Adverse event surveillance in small animal anaesthesia: an intervention-based, voluntary reporting audit.
To develop, test and refine an 'intervention-based' system for the surveillance of adverse events (AEs) during small animal anaesthesia. Prospective, voluntary reporting audit. A total of 1386 consecutive small animal anaesthetics (including 972 dogs and 387 cats). Adverse events were defined as undesirable perianaesthetic events requiring remedial intervention to prevent or limit patient morbidity. Using previous reports, 11 common AEs were selected and 'intervention-based' definitions were devised. A voluntary reporting audit was performed over 1 year at a university teaching hospital. Data on AEs were collected via paper checkbox forms completed after each anaesthetic and were assimilated using an electronic database. Interventions were performed entirely at the discretion of the attending anaesthetist. Comparisons between dogs and cats were made using Fisher's exact tests. Forms were completed for 1114 anaesthetics (a compliance of 80.4%), with 1001 AEs reported in 572 patients. The relative frequency of AEs reported were as follows: arousal or breakthrough pain (14.9%), hypoventilation (13.5%), hypotension (10.3%), arrhythmias (5.8%), hyperthermia/hypothermia (5.0%), airway complications (4.8%), recovery excitation (4.6%), aspiration risk (4.5%), desaturation (2.8%), hypertension (1.7%) and 'other' (3.7%). Canine anaesthetics (57.3%) were more likely to involve AEs than were feline anaesthetics (35.5%, p < 0.01). Escalation in postanaesthetic care was required in 20% of cases where an AE was reported (8% of anaesthetics overall). In 6% of cases (2% overall), this involved management in an intensive care unit. There were six intra-anaesthetic fatalities (0.43%) during this period. The tool was widely accepted, being considered quick and easy to complete, but several semantic, logistical and personnel factors were encountered. Simple intervention-based surveillance tools can be easily integrated into small animal anaesthetic practice, providing a valuable evidence base for anaesthetists. A number of considerations must be addressed to ensure compliance and the quality of data collected.


Designed CVD growth of graphene via process engineering.
Graphene, the atomic thin carbon film with honeycomb lattice, holds great promise in a wide range of applications, due to its unique band structure and excellent electronic, optical, mechanical, and thermal properties. Scientists are researching this star material because of the development of various emerging preparation techniques, among which chemical vapor deposition (CVD) has received the fastest advances in the past few years. For the CVD growth of graphene, the ultimate goal is to achieve the highest quality in the largest scale and lowest cost with a precise control of layer thickness, stacking order, and crystallinity. To meet this goal, researchers need a comprehensive understanding and effective controlling of the growth process, especially to its elementary steps. In this Account, we focus on our recent progresses toward the controlled surface growth of graphene and its two-dimensional (2D) hybrids via rational designs of CVD elementary processes, namely, process engineering. A typical CVD process consists of four main elementary steps: (A) adsorption and catalytic decomposition of precursor gas, (B) diffusion and dissolution of decomposed carbon species into bulk metal, (C) segregation of dissolved carbon atoms onto the metal surface, and finally, (D) surface nucleation and growth of graphene. Absence or enhancement of each elementary step would lead to significant changes in the whole growth process. Metals with certain carbon solubility, such as nickel and cobalt, involve all four elementary steps in a typical CVD process, thus providing us an ideal system for process engineering. The elementary segregation process can be completely blocked if molybdenum is introduced into the system as an alloy catalyst, yielding perfect monolayer graphene almost independent of growth parameters. On the other hand, the segregation-only process of predissolved solid carbons is also capable of high-quality graphene growth. By using a synergetic Cu-Ni alloy, we are able to further enhance the control to such a segregation technique, especially for the thickness of graphene. By designing a cosegregation process of carbon atoms with other elements, such as nitrogen, doped graphene could be synthesized directly with a tunable doping profile. Copper with negligible carbon solubility provides another platform for process engineering, where both carbon dissolution and segregation steps are negligible in the CVD process. Carbon atoms decomposed from precursors diffuse on the surface and build up the thermodynamically stable honeycomb lattice. As a result, graphene growth on copper is self-limited, and formation of multilayer graphene is generally prohibited. Being able to control this process better, as well as the high quality produced, makes copper-based growth the dominating synthesis procedure in the graphene community. We designed a two-temperature zone system to spatially separate the catalytic decomposition step of carbon precursors and the surface graphitization step for breaking this self-limited growth feature, giving high-quality Bernal stacked bilayer graphene via van der Waals epitaxy. We performed the so-called wrinkle engineering by growing graphene on nanostructured copper foil together with a structure-preserved surface transfer. In such a way, we controlled the wrinkling or folding on graphene and further fabricated graphene nanoribbon arrays by self-masked plasma etching. Moreover, by designing a two-step patching growth process on copper, we succeeded in synthesizing the mosaic graphene, a patchwork of intrinsic and nitrogen-doped graphene connected by single crystalline graphene p-n junctions. By following a general concept of process engineering, our work on the designed CVD growth of graphene and its 2D hybrids provides a unique insight of this research field. It enables the precise growth control of graphene together with the in-depth understanding of CVD growth process, which would further stimulate the pace of graphene applications.


Complication of barbotage, acute calcific bursitis treated successfully.
One of the most common causes of chronic shoulder pain that significantly impairs patients' quality of life is rotator cuff calcific tendinitis. The supportive approach of analgesics, anti-inflammatory medications, and physiotherapy is preferred to treat mild symptoms. However, severe intractable symptoms necessitate alternative treatment, and ultrasonography-guided percutaneous barbotage is regarded as an effective treatment technique due to its minimal invasiveness, low cost, and rapid significant pain relief. Post-barbotage complications are reported as acute calcific bursitis along with infection, bleeding, and tendon rupture. In our case, a 41-year-old female presented with a substantial amount of pain shortly after barbotage, which was diagnosed as acute calcific bursitis, a complication of the procedure. Subacromial-subdeltoid steroid injection was used successfully to treat this condition.


QM/MM Studies Reveal How Substrate-Substrate and Enzyme-Substrate Interactions Modulate Retaining Glycosyltransferases Catalysis and Mechanism.
Glycosyltransferases (GTs) catalyze the biosynthesis of glycosidic linkages by transferring a monosaccharide from a nucleotide sugar donor to an acceptor substrate, and they do that with exquisite regio- and stereospecificity. Retaining GTs act with retention of the configuration at the anomeric carbon of the transferred sugar. Their chemical mechanism has been under debate for long as conclusive experimental data to confirm the mechanism have been elusive. In the past years, quantum mechanical/molecular mechanical (QM/MM) calculations have shed light on the mechanistic discussion. Here, we review the work carried out in our group investigating three of these retaining enzymes (LgtC, α3GalT, and GalNAc-T2). Our results support the controversial front-side attack mechanism as the general mechanism for most retaining GTs. The latest structural data are in agreement with these findings. QM/MM calculations have revealed how enzyme-substrate and substrate-substrate interactions modulate the transfer reaction catalyzed by these enzymes. Moreover, they provide an explanation on why in some cases a strong nucleophilic residue is found on the β-face of the sugar, opening the door to a shift toward a double-displacement mechanism.


