Construction of a highly selective and sensitive carbohydrate-detecting biosensor utilizing Computational Identification of Non-disruptive Conjugation sites (CINC) for flexible and streamlined biosensor design.
Fluorescently-labeled solute-binding proteins that alter their fluorescence output in response to ligand binding have been utilized as biosensors for a variety of applications. Coupling protein ligand binding to altered fluorescence output often requires trial and error-based testing of both multiple labeling positions and fluorophores to produce a functional biosensor with the desired properties. This approach is laborious and can lead to reduced ligand binding affinity or altered ligand specificity. Here we report the Computational Identification of Non-disruptive Conjugation sites (CINC) for streamlined identification of fluorophore conjugation sites. By exploiting the structural dynamics properties of proteins, CINC identifies positions where conjugation of a fluorophore results in a fluorescence change upon ligand binding without disrupting protein function. We show that a CINC-developed maltooligosaccharide (MOS)-detecting biosensor is capable of rapid (kon = 20 μM-1s-1), sensitive (sub-μM KD) and selective MOS detection. The MOS-detecting biosensor is modular with respect to the spectroscopic properties and demonstrates portability to detecting MOS released via α-amylase-catalyzed depolymerization of starch using both a stopped-flow and a microplate reader assay. Our MOS-detecting biosensor represents a first-in-class probe whose design was guided by changes in localized dynamics of individual amino acid positions, supporting expansion of the CINC pipeline as an indispensable tool for a wide range of protein engineering applications.


Examination of differential glycoprotein preferences of N-acetylglucosaminyltransferase-IV isozymes a and b.
The N-glycans attached to proteins contain various GlcNAc branches, the aberrant formation of which correlates with various diseases. N-Acetylglucosaminyltransferase-IVa (GnT-IVa or MGAT4A) and Gnt-IVb (or MGAT4B) are isoenzymes that catalyze the formation of the β1,4-GlcNAc branch in N-glycans. However, the functional differences between these isozymes remain unresolved. Here, using cellular and UDP-Glo enzyme assays, we discovered that GnT-IVa and GnT-IVb have distinct glycoprotein preferences both in cells and in vitro. Notably, we show that GnT-IVb acted efficiently on glycoproteins bearing an N-glycan premodified by GnT-IV. To further understand the mechanism of this reaction, we focused on the noncatalytic C-terminal lectin domain, which selectively recognizes the product glycans. Replacement of a nonconserved amino acid in the GnT-IVb lectin domain with the corresponding residue in GnT-IVa altered the glycoprotein preference of GnT-IVb to resemble that of GnT-IVa. Our findings demonstrate that the C-terminal lectin domain regulates differential substrate selectivity of GnT-IVa and GnT-IVb, highlighting a new mechanism by which N-glycan branches are formed on glycoproteins.


Dopamine neuron transplants: electrophysiological unit activity of intrastriatal nigral grafts in freely moving cats.
Fetal transplants of substantia nigra dopamine (DA)-containing neurons into the caudate nucleus in cats produced functional synaptic connections, as measured by behavioral analysis. Electrophysiological activity of single units from the intrastriatal nigral grafts revealed that these neurons displayed some of the characteristic electrophysiological parameters of DA neurons, i.e., long duration action potentials and tri-phasic waveform. However, these neurons discharged at a faster rate and never displayed the characteristic decremental burst pattern seen in intact, non-grafted animals. These data are the first electrophysiological recordings of grafted DA neurons in freely moving animals.


Challenges and strategies for ocular posterior diseases therapy via non-invasive advanced drug delivery.
Posterior segment diseases, such as age-related macular degeneration (AMD) and diabetic retinopathy (DR) are vital factor that seriously threatens human vision health and quality of life, the treatment of which poses a great challenge to ophthalmologists and ophthalmic scientists. In particular, ocular posterior drug delivery in a non-invasive manner is highly desired but still faces many difficulties such as rapid drug clearance, limited permeability and low drug accumulation at the target site. At present, many novel non-invasive topical ocular drug delivery systems are under development aiming to improve drug delivery efficiency and biocompatibility for better therapy of posterior segment oculopathy. The purpose of this review is to present the challenges in the noninvasive treatment of posterior segment diseases, and to propose strategies to tackle these bottlenecks. First of all, barriers to ocular administration were introduced based on ocular physiological structure and behavior, including analysis and discussion on the influence of ocular structures on noninvasive posterior segment delivery. Thereafter, various routes of posterior drug delivery, both invasive and noninvasive, were illustrated, along with the respective anatomical obstacles that need to be overcome. The widespread and risky application of invasive drug delivery, and the need to develop non-invasive local drug delivery with alternative to injectable therapy were described. Absorption routes through topical administration and strategies to enhance ocular posterior drug delivery were then discussed. As a follow-up, an up-to-date research advances in non-invasive delivery systems for the therapy of ocular fundus lesions were presented, including different nanocarriers, contact lenses, and several other carriers. In conclusion, it seems feasible and promising to treat posterior oculopathy via non-invasive local preparations or in combination with appropriate devices.


The Role of Care Coordination: A Qualitative Study of Care Coordinator Perceptions.
Care coordination is important for patients with complex needs; yet, little is known about the factors impacting implementation from the care coordinator perspective. To understand how care coordination implementation differs across clinics and what care coordinators perceive as barriers and facilitators of effective coordination. Nineteen care coordinators from primary care clinics in Minnesota participated in interviews about their perceptions of care coordination. A team of analysts coded interviews using inductive thematic analysis. Four major themes emerged: variety in care coordination implementation; importance of social needs; necessity for leader buy-in; and importance of communication skills. Described differences in care coordination implementation were often logistical, but the implications of these differences were foundational to care coordinator perceived effectiveness.


Cancer and ageing: a nexus at several levels.
Cancer is largely a disease of older people. With the relatively recent expansive growth within the geriatric population, a number of pressing biological and clinical questions that currently remain unanswered need to be addressed. These include what effect age itself has on the development or growth of cancer, and what are the benefits and risks of cancer prevention and treatment for the older person? New insights into the underlying biological processes of ageing provide a frame of reference for addressing these and other related questions.


Pulmonary shunting during alveolar hypoventilation in the dog.
Pulmonary shunting (Qs/Qt) was measured in 16 anesthetized and paralyzed healthy beagle puppies during a control period when the end-expired CO2 concentration was 5 per cent and during alveolar hypoventilation, when the PaCO2 increased to 100 plus or minus 13 torr (mean plus or minus SE) and the ;Ha vecame 7.04 plus or minus 0.04. A constant tidal volume was maintained throughout each study, and hypoventilation was achieved by slowing only the respiratory rate. During the control period Qs/Qt was 2.9 plus or minus 0.3 per cent; during hypoventilation Qs/Qt was 3.2 plus or minus 0.3 per cent (an insignificant difference). The puppies, therefore, responded to anesthesia and alveolar hypoventilation without development of increased pulmonary shunts.


A1 adenosine receptor of rat testis membranes. Purification and partial characterization.
Purification of an A1 adenosine receptor of rat testes was performed using a newly developed affinity chromatography system (Nakata, H. (1989) Mol. Pharmacol. 35, 780-786). The A1 adenosine receptor was solubilized with digitonin from rat testicular membranes and then purified more than 25,000-fold by sequential use of affinity chromatography on xanthine amine congener-immobilized agarose, hydroxylapatite chromatography, re-affinity chromatography on xanthine amine congener-agarose, and finally gel permeation chromatography on TSK-3000SW. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of the final preparation showed a single broad band of Mr 41,000 by autoradiography after radioiodination. This Mr 41,000 peptide was also specifically labeled with an A1 adenosine receptor affinity labeling reagent. A high affinity A1 adenosine receptor antagonist, 8-cyclopentyl-1,3-[3H]dipropylxanthine, bound saturably to the purified receptor with a KD of approximately 1.4 nM. The purified receptor also showed essentially the same specificity for adenosine agonists and antagonists as the unpurified receptor preparations, although the affinities of the purified adenosine receptor for agonists were significantly low compared to those of unpurified receptor preparations indicating that the purified A1 adenosine receptor exists as a low agonist-high antagonist affinity state. Deglycosylation of the purified testis adenosine A1 receptors with endoglycosidase F produced an increase in the mobility of the receptor protein to an apparent Mr 30,000 in sodium dodecyl sulfate-polyacrylamide gel electrophoresis, similar to that of deglycosylated A1 adenosine receptors of rat brain membranes. Peptide maps of the purified testis and brain A1 adenosine receptors using trypsin and V8 protease suggest that these receptors show some structural homologies.


Influence of non-invasive X-ray computed tomography (XRCT) on the microbial community structure and function in soil.
In this study the influence of X-ray computed tomography (XRCT) on the microbial community structure and function in soils has been investigated. Our results clearly indicate that XRCT of soil samples has a strong impact on microbial communities and changes structure and function significantly due to the death of selected microbial groups as a result of the treatment.


Effect of antipsychotics on mitochondrial bioenergetics of rat ovarian theca cells.
Antipsychotics (APs) are widely prescribed drugs, which are well known to cause reproductive adverse effects through mechanisms yet to be determined. The purpose of this study was to investigate the effect of antipsychotics on mitochondrial bioenergetics of rat ovarian theca cells as a possible mechanism of reproductive toxicity. Isolated rat theca interstitial cells (TICs) were treated with two typical (chlorpromazine [CPZ] and haloperidol [HAL]) and two atypical APs (risperidone [RIS] and clozapine [CLZ]). The effects of these APs on TICs bioenergetics (ATP content, mitochondrial complexes I and III activities, oxygen consumption rates (OCRs), mitochondrial membrane potential (MPP) and lactate production) and on steroidogenesis (androstenedione and progesterone synthesis) were investigated. All APs resulted in a concentration-dependent decrease in the ATP content of TICs. All APs at their estimated IC50s (6μM, 21μM, 35μM and 37μM for CPZ, HAL, CLZ and RIS respectively) significantly decreased TICs OCRs (p<0.0001), MPP (p<0.0001) and significantly (p=0.0003) inhibited mitochondrial complex I activity. Only typical APs inhibited complex III (p=0.005). Also, APs at IC50s increased TICs lactate production to varying degrees. All APs used at their IC50s significantly inhibited progesterone (p=0.0022) and androstenedione (p=0.0027) production. Only CPZ was found to inhibit these hormones at the low concentration (1μM). All four antipsychotics seem to inhibit mitochondrial bioenergetics and steroidogenesis in rat's ovarian theca cells. These findings support the hypothesis that APs-induced reproductive toxicity may be through mechanisms involving mitochondrial insult>. Further research is required to establish the link between APs-induced mitochondrial dysfunction and disordered steroidogenesis.


