Risk factors for anesthetic-related complications in pediatric patients with a newly diagnosed mediastinal mass.

BACKGROUND: Pediatric patients with a mediastinal mass can experience severe complications while undergoing anesthesia. Nearly, all published reviews involve either patients with an anterior mediastinal mass or patients with an oncologic disease. AIM: The identification of risk factors for anesthetic-related complications in pediatric patients with any type of mediastinal mass. METHODS: From January 1, 2008 to December 31, 2019, patients with a newly diagnosed mediastinal mass that underwent anesthesia were retrospectively identified. Each patient's medical record was reviewed for presenting symptoms, preprocedure imaging results, the type of anesthetic delivered, and the occurrence of any anesthetic-related complications. A complication was defined as severe hypoxia, severe hypotension, or loss of endtidal carbon dioxide. RESULTS: Eighty-six patients presented with a new mediastinal mass. Six of these patients (7%) had a complication. Complications were no more likely in patients with orthopnea than in patients without orthopnea (P = 1.00; relative risk (RR) = 0.95; 95% CI (0.1, 7.5). Complication rates in patients with anterior, middle, and posterior mediastinal masses were similar, as were complication rates in patients with large, medium, and small masses. Six of the 41 patients (15%) who had tracheal compression had a complication, while none of the 45 patients (0%) who did not have tracheal compression had a complication (p = .0096). Six of the 48 patients (13%) that were intubated had a complication, while none of the 38 patients (0%) who were not intubated had a complication (p = .032). Five of 36 patients (14%) who had mainstem bronchus compression had a complication, while one of 50 patients (2%) who did not have mainstem bronchus compression had a complication (p = .078; RR = 6.9l; 95% CI (0.8, 56.9)). CONCLUSIONS: Anesthetic-related complications were associated with airway compression and endotracheal intubation. The absence of preprocedure orthopnea did not ensure that the anesthetic would be uncomplicated. Complications occurred in similar frequencies in patients with a mediastinal mass of any location or size.

A Stacked Back Side-Illuminated Voltage Domain Global Shutter CMOS Image Sensor with a 4.0 µm Multiple Gain Readout Pixel †.

A backside-illuminated complementary metal-oxide-semiconductor (CMOS) image sensor with 4.0 µm voltage domain global shutter (GS) pixels has been fabricated in a 45 nm/65 nm stacked CMOS process as a proof-of-concept vehicle. The pixel components for the photon-to-voltage conversion are formed on the top substrate (the first layer). Each voltage signal from the first layer pixel is stored in the sample-and-hold capacitors on the bottom substrate (the second layer) via micro-bump interconnection to achieve a voltage domain GS function. The two sets of voltage domain storage capacitor per pixel enable a multiple gain readout to realize single exposure high dynamic range (SEHDR) in the GS operation. As a result, an 80dB SEHDR GS operation without rolling shutter distortions and motion artifacts has been achieved. Additionally, less than -140dB parasitic light sensitivity, small noise floor, high sensitivity and good angular response have been achieved.

Plant cell wall glycosyltransferases: High-throughput recombinant expression screening and general requirements for these challenging enzymes.

Molecular characterization of plant cell wall glycosyltransferases is a critical step towards understanding the biosynthesis of the complex plant cell wall, and ultimately for efficient engineering of biofuel and agricultural crops. The majority of these enzymes have proven very difficult to obtain in the needed amount and purity for such molecular studies, and recombinant cell wall glycosyltransferase production efforts have largely failed. A daunting number of strategies can be employed to overcome this challenge, including optimization of DNA and protein sequences, choice of expression organism, expression conditions, co-expression partners, purification methods, and optimization of protein solubility and stability. Hence researchers are presented with thousands of potential conditions to test. Ultimately, the subset of conditions that will be sampled depends on practical considerations and prior knowledge of the enzyme(s) being studied. We have developed a rational approach to this process. We devise a pipeline comprising in silico selection of targets and construct design, and high-throughput expression screening, target enrichment, and hit identification. We have applied this pipeline to a test set of Arabidopsis thaliana cell wall glycosyltransferases known to be challenging to obtain in soluble form, as well as to a library of cell wall glycosyltransferases from other plants including agricultural and biofuel crops. The screening results suggest that recombinant cell wall glycosyltransferases in general have a very low soluble:insoluble ratio in lysates from heterologous expression cultures, and that co-expression of chaperones as well as lysis buffer optimization can increase this ratio. We have applied the identified preferred conditions to Reversibly Glycosylated Polypeptide 1 from Arabidopsis thaliana, and processed this enzyme to near-purity in unprecedented milligram amounts. The obtained preparation of Reversibly Glycosylated Polypeptide 1 has the expected arabinopyranose mutase and autoglycosylation activities.

