Parent-proxy pediatric CMT quality of life outcome measure: Validation of the Italian version.

BACKGROUND AND AIMS: The parent-proxy reports can offer complementary informations or be the only source of Quality of Life measurement in young children. The aim of this study was to provide and validate the Italian version of the recently published parent-proxy pCMT-QOL for patients aged 8-18 years old, making it available for possible trials in Italian speaking children. METHODS: The English-language instrument was translated and adapted into the Italian language using standard procedures: translation, transcultural adaptation, and back-translation. Parent-proxy pCMT-QOL was administered to parents of patients with a genetic diagnosis of CMT, aged 8-18 years old. All parents were retested 2 weeks later to assess reliability. RESULTS: A total of 21 parents of CMT patients (18 CMT1A, 2 CMT2A, 1 CMT2K) were assessed during their children clinical appointments. The Italian-pCMT-QOL showed a high test-retest reliability; none of the parents had any difficulties with the completion of the questionnaire and no further revisions were necessary after completion. INTERPRETATION: The Italian parent-proxy pCMT-QOL is a reliable, culturally adapted, and comparable version of the original English instrument. This questionnaire will improve the quality of the follow-up and will make it possible to monitor more accurately the severity of the disease in Italian-speaking families.

Sympathetic innervation of the supraclavicular brown adipose tissue: A detailed anatomical study.

BACKGROUND: The supraclavicular fossa is the dominant location for human brown adipose tissue (BAT). Activation of BAT promotes non-shivering thermogenesis by utilization of glucose and free fatty acids and has been the focus of pharmacological and non-pharmacological approaches for modulation in order to improve body weight and glucose homeostasis. Sympathetic neural control of supraclavicular BAT has received much attention, but its innervation has not been extensively investigated in humans. METHODS: Dissection of the cervical region in human cadavers was performed to find the distribution of sympathetic nerve branches to supraclavicular fat pad. Furthermore, proximal segments of the 4th cervical nerve were evaluated histologically to assess its sympathetic components. RESULTS: Nerve branches terminating in supraclavicular fat pad were identified in all dissections, including those from the 3rd and 4th cervical nerves and from the cervical sympathetic plexus. Histology of the proximal segments of the 4th cervical nerves confirmed tyrosine hydroxylase positive thin nerve fibers in all fascicles with either a scattered or clustered distribution pattern. The scattered pattern was more predominant than the clustered pattern (80% vs. 20%) across cadavers. These sympathetic nerve fibers occupied only 2.48% of the nerve cross sectional area on average. CONCLUSIONS: Human sympathetic nerves use multiple pathways to innervate the supraclavicular fat pad. The present finding serves as a framework for future clinical approaches to activate human BAT in the supraclavicular region.

Examining the Molecular Origins of Anomalously High H2O Generation at Oxide-Passivated Metal Surfaces for Plasma Applications.

Elucidating the mechanisms responsible for sub-microsecond desorption of water and other impurities from electrode surfaces at high heating rates is crucial for understanding pulsed-power behavior and optimizing its efficiency. Ionization of desorbed impurities in the vacuum regions may create parallel loads and current loss. Devising methods to limit desorption during the short time duration of pulsed-power will signficantly improve the power output. This problem also presents an exciting challenge to and paradigm for molecular length-scale modeling and theories. Previous molecular modeling studies have strongly suggested that, under high vacuum conditions, the amount of water impurity adsorbed on oxide surfaces on metal electrodes is at a sub-monolayer level, which appears insufficient to explain the observed pulsed-power losses at high current densities. Based on Density Functional Theory (DFT) calculations, we propose that hydrogen trapped inside iron metal can diffuse into iron (III) oxide on the metal surface in sub-microsecond time scales, explaining the extra desorbed inventory. These hydrogen atoms react with the oxide to form Fe(II) and desorbed H2O at elevated temperatures. Cr2O3 is found to react more slowly to form Cr(II). H2 evolution is also predicted to require higher activation energies, so H2 may be evolved at later times than H2O. A one-dimensional diffusion model, based on DFT results, is devised to estimate the water outgassing rate under different conditions. This model explains outgassing above 1 ML for surface temperatures of 1 eV often assumed in pulsed-power systems. Finally, we apply a suite of characterization techniques to demonstrate that when iron metal is heated to 650 Celsius, the dominant surface oxide component becomes alpha-Fe2O3. We propose such specially-prepared samples will lead to convergence between atomic modeling and measurements like temperature-programmed desorption. .