Overexpression of the X-linked ribosomal protein S4 predicts poor prognosis in patients with intrahepatic cholangiocarcinoma.
X-linked ribosomal protein S4 (RPS4X) has previously been reported to be associated with cisplatin resistance and clinical outcome in bladder and ovarian cancer. However, the value of RPS4X as a diagnostic and prognostic marker in intrahepatic cholangiocarcinoma (ICC) has not yet been investigated. The present study evaluated the expression pattern, and diagnostic and prognostic value of RPS4X in patients with ICC. Retrospective analysis was performed for a total of 201 patients with intrahepatic cholangiocarcinoma, and 8 patients with inflammation of the bile duct. Immunohistochemistry was performed using tissue microarrays to characterize the expression profile of RPS4X. Receiver operating characteristic (ROC) curves, the Kaplan-Meier estimator and Cox regression analysis were applied to evaluate the potential diagnostic and prognostic value of RPS4X in ICC. RPS4X was significantly upregulated in ICC tissues compared with the inflamed bile duct tissues. When differentiating ICC from normal controls, ROC analysis of RPS4X gave an area under the curve value of 0.9030 (sensitivity, 82.59%; specificity, 100%). RPS4X expression was significantly positively correlated with serum alkaline phosphatase levels. Survival analysis demonstrated that RPS4X expression levels were an independent prognostic factor for overall survival. Therefore, RPS4X expression levels may serve as a novel diagnostic and prognostic marker in ICC.


CD36 overexpression: a possible etiopathogenic mechanism of atherosclerosis in patients with prediabetes and diabetes.
CD36 is a scavenger receptor located on monocytes which is involved in foam cell transformation. To evaluate CD36 expression under different glycemic states in both healthy subjects and in atherosclerotic patients. In order to evaluate the possible effects of hyperglycemia on CD36 expression in healthy subjects, an in vitro experiment was carried out using monocyte in three different conditions: extreme hyperglycemia (HG), euglycemia (EG) and in the absence of glucose. On the other hand, three groups of atherosclerotic patients were evaluated according to their glycemic conditions: normoglycemic (NG), prediabetic (preDM) and diabetic (DM) patients. CD36 expression (mRNA, non-glycated and glycated protein) was analyzed in monocytes. CD36 mRNA expression in the in vitro experiment peaked at 4 and 24 h under HG conditions. No differences in mRNA levels were found in the EG and control group. The level of non-glycated proteins was higher in HG and EG conditions compared with control group. Glycated protein expression was inhibited by glucose in a sustained manner. In atherosclerotic patients, a significant association was observed when comparing glycated CD36 protein expression in DM with NG patients (p = 0.03). No significant differences were found in mRNA and non-glycated CD36 expression in these patients. Moreover, BMI, insulin, weight and treatment were shown to be related to CD36 expression (mRNA, non-glycated and glycated protein levels, depending of the case) in atherosclerotic patients. Hyperglycemia is an important modulator of CD36 mRNA and non-glycated protein expression in vitro, increasing de novo synthesis in healthy subjects. In atherosclerotic patients, there are progressive increases in CD36 receptors, which may be due to a post-translational stimulus.


Type IV pili are involved in phenotypes associated with Clostridioides difficile pathogenesis.
Clostridioides difficile is a Gram-positive, spore-forming, rod-shaped, obligate anaerobe that is the leading cause of antibiotic-associated diarrhea. Type IV pili (T4P) are elongated appendages on the surface of C. difficile that are polymerized from many pilin proteins. T4P play an important role in C. difficile adherence and particularly in its persistence in the host intestine. Recent studies have shown that T4P promote C. difficile aggregation, surface motility, and biofilm formation, which may enhance its pathogenicity. Additionally, the second messenger cyclic diguanylate increases pilA1 transcript abundance, indirectly promoting T4P-mediated aggregation, surface motility, and biofilm formation of C. difficile. This review summarizes recent advances in C. difficile T4P research and the physiological activities of T4P in the context of C. difficile pathogenesis.


Identification of anthropogenic influences on water quality of rivers in Taihu watershed.
Surface water bodies are progressively subjected to stress as a result of anthropogenic activities. This study assessed and examined the impact of human activities on spatial variation in the water quality of 19 rivers in the Taihu watershed. Concentrations of physicochemical parameters of surface water quality were determined at the mouth of each river during the period of 2000-2004. Multivariate statistical techniques were applied to identify characteristics of the water quality in the studied rivers. The results showed that rivers strongly influenced by household wastewater have the highest concentrations of nutrients (TN and TP). Moreover, rivers in the vicinity of a metropolis presented low dissolved oxygen (DO) levels. However, organic-chemical pollution (petroleum and volatile phenolics) was identified with high localization. Two rivers influenced by sewage from industry and ships were distinguished from other rivers with high values of petroleum. The Taige channel, a river located in Changzhou City that is strongly influenced by wastewater from industry, was characterized with an extraordinarily high value of volatile phenolics. Rivers passing through countries, especially through hilly countries were characterized with high DO contents and low nutrient and organic-chemical pollution, suggesting that agriculture puts less pressure on water quality in adjacent rivers. Therefore, more effort should be made in controlling point pollution to restore water quality in rivers adjacent to cities.


Randomization tests for changing criterion designs.
Randomization tests for alternating treatments designs, multiple baseline designs, and withdrawal/reversal designs are well-established. Recent classifications, however, also mention the "changing criterion design" as a fourth important type of single-case experimental design. In this paper, we examine the potential of randomization tests for changing criterion designs. We focus on the rationale of the randomization test, the random assignment procedure, the choice of the test statistic, and the calculation of randomization test p-values. Two examples using empirical data and an R computer program to perform the calculations are provided. We discuss the problems associated with conceptualizing the changing criterion design as a variant of the multiple baseline design, the potential of the range-bound changing criterion design, experimental control as an all-or-none phenomenon, the necessity of random assignment for the statistical-conclusion validity of the randomization test, and the use of randomization tests in nonrandomized designs.