X-ray diffraction analysis and in vitro characterization of the UAM2 protein from Oryza sativa.

The role of seemingly non-enzymatic proteins in complexes interconverting UDP-arabinopyranose and UDP-arabinofuranose (UDP-arabinosemutases; UAMs) in the plant cytosol remains unknown. To shed light on their function, crystallographic and functional studies of the seemingly non-enzymatic UAM2 protein from Oryza sativa (OsUAM2) were undertaken. Here, X-ray diffraction data are reported, as well as analysis of the oligomeric state in the crystal and in solution. OsUAM2 crystallizes readily but forms highly radiation-sensitive crystals with limited diffraction power, requiring careful low-dose vector data acquisition. Using size-exclusion chromatography, it is shown that the protein is monomeric in solution. Finally, limited proteolysis was employed to demonstrate DTT-enhanced proteolytic digestion, indicating the existence of at least one intramolecular disulfide bridge or, alternatively, a requirement for a structural metal ion.

Transgenic expression of fungal accessory hemicellulases in Arabidopsis thaliana triggers transcriptional patterns related to biotic stress and defense response.

The plant cell wall is an abundant and renewable resource for lignocellulosic applications such as the production of biofuel. Due to structural and compositional complexities, the plant cell wall is, however, recalcitrant to hydrolysis and extraction of platform sugars. A cell wall engineering strategy to reduce this recalcitrance makes use of microbial cell wall modifying enzymes that are expressed directly in plants themselves. Previously, we constructed transgenic Arabidopsis thaliana constitutively expressing the fungal hemicellulases: Phanerochaete carnosa glucurnoyl esterase (PcGCE) and Aspergillus nidulans α-arabinofuranosidase (AnAF54). While the PcGCE lines demonstrated improved xylan extractability, they also displayed chlorotic leaves leading to the hypothesis that expression of such enzymes in planta resulted in plant stress. The objective of this study is to investigate the impact of transgenic expression of the aforementioned microbial hemicellulases in planta on the host arabidopsis. More specifically, we investigated transcriptome profiles by short read high throughput sequencing (RNAseq) from developmentally distinct parts of the plant stem. When compared to non-transformed wild-type plants, a subset of genes was identified that showed differential transcript abundance in all transgenic lines and tissues investigated. Intriguingly, this core set of genes was significantly enriched for those involved in plant defense and biotic stress responses. While stress and defense-related genes showed increased transcript abundance in the transgenic plants regardless of tissue or genotype, genes involved in photosynthesis (light harvesting) were decreased in their transcript abundance potentially reflecting wide-spread effects of heterologous microbial transgene expression and the maintenance of plant homeostasis. Additionally, an increase in transcript abundance for genes involved in salicylic acid signaling further substantiates our finding that transgenic expression of microbial cell wall modifying enzymes induces transcriptome responses similar to those observed in defense responses.

Loss of Inositol Phosphorylceramide Sphingolipid Mannosylation Induces Plant Immune Responses and Reduces Cellulose Content in Arabidopsis.