Influence of accommodation on the anterior and posterior cornea.
To evaluate whether significant change occurs in the topography of the anterior or posterior cornea or in corneal thickness with accommodation. Contact Lens and Visual Optics Laboratory, School of Optometry, Queensland University of Technology, Brisbane, Queensland, Australia. Corneal data were acquired for 12 young, healthy emmetropic subjects under 2 accommodation levels (relaxed and approximately 5.0 diopters) using a rotating Scheimpflug camera. Anterior and posterior corneal elevation and sagittal curvature data as well as pachymetry data were analyzed to evaluate changes associated with accommodation. Corneal elevation data were also analyzed using a recentering procedure designed to minimize the effect of ocular micromovements between measurements. Changes in corneal parameters resulting from accommodation were small and generally insignificant. The mean change in pachymetry was 0.49 microm +/- 2.62 (SD) across an 8.0 mm pachymetry map. No significant change was found in the best-fit corneal spherocylinder (anterior and posterior cornea) (P>.05). Small changes were found in the corneal cylinder axis, indicative of small cyclotorsional eye movements accompanying accommodation. The mean changes in corneal elevation were 0.09 +/- 0.41 microm (anterior cornea) and 0.17 +/- 0.65 microm (posterior cornea). Only 0.50% (anterior cornea) and 0.27% (posterior cornea) of corneal elevation data points exhibited a statistically significant change with accommodation. Recentering the corneal elevation data led to a reduction in the magnitude of change in corneal elevation with accommodation. Small cyclotorsional eye movements occurred with accommodation. When these eye movements were accounted for in data analysis, no consistent significant change in the topography of the cornea was found with accommodation across an 8.0 to 9.0 mm corneal diameter.


Role of hydrogenase 1 of Clostridium pasteurianum in the reduction of metronidazole.
Competition studies between the phosphoroclastic reaction and the metronidazole reduction reaction using dialyzed crude cell-free extracts of Clostridium pasteurianum which were essentially devoid of Hydrogenase 1 activity demonstrated that this enzyme plays an important role in the reduction of metronidazole. To determine further the exact function for Hydrogenase 1 in the reduction of the drug, this enzyme was highly purified from C. pasteurianum. Metronidazole reduction activity copurified with Hydrogenase 1 specific activity throughout the purification procedure. Drug reduction required the presence of an electron carrier and could not be accomplished by the enzyme alone. Ferredoxin, and also the low potential electron carrier dyes, methyl and benzyl viologen, and the flavin coenzymes, FAD and flavin mononucleotide (FMN), could couple the reduction of metronidazole. Hydrogenase 1 activity and its metronidazole reduction activity were inactivated irreversibly in the presence of oxygen. Metronidazole could be reduced only by an electron carrier-Hydrogenase 1 mechanism or directly by sodium dithionite.


Diatom carbon export enhanced by silicate upwelling in the northeast Atlantic.
Diatoms are unicellular or chain-forming phytoplankton that use silicon (Si) in cell wall construction. Their survival during periods of apparent nutrient exhaustion enhances carbon sequestration in frontal regions of the northern North Atlantic. These regions may therefore have a more important role in the 'biological pump' than they have previously been attributed, but how this is achieved is unknown. Diatom growth depends on silicate availability, in addition to nitrate and phosphate, but northern Atlantic waters are richer in nitrate than silicate. Following the spring stratification, diatoms are the first phytoplankton to bloom. Once silicate is exhausted, diatom blooms subside in a major export event. Here we show that, with nitrate still available for new production, the diatom bloom is prolonged where there is a periodic supply of new silicate: specifically, diatoms thrive by 'mining' deep-water silicate brought to the surface by an unstable ocean front. The mechanism we present here is not limited to silicate fertilization; similar mechanisms could support nitrate-, phosphate- or iron-limited frontal regions in oceans elsewhere.


Nanotherapeutics for regeneration of degenerated tissue infected by bacteria through the multiple delivery of bioactive ions and growth factor with antibacterial/angiogenic and osteogenic/odontogenic capacity.
Therapeutic options are quite limited in clinics for the successful repair of infected/degenerated tissues. Although the prevalent treatment is the complete removal of the whole infected tissue, this leads to a loss of tissue function and serious complications. Herein the dental pulp infection, as one of the most common dental problems, was selected as a clinically relevant case to regenerate using a multifunctional nanotherapeutic approach. For this, a mesoporous bioactive glass nano-delivery system incorporating silicate, calcium, and copper as well as loading epidermal growth factor (EGF) was designed to provide antibacterial/pro-angiogenic and osteo/odontogenic multiple therapeutic effects. Amine-functionalized Cu-doped bioactive glass nanospheres (Cu-BGn) were prepared to be 50-60 nm in size, mesoporous, positive-charged and bone-bioactive. The Cu-BGn could release bioactive ions (copper, calcium and silicate ions) with therapeutically-effective doses. The Cu-BGn treatment to human umbilical vein endothelial cells (HUVEC) led to significant enhancement of the migration, tubule formation and expression of angiogenic gene (e.g. vascular endothelial growth factor, VEGF). Furthermore, the EGF-loaded Cu-BGn (EGF@Cu-BGn) showed pro-angiogenic effects with antibacterial activity against E. faecalis, a pathogen commonly involved in the pulp infection. Of note, under the co-culture condition of HUVEC with E. faecalis, the secretion of VEGF was up-regulated. In addition, the osteo/odontogenic stimulation of the EGF@Cu-BGn was evidenced with human dental pulp stem cells. The local administration of the EGF@Cu-BGn in a rat molar tooth defect infected with E. faecalis revealed significant in vivo regenerative capacity, highlighting the nanotherapeutic uses of the multifunctional nanoparticles for regenerating infected/damaged hard tissues.


Conformation and activity of lysozyme on binding to two types of gold nanorods: a comparative study.
The unique morphology of anisotropic rod-shaped gold nanostructures has offered new prospects for biomedical and biosensing applications. This study investigates the interaction of two types of rod-shaped nanostructures, gold nanorods and gold nanorices with lysozyme as a model protein, comparing the probable structural, activity and kinetic stability alterations. Circular dichroism spectropolarimeter revealed that lysozyme retains high fraction of its native conformation in the presence of both nanostructures, with a slight increase in the helical and beta content. Upon the protein adsorption on both types of nanorods, kinetic studies showed maintenance of enzymatic activity, together with increase in the enzymatic affinity and kinetic stability at high temperature. Comparatively, gold nanorice induced better effect on the activity and stability of enzyme than that of gold nanorod. This study might open new insight into potential applications of gold nanorods as nanocarriers for genes and drugs; provided that the toxicological aspect of cationic surfactant-coated nanostructure is taken into consideration.


A change in the trend in dosulepin usage following the introduction of a prescribing indicator but not after two national safety warnings.
The tricyclic antidepressant dosulepin has been associated with an increased risk of toxicity in overdose compared with other antidepressants. In the UK, the MHRA and NICE have issued advice on the prescribing of dosulepin, and a National Prescribing Indicator (NPI) to monitor usage was introduced in Wales in 2011. The aim of this study was to assess whether trends in dosulepin usage in Wales and NE England changed following the two pieces of safety guidance and the introduction of the National Prescribing Indicator in Wales. Primary care dosulepin usage in the 12 months prior to and following MHRA safety advice (in 2007), NICE guideline CG90 (in 2009) and the introduction of the NPI (in 2011) was obtained. Usage was measured using defined daily doses (DDDs) per 1000 prescribing units (PUs). The trends in the 12 months prior to and following the introduction of prescribing advice and the NPI were compared using an autoregressive integrated moving average (ARIMA) model. In Wales, the trend in dosulepin usage did not change significantly prior to and following the MHRA advice: -0·18 and -0·43 DDDs/1000PUs per month, respectively (P = 0·07), or prior to and following NICE CG90: -0·30 and -0·49 DDDs/1000PUs per month, respectively (P = 0·35). In the 12 months prior to and following the introduction of the NPI, the trend was -0·45 and -0·98 DDDs/1000PUs per month, respectively (P = 0·001). In NE England, the trend did not alter significantly following the NICE advice or the introduction of the NPI in Wales. The trend in dosulepin usage in Wales altered significantly following the introduction of the NPI, but not after the other prescribing advice. This association, coupled with the absence of a significant change in NE England over the same period, provided some evidence of the effectiveness of the NPI in prompting a change in prescribing behaviour in Wales.


An extraluminal force transducer for recording contractile activity of the gastrointestinal smooth muscle in the conscious dogs: its construction and implantation.
An extraluminal strain-gauge force transducer has been developed for recording contractile activity of the gastrointestinal smooth muscle. The details of its construction and implantation were described. The transducer unit allows for continuous long-term recordings (more the 5 months) from conscious experimental animals. It could be constructed in any laboratory and easily implanted in animals if it is done in aseptic technique. Since it is sutured onto the outside of the gut, it does not interfere passage of the contents. The only disadvantages are that it only measures muscle contractions and it is hard to determine movements of intraluminal contents especially in the digestive state.


The secret world of shrimps: polarisation vision at its best.
Animal vision spans a great range of complexity, with systems evolving to detect variations in light intensity, distribution, colour, and polarisation. Polarisation vision systems studied to date detect one to four channels of linear polarisation, combining them in opponent pairs to provide intensity-independent operation. Circular polarisation vision has never been seen, and is widely believed to play no part in animal vision. Polarisation is fully measured via Stokes' parameters--obtained by combined linear and circular polarisation measurements. Optimal polarisation vision is the ability to see Stokes' parameters: here we show that the crustacean Gonodactylus smithii measures the exact components required. This vision provides optimal contrast-enhancement and precise determination of polarisation with no confusion states or neutral points--significant advantages. Linear and circular polarisation each give partial information about the polarisation of light--but the combination of the two, as we will show here, results in optimal polarisation vision. We suggest that linear and circular polarisation vision not be regarded as different modalities, since both are necessary for optimal polarisation vision; their combination renders polarisation vision independent of strongly linearly or circularly polarised features in the animal's environment.


Characterization of serum immunoglobulin M of grouper and cDNA cloning of its heavy chain.
Immunoglobulin M (IgM) from the whole serum of grouper fish, Epinephelus coioides was purified by affinity chromatography using protein A-Sepharose column. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) under reducing conditions revealed that the relative molecular masses (Mr) of the equimolar heavy and light chains of IgM were 78,000 and 27,000, respectively. The cDNAs encoding IgM heavy chain comprising its variable (VH) and constant (CH) regions have been cloned and sequenced from a grouper kidney cDNA library by antibody screening method. Five VH (130-142 amino acids) and four CH (450-454 amino acids) families were identified. The variable and constant regions were conserved with their putative domains. All the four constant region domains (CH1-CH2-CH3-CH4) contained each three conserved cysteine residues, which are considered to form the inter- and intra-chain disulfide bridges. There were three carbohydrate acceptor sites in the constant region. In general, the pattern of IgM gene organization seems to resemble that of other teleosts. Moreover, the CH genes in grouper IgM occur as multifamily as reported in Atlantic salmon and common carp.