Binding affinities for sulfonamide inhibitors with human thrombin using Monte Carlo simulations with a linear response method.
The binding of sulfonamide inhibitors to human thrombin is examined to evaluate the viability of calculating free energies of binding, deltaGb, utilizing Monte Carlo (MC) statistical mechanics with a linear response approach. Coulombic and van der Waals energy components determined from MC simulations of the bound and unbound inhibitors solvated in water plus a solvent-accessible surface area term, as an index for cavity formation, were correlated with the free energies of binding for the inhibitor MD-805 and six derivatives. The best correlations yield an average error of 0.8 kcal/mol for the seven binding affinities, which cover an observed range of 6.0 kcal/mol. The MC simulations also provided insights into the interactions occurring in the active site and the origins of variations in deltaGb. Equatorial placement of the carboxylate group at C2 in the piperidine ring of the inhibitors causes electrostatic destabilization with the side chain of Glu-H192, while axial disposition of the C4-methyl group reduces favorable hydrophobic interactions in the P-pocket of the enzyme.


Genetic structure of avian myeloblastosis virus, released from transformed myeloblasts as a defective virus particle.
Chicken myeloblasts transformed by avian myeloblastosis virus (AMV) in the absence of nondefective helper virus (termed nonproducer cells) were found to release a defective virus particle (DVP) that contains avian tumor viral gag proteins but lacks envelope glycoprotein and a DNA polymerase. Nonproducer cells contain a Pr76 gag precursor protein and also a protein that is indistinguishable from the Pr180 gag-pol protein of nondefective viruses. The RNA of the DVP is 7.5 kilobases (kb) long and is 0.7 kb shorter than the 8.2-kb RNAs of the helper viruses of AMV, MAV-1 and MAV-2. Comparisons based on RNA.cDNA hybridization and mapping of RNase T1-resistant oligonucleotides indicated that DVP RNA shares with MAV RNAs nearly isogenic 5'-terminal gag and pol-related sequences of 5.3 kb and a 3'-terminal c-region of 0.7 kb that is different from that found in other avian tumor viruses. Adjacent to the c-region, DVP RNA contains a contiguous specific sequence of 1.5 kb defined by 14 specific oligonucleotides. Except for two of these oligonucleotides that map at its 5' end, this sequence is unrelated to any sequences of nondefective avian tumor viruses of four different envelope subgroups as well as to the specific sequences of fibroblast-transforming avian acute leukemia and sarcoma viruses of four different RNA subgroups. The specific sequence of the DVP RNA is present in infectious stocks of AMV from this and other laboratories in an AMV-transformed myeloblast line from another laboratory, and it is about 70% related to nucleotide sequences of E26 virus, an independent isolate of an AMV-like virus. Preliminary experiments show DVP to be leukemogenic if fused into susceptible cells in the presence of helper virus. We conclude that DVP RNA is the leukemogenic component of infectious AMV and that its specific sequence, termed AMV, may carry genetic information for oncogenicity. Thus we have found here a transformation-specific RNA sequence, unrelated to helper virus, in a highly oncogenic virus that does not transform fibroblasts.


Counterion Exchange in Peptide-Complexed Core-Shell Microgels.
The complexation of polyvalent macroions with oppositely charged polyelectrolyte microgels can lead to core-shell structures. The shell is believed to be highly deswollen with a high concentration of counter-macroions. The core is believed to be relatively free of macroions but under a uniform compressive stress due to the deswollen shell. We use cryo-scanning electron microscopy (SEM) with X-ray microanalysis to confirm this understanding. We study poly(acrylic acid) (PAA) microgels which form a core-shell structure when complexed with a small cationic antimicrobial peptide (L5). We follow the spatial distribution of polymer, water, Na counterions, and peptide based on the characteristic X-ray intensities of C, O, Na, and N, respectively. Frozen-hydrated microgel suspensions include buffers of known composition from which calibration curves can be generated and used to quantify both the microgel water and sodium concentrations, the latter with a minimum quantifiable concentration less than 0.048 M. We find that as-synthesized PAA microgels are enriched in Na relative to the surrounding buffer as anticipated from established ideas of counterion shielding of electrostatic charge. The shell in L5-complexed microgels is depleted in Na and enriched in peptide and contains relatively little water. Our measurements furthermore show that shell/core interface is diffuse over a length scale of a few micrometers. Within the limits of detection, the core Na concentration is the same as that in as-synthesized microgels, and the core is free of peptide. The core has a slightly lower water concentration than as-synthesized controls, consistent with the hypothesis that the core is under compression from the shell.


Electron Transport Lipids Fold Within Membrane-Like Interfaces.
Lipoquinones, such as ubiquinones (UQ) and menaquinones (MK), function as essential lipid components of the electron transport system (ETS) by shuttling electrons and protons to facilitate the production of ATP in eukaryotes and prokaryotes. Lipoquinone function in membrane systems has been widely studied, but the exact location and conformation within membranes remains controversial. Lipoquinones, such as Coenzyme Q (UQ-10), are generally depicted simply as "Q" in life science diagrams or in extended conformations in primary literature even though specific conformations are important for function in the ETS. In this study, our goal was to determine the location, orientation, and conformation of UQ-2, a truncated analog of UQ-10, in model membrane systems and to compare our results to previously studied MK-2. Herein, we first carried out a six-step synthesis to yield UQ-2 and then demonstrated that UQ-2 adopts a folded conformation in organic solvents using 1H-1H 2D NOESY and ROESY NMR spectroscopic studies. Similarly, using 1H-1H 2D NOESY NMR spectroscopic studies, UQ-2 was found to adopt a folded, U-shaped conformation within the interface of an AOT reverse micelle model membrane system. UQ-2 was located slightly closer to the surfactant-water interface compared to the more hydrophobic MK-2. In addition, Langmuir monolayer studies determined UQ-2 resided within the monolayer water-phospholipid interface causing expansion, whereas MK-2 was more likely to be compressed out and reside within the phospholipid tails. All together these results support the model that lipoquinones fold regardless of the headgroup structure but that the polarity of the headgroup influences lipoquinone location within the membrane interface. These results have implications regarding the redox activity near the interface as quinone vs. quinol forms may facilitate locomotion of lipoquinones within the membrane. The location, orientation, and conformation of lipoquinones are critical for their function in generating cellular energy within membrane ETS, and the studies described herein shed light on the behavior of lipoquinones within membrane-like environments.


