sizing flow. Since the possible performance degradation has been considered, no redesign cycles and reserved design margins are required in the proposed sizing flow, which significantly reduces the design overhead. As shown in the experimental results, the design retargeting can be finished in one second by using the proposed flow, which demonstrates the feasibility and efficiency of this approach.

The present study was performed to evaluate the 5-year status of immediately placed implants subjected to maxillary sinus elevation without grafting.Implants were placed in 2004 and 2005. A minimum of 3 mm of residual bone height (RBH) was required. All implants were placed with a sinus elevation performed through a lateral approach by the trap-door, open-window method without placement of any grafting material. Regular follow-up included oral hygiene instruction, periodontal charting, panoramic radiographs, and cone beam computed tomographic scans. The gained bone height (GBH) in the sinus, peri-implant sulcus depth, and marginal bone loss were analyzed statistically.Forty-four patients (26 men, 18 women) with an average age of 58 years received 80 implants, which were followed for 5 years after prosthesis delivery. No patients developed sinusitis or other complications leading to implant loss. The average RBH was 5.06 ± 1.51 mm and the average intrasinus implant length was 7.77 ± 1.69 mm. Survival rates for the implants were 100% after 2 and 5 years. Average GBH was 7.24 ± 1.83 mm at 2 years (range, 3 to 12 mm) and 7.44 ± 1.94 mm at 5 years (P.05). The average peri-implant sulcus depths were 2.5 ± 0.4 mm at 2 years and 3.1 ± 0.5 mm at 5 years (P.05). The mean peri-implant marginal bone loss was 1.3 ± 0.3 mm at 2 years and 2.1 ± 0.5 mm at 5 years (P.05).New bone formation in the sinus was confirmed, and good survival of implants with maxillary sinus elevation by the lateral approach without grafting was observed after 5 years. Attention should be focused on oral hygiene maintenance to ensure peri-implant gingival health.

We applied a multiplex polymerase chain reaction (PCR) and culture to detect Streptococcus pneumoniae and detected 3 other respiratory pathogens--Haemophilus influenzae, Moraxella catarrhalis, and Alloiococcus otitidis--simultaneously by PCR, in the nasopharynx of 386 children aged under 5 y. S. pneumoniae was the most common pathogen carried by children in all age groups, with the rate ranging from 15.8% in children aged 3-4 y to 28.6% in children aged 2-3 y. H. influenzae and M. catarrhalis showed similar carriage rates across all the age groups. Only 2 young children (0.5%) carried A. otitidis. Higher carriage of S. pneumoniae was found in children who had not received the heptavalent pneumococcal conjugate vaccine (PCV7). Cefotaxime non-susceptibility was high (51.4%) in S. pneumoniae nasopharyngeal isolates. Serotype 6B was the most common in fully immunized carriers and also in those who received catch-up immunization. Due to low PCV7 coverage in Taiwan, the carriage of vaccine and non-vaccine serotypes of S. pneumoniae in children remains common.

Zoledronic acid (ZOL) was shown earlier to prolong survival in animal models of lung cancer. The aim of this study was to examine whether alteration of intracellular cyclins, cyclin-dependent kinases, cyclin-dependent kinase inhibitors, retinoblastoma, and Ras protein expression and E2F localization are among the possible antilung cancer mechanisms driven by ZOL. Furthermore, we used geranylgeraniol to test whether the mevalonate pathway is involved in the antitumor effects of ZOL against lung cancer. Line-1 cells, a murine lung adenocarcinoma cell line, were examined. ZOL significantly slowed the growth of these cells both in vitro and in vivo. The ZOL-treated cells typically arrested at the S/G2/M phase of the cell cycle, accompanied by increased intracellular levels of cyclin A, B1, and CDC2 and decreased levels of cyclin D, p21, p27, phosphorylated retinoblastoma, and Ras. In addition, ZOL affected the distribution of E2F. When geranylgeraniol was added to the ZOL-treated cells, either in vitro or in vivo, tumor growth, cell-cycle progression, the expression of certain cyclins, and cyclin-related regulatory proteins were partially returned to that of untreated controls. Therefore, ZOL elicits cell-cycle prolongation that seems to be associated with alterations in the levels of certain cyclins and cyclin-related regulatory proteins. Furthermore, the mevalonate pathway regulates ZOL-induced murine lung cancer inhibition both in vitro and in vivo.