Glycosylinositol phosphorylceramides (GIPCs) are a class of glycosylated sphingolipids found in plants, fungi, and protozoa. These lipids are abundant in the plant plasma membrane, forming ∼25% of total plasma membrane lipids. Little is known about the function of the glycosylated headgroup, but two recent studies have indicated that they play a key role in plant signaling and defense. Here, we show that a member of glycosyltransferase family 64, previously named ECTOPICALLY PARTING CELLS1, is likely a Golgi-localized GIPC-specific mannosyl-transferase, which we renamed GIPC MANNOSYL-TRANSFERASE1 (GMT1). Sphingolipid analysis revealed that the Arabidopsis thaliana gmt1 mutant almost completely lacks mannose-carrying GIPCs. Heterologous expression of GMT1 in Saccharomyces cerevisiae and tobacco (Nicotiana tabacum) cv Bright Yellow 2 resulted in the production of non-native mannosylated GIPCs. gmt1 displays a severe dwarfed phenotype and a constitutive hypersensitive response characterized by elevated salicylic acid and hydrogen peroxide levels, similar to that we previously reported for the Golgi-localized, GIPC-specific, GDP-Man transporter GONST1 (Mortimer et al., 2013). Unexpectedly, we show that gmt1 cell walls have a reduction in cellulose content, although other matrix polysaccharides are unchanged.

Effect of systemic antibiotics on clinical and patient-reported outcomes of implant therapy - a multicenter randomized controlled clinical trial.

OBJECTIVES: To determine the effect of various systemic antibiotic prophylaxis regimes on patient-reported outcomes and postsurgical complications in patients undergoing conventional implant installation. MATERIAL AND METHODS: Three hundred and twenty-nine healthy adults in need of conventional implant installation were randomly assigned to one of four groups: (i) preoperatively 2 g of amoxycillin 1 h before surgery (positive control, PC), (ii) postoperatively 2 g of amoxycillin immediately following surgery (test 1, T1), (iii) preoperatively 2 g of amoxycillin 1 h before and 500 mg thrice daily on days 2 and 3 after surgery (test 2, T2), (iv) preoperatively 2 g of placebo 1 h before surgery (negative control, NC). Subjects were examined clinically by blinded examiners over 8 weeks after implant installation. In addition, Visual Analogue Scales (VAS) for pain, swelling, bruising and bleeding were obtained over 14 days. ANOVA was performed for the VAS. Chi-square tests were applied for postsurgical complications. RESULTS: All VAS scores were low for all groups and decreased over time (P < 0.001). There were no significant differences for the VAS scores between the various groups at any time point (P > 0.05). There was only a significant difference in flap closure at week 4, where NC had 5% of the subjects not achieving complete wound closure compared to 0% for the three other groups (P = 0.01), with no other significant differences for any postsurgical complications (P > 0.05). CONCLUSION: For standard single implant placement, prophylactic systemic antibiotics either before or after, or before and after the surgical procedure do not improve patient-reported outcomes or prevalence of postsurgical complications.

Towards practical time-of-flight secondary ion mass spectrometry lignocellulolytic enzyme assays.

BACKGROUND: Time-of-Flight Secondary Ion Mass Spectrometry (ToF-SIMS) is a surface sensitive mass spectrometry technique with potential strengths as a method for detecting enzymatic activity on solid materials. In particular, ToF-SIMS has been applied to detect the enzymatic degradation of woody lignocellulose. Proof-of-principle experiments previously demonstrated the detection of both lignin-degrading and cellulose-degrading enzymes on solvent-extracted hardwood and softwood. However, these preliminary experiments suffered from low sample throughput and were restricted to samples which had been solvent-extracted in order to minimize the potential for mass interferences between low molecular weight extractive compounds and polymeric lignocellulose components. RESULTS: The present work introduces a new, higher-throughput method for processing powdered wood samples for ToF-SIMS, meanwhile exploring likely sources of sample contamination. Multivariate analysis (MVA) including Principal Component Analysis (PCA) and Multivariate Curve Resolution (MCR) was regularly used to check for sample contamination as well as to detect extractives and enzyme activity. New data also demonstrates successful ToF-SIMS analysis of unextracted samples, placing an emphasis on identifying the low-mass secondary ion peaks related to extractives, revealing how extractives change previously established peak ratios used to describe enzyme activity, and elucidating peak intensity patterns for better detection of cellulase activity in the presence of extractives. The sensitivity of ToF-SIMS to a range of cellulase doses is also shown, along with preliminary experiments augmenting the cellulase cocktail with other proteins. CONCLUSIONS: These new procedures increase the throughput of sample preparation for ToF-SIMS analysis of lignocellulose and expand the applications of the method to include unextracted lignocellulose. These are important steps towards the practical use of ToF-SIMS as a tool to screen for changes in plant composition, whether the transformation of the lignocellulose is achieved through enzyme application, plant mutagenesis, or other treatments.