Blueberry and cardiovascular disease risk factors: A systematic review and meta-analysis of randomized controlled trials.

BACKGROUND AND AIMS: Cardiovascular disease (CVD) remains an important cause of mortality and morbidity that can be prevented by the consumption of healthy foods. These include blueberry, a dark coloured berry containing extremely high amounts of functional ingredients. We therefore examined the extent to which supplementation with blueberry effects on CVD risk indices. METHODS: We searched the ISI Web of Science, Scopus, PubMed and Cochrane Library on March 2020 and checked reference lists from primary studies and review articles for any additional studies. No language restrictions were applied. All randomized and controlled clinical trials (RCTs) using blueberry supplements to modify CVD risk factors were included in our analysis. RESULTS: Mean Difference (MD) was pooled using a random effects model and 11 studies were included in the final analysis. Pooled effect size showed that supplementation with blueberry had a small insignificant effect in reducing plasma triglycerides (MD = -0.27 mmol/l; 95 % CI: -0.57, 0.17, p = 0.06). Although current study found no differences between blueberry and control groups for any other outcomes, subgroup analysis suggested a favourable impact of blueberry on reducing body weight. Significant weight loss was indicated from studies longer with a follow up of more than 6 weeks or with blueberry powder or freeze-dried blueberry. CONCLUSION: Current evidence is insufficient to show a benefit of blueberry supplements in modifying CVD risk factors across a variety of adult populations. Robust data and larger studies are required to assess potential effects.

Effect of flaxseed supplementation on lipid profile: An updated systematic review and dose-response meta-analysis of sixty-two randomized controlled trials.

Raised plasma lipids are one the most important risk factors for cardiovascular disease. Flaxseed contains considerable amounts of α-linolenic acid, phenolic compounds, and lignans, which each have the capacity to reduce circulating lipid concentrations. This study aimed to systematically review current evidence to identify the potential effects of flaxseed supplementation on blood lipid profiles using a meta-analysis of randomized controlled trials (RCTs). PubMed, Scopus, Web of Science, and Google Scholar databases were searched for publications between January 1900 and May 2019. Weighted mean differences (WMDs) were analyzed using a random-effects model. The Cochrane Collaboration tool was also used to assess the risk of bias of the studies included. Sixty-two RCTs with a total of 3772 participants met the eligibility criteria. Our analysis showed that flaxseed supplementation significantly reduced total cholesterol (TC) (WMD = -5.389 mg/dL; 95% CI: -9.483, -1.295, p = 0.010), triglyceride (TG) (WMD = -9.422 mg/dL; 95% CI: -15.514, -3.330, p = 0.002), and low-density lipoprotein cholesterol (LDL-C) (WMD = -4.206 mg/dl; 95% CI: -7.260, -1.151, p = 0.007) concentrations. However, it had no effects on high-density lipoprotein cholesterol (WMD = 0.047 mg/dl; 95% CI: -0.777, 0.872, p = 0.910). This meta-analysis suggested that flaxseed supplementation improves serum TC, TG, and LDL-C, which could delay the progression of heart disease. Further studies with large-scale and better design are now needed to confirm these results.

Effect of flaxseed supplementation on markers of inflammation and endothelial function: A systematic review and meta-analysis.