Population pharmacokinetics of APOMINE: a meta-analysis in cancer patients and healthy males.
1) To characterize the population pharmacokinetics of apomine in healthy males and in male and female patients with solid tumours and 2) to understand more fully the influence of induction and between- and within-subject variability on exposure to drug using Monte Carlo simulation. Apomine was administered once- or twice-daily with or without food in single and multiple oral doses of 30-2100 mg to healthy males (n = 19) and patients with solid tumours (n = 19). The data were divided into model development and validation sets. Models were developed using standard population methods. These were the identification of an appropriate base model, calculation of the empirical Bayes estimates of the primary pharmacokinetic parameters, covariate screening, forward stepwise addition of covariates using the likelihood ratio test as a model selection criteria, and backwards elimination to obtain the final model. To study the influence of data from individual subjects, the model development dataset was subjected to the delete-1 jack-knife and the final model was fitted to each jack-knifed dataset. Principal components analysis of the jack-knifed matrix of model parameters identified two influential subjects who were removed from the dataset, and the final model contained data from the remaining subjects. Model validation was examined using goodness of fit statistics and relative error measures using independent datasets from cancer patients. The model provided a reasonable approximation to the pharmacokinetic measurements in the validation datasets. Computer simulations were undertaken to understand further the pharmacokinetics of apomine in otherwise healthy females, a population not yet studied. Apomine pharmacokinetics were complex and consistent with a two-compartment model with a lag-time. Apparent oral clearance at baseline and apparent volume of distribution at steady-state were larger in healthy males than in cancer patients (41 ml h(-1) and 14.1 l vs 10 ml h(-1) and 8.9 l, respectively, for a 75 kg person). Clearance was time-variant showing a maximal increase with full induction of 320 ml h(-1), independent of patient type. The time to reach 50% maximal induction was about 2 days. The fraction of drug absorbed was relatively constant at doses less than 100-200 mg once daily but decreased at higher doses. Food also decreased relative bioavailability by 36%. Patient characteristics had no effect on apomine pharmacokinetics except for weight, which was proportional to the volume of the central compartment. Between-subject variability (68% for clearance, 30% for central volume, and 141% for peripheral volume) was moderate to large and independent of patient type. Inter-occasion variability was small (18% for both clearance and central volume). Residual variability was modelled with an additive and proportional error model. Cancer patients had slightly higher plasma concentrations than healthy males but this difference was probably not clinically significant. Steady-state was reached in about 3-4 days after once-daily drug administration. The half-life of apomine after three weeks of once-daily dosing was 41 h in cancer patients and 32 h in healthy males. A population model for apomine has been developed has been developed that characterizes its pharmacokinetics in cancer patients and healthy subjects under a variety of conditions.


Carbamate kinase of Lactobacillus buchneri NCDO110. II. Kinetic studies and reaction mechanism.
The participation of Mg2+ or Mn2+ nucleoside diphosphates in the reverse reaction catalyzed by purified carbamate kinase (ATP:carbamate phosphotransferase, EC 126.96.36.199) of Lactobacillus buchneri NCDO110 was studied. The results of initial velocity studies have indicated that Mn2+ ADP is as effective as a substrate as Mg2+ ADP is. Product inhibition studies have revealed that the enzyme has two distinct sites, one for nucleoside diphosphate and the other for carbamyl phosphate. The reaction of the enzyme with the substrates is of the random type.


Cuspal flexure, depth-of-cure, and bond integrity of bulk-fill composites.
Evaluate cuspal flexure caused by polymerization shrinkage stress, bond integrity, and depth-of-cure of bulk-fill composites. Twenty-eight extracted permanent molars were mounted in stainless steel rings. Slot-shaped Class II mesio-occlusal-distal preparations (four mm deep) were restored with Filtek Supreme Ultra (control; two two-mm increments) and bulk-fill composites (Tetric EvoCeram, Venus, Filtek Bulk Fill). The teeth were digitized using a 3D scanner before and after restoration. Before- and after-restoration scans were aligned, and cuspal flexure was calculated. Bond integrity along occlusal interfaces was assessed by dye penetration and measured after overnight immersion in basic fuchsin dye and cross-sectioning. Depth-of-cure was determined on the cross-sections using Vickers hardness. Statistical analysis was performed with one-way analysis of variance. All composites caused inward cuspal flexure (10.4 to 13.6 μm). No statistical difference in flexure was found among the composites (P=.07). No significant difference in microhardness at any restoration depths was found for any composite (.35<P<.99). Dye penetration showed comparable bonding along occlusal interfaces for the tested composites. Bulk-fill composites cured all the way through four-mm restoration depths. Cuspal flexure, caused by polymerization shrinkage, and bond integrity were not different between teeth restored with the bulk-fill and conventional incrementally placed restorative composites.


Bisphosphonates inhibit stromelysin-1 (MMP-3), matrix metalloelastase (MMP-12), collagenase-3 (MMP-13) and enamelysin (MMP-20), but not urokinase-type plasminogen activator, and diminish invasion and migration of human malignant and endothelial cell lines.
Bisphosphonates (clodronate, alendronate, pamidronate and zoledronate) at therapeutically attainable non-cytotoxic concentrations inhibited MMP-3, -12, -13 and -20 as well as MMP-1, -2, -8 and -9, but not urokinase-type plasminogen activator (uPA), a serine proteinase and a pro-MMP activator. Dose-dependent inhibition was shown by three independent MMP assays. The inhibition was reduced in the presence of an increased concentration of Ca(2+) when compared to physiologic Ca(2+) concentration. Alendronate inhibited the in vitro invasion (Matrigel) of human HT1080 fibrosarcoma and C8161 melanoma cells, and the random migration of these malignant and endothelial cell lines capable of expressing MMPs and uPA. The concentration of alendronate required to inhibit 50% of the activity (IC(50)=40-70 microM) of MMPs corresponded to the IC(50) of down-regulation of in vitro invasion and migration. The ability of bisphosphonates to down-regulate the in vitro invasion and random migration was comparable or slightly better in relation to the selective gelatinase inhibitor CTTHWGFTLC peptide. Alendronate but not CTTHWGFTLC peptide promoted the adhesion of HT1080 fibrosarcoma and C8161 melanoma cell lines on fibronectin. Bisphosphonates are broad-spectrum MMP inhibitors and this inhibition involves cation chelation. Bisphosphonates further exert antimetastatic, anti-invasive and cell adhesion-promoting properties, which may prevent metastases not only into hard tissues but also to soft tissues.