Tail tip proteins related to bacteriophage λ gpL coordinate an iron-sulfur cluster.

The assembly of long non-contractile phage tails begins with the formation of the tail tip complex (TTC). TTCs are multi-functional protein structures that mediate host cell adsorption and genome injection. The TTC of phage λ is assembled from multiple copies of eight different proteins, including gpL. Purified preparations of gpL and several homologues all displayed a distinct reddish color, suggesting the binding of iron by these proteins. Further characterization of the gpL homologue from phage N15, which was most amenable to in vitro analyses, showed that it contains two domains. The C-terminal domain was demonstrated to coordinate an iron-sulfur cluster, providing the first example of a viral structural protein binding to this type of metal group. We characterized the iron-sulfur cluster using inductively coupled plasma-atomic emission spectroscopy, absorbance spectroscopy, and electron paramagnetic resonance spectroscopy and found that it is an oxygen-sensitive [4Fe-4S](2+) cluster. Four highly conserved cysteine residues were shown to be required for coordinating the iron-sulfur cluster, and substitution of any of these Cys residues with Ser or Ala within the context of λ gpL abolished biological activity. These data imply that the intact iron-sulfur cluster is required for function. The presence of four conserved Cys residues in the C-terminal regions of very diverse gpL homologues suggest that utilization of an iron-sulfur cluster is a widespread feature of non-contractile tailed phages that infect Gram-negative bacteria. In addition, this is the first example of a viral structural protein that binds an iron-sulfur cluster.

Constitutive expression of a fungal glucuronoyl esterase in Arabidopsis reveals altered cell wall composition and structure.

A family 15 carbohydrate esterase (CE15) from the white-rot basidiomycete, Phanerochaete carnosa (PcGCE), was transformed into Arabidopsis thaliana Col-0 and was expressed from the constitutive cauliflower mosaic virus 35S promoter. Like other CE15 enzymes, PcGCE hydrolyzed methyl-4-O-methyl-d-glucopyranuronate and could target ester linkages that contribute to lignin-carbohydrate complexes that form in plant cell walls. Three independently transformed Arabidopsis lines were evaluated in terms of nine morphometric parameters, total sugar and lignin composition, cell wall anatomy, enzymatic saccharification and xylan extractability. The transgenic lines consistently displayed a leaf-yellowing phenotype, as well as reduced glucose and xylose content by as much as 30% and 35%, respectively. Histological analysis revealed 50% reduction in cell wall thickness in the interfascicular fibres of transgenic plants, and FT-IR microspectroscopy of interfascicular fibre walls indicated reduction in lignin cross-linking in plants overexpressing PcGCE. Notably, these characteristics could be correlated with improved xylose recovery in transgenic plants, up to 15%. The current analysis represents the first example whereby a fungal glucuronoyl esterase is expressed in Arabidopsis and shows that the promotion of glucuronoyl esterase activity in plants can alter the extent of intermolecular cross-linking within plant cell walls.

Association of BACE1 Gene Polymorphism with Cerebellar Volume but Not Cognitive Function in Normal Individuals.