OBJECTIVES: The rationale for the current study was to evaluate the efficacy of flaxseed supplementation on important adhesion molecules and inflammatory cytokines in adults. METHODS: We conducted searches of published literature in PubMed, Scopus, Web of Science, and Google Scholar databases from inception until May 2019. All randomized controlled trials (RCTs) which investigated the effects of flaxseed supplementation on the circulating concentrations of C-reactive protein (CRP), interleukin-6 (IL-6), tumor necrosis factor-alpha (TNF-α), vascular cell adhesion protein 1 (VCAM-1), E-selectin, and intercellular adhesion molecule 1 (ICAM-1) were included in our analysis. Results were summarized using weighted mean differences (WMDs) by random-effects model. RESULTS: Forty eligible RCTs, including 2520 participants were identified. The results of the meta-analysis revealed flaxseed supplementation reduced the concentrations of CRP (WMD = -0.387 mg/L; 95% CI: -0.653, -0.121, p = 0.004), IL-6 (WMD = -0.154 pg/Ml; 95% CI: -0.299, -0.010, p = 0.036), and VCAM-1 (WMD = -22.809 ng/ml; 95% CI: -41.498, -4.120, p = 0.017) but had no significant effect on TNF-α (WMD = -0.077 pg/mL; 95% CI: -0.317, 0.163, p = 0.530), ICAM-1 (WMD = -8.610 ng/ml; 95% CI: -21.936, 4.716, p = 0.205), and E-selectin (WMD = -1.427 ng/ml; 95% CI: -4.074, 1.22, p = 0.291). CONCLUSIONS: These findings showed that flaxseed supplementation may improve some circulating concentrations of specific adhesion molecules and inflammatory cytokines. However, well-designed trials are needed to confirm the range of non-significant and/or equivocal findings.

WWOX somatic ablation in skeletal muscles alters glucose metabolism.

OBJECTIVE: WWOX, a well-established tumor suppressor, is frequently lost in cancer and plays important roles in DNA damage response and cellular metabolism. METHODS: We re-analyzed several genome-wide association studies (GWAS) using the Type 2 Diabetes Knowledge Portal website to uncover WWOX's association with metabolic syndrome (MetS). Using several engineered mouse models, we studied the effect of somatic WWOX loss on glucose homeostasis. RESULTS: Several WWOX variants were found to be strongly associated with MetS disorders. In mouse models, somatic ablation of Wwox in skeletal muscle (WwoxΔSKM) results in weight gain, glucose intolerance, and insulin resistance. Furthermore, WwoxΔSKM mice display reduced amounts of slow-twitch fibers, decreased mitochondrial quantity and activity, and lower glucose oxidation levels. Mechanistically, we found that WWOX physically interacts with the cellular energy sensor AMP-activated protein kinase (AMPK) and that its loss is associated with impaired activation of AMPK, and with significant accumulation of the hypoxia inducible factor 1 alpha (HIF1α) in SKM. CONCLUSIONS: Our studies uncover an unforeseen role of the tumor suppressor WWOX in whole-body glucose homeostasis and highlight the intimate relationship between cancer progression and metabolic disorders, particularly obesity and type-2 diabetes. SUBJECT AREAS: Genetics, Metabolic Syndrome, Diabetes.

Association of p16 expression with prognosis varies across ovarian carcinoma histotypes: an Ovarian Tumor Tissue Analysis consortium study.