Urinary and hemodynamic responses to blood volume changes in fetal sheep.
Because of the potential importance of the fetal kidney in regulating blood volume and amniotic fluid volume, and their composition, we studied renal and cardiovascular responses to changes in blood volume in 12 chronically-catheterized fetal sheep at 110-135 days gestation. During the hour following volume expansion with 50 ml of maternal whole blood, blood pressure rose by 14%, cardiac output by 11%, renal blood flow by 68% and urine output by 36%. Packed red cell volume rose from 31.3 to 34.0%, suggesting a net fluid loss from the circulation through the kidneys, as well as through peripheral and placental capillaries. During the hour following volume depletion by haemorrhage of 70-120 ml, blood pressure fell by 17%, cardiac output by 29%, renal flow by 46%, glomerular filtration rate by 19%, and urine output by 76%, compared to the preceding phase of the study. The urine-to-plasma osmolality ratio approached unity and urine sodium and U/Pcreatinine increased sharply. Following haemorrhage, fluid rapidly entered the circulation as evidenced by falling packed red cell volume and plasma protein concentration. The data support the hypothesis that the fetal kidney, although immature in some respects, effectively responds to maintain blood volume in the fetal sheep.


Recession of the soft tissue margin at oral implants. A 2-year longitudinal prospective study.
The aim of this study was to longitudinally evaluate alterations in the position of the periimplant soft tissue margin, occurring during a 2-year period after insertion of fixed prostheses. 41 patients, subjected to treatment with partial or full-arch implant supported fixed prostheses based on a total of 163 standard Brånemark oral implants, were included in the study. A baseline examination was performed at time of insertion of the prosthetic construction and involved assessments of plaque, mucositis, probing depth, bleeding on probing marginal soft tissue level, width of masticatory mucosa and marginal soft tissue mobility. All patients were re-examined after 6 months, and after 1 and 2 years, respectively. A regression analysis was performed to evaluate if alterations, occurring in the position of the periimplant soft tissue margin during the 2 years, could be predicted by various variables recorded at time of the installation of the fixed prosthesis. The descriptive analysis showed a slight decrease in mean probing depth (0.2 mm) and width of masticatory mucosa (0.3 mm) during the follow-up period. Apical displacement of the soft tissue margin mainly took place during the first 6 months of observation. Lingual sites in the mandible showed the most pronounced soft tissue recession, decrease of probing depth, and decrease of width of masticatory mucosa. The statistical analysis revealed that lack of masticatory mucosa and mobility of the periimplant soft tissue at time of bridge installation were poor predictors of soft tissue recession occurring during the 2 years of follow-up, but indicated in greater amount of recession in women than men, in the mandible than in the maxilla, at lingual than at facial sites and with increased initial probing depth. It was suggested that the recession of the periimplant soft tissue margin mainly may be the result of a remodelling of the soft tissue in order to establish "appropriate biological dimensions" of the periimplant soft tissue barrier, i.e., the required dimension of epithelial-connective tissue attachment in relation to the faciolingual thickness of the supracrestal soft tissue.


Upregulation of D site of albumin promoter binding protein in the brain of patients with intractable epilepsy.
The mechanisms that underlie the pathogenesis of intractable epilepsy (IE) remain to be elucidated. The present study aimed to investigate the expression of D site of albumin promoter binding protein (DBP) and mitogen‑activated protein kinases (MAPKs) in the temporal lobes of patients with IE, in order to examine the possible roles of DBP in the pathogenesis of IE. The expression of DBP and MAPK was detected by immunohistochemistry and double‑label immunofluorescence staining against DBP/MAPK in 35 patients with IE, and the data were compared with those of the 15 controls. The results demonstrated that DBP expression in IE group (0.31±0.03) was significantly higher compared with that in the controls (0.18±0.02; P<0.05) and MAPK expression in the IE group (0.19±0.03) was also higher compared with that in the controls (0.12±0.02; P<0.05). DBP and MAPK were mainly expressed in the cytoplasm of neurons and the double‑label immunofluorescence staining demonstrated that DBP and MAPK expression occurred in the same neurons. Therefore, the expression of DBP and MAPK in epilepsy patients was upregulated, suggesting a possible pathogenetic role in IE.


Sepsis-related mortality in the Czech Republic: multiple causes of death analysis.
Sepsis represents an increasingly frequent complication in the survival of the hospitalized persons. The nation-wide burden of the sepsis-related mortality in the Czech Republic has not yet been analysed. The present study aims to assess the trends and the disparities in the sepsis-related mortality in the Czech Republic for the period 1998-2011 using multiple causes of death reported on the death certificates. Individual death records provided by the Institute for Health Information and Statistics were used to identify sepsis-related deaths based on the codes of the 10th revision of the International classification of the diseases (ICD-10). Frequencies, age-standardized rates and comorbidity patterns were studied. A total of 41,425 cases (3%) out of 1,456,539 deaths within the period 1998-2011 were associated with sepsis. In 2011, sepsis was reported in 8% of all in-hospital deaths. Sepsis is the most likely to be reported as immediate cause of death (66%), which results in its considerable underestimation in the underlying cause of death statistics. The sepsis-related mortality almost tripled between 1998 and 2011, and the most of the increase is attributable to persons aged 65 and over. In 44% of cases, circulatory disease or cancer was reported as the underlying cause of death. Significant associations with sepsis were however found for infectious diseases, diseases of skin, and metabolic or musculoskeletal disorders. Multiple cause of death analysis of sepsis-related mortality revealed that sepsis represents a growing burden related to the population ageing and increased prevalence of complications of chronic diseases. The observed upward trends, as well as the expected continuation of the ageing process, may result in further increase of sepsis-related mortality. Preventive measures in the clinical management of sepsis are recommended.


NETQUANT: Automated Quantification of Neutrophil Extracellular Traps.
Neutrophil extracellular traps (NETs) that are extensive webs of DNA covered with antimicrobial proteins into the extracellular environment during infection or inflammation as a part of their defense arsenal. Image acquisition of fluorescently labeled NETs and subsequent image-based quantification is frequently used to analyze NET formation (NETosis) in response to various stimuli. However, there are important limitations in the present methods for quantification. Manual methods tend to be error-prone, tedious, and often quite subjective, whereas the software-rooted options are either semi-automatic or difficult to operate. Here, we present an automated and uncomplicated approach for quantifying NETs from fluorescence images, built as a freely available app for MATLAB®. It is based on detection of a set of clearly defined parameters, all related to the biological manifestation of NETs and allowing for single-cell resolution quantification and analysis.