AIMS: ß-Site amyloid precursor protein (APP)-cleaving enzyme 1 (BACE1) is a biological and positional candidate gene for Alzheimer's disease (AD). Previous studies found that BACE1-null mice had impaired performance on cognition and neurodegeneration during the aging process. Additionally, a synonymous polymorphism of BACE1 (rs638405) in exon 5 has been reported to be associated with risk for AD. We hypothesized that this BACE1 gene variant might influence regional brain volumes and cognitive tests in normal individuals. METHODS: Participants were 330 normal volunteers between 21 and 92 years of age (mean age 56.3 ± 22.0 years; 191 males, 139 females). Cognitive tests (the Mini-Mental State Examination and Digit Spans), magnetic resonance imaging, and genotyping of BACE1 rs638405 were examined for each subject. The differences in regional gray matter (GM) volumes between G homozygotes and C-allele carriers were tested using optimized voxel-based morphometry. RESULTS: Compared to C-allele carriers, G homozygotes exhibited significantly larger GM volumes in the left cerebellar culmen and right cerebellar lingual area, but no significant differences on cognitive function tests. CONCLUSION: The findings suggest that the BACE1 rs638405 polymorphism may affect cerebellar morphology, but not cognitive function in healthy humans.

A quantitative scale for the extent of conjugation of the amide bond. Amidity percentage as a chemical driving force.

The amide bond may be considered as one of the most important chemical building blocks, playing an important role not only in living organisms but in organic chemistry as well. The exact description and precise quantification of the amide bond strength is difficult, requiring a particular type of theoretical investigation. The present paper suggests a novel, yet simple, method toward quantifying amide bond strength on a linear scale, defined as the "amidity scale". This is achieved using the computed enthalpy of hydrogenation (DeltaHH2) of the compound examined. In the present conceptual work, the DeltaHH2 value for dimethylacetamide is used to define perfect amidic character (amidity=+100%), while azaadamantane-2-on represents complete absence of amidic character (amidity=0%). The component DeltaHH2 values were computed at differing levels of theory, providing a computational and quasi-"method-independent" measure of amidity. A total of 29 well-known amides were examined to demonstrate the "scoring" accuracy of this methodology. For the compounds examined, a correlation has been made between the computed amidity percentage and their common COSNAR resonance energy values, proton affinities, and reactivity in a nucleophilic addition reaction. Selected chemical reactions were also studied. It has been shown that the change of the amidity value, during acyl transfer reactions, represents a thermodynamic driving force for the reaction.

Comparison of acellular dermal graft and palatal autograft in the reconstruction of keratinized gingiva around dental implants: a case report.

The use of autogenous gingival grafts has proved to be an effective and predictable way to increase the amount of keratinized gingiva. However, discomfort and pain at the donor site are unavoidable. Acellular dermal matrix (ADM) allograft can be used as a donor tissue to eliminate the need for another surgical site and alleviate pain and trauma. The purpose of this study was to evaluate the effectiveness of ADM allograft in increasing the width of keratinized gingiva around dental implants. A patient with inadequate keratinized gingiva around dental implants in maxillary and mandibular anterior regions received either an ADM graft or palatal autograft by random allocation. The width of keratinized gingiva and other clinical periodontal parameters were recorded initially and at 3 and 6 months after surgery. Both grafts provided satisfactory results. The width of keratinized tissues was increased by using the ADM allograft, but by a lesser amount than seen with the autogenous gingival graft.

Double laterally-rotated bilayer flap operation for treatment of gingival recession.

BACKGROUND AND PURPOSE: Many root coverage techniques, such as lateral pedicle flap, double papilla flap, free gingival graft, connective tissue graft, and guided tissue regeneration (GTR) procedure have been widely applied in clinical dental practice. Although they all provide various degrees of successful root coverage, some procedures are technique-sensitive and others require a 2-stage operation. METHODS: In this study, we describe a new technique with a double laterally-rotated bilayer flap for root coverage. Briefly, 2 partial-thickness flaps were prepared, one on each side of the gingival recession, and then laterally repositioned (with mutual overlapping) to cover the root exposure. After thorough planing of the exposed root surfaces, root conditioning with saturated citric acid solution (pH = 1.0) was performed. One partial-thickness flap was de-epithelialized and then rotated to cover the citric acid-conditioned root surface. Another partial-thickness flap was further rotated to cover the de-epithelialized flap. Both flaps were sutured to the recipient periosteal bed. This new root coverage technique was used to treat 2 teeth with gingival recession. RESULTS: The results demonstrated that this new technique provides satisfactory root coverage of gingival recession and improves root dentin hypersensitivity of treated teeth. The main advantages of this technique are simple 1-stage surgery, esthetic color matching of the soft tissue, and a good blood supply of the reflected flap. CONCLUSIONS: We conclude that the double laterally-rotated bilayer flap operation may provide a new alternative for the treatment of gingival recession.