We aimed to validate the prognostic association of p16 expression in ovarian high-grade serous carcinomas (HGSC) and to explore it in other ovarian carcinoma histotypes. p16 protein expression was assessed by clinical-grade immunohistochemistry in 6525 ovarian carcinomas including 4334 HGSC using tissue microarrays from 24 studies participating in the Ovarian Tumor Tissue Analysis consortium. p16 expression patterns were interpreted as abnormal (either overexpression referred to as block expression or absence) or normal (heterogeneous). CDKN2A (which encodes p16) mRNA expression was also analyzed in a subset (n = 2280) mostly representing HGSC (n = 2010). Association of p16 expression with overall survival (OS) was determined within histotypes as was CDKN2A expression for HGSC only. p16 block expression was most frequent in HGSC (56%) but neither protein nor mRNA expression was associated with OS. However, relative to heterogeneous expression, block expression was associated with shorter OS in endometriosis-associated carcinomas, clear cell [hazard ratio (HR): 2.02, 95% confidence (CI) 1.47-2.77, p < 0.001] and endometrioid (HR: 1.88, 95% CI 1.30-2.75, p = 0.004), while absence was associated with shorter OS in low-grade serous carcinomas (HR: 2.95, 95% CI 1.61-5.38, p = 0.001). Absence was most frequent in mucinous carcinoma (50%), and was not associated with OS in this histotype. The prognostic value of p16 expression is histotype-specific and pattern dependent. We provide definitive evidence against an association of p16 expression with survival in ovarian HGSC as previously suggested. Block expression of p16 in clear cell and endometrioid carcinoma should be further validated as a prognostic marker, and absence in low-grade serous carcinoma justifies CDK4 inhibition.

High-definition infrared thermography of ice nucleation and propagation in wheat under natural frost conditions and controlled freezing.

MAIN CONCLUSION: An extremely high resolution infrared camera demonstrated various freezing events in wheat under natural conditions. Many of those events shed light on years of misunderstanding regarding freezing in small grains. Infrared thermography has enhanced our knowledge of ice nucleation and propagation in plants through visualization of the freezing process. The majority of infrared analyses have been conducted under controlled conditions and often on individual organs instead of whole plants. In the present study, high-definition (1280 × 720 pixel resolution) infrared thermography was used under natural conditions to visualize the freezing process of wheat plants during freezing events in 2016 and 2017. Plants within plots were found to freeze one at a time throughout the night and in an apparently random manner. Leaves on each plant also froze one at a time in an age-dependent pattern with oldest leaves freezing first. Contrary to a common assumption that freezing begins in the upper parts of leaves; freezing began at the base of the plant and spread upwards. The high resolution camera used was able to verify that a two stage sequence of freezing began within vascular bundles. Neither of the two stages was lethal to leaves, but a third stage was demonstrated at colder temperatures that was lethal and was likely a result of dehydration stress; this stage of freezing was not detectable by infrared. These results underscore the complexity of the freezing process in small grains and indicate that comprehensive observational studies are essential to identifying and selecting freezing tolerance traits in grain crops.

Activation of Fibrinolysis, But Not Coagulation, During End-Ischemic Ex Situ Normothermic Machine Perfusion of Human Donor Livers.

BACKGROUND: Ex situ normothermic machine perfusion (NMP) can be performed after traditional static cold preservation to assess graft function and viability before transplantation. It is unknown whether this results in activation of coagulation and fibrinolysis, as may occur upon graft reperfusion in vivo. METHODS: Twelve donor livers declined for transplantation underwent 6 hours of end-ischemic NMP using a heparinized plasma-based perfusion fluid. Concentration of prothrombin fragment F1 + 2 (marker of coagulation activation), D-dimer, plasmin-antiplasmin complex, tissue plasminogen activator and plasminogen activator inhibitor-1 (markers for fibrinolysis) and alanine aminotransferase (ALT) (marker of ischemia-reperfusion [I/R] injury) were measured in perfusion fluid at regular intervals. Liver biopsies were examined for the presence of fibrin, using light microscopy after Maurits, Scarlet and Blue staining. RESULTS: No significant increase in prothrombin F1 + 2 was noted during NMP. D-dimer and plasmin-antiplasmin complex levels increased soon after start of NMP and D-dimer concentrations correlated significantly with levels of tissue plasminogen activator. In livers displaying good function during NMP, perfusate levels of ALT and D-dimers were low (≤3500 ng/mL), whereas significantly higher D-dimer levels (>3500 ng/mL) were in found in livers with poor graft function. Activation of fibrinolysis correlated significantly with the degree of I/R injury, as reflected by ALT levels. CONCLUSIONS: End-ischemic ex situ NMP results in activation of fibrinolysis, but not of coagulation. Markers of fibrinolysis activation correlate significantly with markers of I/R injury. High concentrations of D-dimer early after start of NMP can be considered a marker of severe I/R injury and a predictor of poor liver graft function.