Boolean modeling identifies Greatwall/MASTL as an important regulator in the AURKA network of neuroblastoma.
Aurora Kinase A (AURKA) is often overexpressed in neuroblastoma (NB) with poor outcome. The causes of AURKA overexpression in NB are unknown. Here, we describe a gene regulatory network consisting of core regulators of AURKA protein expression and activation during mitosis to identify potential causes. This network was transformed to a dynamic Boolean model. Simulated activation of the serine/threonine protein kinase Greatwall (GWL, encoded by MASTL) that attenuates the pivotal AURKA inhibitor PP2A, predicted stabilization of AURKA. Consistent with this notion, gene set enrichment analysis showed enrichment of mitotic spindle assembly genes and MYCN target genes in NB with high GWL/MASTL expression. In line with the prediction of GWL/MASTL enhancing AURKA, elevated expression of GWL/MASTL was associated with NB risk factors and poor survival of patients. These results establish Boolean network modeling of oncogenic pathways in NB as a useful means for guided discovery in this enigmatic cancer.


Cryptotanshinone and dihydrotanshinone I exhibit strong inhibition towards human liver microsome (HLM)-catalyzed propofol glucuronidation.
Danshen is one of the most famous herbs in the world, and more and more danshen-prescribed drugs interactions have been reported in recent years. Evaluation of inhibition potential of danshen's major ingredients towards UDP-glucuronosyltransferases (UGTs) will be helpful for understanding detailed mechanisms for danshen-drugs interaction. Therefore, the aim of the present study is to investigate the inhibitory situation of cryptotanshinone and dihydrotanshinone I towards UGT enzyme-catalyzed propofol glucuronidation. In vitro the human liver microsome (HLM) incubation system was used, and the results showed that cryptotanshinone and dihydrotanshinone I exhibited dose-dependent inhibition towards HLM-catalyzed propofol glucuronidation. Dixon plot and Lineweaver-Burk plot showed that the inhibition type was best fit to competitive inhibition type for both cryptotanshinone and dihydrotanshinone I. The second plot using the slopes from the Lineweaver-Burk plot versus the concentrations of cryptotanshinone or dihydrotanshinone I was employed to calculate the inhibition parameters (Ki) to be 0.4 and 1.7μM, respectively. Using the reported maximum plasma concentration (Cmax), the altered in vivo exposure of propofol increased by 10% and 8.2% for the co-administration of dihydrotanshinone I and cryptotanshinone, respectively. All these results indicated the possible danshen-propofol interaction due to the inhibition of dihydrotanshinone I and cryptotanshinone towards the glucuronidation reaction of propofol.


A fully digital protocol to provide a fixed interim complete denture for immediate loading for a completely edentulous patient: A dental technique.
Complete arch immediate-loading implant-supported prostheses can represent a major challenge for the patient and the dental team. Obtaining stable references and an accurate occlusal record after implant placement to provide an interim prosthesis is a difficult task and can deviate from the initial treatment plan. The described technique presents a fully digital protocol to provide an immediate complete arch implant-supported fixed interim prosthesis for edentulous patients by making postoperative cone beam computed tomography and intraoral digital scans that correlate with the previous plan.


Magnetic dispersive solid phase extraction based on carbonized cellulose-ferromagnetic nanocomposite for screening phthalate esters in aqueous samples.
In this work, a sorbent of the carbonized cellulose-ferromagnetic nanocomposite has been proposed for the magnetic dispersive solid phase extraction of some plasticizers in aqueous samples. Carbonized cellulose nanoparticles were prepared by treatment of cellulose filter paper with concentrated sulfuric acid and then loaded on Fe3O4 nanoparticles using coprecipitation. This sorbent is compatible with aqueous samples and can be considered as a viable sorbent for extraction of plasticizers from aqueous samples. In this study, magnetic dispersive solid phase extraction is followed by a dispersive liquid-liquid microextraction method. This combination makes the proposed approach as an efficient clean-up method with high enrichment factors for the selected analytes. The enriched analytes are monitored by gas chromatography equipped with a flame ionization detector. Parameters affecting the method efficiency were investigated in details. Under the optimized extraction conditions, limits of detection could reach up to of 0.15-0.50 µg L-1. The satisfactory enrichment factors of 286-403 were obtained, and the extraction recoveries were found to be in the range of 57-80%. Relative standard deviations were in the range of 3-7% for intra-day and inter-day precisions for six replicate extractions at 25 µg L-1 of each plasticizer. Calibration curves were linear in wide ranges with coefficients of determination ≥ 0.995. Eventually, efficiency of the prepared sorbent was confirmed by the extraction of some plasticizers from real samples including fruit juices, mineral water, injection solution, cola, and yoghourt drink packed in plastic containers.


Quality of websites of obstetrics and gynecology departments: a cross-sectional study.
The internet has become an easily accessible and widely used source of healthcare information. There are, however, no standardized or commonly accepted criteria for the quality of Obstetrics and Gynecology websites. In this study, we aimed to evaluate the quality of websites of Obstetrics and Gynecology departments in German-speaking countries and to compare websites nationally and internationally. We scored 672 websites from Germany (n = 566), Austria (n = 57), and Switzerland (n = 49) using the objective criteria: Google search rank (2 items), technical aspects (11 items), navigation (8 items), and content (6 items) for a 26 point score. Scores were compared nationally and internationally. Multivariable regression models assessed good quality scores (≥50% of maximum) as the dependent variables and country, academic affiliation, being member of a healthcare consortium, confessional affiliation, and content management system (CMS) use as independent variables. The mean score of websites was 13.8 ± 3.3. 4.2% were rated as good (≥75% of maximum), 61.8% as fair (≥50% of maximum). German (14.0 ± 3.2) and Swiss (13.8 ± 4.0) websites scored significantly higher compared to Austrian websites (11.6 ± 2.5) (P < 0.001 and P = 0.005, respectively). Within Germany, academic had higher scores than non-academic departments (14.9 ± 3.2 vs. 13.7 ± 3.1, P < 0.001). Single institutions had higher scores compared to healthcare consortium institutions (14.1 ± 3.2 vs. 13.2 ± 2.6, P = 0.003). Departments in Northern and Southern states had higher scores compared to Eastern states (14.4 ± 3.2 and 14.2 ± 3.2 vs. 13.0 ± 3.0, P < 0.001). In multivariate regression models, all subscores (all: P < 0.001) independently predicted a website's reaching a good quality score, with navigation subscore as strongest predictor. Affiliations were predictors for some good individual subscores, but not for others. High content subscore was associated with good Google search rank, technical aspects, and navigation subscores. The quality of websites of Obstetrics and Gynecology departments varies widely. We found marked differences depending on country, affiliation, and region.


The effect of maternal smoking on birth weight and the subsequent health of the child.
The effect of maternal smoking during pregnancy on birth weight was studied in 12,068 births, the mothers in 1819 of which were regarded as smokers. The children of the smokers were compared with those of controls of similar age, parity, marital status and place of residence. Maternal smoking reduced birth weight in a dose-related manner. Birth weight was least affected among young, primiparous mothers who smoked only slightly, a difference which was, however, entirely caused by those mothers who stopped smoking for the last 3 mth of pregnancy, and this subgroup showed similar figures for postneonatal mortality and morbidity up to the age of 5 to those of their controls. Postneonatal mortality was higher in the total group of the smokers than among their controls, the primiparous or young mothers not differing in this respect from the others. Morbidity up to the age of 5 was significantly higher (p less than 0.001) among the children of the smokers, the children of the primiparas and young mothers being affected in a similar way to the others. The low birth weight infants of the smokers had a higher mean birth weight and lower perinatal mortality than the low birth weight infants of the controls, but morbidity up to age of 5 and postneonatal mortality were higher among the smokers in respect of both low birth weight infants and others.


Early and Short-term Triiodothyronine Supplementation Prevents Adverse Postischemic Cardiac Remodeling: Role of Transforming Growth Factor-β1 and Antifibrotic miRNA Signaling.
Activation of transforming growth factor (TGF)-β1 signaling in the ischemia/reperfusion (I/R) injured myocardium leads to dysregulation of miR-29-30-133, favoring the profibrotic process that leads to adverse cardiac remodeling (CR). We have previously shown that timely correction of the postischemic low-T3 syndrome (Low-T3S) exerts antifibrotic effects, but the underlying molecular players are still unknown. Here we hypothesize that a prompt, short-term infusion of T3 in a rat model of post I/R Low-T3S could hamper the early activation of the TGFβ1-dependent profibrotic cascade to confer long-lasting cardioprotection against adverse CR. Twenty-four hours after I/R, rats that developed the Low-T3S were randomly assigned to receive a 48-h infusion of 6 μg/kg/d T3 (I/R-L+T3) or saline (I/R-L) and sacrificed at 3 or 14 d post-I/R. Three days post-I/R, Low-T3S correction favored functional cardiac recovery. This effect was paralleled by a drop in TGFβ1 and increased miR-133a, miR-30c and miR-29c in the infarcted myocardium. Consistently, connective transforming growth factor (CTGF) and matrix metalloproteinase-2(MMP-2), validated targets of the above miRNAs, were significantly reduced. Fourteen days post-I/R, the I/R-L+T3 rats presented a significant reduction of scar size with a better preservation of cardiac performance and LV chamber geometry. At this time, TGFβ1 and miR-29c levels were in the normal range in both groups, whereas miR-30c-133a, MMP-2 and CTGF remained significantly altered in the I/R group. In conclusion, the antifibrotic effect exerted by T3 in the early phase of postischemic wound healing triggers a persistent cardioprotective response that hampers the progression of heart dysfunction and adverse CR.


Determination of free and total phenylacetic and p- and m-hydroxyphenylacetic acids in human urine by liquid chromatography with fluorimetric detection.
A highly sensitive and rapid liquid chromatographic method for the determination of free and total phenylacetic and p- and m-hydroxyphenylacetic acids in human urine is described. After extraction of urine with diethyl ether, these acids and phenylpropionic acid (internal standard) are converted into the corresponding fluorescent derivatives by treatment with 3-bromomethyl-6,7-dimethoxy-1-methyl-2(1H)-quinoxalinone in the presence of potassium hydrogen carbonate and 18-crown-6 in acetonitrile. The derivatives are separated on a reversed-phase column (Radial-Pak cartridge C18) with aqueous 65% (v/v) methanol and detected fluorimetrically. The detection limits for phenylacetic and p- and m-hydroxyphenylacetic acids are 5, 30 and 100 fmol, respectively, at a signal-to-noise ratio of 5 in a 20-microliter injection volume. This sensitivity permits precise determination of the free and total acids in 20 microliter of normal human urine.