Molecular cloning and characterization of Bifidobacterium bifidum 1,2-alpha-L-fucosidase (AfcA), a novel inverting glycosidase (glycoside hydrolase family 95).
A genomic library of Bifidobacterium bifidum constructed in Escherichia coli was screened for the ability to hydrolyze the alpha-(1-->2) linkage of 2'-fucosyllactose, and a gene encoding 1,2-alpha-l-fucosidase (AfcA) was isolated. The afcA gene was found to comprise 1,959 amino acid residues with a predicted molecular mass of 205 kDa and containing a signal peptide and a membrane anchor at the N and C termini, respectively. A domain responsible for fucosidase activity (the Fuc domain; amino acid residues 577 to 1474) was localized by deletion analysis and then purified as a hexahistidine-tagged protein. The recombinant Fuc domain specifically hydrolyzed the terminal alpha-(1-->2)-fucosidic linkages of various oligosaccharides and a sugar chain of a glycoprotein. The stereochemical course of the hydrolysis of 2'-fucosyllactose was determined to be inversion by using (1)H nuclear magnetic resonance. The primary structure of the Fuc domain exhibited no similarity to those of any glycoside hydrolases (GHs) but showed high similarity to those of several hypothetical proteins in a database. Thus, it was revealed that the AfcA protein constitutes a novel inverting GH family (GH family 95).


Vitamin D3 supplementation reduces serum markers of bone resorption and muscle damage in female basketball players with vitamin D inadequacy.
This study aimed to investigate the effect of vitamin D3 supplementation at a dose of 4,000 IU/day for 6 weeks on serum levels of: 25-hydroxyvitamin D [25(OH)D], bone turnover [osteocalcin and carboxy-terminal telopeptides of crosslinks of type I collagen (CTx-I)], and muscle damage [lactate dehydrogenase (LDH) and creatine kinase (CK)] in middle adolescent (15-18 years) and late-adolescent to early adulthood (19-30 years) female basketball players with inadequate vitamin D status. Participants (N=24) were randomly assigned in a double-blind manner into a vitamin D or placebo group. Data were analyzed using separate 2×2x2 mixed ANOVAs with one within-subjects factor (time) and two between-subjects factors (condition and age). When a significant interaction was observed, the intra-group changes were assessed using paired t tests and Hedge's g. Paired t-tests comparing intra-group changes in the vitamin D condition revealed a non-significant large improvement in 25(OH)D (p = 0.06; g = 0.86), non-significant small decreases in CTx-I (p = 0.13; g = -0.22) and CK (p = 0.07; g = -0.26), as well as a significant moderate decrease in LDH (p = 0.004; g = -0.74). Paired t-tests comparing intra-group changes in the placebo condition revealed a significant moderate decline in 25(OH)D (p <0.001; g = -0.77), as well as significant small increases in CTx-I (p = 0.04; g = 0.47) and CK (p = 0.04; g = 0.36). Vitamin D3 supplementation at 4,000 IU/day could be effective in reducing bone resorption and muscle damage in female basketball players with inadequate baseline vitamin D, irrespective of age.


Pelvic MRI in children with Crohn disease and suspected perianal involvement.
MRI is an important imaging tool in evaluation of adult and pediatric patients with Crohn disease. Pelvic MRI, in particular, has become the method of choice to evaluate for perianal fistulas and associated complications of Crohn disease. MRI can define the extent and location of perianal fistulas and abscesses, as well as provide critical information for operative management. In this pictorial essay, we describe useful MRI techniques for evaluation of perianal complications in pediatric patients with Crohn disease. We review pertinent anatomy and illustrate typical examples of perianal fistulas with and without abscess. We show one case of clinically suspected perianal fistula that was actually a pilonidal sinus.


Prognostic value and the changes of plasma levels of secretory type II phospholipase A2 in patients with coronary artery disease undergoing percutaneous coronary intervention.
To evaluate the serial changes of plasma secretory type II phospholipase A(2)(sPLA(2)), C-reactive protein (CRP) and cardiac injury markers in coronary artery disease (CAD) patients undergoing percutaneous coronary intervention (PCI) and their prognostic impacts. Plasma levels of sPLA(2), CRP, creatine kinase (CK), CK-MB and troponin-T were measured in 247 consecutive CAD patients receiving PCI procedure and 100 control subjects without CAD. In CAD group, serial blood samples were taken before coronary angiography, after coronary angiography, immediately after PCI, 24-h and 48-h after PCI. The sPLA(2)and CRP levels did not change after coronary angiography. The level of sPLA(2)significantly increased immediately after PCI. Creatine kinase and cardiac injury markers did not rise immediately after PCI, but elevated significantly at 24h after intervention. After a 2-year follow up, increased sPLA(2)(>450 ng/dl) after PCI, smoking and diabetes mellitus were the independent risk factors for subsequent coronary events (odds ratios 2.1, 2.3 and 3.1, respectively) in patients with CAD. The present study showed that PCI might cause immediate elevation of circulating levels of sPLA(2)following the mechanical disruption of coronary plaque, and the elevated level of sPLA(2)had significant prognostic impact.


Prognostic value of the 12-lead surface electrocardiogram in sarcomeric hypertrophic cardiomyopathy: data from the REMY French register.
To identify independent electrocardiogram (ECG) predictors of long-term clinical outcome based on standardized analysis of the surface ECG in a large multicentre cohort of patients with sarcomeric hypertrophic cardiomyopathy (HCM). Retrospective observational study from the REMY French HCM clinical research observatory. Primary endpoint was a composite of all-cause mortality, major non-fatal arrhythmic events, hospitalization for heart failure (HF), and stroke. Secondary endpoints were components of the primary endpoint. Uni- and multivariable Cox proportional hazard regression analysis was performed to identify independent predictors. Among 994 patients with HCM, only 1.8% had a strictly normal baseline ECG. The most prevalent abnormalities were inverted T waves (63.7%), P-wave abnormalities (30.4%), and abnormal Q waves (25.5%). During a mean follow-up of 4.0 ± 2.0 years, a total of 272 major cardiovascular events occurred in 217 patients (21.8%): death or heart transplant in 98 (9.8%), major arrhythmic events in 40 (4.0%), HF hospitalization in 115 (11.6%), and stroke in 23 (2.3%). At multivariable analysis using ECG covariates, prolonged QTc interval, low QRS voltage, and PVCs of right bundle branch block pattern predicted worse outcome, but none remained independently associated with the primary endpoint after adjustment on main demographic and clinical variables. For secondary endpoints, abnormal Q waves independently predicted all-cause death [hazard ratio (HR) 2.35, 95% confidence interval (CI) 1.23-4.47; P = 0.009] and prolonged QTc the risk of HF hospitalization (HR 1.006, 95% CI 1.001-1.011; P = 0.024). The 12-lead surface ECG has no independent value to predict the primary outcome measure in patients with HCM. The 12-lead surface ECG has been widely used as a screening tool in HCM but its prognostic value remains poorly known. The value of baseline surface ECG to predict long-term clinical outcomes was studied in a cohort of 994 patients with sarcomeric HCM. The surface ECG has no significant additional value to predict outcome in this patient population.