Quantitative body mass characterization before and after head and neck cancer radiotherapy: A challenge of height-weight formulae using computed tomography measurement.

OBJECTIVES: We undertook a challenge to determine if one or more height-weight formula(e) can be clinically used as a surrogate for direct CT-based imaging assessment of body composition before and after radiotherapy for head and neck cancer (HNC) patients, who are at risk for cancer- and therapy-associated cachexia/sarcopenia. MATERIALS AND METHODS: This retrospective single-institution study included 215 HNC patients, treated with curative radiotherapy between 2003 and 2013. Height/weight measures were tabulated. Skeletal muscle mass was contoured on pre- and post-treatment CT at the L3 vertebral level. Three common lean body mass (LBM) formulae (Hume, Boer, and James) were calculated, and compared to CT assessment at each time point. RESULTS: 156 patients (73%) had tumors arising in the oropharynx and 130 (61%) received concurrent chemotherapy. Mean pretreatment body mass index (BMI) was 28.5±4.9kg/m(2) in men and 27.8±8kg/m(2) in women. Mean post-treatment BMI were 26.2±4.4kg/m(2) in men, 26±7.5kg/m(2) in women. Mean CT-derived LBM decreased from 55.2±11.8kg pre-therapy to 49.27±9.84kg post-radiation. Methods comparison revealed 95% limit of agreement of ±12.5-13.2kg between CT and height-weight formulae. Post-treatment LBM with the three formulae was significantly different from CT (p<0.0001). In all instances, no height-weight formula was practically equivalent to CT within±5kg. CONCLUSION: Formulae cannot accurately substitute for direct quantitative imaging LBM measurements. We therefore recommend CT-based LBM assessment as a routine practice of head and neck cancer patient body composition.

Beam path toxicity in candidate organs-at-risk: assessment of radiation emetogenesis for patients receiving head and neck intensity modulated radiotherapy.

BACKGROUND: To investigate potential dose-response relationship between radiation-associated nausea and vomiting (RANV) reported during radiotherapy and candidate nausea/vomiting-associated regions of interest (CNV-ROIs) in head and neck (HNC) squamous cell carcinomas. METHODS AND MATERIAL: A total of 130 patients treated with IMRT with squamous cell carcinomas of head and neck were evaluated. For each patient, CNV-ROIs were segmented manually on planning CT images. Clinical on-treatment RANV data were reconstructed by a review of the records for all patients. Dosimetric data parameters were recorded from dose-volume histograms. Nausea and vomiting reports were concatenated as a single binary "Any N/V" variable, and as a "CTC-V2+" variable. RESULTS: The mean dose to CNV-ROIs was higher for patients experiencing RANV events. For patients receiving IMRT alone, a dose-response effect was observed with varying degrees of magnitude, at a statistically significant level for the area postrema, brainstem, dorsal vagal complex, medulla oblongata, solitary nucleus, oropharyngeal mucosa and whole brain CNV-ROIs. CONCLUSION: RANV is a common therapy-related morbidity facing patients receiving HNC radiotherapy, and, for those receiving radiotherapy-alone, is associated with modifiable dose to specific CNS structures.