Loss of delta6-desaturase activity leads to impaired docosahexaenoic acid synthesis in Y-79 retinoblastoma cells.
We have studied the synthesis of docosahexaenoic acid (22:6 n-3) from linolenic acid (18:3 n-3) and eicosapentaenoic acid (20:5 n-3) in Y-79 human retinoblastoma cells and in a derived cell line, Y-79a, which has lost delta6-desaturase activity. Whereas Y-79 cells efficiently converted both radiolabeled 18:3 n-3 and 20:5 n-3 to 22:6 n-3, no synthesis of this fatty acid was observed in Y-79a cells from either 18:3 n-3 or 20:5 n-3. Y-79a cells elongated 18:3 n-3 to 20:3 n-3, which was further converted to 5,11,14,17-20:4 through a delta5-desaturation, elongated 20:5 n-3 to 22:5 n-3 and 24:5 n-3, and retained the ability for retroconversion. These results underscore the key role of delta6-desaturase in the synthesis of 22:6 n-3 and stress the importance of an adequate supply of this fatty acid in situations of reduced delta6-desaturase activity, such as in perinatal period, to prevent a 22:6 n-3 deficiency status.


Cerebellar hemispheric agenesis.
A case report is reported of bilateral cerebellar hemispheric agenesis which was associated with secondary degeneration of cerebellofugal and cerebellopetal tracts. Somatotopic correlations between the cerebellar and the medullary olive lesions were obvious: preserved dorsal accessory olives pattern correlated with the spared vermis and normal medial accessory olives with the spared flocculi. Cerebellopetal degeneration was more difficult to analyse. The relation of cerebellar agenesis with basal ganglia abnormalities and microcephaly is discussed.


Three way ANOVA for emulsion of carotenoids extracted in flaxseed oil from carrot bio-waste.
The juice expelled from carrot, a globally produced root vegetable, leavesbehind carrot pomace (a bio- and horticultural waste) which is potentially rich source of micro-nutrients and carotenoids.However, it is discarded as waste or used as animal feed. It holds potential to be channelized to food chain by a couple of technological interventions. In this regard, present work was aimed at preparing stable emulsion based delivery system for 'green' carotenoids extracted from carrot-pomace in flaxseed oil (a green solvent), and at maximizing the amount of core material so that the resultant emulsion can potentially be used as a source of both carotenoids and omega-3 fatty acid of flaxseed oil origin. The study used natural emulsifier. Preparation of oil-in-water emulsion was optimized using 33 factorial experiment by varying levels of extract containing carotenoid (30-40%), whey protein concentrates (WPC-80) and lactose. The optimized emulsion (CREm) was selected on the basis of particle size, zeta potential, color values (L*, a*, b*) and viscosity statistically analyzed via three-way ANOVA using Proc GLM of SAS 9.3 (described in detail in this paper); the respective values of these parameters being 120.03 ± 8.20 nm, -16.57 ± 0.49 mV, 75.11 ± 0.04, 9.66 ± 0.32, 50.29 ± 0.62, and 0.124 ± 0.0115 Pa.s for CREm. CREm contained 35% flaxseed oil, 10% WPC-80 and 5% lactose and showed good centrifugal and gravitational stability (15 days). It was analyzed for total carotenoid content, antioxidant activities (ABTS (2,2-azinobis-(3-ethylbenzthiazoline-6sulfonic acid), DPPH (2,2-Diphenyl-1-picrylhydrazyl) and FRAP (Ferric reducing antioxidant power assay)) and microstructure.


Going to the roots of reduced magnesium dietary intake: A tradeoff between climate changes and sources.
Magnesium is essential in plants where it is associated with chlorophyll pigments and serves as a cofactor of enzymes implicated in photosynthesis and metabolism. It is an essential nutrient for animals, involved in hundreds metabolic reaction and crucial for the biological activity of ATP. Not surprisingly, magnesium deficiency is detrimental for the health of plants and animals. In humans, subclinical magnesium deficiency is common and generates chronic inflammation, which is the common denominator of a wide range of mental and physical health problems from metabolic diseases to cognitive impairment, from osteopenia and sarcopenia to depression. It is ascertained that magnesium content in fruits and vegetables dropped in the last fifty years, and about 80% of this metal is lost during food processing. As a consequence, a large percentage of people all over the world does not meet the minimum daily magnesium requirement. In this scoping review, we summarize how agronomic and environmental factors, including global warming, affect magnesium content and availability in the soil and, consequently, in the food chain, with the aim of attracting the interest of botanists, agronomists, animal and human nutritionists and physicians to work on a strategy that grants adequate magnesium intake for everybody.


Structural mechanism of TRPM7 channel regulation by intracellular magnesium.
Zn2+, Mg2+ and Ca2+ are essential divalent cations implicated in many metabolic processes and signalling pathways. An emerging new paradigm is that the organismal balance of these cations predominantly depends on a common gatekeeper, the channel-kinase TRPM7. Despite extensive electrophysiological studies and recent cryo-EM analysis, an open question is how the channel activity of TRPM7 is activated. Here, we performed site-directed mutagenesis of mouse TRPM7 in conjunction with patch-clamp assessment of whole-cell and single-channel activity and molecular dynamics (MD) simulations to show that the side chains of conserved N1097 form an inter-subunit Mg2+ regulatory site located in the lower channel gate of TRPM7. Our results suggest that intracellular Mg2+ binds to this site and stabilizes the TRPM7 channel in the closed state, whereas the removal of Mg2+ favours the opening of TRPM7. Hence, our study identifies the structural underpinnings through which the TRPM7 channel is controlled by cytosolic Mg2+, representing a new structure-function relationship not yet explored among TRPM channels.


Iron metabolism disorder regulated by BMP signaling in hypoxic pulmonary hypertension.
Unexplained iron deficiency is associated with poorer survival in patients with pulmonary hypertension (PH). Bone morphogenetic protein (BMP) signaling and BMP protein type II receptor (BMPR2) expression are important in the pathogenesis of PH. BMP6 in hepatocytes is a central transcriptional regulator of the iron hormone hepcidin that controls systemic iron balance. This study aimed to investigate the effects of BMP signaling on iron metabolism and its implication in hypoxia-induced PH. PH was induced in Sprague-Dawley Rats under hypoxia for 4 weeks. Compared with the control group, right ventricular systolic pressure and right ventricle hypertrophy index were both markedly increased, and serum iron level was significantly decreased with iron metabolic disorder in the hypoxia group. In cultured human pulmonary artery endothelial cells (HPAECs), hypoxia increased oxidative stress and apoptosis, which were reversed by supplementation with Fe agent. Meanwhile, iron chelator deferoxamine triggered oxidative stress and apoptosis in HPAECs, and treatment with antioxidant alleviated iron-deficiency-induced apoptosis by reducing reactive oxygen species production. Expression of hepcidin, BMP6 and hypoxia-inducible factor (HIF)-1α were significantly upregulated, while expression of BMPR2 was downregulated in hepatocytes in the hypoxia group, both in vivo and in vitro. Expression of hepcidin and HIF-1α were significantly increased by BMP6, while pretreatment with siRNA-BMPR2 augmented the enhanced expression of hepcidin and HIF-1α induced by BMP6. Iron deficiency promoted oxidative stress and apoptosis in HPAECs in hypoxia-induced PH, and enhanced expression of hepcidin regulated by BMP6/BMPR2 signaling may contribute to iron metabolic disorder.


Why the VA matters: Resident education, research and patient care.
The Veterans' Health Administration (VHA) has had a long and storied relationship with academic medicine and particularly academic surgery throughout its history. Since the initial inception of the Veterans' Health Administration in 1946 there have been relationships between medical schools and the VHA to provide care to our veterans as well as provide a fertile training environment for the residents that will enhance their overall training experience and prepare them to the provide health care for the nation as a whole. At this point in our history that relationship is in jeopardy. The problems facing the Veterans' Health Administration are well known and seemingly are proving to be an insurmountable obstacle for continuation of this relationship. It is my intention in this lecture honoring Dr. Organ, who was committed to surgical education as well as the surgical care of our veterans to demonstrate why the Veterans' Health Administration is more important than ever to the well-being of our residents and graduate medical education as a whole.


The nasopharyngeal bacterial flora in the sudden infant death syndrome.
The nasopharyngeal bacterial flora in babies who had died of the sudden infant death syndrome (SIDS) (n = 46) and in healthy infants aged 2 weeks to 6 months (n = 46) is described. Of those who had died, 41.3% carried Staphylococcus aureus (95% confidence limits: 27.3-55.3%) compared with 28.3% of healthy infants (95% confidence limits: 15.3-41.3%). The isolation rate of streptococci was 78.3% in cases (95% confidence limits: 66.4-90.2%) and 32.6% in healthy infants (95% confidence limits: 19.1-46.1%) (significant difference P less than 0.0001). Enterobacteria were isolated from 45.6% of cases (95% confidence limits: 31.2-60%) but only 2.2% of healthy infants (95% confidence limits 0-6.4%) (significant difference, P less than 0.0001). These results indicate a disordered nasopharyngeal flora in SIDS. They also provide baseline data for investigating the hypothesis that common bacterial toxins are involved in the pathogenesis of SIDS.


How informed is consent in a modern ENT department.
The Department of Health issued a model consent form for use throughout the NHS from the 1st April 2002. Details of benefits and serious or frequent risks should be included on the form. We undertook a study to identify what proportion of complications from otolaryngology procedures were being recorded. Local morbidity and mortality records and case notes were examined from March to November 2004. Complications were identified and recorded. The consent forms for these operations were examined to identify if these complications had been recorded on the form prior to surgery. Complications were classified as "serious", causing significant morbidity or increasing length of hospital stay or "frequent", occurring in 1% or more of cases. A total of 2,978 operations were performed between March and November 2004. Seventy complications were identified in 60 patients (2% of operated patients). Twenty-three complications were not recorded on the consent forms in 20 (33%) patients. A total of 67% of all the complications were documented on the form as potential problems resulting from planned operations. Seven (74%) of complications that occurred but were not recorded on the consent forms were judged as "serious" or "frequent". A significant proportion of serious or frequent complications are not being documented on the national consent forms before otolaryngological procedures and may not have been discussed. This may reflect a lack of openness during the consent process. In the current medical climate, this has serious ethical and medico-legal ramifications. It may also reflect a problem with the form and a need for a re-think of its design.