Teratological evaluation of gamma-glycidoxypropyltrimethoxysilane in rats.
The embryotoxic and teratogenic potential of gamma- glycidoxypropyltrimethoxysilane ( GPTS ) was evaluated in rats. Pregnant Sprague-Dawley rats were administered 0, 50, 500, or 1,000 mg/kg/day of GPTS by gavage on days 6 through 15 of gestation. No treatment related signs of toxicity, behavioral alterations or mortalities were observed in any of the pregnant animals. There was no evidence of adverse effects in mean maternal body weight, liver weight or food consumption of the treated females. The number of implantation sites, number of live fetuses per litter, the mean litter size, the sex ratio, the fetal body weight or the crown-rump length were not affected by treatment. The incidence of resorptions among the total fetal population was not altered by the administration of GPTS to pregnant rats, indicating that the test material is not embryolethal in rats at the tested dose levels. Few scattered incidences of fetal alterations in the external, soft tissue or skeletal examinations were seen both among treated and untreated litters, however, no single alteration was observed in treated litters at an incidence which was significantly different from the control. In conclusion GPTS was not embryotoxic or teratogenic in rats at dose levels up to 1000 mg/kg/day.


Methodologies for bacterial ribonuclease characterization using RNA-seq.
Bacteria adjust gene expression at the post-transcriptional level through an intricate network of small regulatory RNAs and RNA-binding proteins, including ribonucleases (RNases). RNases play an essential role in RNA metabolism, regulating RNA stability, decay, and activation. These enzymes exhibit species-specific effects on gene expression, bacterial physiology, and different strategies of target recognition. Recent advances in high-throughput RNA sequencing (RNA-seq) approaches have provided a better understanding of the roles and modes of action of bacterial RNases. Global studies aiming to identify direct targets of RNases have highlighted the diversity of RNase activity and RNA-based mechanisms of gene expression regulation. Here, we review recent RNA-seq approaches used to study bacterial RNases, with a focus on the methods for identifying direct RNase targets.


Contribution of a Streptococcus mutans antigen expressed by a Salmonella vector vaccine in dendritic cell activation.
A Salmonella vector vaccine expressing the saliva-binding region (SBR) of the adhesin AgI/II of Streptococcus mutans has been shown to induce a mixed Th1/Th2 anti-SBR immune response in mice and to require Toll-like receptor 2 (TLR2), TLR4, and MyD88 signaling for the induction of mucosal anti-SBR antibody responses. Since dendritic cells (DC) are critical in innate and adaptive immunity, the present study assessed the role of SBR expression by the vector vaccine in DC activation. Bone marrow-derived DC from wild-type and TLR2, TLR4, and MyD88 knockout mice were stimulated with Salmonella vector BRD509, the SBR-expressing Salmonella vector vaccine BRD509(pSBRT7), or SBR protein, and the DC responses to different stimuli were compared by assessing costimulatory molecule expression, cytokine production, and signaling pathways. The DC response to both BRD509(pSBRT7) and BRD509 was dependent mainly on TLR4. BRD509(pSBRT7) and BRD509 induced upregulation of CD80, CD86, CD40, and major histocompatibility complex class II (MHC II) expression. Lower levels of interleukin-10 (IL-10) and IL-12p40 were produced by BRD509(pSBRT7)-stimulated DC than by BRD509-stimulated DC. Furthermore, BRD509(pSBRT7)-stimulated DC showed decreased p38 phosphorylation compared to that induced by DC stimulated with BRD509. However, BRD509(pSBRT7)-treated DC produced a higher level of IL-6 than BRD509-stimulated cells. The low IL-12p40 and high IL-6 cytokine profile expressed by BRD509(pSBRT7)-stimulated DC may represent a shift toward a Th2 response, as suggested by the increased expression in Jagged-1. These results provide novel evidence that a heterologous protein expressed by a Salmonella vector vaccine can differentially affect DC activation.


Evaluating medical technology in the context of a fiscal crisis: the case of New Zealand.
Planning for health services in New Zealand, and the implications of increasing use of high technology, fail to take account of the mounting crisis in the country's economy. Social policy that ignores structural problems in the economy or the system of medical care is unrealistic. A strategy is advanced to determine what resources are likely to be available, and how they can be allocated to ensure value for money. The situation is not unique to New Zealand.


CAMPR3: a database on sequences, structures and signatures of antimicrobial peptides.
Antimicrobial peptides (AMPs) are known to have family-specific sequence composition, which can be mined for discovery and design of AMPs. Here, we present CAMPR3; an update to the existing CAMP database available online at www.camp3.bicnirrh.res.in. It is a database of sequences, structures and family-specific signatures of prokaryotic and eukaryotic AMPs. Family-specific sequence signatures comprising of patterns and Hidden Markov Models were generated for 45 AMP families by analysing 1386 experimentally studied AMPs. These were further used to retrieve AMPs from online sequence databases. More than 4000 AMPs could be identified using these signatures. AMP family signatures provided in CAMPR3 can thus be used to accelerate and expand the discovery of AMPs. CAMPR3 presently holds 10247 sequences, 757 structures and 114 family-specific signatures of AMPs. Users can avail the sequence optimization algorithm for rational design of AMPs. The database integrated with tools for AMP sequence and structure analysis will be a valuable resource for family-based studies on AMPs.


Visual short-term memory through the lifespan: Preserved benefits of context and metacognition.
Visual short-term memory (VSTM) ability falls throughout the life span in healthy adults. Using a continuous report task, in a large, population-based sample, we first confirmed that this decline affects the quality and quantity of reported memories as well as knowledge of which item went where. Visual and sensorimotor precision also worsened with advancing age, but this did not account for the reduced memory performance. We then considered two strategies that older individuals might be able to adopt, to offset these memory declines: the use of contextual encoding, and metacognitive monitoring of performance. Context and metacognitive awareness were both associated with significantly better performance, however these effects did not interact with age in our sample. This suggests that older adults retain their capacity to boost memory performance through attention to external context and monitoring of their performance. Strategies that focus on taking advantage of these preserved abilities may therefore help to maintain VSTM performance with advancing age. The article reports on analysis of the Cambridge Centre for Ageing and Neuroscience (Cam-CAN) data. (PsycINFO Database Record


Protein carbamylation: in vivo modification or in vitro artefact?
Carbamylation (carbamoylation) of lysine residues and protein N-termini is a nonenzymatic PTM that has been related to protein ageing. In contrast to other PTM, such as phosphorylation, carbamylation can be artificially introduced during sample preparation with urea, thus affecting studies directed toward in vivo carbamylation. In aqueous solution, urea-commonly used for denaturing proteins-is in equilibrium with ammonium and isocyanate. Under alkaline conditions, the latter can react with primary amines of free N-termini and ε-amine groups of lysines to form carbamyl derivatives. Despite being a relatively slow process, which is accelerated at elevated temperatures, prolonged incubation of protein/peptide samples in urea buffers can induce undesired carbamylation, hampering not only the proteolytic digestion with trypsin and peptide identification by MS, but also interfering with stable isotope-labeling techniques such as iTRAQ, tandem mass tags, and isotope-coded protein labeling. Here, we evaluated the extent of urea-induced carbamylation under commonly used sample preparation conditions. From our results, we can deduce that carbamylation occurs in all cases involving urea, however with varying degree: e.g. carbamidomethylation in the presence of 8.0 M urea induced carbamylation of 17% of N-termini and 4% of Lys residues. Additionally, researching a recently published large-scale dataset revealed a high degree of urea-induced carbamylation in current proteomic samples.