Modulation of peripheral Na(+) channels and neuronal firing by n-butyl-p-aminobenzoate.

n-butyl-p-aminobenzoate (BAB), a local anesthetic, is administered epidurally in cancer patients to treat pain that is poorly controlled by other drugs that have a number of adverse effects. The purpose of the study was to unravel the mechanisms underlying the apparent selective pain suppressant effect of BAB. We used the whole-cell patch-clamp technique to record Na(+) currents and action potentials (APs) in dissociated, nociceptive dorsal root ganglion (DRG) cells from rats, two types of peripheral sensory neuron Na(+) channels (Nav1.7 and Nav1.8), and the motor neuron-specific Na(+) channel (Nav1.6) expressed in HEK293 cells. BAB (1-100µM) inhibited, in a concentration-dependent manner, the depolarization evoked repetitive firing in DRG cells, the three types of Na(+) current expressed in HEK293 cells, and the TTXr Na(+) current of the DRG neurons. BAB induced a use-dependent block that caused a shift of the inactivation curve in the hyperpolarizing direction. BAB enhanced the onset of slow inactivation of Nav1.7 and Nav1.8 currents but not of Nav1.6 currents. At clinically relevant concentrations (1-100µM), BAB is thus a more potent inhibitor of peripheral TTX-sensitive TTXs, Nav1.7 and TTX-resistant NaV1.8 Na(+) channels than of motor neuron axonal Nav1.6 Na(+) channels. BAB had similar effects on the TTXr Na(+) channels of rat DRG neurons and Nav1.8 channels expressed in HEK293 cells. The observed selectivity of BAB in treating cancer pain may be due to an enhanced and selective responsiveness of Na(+) channels in nociceptive neurons to this local anesthetic.

Involvement of the CD200 receptor complex in microglia activation in experimental glaucoma.

The interaction of the myeloid restricted molecule CD200R with its widely expressed ligand CD200 is involved in the down-regulation of microglia activation. In the present study, we examined the involvement of CD200R in microglia activation in experimental ocular hypertension to determine the role of microglia activation in retinal ganglion cell (RGC) death, the key pathological event in glaucoma. Experimental glaucoma was induced in adult Brown Norway rats by sclerosis of the episcleral veins with the injection of hypertonic saline. Immunohistochemical methods were used to determine the involvement of microglia using GFAP, CD45, OX42 and OX41 and the involvement of CD200 and CD200R in the optic nerve head. Our data demonstrate the increased presence of microglia within the optic nerve head during ocular hypertension, identified by positive staining with OX42 and OX41. The peak of microglia correlates with peak in RGC death at days 20-27 (T3) post OHT induction. In addition, CD200 and CD200R positive cells were increased in ocular hypertensive eyes. Increased expression of CD200 was detected in the early phase (days 1-7; T1) of OHT and decreased over time, whilst the expression of CD200R was detected in the middle phase (days 20-27; T3) of OHT, correlating with the increase in microglia markers. Changes in the expression of CD200R/CD200 occur early in experimental glaucoma and precede the peak in microglia infiltration and RGC death, suggesting that CD200R-positive microglia play an important role in the initiation of RGC death during OHT, indicating a potential area for therapeutic intervention in treating glaucoma.

Nonactin biosynthesis: unexpected patterns of label incorporation from 4,6-dioxoheptanoate show evidence of a degradation pathway for levulinate through propionate in Streptomyces griseus.

The polyketide nonactin, a polyketide possessing antitumor and antibacterial activity, is produced by an unusual biosynthesis pathway in Streptomyces griseus that uses both enantiomers of the nonactin precursor, nonactic acid. Despite many studies with labeled precursors, much of the biosynthesis pathway remains unconfirmed, particularly the identity of the last achiral intermediate in the pathway, which is believed to be 4,6-diketoheptanoyl-CoA. We set out to confirm the latter hypothesis with feeding studies employing [4,5-(13)C(2)]-, [5,6-(13)C(2)]-, and [6,7-(13)C(2)]-4,6-diketoheptanoate thioester derivatives. In each case the isotopic label was incorporated efficiently into nonactin; however, at positions inconsistent with the currently accepted biosynthesis pathway. To resolve the discrepancy, we conducted additional feeding studies with a [3,4-(13)C(2)]levulinate thioester derivative and again observed efficient label incorporation. The latter result was intriguing, as levulinate is not an obvious precursor to nonactin. Levulinate, however, is known to be efficiently degraded into propionate even though the pathway for the conversion is not known. On the basis of both our levulinate and diketoheptanoate isotope incorporation data we can now postulate a pathway from levulinate to propionate that can also account for the conversion of 4,6-diketoheptanoate into levulinate in S. griseus.