Tryptophan-containing dipeptide derivatives as potent PPARgamma antagonists: design, synthesis, biological evaluation, and molecular modeling.
The discovery of peroxisome proliferator-activated receptor gamma (PPARgamma) antagonists (also termed "selective PPARgamma modulators, SPPARgammaM") is now of a great interest in the treatment of diabetes and obesity. The structure of compound 1a (G3335, Fig. 1), a novel class of PPARgamma antagonist, is entirely different from that of other reported PPARgamma antagonists. A series of 35 novel analogues (1b-l, 9a-d, 13a-t) were designed, synthesized and evaluated against the agonistic effects exerted by rosiglitazone. These results indicated that most functional groups of 1a were conserved, and six new compounds (1b, 1c, and 9a-d) exhibited strong PPARgamma antagonistic activities (IC(50) values of 5.2-25.8 microM) against 10 microM rosiglitazone in the promotion of the PPARgamma-LBD-CBP (ligand-binding domain and cAMP-response-element binding protein) interaction as investigated by yeast two-hybrid technology based assay. Molecular modeling studies for compounds 1a-d, 1h, 9c-d, and 13a were also presented.


Local regulation of blood flow in the feline jejunum. A possible role for endoluminally released substance P.
Serotonin and substance P of gastrointestinal origin have been measured by radioimmunoassay in the bowel lumen under basal and stimulated conditions. To investigate the possibility that local blood flow may be influenced by these endoluminal hormones, 26 cats were studied with exogenous serotonin and substance P infused endoluminally into isolated proximal jejunal segments in vivo. Regional blood flow was measured by using the radioactive microsphere technique before, during, and after the endoluminal instillation of two doses of substance P (3.9 and 30 ng/min) or serotonin (0.9 and 21 micrograms/min). Neither dose of substance P changed systemic blood pressure. Substance P at the low dose caused an increase in blood flow to the experimental jejunal mucosa (from 53 +/- 10 ml/min per 100 g to 102 +/- 20 ml/min per 100 g, P less than 0.01). The higher dose of endoluminal substance P similarly increased blood flow to the experimental jejunal mucosal fraction, and also increased blood flow to the experimental jejunal muscularis fraction (from 17 +/- 3 ml/min per 100 g to 23 +/- 3 ml/min per 100 g, P less than 0.02). Serotonin increased blood flow to the experimental jejunal muscularis only at the high dose (17 +/- 4 ml/min per 100 g to 25 +/- 4 ml/min per 100 g tissue, P less than 0.01). These results provide evidence for a dose-related local effect of endoluminal substance P on gastrointestinal blood flow.


Controlled release from thermo-sensitive PNVCL-co-MAA electrospun nanofibers: The effects of hydrophilicity/hydrophobicity of a drug.
The thermo-sensitive copolymer poly(N-vinylcaprolactam-co-methacrylic acid) (PNVCL-co-MAA) was synthesized by free radical polymerization and the resulting nanofibers were fabricated using an electrospinning process. The molecular weight of the copolymer was adjusted by varying the content of methacrylic acid (MAA) while keeping that of N-vinylcaprolactam (NVCL) constant. Hydrophilic captopril and hydrophobic ketoprofen were used as model drugs, and PNVCL-co-MAA nanofibers were used as the drug carrier to investigate the effects of drug on its release properties from nanofibers at different temperatures. The results showed that slow release over several hours was observed at 40°C (above the lower critical solution temperature (LCST) of PNVCL-co-MAA), while the drugs exhibited a burst release of several seconds at 20°C (below the LCST). Drug release slowed with increasing content of the hydrophobic monomer NVCL. The hydrophilic captopril was released at a higher rate than the hydrophobic ketoprofen. The drug release characteristics were dependent on the temperature, the portion of hydrophilic groups and hydrophobic groups in the copolymer and hydrophilicity/hydrophobicity of drug. Study on the mechanism of release showed that Korsmeyer-Peppas model as a major drug release mechanism. Given these results, the PNVCL-co-MAA copolymers are proposed to have useful applications in intellectual drug delivery systems.


Are endoscopic changes predictive of histologic esophagitis in children?
Controversy exists regarding the value of gross mucosal changes of the esophagus at the time of endoscopy in predicting histologic esophagitis. Accordingly, we reviewed the records of 279 pediatric patients who underwent both an upper endoscopy and simultaneous grasp esophageal biopsy. Of those patients with no gross mucosal abnormalities, 30% had evidence of esophagitis on biopsy. Similarly, there was a poor association between the presence of endoscopic findings and histologic esophagitis. We conclude that gross esophageal mucosal changes are poor predictors of histologic esophagitis and that endoscopic evaluation alone is inadequate for children and adolescents in whom esophagitis is suspected. Esophageal biopsies should be obtained in all such patients, even when no gross mucosal abnormalities are found.


Primary infiltrating ductal carcinoma of the axillary breast with metastasis to the contralateral chest wall.
Primary infiltrating ductal carcinoma of the axillary breast is rare and has a high frequency of lymph node (LN) involvement. We report a woman with primary infiltrating ductal carcinoma arising from the right axillary breast with metastasis to the contralateral chest wall. Excisional biopsy of the left chest wall nodule and the right axillary mass was carried out and both showed invasive ductal carcinomas histologically. The lesion of the right axillary mass arose from the breast tissue, rather than the LN. Further surgery proved the right axillary LN metastasis. After further review, a primary infiltrating ductal carcinoma of the right axillary breast with metastasis to axillary LNs and contralateral chest wall was diagnosed. The patient also received chemotherapy and radiation and there was no evidence of tumor recurrence after treatment. The present report demonstrated a rare case with uncommon manifestation. Lesions of uncertain origin around the periphery of the breast should be suspected for breast carcinoma.


Decreased activity and increased aggregation of brain calcineurin during aging.
Age-related decline in strength of synaptic transmission and memory formation has been attributed to age-associated increases in the activity of calcineurin (Cn) in hippocampus neurons. In the present study, we examined how brain Cn activity, Cn subunit levels, and Cn protein oxidation were changing during the aging process. Cn activity decreased with advancing age in three brain subcellular fractions, homogenate, cytosol, and synaptic membranes, obtained from F344/BNF1 rats of 5-6, 22-24, and 34-36 months of age. Cn activity also decreased during aging in homogenate, cytosol, and a nerve ending-enriched fraction from the hippocampus. Cn protein levels in homogenate and cytosol, as determined by the immune reactivity of its subunits A and B, were not altered during aging. But, in synaptic membranes, there was an age-related decrease in CnA levels, but not of CnB. Another important observation was that of an oxidative modification of CnA, not CnB, with increasing age. Such modification caused the formation of large aggregates of CnA. Aggregate formation was due to SH-group oxidation as the monomeric form of CnA was recovered upon disulfide reduction of the proteins with dithiothreitol. The age-related formation of aggregates of the catalytic subunit of Cn was suggestive of a correlation between aggregate formation and diminished enzyme activity. The loss of Cn activity may alter signal transduction at synapses during the aging process.


[Efficiency of Bakri intra uterine tamponade balloon use in postpartum hemorrhage: Before and after study].
To assess the impact of the introduction of intrauterine balloon tamponade on second-line management practices for post-partum hemorrhage (PPH). This retrospective study was conducted in a tertiary care centre where arterial embolisation was not available. Two periods were compared, before (2008-2010) and after (2015-2017) the introduction of an intrauterine tamponade balloon protocol using the Bakri balloon. All consecutive patients with PPH refractory to sulprostone in the two periods were included. Outcomes studied were the incidence of invasive procedures (conservative surgery, embolisation, hysterectomy) as well as transfer to centres with arterial embolisation facilities. A total of 109 patients had refractory PPH, 44 in the first period and 65 in the second period. The proportion of all PPH cases which required sulprostone did not differ between the periods (24.2 and 29.3%, respectively, P=0.25). There were significant decreases between the first and the second period in the proportion requiring a transfer to an embolisation centre (79.6% during period 1, vs. 6.2% respectively, P<0.001), embolisation (20.6% vs. 0%, P<0.001), uterine compression sutures (13.6% vs. 3.1% P=0.04) and surgical management (22.7% vs. 7.7%, P=0.025). The decrease in the incidence of hysterectomy did not reach statistical significance (11,4 vs. 1,5%, P=0.095). Among 29 cases of PPH following cesarean sections (14 in the first period and 15 in the second period), we observed a significant decrease in transfer rates (66.7% vs. 0%, P<0.01), as well as decreases in embolisation (26.7% vs. 0%, P=0.10) and invasive procedures (60% vs. 28.7%, P=0.089), which did not reach statistical significance. The introduction of intrauterine tamponade was associated with a decrease in the need for invasive procedures. In a centre without access to on-site arterial embolisation, transfer rates were reduced improving patient comfort and reducing costs.


How the R1 replication control system responds to copy number deviations.
The copy number of R1 and of a repA-lacZ gene fusion was increased above normal by coupling to a plasmid which is present at a fivefold higher copy number at 30 degrees C. This carrier plasmid is deficient in replication at 42 degrees C, and it was thus possible after a temperature shift to analyze the response to the increased plasmid concentration of the R1 replication control system. Both the frequency of replication per plasmid molecule and the rate of repA expression per gene copy were reduced under these conditions, and the data strongly suggest that there is an inverse proportionality between the specific rate of plasmid replication viz repA expression and the copy number/gene dosage of the plasmid.


Use of allogeneic bone graft in maxillary reconstruction for installation of dental implants.
The aim of this study is to evaluate the efficacy of the application of allogenous bone at the maxillomandibular reconstructions for future rehabilitation with dental implants. The patients were submitted to reconstruction of maxilla, using allogeneic bone grafts, in 3 different techniques: onlay grafts for lateral ridge augmentation, onlay and particulate bone for sinus lift grafting, and particulate alone for sinus lift grafts. Clinical and radiographic control was done at the postoperative phase for at least 8 months, until the patient could be submitted to the installation of dental implants. The results showed success in the majority of the cases, and dental implants could be installed. This can be considered an excellent alternative when compared with the use of autogenous grafts; because handling is easier, there is a great amount of material available and a possibility of using local anesthesia, and consequently there is a reduction of patient morbidity.