The AFM combined nanoindentation was performed to observe the ultrastructure of enamel rod from various section plans and positions while probing their mechanical and tribological properties of the area. The nanohardness and the elastic modulus of the head region of the enamel rods are significantly higher than that of the tail region and the axial-sectional plane. Both nanohardness and elastic modulus gradually decrease from enamel surface toward dentino-enamel junction. Such a variation correlates well with the decreasing trend of calcium composition from our element analysis. The friction coefficient and nanowear of the enamel showed an inversed trend to the hardness with respect to their relative topological position in the long axis of enamel rod toward DEJ. The relationship between the nanowear depth and the distance from the outer enamel surface to DEJ presented exponential function. The results presented clarify the basic nanomechanical and nanotribological properties of human enamel rods and provide a useful reference for the future development of dental restorative materials.

Glass ionomers have been used for perforation repair and retrograde filling where biointegration with periodontal tissue is required. Collagen has been demonstrated to promote cellular adhesion and enhance mineral tissue compressive strength. It was hypothesized that an appropriate concentration of collagen integrated into glass ionomer may improve both bio-compatibility and the mechanical properties of the material. By SEM and AFM, we discovered 70-nm granules appearing on the surfaces of glass-ionomer/collagen hybrids. Acid-etching revealed irregularly shaped particles interlinked by membrane-like sheets on the surface of the material with the typical 70-nm granules. WST-1 assay showed that acid-etching significantly enhanced the viability of attached gingival fibroblasts. However, the glass-ionomer/collagen hybrids’ combined surface-etching outperformed other groups. The glass-ionomer/collagen hybrids presented enhanced compressive strength when integrated with 0.01% collagen, while higher concentrations of collagen compromised their mechanical property. In summary, collagen improved both the mechanical and biocompatible properties of glass ionomers. Further in vivo study is warranted.

Calcium-fluoride-like deposits play a key role in caries prevention by topical fluoride. Previous microhardness analyses have introduced errors due to a substrate effect, and thereby could not substantiate the early loss of these deposits. To address this question, we applied Atomic Force Microscopy (AFM) and a nano-indentation technique in this study to characterize the nano-mechanical properties and topographic structure of enamel surfaces following topical fluoride treatment. The deposits were found to have a low nano-hardness and a high nano-wear depth, which explains the early loss of calcium-fluoride-like deposits. However, a 22% increase in the fluoride concentration could still be detected on the treated enamel surface following the removal of the surface deposits, justifying the long-term effectiveness of topical fluoride treatment.

Nordihydroguaiaretic acid (NDGA) is widely used as a pharmacological tool to inhibit lipoxygenases; however, recent evidence suggests that it increases renal intracellular [Ca2+]i via novel mechanisms. Here the effect of NDGA on Ca2+ signaling in MG63 osteoblastic cells was explored using fura-2 as a Ca2+ indicator. NDGA (2-50 microM) increased [Ca2+]i in a concentration-dependent manner. The signal comprised an initial rise and an elevated phase over a time period of 4 min. Removing extracellular Ca2+ reduced 2-50 microM NDGA-induced signals by 62+/-2%. After incubation with 50 microM NDGA in Ca2+-free medium for several minutes, addition of 3 mM CaCl2 induced an increase in [Ca2+]i. NDGA (50 microM)-induced [Ca2+]i increases were not changed by pretreatment with 10 microM of verapamil, diltiazem, nifedipine, nimodipine and nicardipine. In Ca2+-free medium, pretreatment with the endoplasmic reticulum Ca2+ pump inhibitor thapsigargin (1 microM) inhibited 50 microM NDGA-induced [Ca2+]i increases by 69+/-3%. Inhibition of phospholipase C with 2 microM U73122 had little effect on 50 microM NDGA-induced Ca2+ release. Several other lipoxygenase inhibitors had no effect on basal [Ca2+]i. At a concentration that did not increase basal [Ca2+]i, NDGA (1 microM) did not alter 10 microM ATP- or 1 microM thapsigargin-induced [Ca2+]i increases. Alteration of protein kinase C activity with 1 nM phorbol 12-myristate 13-acetate or 2 microM GF 109203X did not affect 50 microM NDGA-induced [Ca2+]i increases. Together, the results show that NDGA increased [Ca2+]i in osteoblasts in a lipoxygenase-independent manner, by releasing stored Ca2+ in a fashion independent of phospholipase C activity, and by causing Ca2+ influx.