Drosophila CtBP regulates proliferation and differentiation of eye precursors and complexes with Eyeless, Dachshund, Dan, and Danr during eye and antennal development.

Specification factors regulate cell fate in part by interacting with transcriptional co-regulators like CtBP to regulate gene expression. Here, we demonstrate that CtBP forms a complex or complexes with the Drosophila melanogaster Pax6 homolog Eyeless (Ey), and with Distal antenna (Dan), Distal antenna related (Danr), and Dachshund to promote eye and antennal specification. Phenotypic analysis together with molecular data indicate that CtBP interacts with Ey to prevent overproliferation of eye precursors. In contrast, CtBP,dan,danr triple mutant adult eyes have significantly fewer ommatidia than CtBP single or dan,danr double mutants, suggesting that the CtBP/Dan/Danr complex functions to recruit ommatidia from the eye precursor pool. Furthermore, CtBP single and to a greater extent CtBP,dan,danr triple mutants affect the establishment and maintenance of the R8 precursor, which is the founding ommatidial cell. Thus, CtBP interacts with different eye specification factors to regulate gene expression appropriate for proliferative vs. differentiative stages of eye development.

Synergistic cooperation between two ClpB isoforms in aggregate reactivation.

Bacterial AAA+ ATPase ClpB cooperates with DnaK during reactivation of aggregated proteins. The ClpB-mediated disaggregation is linked to translocation of polypeptides through the channel in the oligomeric ClpB. Two isoforms of ClpB are produced in vivo: the full-length ClpB95 and ClpB80, which does not contain the substrate-interacting N-terminal domain. The biological role of the truncated isoform ClpB80 is unknown. We found that resolubilization of aggregated proteins in Escherichia coli after heat shock and reactivation of aggregated proteins in vitro and in vivo occurred at higher rates in the presence of ClpB95 with ClpB80 than with ClpB95 or ClpB80 alone. Combined amounts of ClpB95 and ClpB80 bound to aggregated substrates were similar to the amounts of either ClpB95 or ClpB80 bound to the substrates in the absence of another isoform. The ATP hydrolysis rate of ClpB95 with ClpB80, which is linked to the rate of substrate translocation, was not higher than the rates measured for the isolated ClpB95 or ClpB80. We postulate that a reaction step that takes place after substrate binding to ClpB and precedes substrate translocation is rate-limiting during aggregate reactivation, and its efficiency is enhanced in the presence of both ClpB isoforms. Moreover, we found that ClpB95 and ClpB80 form hetero-oligomers, which are similar in size to the homo-oligomers of ClpB95 or ClpB80. Thus, the mechanism of functional cooperation of the two isoforms of ClpB may be linked to their heteroassociation. Our results suggest that the functionality of other AAA+ ATPases may be also optimized by interaction and synergistic cooperation of their isoforms.

Esophagus tissue engineering: hybrid approach with esophageal epithelium and unidirectional smooth muscle tissue component generation in vitro.