Malnutrition in elective surgery: How traditional markers might be failing surgeons and patients.
Serologic and anthropometric measures are commonly used as surrogate markers of nutritional status in clinical practice. In 2012, leading dietetic organizations published a standard definition of malnutrition based on clinical characteristics. We hypothesize that surrogate markers underrecognize clinical malnutrition and do not accurately identify patients at risk for adverse outcomes. A single-institution cohort study of elective surgical inpatients from August 2015 to November 2017. Nutritional assessment was completed by trained registered dietitians using leading dietetic guidelines. Multivariable logistic regression was used to determine the association between malnutrition and perioperative outcomes. Among 953 elective surgical admissions, 456 underwent full clinical nutritional assessment. Of these, 202 (44.3%) met malnutrition criteria. In addition, 20.3% of patients with clinical malnutrition were underweight (<18.5 kg/m2) and 38.1% had a serum albumin <3.0 g/dL. Compared with nonmalnourished patients, those with clinical malnutrition had higher rates of any complication (46.5% vs 37.8%, P = .06), overall infectious complications (26.2% vs 14.6%, P = .002), surgical site infections (9.4% vs 3.9%, P = .02), and mortality (8.9% vs 1.9%, P = .001). Clinical malnutrition was associated with death (odds ratio 3.99; 95% confidence interval, 1.27-12.54), overall infectious complication (odds ratio 1.77; 95% confidence interval, 1.07-2.94), and surgical site infections (odds ratio 2.65; 95% confidence interval, 1.12-6.22). In this cohort of elective surgical patients, traditional markers failed to identify malnutrition in a substantial portion of patients who met clinical malnutrition criteria. Clinical malnutrition assessment is effective in identifying patients who may be at risk for suboptimal outcomes. Surgeons should implement clinical nutritional assessment and factor that information into their preoperative evaluation and management of elective surgical patients.


A biocompatible bacterial cellulose/tannic acid composite with antibacterial and anti-biofilm activities for biomedical applications.
Biofilm-associated infections are in a high rate of recurrence and biofilms show formidable resistance to current antibiotics, making them a growing challenge in biomedical field. In this study, a biocompatible composite was developed by incorporating tannic acid (TA) and MgCl2 to bacterial cellulose (BC) for antimicrobial and anti-biofilm purposes. The morphology was investigated by scanning electron microscopy (SEM), and chemical structure were characterized by Fourier transform infrared spectroscopy (FTIR) and X-ray photoelectron spectra (XPS). In vitro release profiles of tannic acid revealed that the Mg2+ cross-links help impede the release of TA from BC matrix, while composite BC-TA lacked Mg2+ ionic cross-links, thus more TA was released from the hydrogel. The BC-TA-Mg composites also displayed strong antibacterial activity against S. aureus, E. coli and P. aeruginosa. Moreover, the composites significantly reduced biofilm formation of S. aureus and P. aeruginosa after 24 h incubation by ∼80% and ∼87%, respectively. As a consequence, the BC-TA-Mg composites are a very promising material for combating biofilm-associated infections in biomedical and public health fields.


Effects of gentamicin inducing readthrough premature stop Codons: A study of alpha-L-iduronidase nonsense variants in COS-7 Cells.
Mucopolysaccharidosis type I Hurler syndrome (MPS IH) is a severe lysosomal storage disorder caused by alpha-l-iduronidase (IDUA) deficiency. Premature truncation mutations (PTC) are the most common (50%-70%) type of IDUA mutations and correlate with MPS IH. Nonsense suppression therapy is a therapeutic approach that aims to induce stop codon readthrough. The different ability of gentamicin to bind mutant mRNA in readthrough is determined by nucleotide sequence (PTC context: UGA > UAG > UAA) and inserted amino acid including the nucleotide position +4 of the PTC, as well as the mRNA secondary structure. We used COS-7 cells to investigate the functional characteristics of p.Q500X and p.R619X, IDUA variants and the effects of gentamicin in inducing stop codon readthrough of seven IDUA variants including p.Q500X, p.R619X, p.Q70X, p.E299X, p.W312X, p.Q380X, and p.W402X. Moreover, we performed prediction of RNA secondary structure using the online tool RNAfold. We found that cells treated with gentamicin showed significantly enhanced full-length IDUA expression and restored IDUA activity, in a dose-dependent manner, only in cells expressing cDNA with W312X, Q380X, W402X, and R619X. Among the readthrough-responsive variants, we observed UGA PTC in W312X, W402X and R619X; and UAG PTC with C at nucleotide +4 in Q380X. Changes of RNA secondary structure were noted only in mutants with readthrough-responsive variants including W312X, Q380X, W402X, and R619X. Additional preclinical studies of selected PTCs with potential readthrough, using drugs with less oto-nephrotoxicity, in patient's skin fibroblasts and animal model are necessary for the premise of personalized medicine.


Remodelling of mandibular condylar head after fixation of fractures with ultrasound activated resorbable pins: A retrospective case series.
The aim of this study was to assess whether the resorption of poly-lactic acid pins, used for condylar head fracture fixation, adversely affect remodelling of the condylar head. A retrospective review of patients was performed that underwent ORIF of CHF with ultrasound activated resorbable poly-lactic acid pins and had CT/CBCT scans of the condyle at least 18 months after surgery, at which point the fixation material was expected to be resorbed. We reviewed the size, shape and position of the condylar head and compared this to the normal side and compared this to the results of conservative management and ORIF with titanium screws in the literature. Most patients had condylar heads normally positioned and shaped, with minimal bone changes. The position of the condyle in the fossa, its shape and bone changes compare favourably with previous studies on CHF managed conservatively or with titanium screw fixation. There was no radiographic evidence of the pins after 18 months. In conclusion, ultrasound activated resorbable pins can provide suitable fixation for ORIF of condylar head fractures, avoiding the need for screw removal, and there was no evidence that the resorption process adversely affected the remodelling of the condylar head.


Characteristics and anxiety symptom presentation associated with autism spectrum traits in youth with anxiety disorders.
There is limited information about the nature of anxiety among youth with symptoms of autism spectrum disorder (ASD). The present study examined (a) differences in the clinical characteristics of anxious youth with and without symptoms of ASD and (b) the symptoms of anxiety that best distinguish between these groups. Results indicated that anxious youth with elevated ASD symptoms had significantly more diagnoses (e.g., specific phobias), and were more likely to meet diagnostic criteria for social phobia (and list social concerns among their top fears) than youth without elevated ASD symptoms. At the symptom level, severity of interpersonal worry based on parent report and severity of fear of medical (doctor/dentist) visits based on youth report best differentiated ASD status. The findings inform diagnostic evaluations, case conceptualization, and treatment planning for youth with anxiety disorders and ASD symptoms.


Anastomosing tubular arrangement of dog exocrine pancreas.
Wax reconstructions of zymogen granule-containing cells in dog show the arrangement of the exocrine pancreas to be a branching, anastomosing system of tubules which vary in diameter and end blindly. This arrangement is not that of a true acinar gland.


HDL- and LDL-cholesterol in normal weight healthy children 13 years of age.
From 108 healthy 13 years old school children who took part on a longitudinal study in regard to serum lipids and lipoproteins 64 with normal body weight were examined. Lipoproteins were estimated by means of preparative ultracentrifuge and polyanionprecipitation (Heparin and Manganese-chloride) according to the Lipid Research Clinics Methods, NIH, Bethesda. Mean values and S.D. were calculated as: Total cholesterol: 155 +/- 29, 150 +/- 35 mg/dl; LDL-cholesterol 98 +/- 21 and 96 +/- 34 mg/dl; HDL-cholesterol 44 +/- 13 and 44 +/- 13 mg/dl for males and females respectively. The few data from the literature showing wide variations in LDL- and HDL-cholesterol concentrations are compared with our results; methodological considerations indicate that for further epidemiological studies concerning the possible risk factors for premature atherosclerosis standaradized laboratory tests should be provided.


Sericin-binded-deprotenized natural rubber film containing chitin whiskers as elasto-gel dressing.
Here, we aims to demonstrate a simple concept in biomaterials design by using natural resources solely as raw materials to fabricate elastic wound care dressing. Elasto-gel films comprise of silk sericin (SRC), natural rubber (NR), and chitin whisker (CTW) were developed. A glue-like protein SRC found in silk cocoons is beneficial for the treatment of wounds due to its superior skin moisturizing ability. However, the pure SRC film is generally difficult to be fabricated because of its weak structural feature. This limitation was overcome by using NR as a binder which consecutively rendered elasticity and strength of the films. CTW was chosen as another component to promote ability of the films for tissue restoration. Before the film formation, protein in the natural rubber latex (NRL) was removed to avoid allergic and cytotoxic problems. The enzyme-treated NR/SRC (ETNR/SRC) films having different blend compositions were fabricated by solution casting technique. The highest amount of the SRC to gain an easy to handle ETNR/SRC film was 30%. The ETNR/SRC/CTW films having 20% SRC were fabricated and studied in comparison. Essential properties of the films as elastic wound care dressings were investigated and effect of the materials chemistry on the observed properties were discussed.


[Apoptosis of endothelial cells in alteration of microvascular permeability in lung during sepsis].
To investigate the apoptosis of endothelial cells in the alteration of microvascular permeability in lung during sepsis. Twenty-four male NIH mice were subjected to cecal ligation and puncture (CLP) or sham operation (SO). The microvascular endothelial cells of the lungs were harvested at 3 hours and 12 hours after operation. The apoptosis of endothelial cells (VIII factor related antigen positive cells) in the lungs was determined by 2-color flow cytometry (FITC-labeled VIII factor related antigen and PI) and Apop Tag) in situ apoptosis detection kit in situ. The apoptosis related gene (bcl-2) was detected by RT-PCR. The microvascular permeability in lung tissue was also examined. The apoptosis of microvascular endothelial cells was increased significantly at 12 hours after CLP (5.03 +/- 0.92 vs. 3.48 +/- 1.21, P < 0.05). The increased apoptosis was paralleled with the decrease in bcl-2 gene expression. The microvascular permeability in lung tissue was slightly increased at 3 hours after CLP, but significantly increased at 12 hours after CLP (0.106 +/- 0.008 vs 0.047 +/- 0.003, P < 0.01). Apoptosis in microvascular endothelial cells seems to be an important reason for the alteration of microvascular permeability.


Thesaurin a, the major protein of Xenopus laevis previtellogenic oocytes, present in the 42 S particles, is homologous to elongation factor EF-1 alpha.
We have purified in SDS X.laevis thesaurin a (Mr 50,000) which is part of the 42 S storage particles. Its N-terminal amino acid is blocked and several peptides obtained by V8 protease treatment were purified and sequenced. As expected from one of the functional roles of the 42 S particles (tRNA binding, protection against deacylation and exchange with the ribosome), the amino acid sequence of thesaurin a was found to be closely related to that of the elongation factor EF-1 alpha. We suggest that all three proteins involved in 5 S RNA and tRNA storage in previtellogenic oocytes, TFIIIA, thesaurin a and thesaurin b, have a dual function: storage and a role in transcription or in protein synthesis.