Polymer-assisted regeneration therapy with Atrisorb barriers in human periodontal intrabony defects.

AIM: This study compared clinical results of 40 periodontal osseous defects treated by two types of absorbable barrier materials. MATERIAL AND METHODS: Thirty patients (23 males and seven females) suffering from moderate to advanced periodontitis (with comparable osseous defects) were randomly assigned to receive either Atrisorb barrier (n = 22; group A) or Resolut XT barrier (n = 18; group B) therapy. Periodontal phase I treatment and oral hygiene instruction were performed before periodontal surgery. Papillary preservation, partial thickness flap, citric acid root conditioning, and decortication procedures were applied during the operation. Bone defects were filled with demineralized freeze-dried bone allograft and minocycline mixture (4:1 ratio). Postoperative care included 0.10% chlorhexidine rinse daily and antibiotic medication for 2 weeks. Clinical assessments including probing depth (PD), clinical attachment level (CAL), gingival recession (GR), plaque index (PII), gingival index (GI), and radiographic examinations were taken at the baseline, preoperatively and at 3 and 6 months after regenerative surgery. RESULTS: Six months following therapy, both Atrisorb and Resolut XT groups had achieved comparable clinical improvement in pocket reduction (3.9 versus 4.4 mm), attachment tissue gain (clinical attachment gain; 3.5 versus 3.6 mm), and reduction in the GI and in the PII. Within-group comparisons showed significant attachment gain and pocket reduction between baseline data and those at both 3 and 6 months postoperatively (p < 0.01). There were no statistically significant differences in any measured data between groups A and B. CONCLUSIONS: The results of this study indicate that a comparable and favorable regeneration of periodontal defects can be achieved with both Atrisorb and Resolut XT barriers. Further long-term study and histologic observations of tissue healing are needed to evaluate whether Atrisorb is promising for clinical use.

Autologous transplantation of gingival fibroblast-like cells and a hydroxylapatite complex graft in the treatment of periodontal osseous defects: cell cultivation and long-term report of cases.

Autogenous cell transplantation via hydroxylapatite (HA) vehicle has been reported to have beneficial effects on the treatment of human periodontal osseous defects. The aim of this study was to explore the possibility of using gingival fibroblast-like cells in the therapy of osseous defects caused by inflammatory periodontitis by reporting long-term results of gingival fibroblast-coated hydroxylapatite (GF-HA) grafting for healing these defects. Gingival fibroblasts were cultured from healthy gingivae of treated subjects. Growth of cells on HA particles was established in vitro, and then the GF-HA complex was transplanted into the periodontal osseous defects. Clinical parameters of gingival and plaque indices, probing depth, and periapical x-ray were monitored at baseline and at various periods from 50 months to 6 years after surgery. Grafting with only HA in the osseous defects of the same patient was used for comparison. The present study shows that GF-HA-treated sites could achieve marked pocket reduction and probing attachment gain at reentry and later recalls. Good clinical bone filling of osseous defects in GF-HA-treated sites was also demonstrated in periapical radiographs (increased bone height and reappearance of the crestal cortex) and in some reentry sites. One HA-treated site was filled with connective tissue only, and the absence of new bone formation was noted during a reentry operation. Another HA-treated site exhibited a comparable increase in radiographic density, while part of HA particles were gradually lost in longer recalls. These limited observations conclude that GF-HA grafting may provide a treatment modality leading to regeneration of periodontal tissues in periodontitis-affected osseous defects. Further studies including more cases and demonstration of the deposition of differentiated periodontal tissues are necessary before further application of this therapy.