PURPOSE: The aim of this study was to engineer the two main components of the esophagus in vitro: (a) esophageal epithelium and (b) smooth muscle tissue. Furthermore, (a) survivability of esophageal epithelial cells (EEC) on basement membrane matrix (BMM)-coated scaffolds and (b) oriented smooth muscle tissue formation on unidirectional BMM-coated collagen scaffolds was investigated. METHODS: Both EEC and smooth muscle cells (SMC) were sourced from Sprague-Dawley rats. The EEC were maintained in vitro and seeded onto BMM-coated 2-D collagen scaffolds. Similarly, smooth muscle cells were obtained using an explants technique and seeded on unidirectional 3-D BMM-coated collagen scaffolds. Cell-polymer constructs for EEC and SMC were maintained in vitro for 8 weeks. RESULTS: Protocols to obtain higher yield of EEC were established. EEC formed a layer of differentiated epithelium after 14 days. EEC survivability on polymers was observed up to 8 weeks. Unidirectional smooth muscle tissue strands were successfully engineered. CONCLUSION: Esophageal epithelium generation, survivability of EEC on BMM-coated scaffolds, and engineering of unidirectional smooth muscle strands were successful in vitro. The hybrid approach of assembling individual tissue components in vitro using BMM-coated scaffolds and later amalgamating them to form composite tissue holds promises in the tissue engineering of complex organ systems.

Management strategies in foreign-body aspiration.

OBJECTIVE: The aim of this study is to outline a management algorithm to ensure effective teamwork in decreasing morbidity and mortality in pediatric Foreign-Body Aspirations (FBA). Furthermore, the role of flexible bronchoscopy when compared to rigid bronchoscopy in FBA was evaluated. METHODS: Charts of patients with suspected FBA from October 1999 to September 2006 were reviewed and data with regards to the history, presenting symptoms, diagnostics and therapeutic tactics, was collected. RESULTS: A total of 77 children with suspicion of FBA were managed in the 7 year period. Bronchoscopies were performed in 63 patients and in 26 foreign-bodies (FB) were found and extracted. At referral, 53 patients did not present acute respiratory symptoms, but had a positive history of FBA, and in 13 FB were found. Despite negative chest x-rays in 55 patients, FB were found in 8. Rigid bronchoscopy was performed in 53 and flexible in 10 patients. In 3 out of 10 patients who had undergone flexible bronchoscopy a FB was identified, the extraction of which was performed using a rigid bronchoscope. CONCLUSION: Clinical and radiological findings in children with typical history of suspected FBA are not enough to confirm the presence of FB. Successful management with an extremely low rate of morbidity and no mortality was observed using the algorithm used at our center. Flexible bronchoscopy reduces the chances of airway tract injury; however a rigid bronchoscope is necessary for FB removal.

Bioavailability of [2-(14)C]quercetin-4'-glucoside in rats.

[2-(14)C]quercetin-4'-glucoside (4 mg/kg body weight) was fed by gavage to rats housed in metabolic cages, and over an ensuing 72 h period, radiolabeled products in body tissues, plasma, feces, and urine were monitored by high-performance liquid chromatography with online radioactivity and MS2 detection. One and 6 h after ingestion, while in the small intestine, the flavonol glucoside was converted to glucuronide and methylated and sulfated derivatives of quercetin, but only trace amounts of these metabolites were excreted in urine. On entering the cecum and the colon, the flavonol metabolites declined as they were converted to phenolic acids, principally 3-hydroxyphenylacetic acid and 3,4-dihydroxyphenylacetic acid, by the colonic microflora. Feces contained mainly 3-hydroxyphenylacetic acid. Urine collected 0-12 and 0-24 h after ingestion contained radiolabeled hippuric acid and 3-hydroxyphenylacetic acid. 14C-Hippuric acid declined markedly in the 24-48 and 48-72 h urine samples, and there was a concomitant increase in labeled benzoic acid. There was minimal accumulation of radioactivity in plasma, despite a 69% recovery of label in urine over the 72 h period, and likewise, very little radioactivity was detected in body tissues out with the gastrointestinal tract. This is reflected in the fact that 72 h after ingestion 96% of the ingested radioactivity was recovered in feces, urine, and the cage washes, which comprise a mixture of urine and feces. The study reveals that as it passes through the gastrointestinal tract, almost all of the of [2-(14)C]quercetin-4'-glucoside is converted to phenolic acids, compounds not monitored in previous flavonol bioavailability studies with model animal systems, some of which have used exceedingly high doses of the aglycone quercetin (500 mg/kg body weight), which is not a normal dietary component.