N-terminal proBNP and mortality in hospitalised patients with heart failure and preserved vs. reduced systolic function: data from the prospective Copenhagen Hospital Heart Failure Study (CHHF).
Preserved systolic function among heart failure patients is a common finding, a fact that has only recently been fully appreciated. The aim of the present study was to examine the value of NT-proBNP to predict mortality in relation to established risk factors among consecutively hospitalised heart failure patients and secondly to characterise patients in relation to preserved and reduced systolic function. At the time of admission 2230 consecutively hospitalised patients had their cardiac status evaluated through determinations of NT-proBNP, echocardiography, clinical examination and medical history. Follow-up was performed 1 year later in all patients. 161 patients fulfilled strict diagnostic criteria for heart failure (HF). In this subgroup of patients 1-year mortality was approximately 30% and significantly higher as compared to the remaining non-heart failure population (approx. 16%). Using univariate analysis left ventricular ejection fraction (LVEF), New York Heart Association classification (NYHA) and plasma levels of NT-proBNP all predicted mortality independently. However, regardless of systolic function, age and NYHA class, risk-stratification was provided by measurements of NT-proBNP. Having measured plasma levels of NT-proBNP, LVEF did not provide any additional prognostic information on mortality among heart failure patients (multivariate analysis). The results show that independent of LVEF, measurements of NT-proBNP add additional prognostic information. It is concluded that NT-proBNP is a strong predictor of 1-year mortality in consecutively hospitalised patients with heart failure with preserved as well as reduced systolic function.


Predictability of connective tissue grafts for root coverage: clinical perspectives and a review of the literature.
Several techniques using subepithelial connective tissue for root coverage have recently been described in the periodontal literature. The predictability of such procedures, however, varies among the different reports. The factors responsible for these differences are not fully understood. The purpose of this article is to review and comment on factors that may enhance the predictability of root coverage.


Absence of positional change in pulmonary diffusing capacity in systemic sclerosis.
Patients with systemic sclerosis frequently have pulmonary function abnormalities, and recent evidence suggests that pulmonary vascular involvement is a common manifestation. To test the hypothesis that patients with systemic sclerosis have impaired ability to recruit or distend the pulmonary vascular bed, the postural change in the coefficient of carbon monoxide diffusing capacity was measured in 11 patients with systemic sclerosis, and the results were compared with results from age-, smoking-, and sex-matched control subjects with rheumatoid arthritis and with results from healthy subjects. In normal subjects and patients with rheumatoid arthritis increased, the coefficient of diffusion by 9.4 percent (p less than 0.005) and 8.4 percent (p less than 0.01), respectively, when they moved from the sitting to the supine position. In contrast, patients with systemic sclerosis did not show a significant increase in coefficient of diffusion, even those who had otherwise normal pulmonary function. Regression analyses showed that the change in coefficient of diffusion decreased with increasing age (r = -0.57) in normal subjects, and that the change in coefficient of diffusion was a function of the percent predicted forced vital capacity, both in patients with systemic sclerosis (r = 0.59) and in those with rheumatoid arthritis (r = 0.70). Thus, these findings indicate that patients with systemic sclerosis have a nondistensible pulmonary capillary bed and that the absence of positional change in the coefficient of diffusion in systemic sclerosis is a subtle indicator of pulmonary involvement.


Design, synthesis, and structure-activity relationships of dihydrofuran-2-one and dihydropyrrol-2-one derivatives as novel benzoxazin-3-one-based mineralocorticoid receptor antagonists.
Dihydrofuran-2-one and dihydropyrrol-2-one derivatives were identified as novel, potent and selective mineralocorticoid receptor (MR) antagonists by the structure-based drug design approach utilizing the crystal structure of MR/compound complex. Introduction of lipophilic substituents directed toward the unfilled spaces of the MR and identification of a new scaffold, dihydropyrrol-2-one ring, led to potent in vitro activity. Among the synthesized compounds, dihydropyrrol-2-one 11i showed an excellent in vitro activity (MR binding IC50=43nM) and high selectivity over closely related steroid receptors such as the androgen receptor (AR), progesterone receptor (PR) and glucocorticoid receptor (GR) (>200-fold for AR and PR, 100-fold for GR).


Gaussian private quantum channel with squeezed coherent states.
While the objective of conventional quantum key distribution (QKD) is to secretly generate and share the classical bits concealed in the form of maximally mixed quantum states, that of private quantum channel (PQC) is to secretly transmit individual quantum states concealed in the form of maximally mixed states using shared one-time pad and it is called Gaussian private quantum channel (GPQC) when the scheme is in the regime of continuous variables. We propose a GPQC enhanced with squeezed coherent states (GPQCwSC), which is a generalization of GPQC with coherent states only (GPQCo) [Phys. Rev. A 72, 042313 (2005)]. We show that GPQCwSC beats the GPQCo for the upper bound on accessible information. As a subsidiary example, it is shown that the squeezed states take an advantage over the coherent states against a beam splitting attack in a continuous variable QKD. It is also shown that a squeezing operation can be approximated as a superposition of two different displacement operations in the small squeezing regime.


The influence of water on CO2 exchange in the lichen Parmelia praesignis Nyl.
Net photosynthetic rates for the lichen Parmelia praesignis Nyl. were obtained as a function of 5 light levels, 5 temperature levels, and of water content as thalli dried from saturated conditions. Data are described as second order polynomials in the light, and as saturation curves in the dark. Rates in the light were depressed at high water contents reaching maximal rates between 110% and 180% water content and declining as thalli dried. Physiological parameters were derived from the drying curves to investigate temperature and light interactions. Dark respiration parameters are the maximal rate, the water content where the rate is half-maximal, the water content at which respiration is zero, and the maximal water efficiency. In the light, parameters are the maximal net photosynthetic rate, the water content at the maximal rate, the water compensation point, the maximal water efficiency, and the sensitivity of net photosynthesis to change in water content.Values of half-maximal rate water contents for respiration were found to increase as temperatures increased. The greatest maximal net photosynthetic rate occurred at higher temperatures as the light intensity increased. In the light, maximal water efficiency and the sensitivity to changes in water content were generally maximal at temperatures yielding the greatest maximal net photosynthetic rates.


Formation of reactive 1-nitropyrene metabolites by lung microsomes and isolated lung cells.
The metabolism and activation of 1-nitropyrene (1-NP) to reactive intermediates by lung microsomes and isolated lung cells was studied. Mutagenicity of 1-NP metabolites was assayed in Salmonella typhimurium TA98NR, a strain lacking a major component of nitroreductase activity. In the presence of NADPH, microsomes from rabbit, rat and hamster lung metabolized 1-NP to mutagenic products to a similar degree. Pretreatment with a mixture of polychlorinated biphenyls (PCB) decreased the formation of mutagenic metabolites by rabbit lung microsomes, but did not affect the production of mutagens by rat or hamster lung microsomes. 3H-1-NP was metabolized to covalently bound protein products at a rate of 82 and 10 pmol/mg by rabbit and hamster lung microsomes, respectively, whereas no binding was detected in rat lung microsomes. PCB-pretreatment increased covalent protein binding of 3H-1-NP in lung microsomes from hamster and rat, but decreased the binding in rabbit lung microsomes. High performance liquid chromatography analysis indicated that 3H-1-NP was readily converted to ring-hydroxylated products by rabbit and hamster lung microsomes; the rate was much lower with rat lung microsomes. 3H-1-NP was activated to metabolites that covalently bound to protein in isolated rabbit lung cells, with the following rates being observed: Clara cells greater than lung digest greater than type II cells. In contrast, covalent protein binding in cells isolated from rat lung was very low. 1-NP was not activated to products mutagenic for S. typhimurium TA 98NR when co-incubated with cells isolated either from rabbit or rat lung.


Differing patterns in thermal injury incidence and hospitalisations among 0-4 year old children from England.
To describe patterns in thermal injury incidence and hospitalisations by age, gender, calendar year and socioeconomic status among 0-4 year olds in England for the period 1998-2013. 708,050 children with linked primary care and hospitalisation data from the Clinical Practice Research Datalink (CPRD) and Hospital Episode Statistics (HES), respectively. Incidence rates of all thermal injuries (identified in CPRD and/or HES), hospitalised thermal injuries, and serious thermal injuries (hospitalised for ≥72h). Adjusted incidence rate ratios (IRR) and 95% confidence intervals (95%CI), estimated using Poisson regression. Incidence rates of all thermal injuries, hospitalised thermal injuries, and serious thermal injuries were 59.5 per 10,000 person-years (95%CI 58.4-60.6), 11.3 (10.8-11.8) and 2.15 (1.95-2.37), respectively. Socioeconomic gradients, between the most and least deprived quintiles, were steepest for serious thermal injuries (IRR 3.17, 95%CI 2.53-3.96). Incidence of all thermal injuries (IRR 0.64, 95%CI 0.58-0.70) and serious thermal injuries (IRR 0.44, 95%CI 0.33-0.59) reduced between 1998/9 and 2012/13. Incidence rates of hospitalised thermal injuries did not significantly change over time. Incidence of all thermal injuries and those hospitalised for ≥72h reduced over time. Steep socioeconomic gradients support continued targeting of preventative interventions to those living in the most deprived areas.


Author Correction: Systems of mechanized and reactive droplets powered by multi-responsive surfactants.
In this Letter, the top structure of the right panel of Fig. 1a should be that of cis-9-octadecenoic acid, not trans-9-octadecenoic acid. Please see the accompanying Author Correction. This error has not been corrected online.


Effect of furosemide on hemodynamics and lung water in acute pulmonary edema secondary to myocardial infarction.
Hemodynamic studies were carried out in 19 patients with left ventricular failure complicating acute myocardial infarction. Fourteen patients were studied before and after the intravenous administration of 0.5 mg/kg of furosemide, and five patients served as a control group. Serial measurements included intracardiac pressures, cardiac output and lung water by a double isotope technique. A significant reduction was noted in right atrial (P less than 0.005), pulmonary arterial (P less than 0.0005) and pulmonary wedge pressures (P less than 0.0005) after administration of furosemide. Only the change in right atrial pressure was significantly different from that in the control group (P less than 0.05). Lung water was not changed in 4 patients studied 2 hours after administration of furosemide but was significantly changed in the remaining 10 patients studied 4 to 24 hours after furosemide (P = 0.0001). This change was also significantly different from values in the control group (P less than 0.05). The patients with no reduction in excess lung water also had a smaller reduction in pulmonary wedge pressure and a lower pretreatment stroke work index than the other patients. The mobilization of excess lung water in patients with acute myocardial infarction complicated by left ventricular failure has several features. Despite a prompt diuresis, the reduction in lung water is delayed for at least several hours after the administration of furosemide and may be related to the degree of left ventricular dysfunction. Venodilation may be a major result of treatment with furosemide.


Quantification of Pseudomonas aeruginosa hydrogen cyanide production by a polarographic approach.
Pseudomonas aeruginosa is an opportunistic pathogen responsible for numerous infections acquired in hospital especially in persons whose immune systems are weakened, such as with patient suffering from AIDS or cystic fibrosis. This bacterium produces a great diversity of virulence factors among them hydrogen cyanide (HCN) which is one of the most potent and toxic. A precise quantification of HCN or CN(-) ion is essential to understand the involvement of this toxin in the pathogenesis of P. aeruginosa. In the present study, we present a new technique based on a polarographic approach to measure the production kinetics of HCN/CN(-) by P. aeruginosa strains, in several media commonly used in microbiology labs. The method was validated using mutants (hcnB- and hcnC-) which are unable to produce detectable HCN/CN(-). The kinetics of HCN/CN(-) production by P. aeruginosa in Luria Bertani (LB) medium showed a parabolic shape with a peak observed at 4, 5 and 8h for strains PA14, PAO1 and MPAO1, respectively. When bacteria were grown in ordinary nutrient broth (ONB) 2.5% medium, a less adapted medium for bacterial growth, the general profile of the kinetics was conserved but peak production was delayed (10 and 12h for PAO1 and MPAO1, respectively). When the bacteria were cultured in minimum medium MMC, bacterial growth was particularly slow and HCN/CN(-) production was markedly reduced. Taken together, this new polarographic method appears as a useful technique to detect and quantify HCN/CN(-) in routine media where the bacteria can express and regulate high amounts of toxins. With this method, we demonstrate that HCN/CN(-) production by P. aeruginosa is maximal at the end of the exponential growth phase and depends on the richness of the growth medium used.


Autoradiographic localization of 24R,25-dihydroxyvitamin D3 in epiphyseal cartilage.
There is emerging evidence for specific binding sites and biologic action for 24,25(OH)2D3 in the epiphyseal cartilage. The present study was undertaken to identify the target cells of 24,25(OH)2D3 in the epiphyses of rat bone using an autoradiographic technique. Pieces of epiphyseal cartilage obtained from 4-day-old vitamin D-deficient rats were incubated for 15 or 60 min with [3H]-24,25(OH)2D3 in the presence or absence of 100-fold excess of 25(OH)D3, 1,25(OH)2D3, or 24R,25(OH)2D3. The pieces were prepared for autoradiographic study by a new modified fixation method. Cytoplasmic and nuclear concentration of radioactivity was observed in all cell layers of the epiphyseal cartilage except for the hypertrophic cartilage zone. The highest concentration of radioactivity was seen in the proliferating chondroblasts of the columnar zone. After 15 min of incubation the radioactivity was seen mainly in the cell membrane and cytoplasm, whereas at 60 min radioactivity was also prominent in the nuclei. The competition with excess of cold metabolites revealed that only 24R,25(OH)2D3 caused a significant decrease in cytoplasmic and nuclear radioactivity. These data support the biochemical studies showing that the epiphyseal cartilage is a target tissue for 24,25(OH)2D3.


Which persistent organic pollutants in the rivers of the Bohai Region of China represent the greatest risk to the local ecosystem?
Freshwater aquatic organisms can be exposed to hundreds of persistent organic pollutants (POPs) discharged by natural and anthropogenic activities. Given our limited resources it is necessary to identify, from the existing evidence, which is the greatest threat so that control measures can be targeted wisely. The focus of this study was to rank POPs according to the relative risk they represent for aquatic organisms in rivers in the Bohai Region, China. A list of 14 POPs was compiled based on the available data on their presence in these rivers and ecotoxicological data. Those that were widely detected were benzo[a]pyrene, p,p'-DDE, p,p'-DDT, endrin, fluoranthene, heptachlor, hexabromocyclododecane, hexachlorobenzene, α-hexachlorocyclohexane, γ-hexachlorocyclohexane, naphthalene, perfluorooctanoic acid, perfluorooctane sulfonate and phenanthrene. Effect concentrations were compiled for Chinese relevant and standard test species and compared with river aqueous concentrations. Only bed-sediment concentrations were available so water levels were calculated based on the known local sediment organic carbon concentration and the Koc. The POPs were ranked on the ratio between the median river and median effect concentrations. Of the POPs studied, fluoranthene was ranked as the highest threat, followed by phenanthrene, naphthalene and p,p'-DDE. The risk from p,p'-DDE may be magnified due to being highly bioaccumulative. However, the greatest overlap between river concentrations and effect levels was for lindane. Overall, fish was the most sensitive species group to the risks from POPs. Hotspots with the highest concentrations and hence risk were mainly associated with watercourses draining in Tianjin, the biggest city in the Bohai Region.


Optimization of annealing conditions for Ag/p-GaN ohmic contacts.
The electrical and optical properties of Ag/p-GaN contacts have been investigated as a function of the annealing temperature, oxygen concentration, and annealing time. Specific contact resistance (ρ c) values as low as 1.2 × 10-4 Ω·cm2 were obtained from the Ag/p-GaN contact annealed at 400 °C for 60 s in ambient O2/N2 (1:10). We found that the participation of oxygen improves the formation of ohmic contacts. Oxygen might remove the H in Mg-H complexes to activate the Mg acceptors and enhance Ga out-diffusion to form an Ag-Ga solid solution. We also found that the reflectivity of the Ag layer decreases with increasing annealing temperature in the O2-containing ambient environment. Thus, an optimal annealing condition of Ag/p-GaN for blue and green LEDs is suggested based on these results. We also used the suggested annealing conditions to form ohmic contacts on DUV LEDs and achieved good electrical performance. The forward voltages of UVC LEDs fabricated with annealed Ag contacts were 6.60 V (7.66 V) at a 40 mA (100 mA) injection current.


Dissecting coffee seeds metabolome in context of genotype, roasting degree, and blending in the Middle East using NMR and GC/MS techniques.
With a favored taste and various bioactivities, coffee has been consumed as a daily beverage worldwide. The current study presented a multi-faceted comparative metabolomics approach dissecting commercially available coffee products in the Middle East region for quality assessment and functional food purposes using NMR and GC/MS platforms. NMR metabolites fingerprinting led to identification of 18 metabolites and quantification (qNMR) of six prominent markers for standardization purposes. An increase of β-ethanolamine (MEA) reported for the first time, 5-(hydroxymethyl) furfural (5-HMF), concurrent with a reduction in chlorogenic acid, kahweol, and sucrose levels post roasting as revealed using multivariate data analyses (MVA). The diterpenes kahweol and cafestol were identified in green and roasted Coffea arabica, while 16-O-methyl cafestol in roasted C. robusta. Moreover, GC/MS identified a total of 143 metabolites belonging to 15 different chemical classes, with fructose found enriched in green C. robusta versus fatty acids abundance, i.e., palmitic and stearic acids in C. arabica confirming NMR results. These potential results aided to identify novel quality control attributes, i.e., ethanolamine, for coffee in the Middle East region and have yet to be confirmed in other coffee specimens.


Stress response mechanism of wastewater biological nitrogen removal systems to environmentally realistic concentrations of tire wear particles: Contribution of leachable additives.
The study quantified the biological nitrogen removal performance, microbial metabolism, microbial community structure, and antioxidant system in a sequencing batch reactor under long-term exposure to 0.1 and 1 mg/L tire wear particles (TWPs), and determined the contribution of leachable additives to the biotoxicity of TWPs. The results showed that long-term exposure to 0.1 and 1 mg/L TWPs inhibited both the nitrification and denitrification processes, reducing ammonia nitrogen (NH4+-N) and total nitrogen (TN) removal efficiency. The TWP leachate (TWPL) primarily contributed to the denitrification inhibition by TWPs, potentially due to the high concentration of zinc ions in the leachable additive. Furthermore, both TWP and TWPL inhibit nitrogen conversion, with TWP inhibiting the generation and transfer of electrons, while TWPL only negatively affects the electron transfer process. This study presents novel insights into the impact of TWPs on biological nitrogen removal, underscoring its broader implications for the geochemical nitrogen cycle.


Trabecular bone anisotropy and orientation in an Early Pleistocene hominin talus from East Turkana, Kenya.
Among the structural properties of trabecular bone, the degree of anisotropy is most often found to separate taxa with different habitual locomotor modes. This study examined the degree of anisotropy, the elongation, and primary orientation of trabecular bone in the KNM-ER 1464 Early Pleistocene hominin talus as compared with extant hominoid taxa. Modern human tali were found to have a pattern of relatively anisotropic and elongated trabeculae on the lateral aspect, which was not found in Pan, Gorilla, Pongo, or KNM-ER 1464. Trabecular anisotropy in the fossil talus most closely resembled that of the African apes except for a region of high anisotropy in the posteromedial talus. The primary orientation of trabeculae in the anteromedial region of KNM-ER 1464 was strikingly different from that of the great apes and very similar to that of modern humans in being directed parallel to the talar neck. These results suggest that, relative to that of modern humans, the anteromedial region of the KNM-ER 1464 talus may have transmitted body weight to the midfoot in a similar manner while the lateral aspect may have been subjected to more variable loading conditions.


Real-time analysis of heart rate variability during aerobic exercise in patients with cardiovascular disease.
Exercise therapy for cardiovascular disease (CVD) is mainly evaluated based on the heart rate (HR) or Borg scale. However, these indices can be unreliable depending on the patient's medication or their subjective decisions; thus, alternative methods are required for easier and safer implementation of aerobic exercise. Here, we examined whether real-time analysis of HR variability (HRV) can help maintain exercise intensity at the ventilatory threshold (VT) during exercise. Twenty-eight patients with CVD treated at Keio University Hospital between August 2018 and March 2020 were enrolled. Initially, oxygen uptake (VO2) and HR at the VT were determined using the cardiopulmonary exercise test. Patients then performed aerobic exercise on a stationary bicycle for 30 min while a parameter of HRV, the high-frequency (HF) component, was monitored in real time using an electrocardiograph. The work rate during exercise was adjusted every 2 min to maintain the HF range between 5 and 10. The VO2 and HR values, recorded every 2 min during exercise, were compared with those at VT. The Bland-Altman method was used to confirm similarity. VO2 and HR during exercise were closely correlated with those at VT (e.g., 19 min after exercise initiation; VO2: r = 0.647, HR: r = 0.534). The Bland-Altman plot revealed no bias between the mean values (e.g., 19 min; VO2: -0.22 mL/kg/min; HR: -0.07/min). Real-time HRV analysis with electrocardiograph alone during exercise can provide continuous and non-invasive exercise intensity measurements at VT, promoting safer and effective exercise strategies.


Work in progress. Occurence of phospholipase A1 and A2 in human decidua.
Phospholipase A2, an enzyme which may regulate the formation of polyunsaturated fatty acids utilized for prostaglandin synthesis, was found to have significant higher activity in decidual than in myometrial tissue. The major part of phospholipase A2 in the decidua had an acid pH optimum, which indicates that most of the enzyme is stored in the lysosomes of this tissue. These findings, together with previous observations, lend further support to the view that lysosomal phospholipase A2 released within decidual cells might be a trigger of abortion and parturition.