Microplastic Contamination in Karst Groundwater Systems.
Groundwater in karst aquifers constitutes about 25% of drinking water sources globally. Karst aquifers are open systems, susceptible to contamination by surface-borne pollutants. In this study, springs and wells from two karst aquifers in Illinois, USA, were found to contain microplastics and other anthropogenic contaminants. All microplastics were fibers, with a maximum concentration of 15.2 particles/L. The presence of microplastic was consistent with other parameters, including phosphate, chloride and triclosan, suggesting septic effluent as a source. More studies are needed on microplastic sources, abundance, and impacts on karst ecosystems.


Data for Genetic Analysis Workshop 18: human whole genome sequence, blood pressure, and simulated phenotypes in extended pedigrees.
Genetic Analysis Workshop 18 (GAW18) focused on identification of genes and functional variants that influence complex phenotypes in human sequence data. Data for the workshop were donated by the T2D-GENES Consortium and included whole genome sequences for odd-numbered autosomes in 464 key individuals selected from 20 Mexican American families, a dense set of single-nucleotide polymorphisms in 959 individuals in these families, and longitudinal data on systolic and diastolic blood pressure measured at 1-4 examinations over a period of 20 years. Simulated phenotypes were generated based on the real sequence data and pedigree structures. In the design of the simulation model, gene expression measures from the San Antonio Family Heart Study (not distributed as part of the GAW18 data) were used to identify genes whose mRNA levels were correlated with blood pressure. Observed variants within these genes were designated as functional in the GAW18 simulation if they were nonsynonymous and predicted to have deleterious effects on protein function or if they were noncoding and associated with mRNA levels. Two simulated longitudinal phenotypes were modeled to have the same trait distributions as the real systolic and diastolic blood pressure data, with effects of age, sex, and medication use, including a genotype-medication interaction. For each phenotype, more than 1000 sequence variants in more than 200 genes present on the odd-numbered autosomes individually explained less than 0.01-2.78% of phenotypic variance. Cumulatively, variants in the most influential gene explained 7.79% of trait variance. An additional simulated phenotype, Q1, was designed to be correlated among family members but to not be associated with any sequence variants. Two hundred replicates of the phenotypes were simulated, with each including data for 849 individuals.


The role of intracellular redox imbalance in nanomaterial induced cellular damage and genotoxicity: a review.
The terms oxidative stress, free radical generation, and intracellular antioxidant protection have become part of everyday nanotoxicology terminology. In recent years, an ever increasing number of in vitro and in vivo studies have implicated disruptions to the redox balance and oxidative stress as one of the main contributors to nanomaterial (NM) induced adverse effects. One of the most important and widely investigated of these effects is genotoxicity. In general, systems that defend an organism against oxidative damage to DNA are very complex and include prevention of reactive oxygen species (ROS) production, neutralizing ROS (scavengers), enzymatic nucleotide pool sanitation, and DNA repair. This review discusses the importance of the maintenance of the redox balance in this context before examining studies that have investigated engineered NM induced redox imbalance and genotoxicity. Furthermore, we identify data gaps, and highlight a number of issues that exist with the methodologies that are routinely utilized to investigate intracellular ROS production or anti-oxidant depletion. We conclude that for a large number of engineered NM types changes in the redox balance toward oxidative stress are normally associated with DNA damage.


Adenosylhomocysteinase from yellow lupin seeds. Purification and properties.
Adenosylhomocysteinase from yellow lupin seeds (Lupinus luteus) has been purified to homogeneity. Active enzyme, Mr = 110000, consists of two probably identical subunits with Mr = 55000 as judged by gel filtration and dodecyl sulphage/polyacrylamide gel electrophoresis in the presence of 2-mercaptoethanol. The isoelectric point of the enzyme was shown to be 4.9 +/- 0.1. It was demonstrated by disc and pore gradient electrophoresis that the most purified fraction formed multimers. The enzyme shows optimum activity at pH 8.5-9.0. Km values are 2.3 micrometer, 4.6 mM and 12 micrometer for adenosine, DL-homocysteine and S-adenosyl-L-homocysteine, respectively. The energy of activation for S-adenosylhomocysteine synthesis was estimated as 14.4 kcal/mol (60.2 kJ/mol) and temperature coefficient as 2.4. The equilibrium constant for the hydrolysis of S-adenosylhomocysteine amounts to 5 X 10(-7) M. Anti-sulfhydryl reagents such as p-hydroxymercuribenzoate and N-ethylmaleimide acted as irreversible inhibitors. The enzyme exhibits high specificity for homocysteine whereas some of the rare nucleosides tested could substitute for adenosine.


Distal skeletal tibia assessed by HR-pQCT is highly correlated with femoral and lumbar vertebra failure loads.
Dual energy X-ray absorptiometry (DXA) is the standard for assessing fragility fracture risk using areal bone mineral density (aBMD), but only explains 60-70% of the variation in bone strength. High-resolution peripheral quantitative computed tomography (HR-pQCT) provides 3D-measures of bone microarchitecture and volumetric bone mineral density (vBMD), but only at the wrist and ankle. Finite element (FE) models can estimate bone strength with 86-95% precision. The purpose of this study is to determine how well vBMD and FE bone strength at the wrist and ankle relate to fracture strength at the hip and spine, and to compare these relationships with DXA measured directly at those axial sites. Cadaveric samples (radius, tibia, femur and L4 vertebra) were compared within the same body. The radius and tibia specimens were assessed using HR-pQCT to determine vBMD and FE failure load. aBMD from DXA was measured at the femur and L4 vertebra. The femur and L4 vertebra specimens were biomechanically tested to determine failure load. aBMD measures of the axial skeletal sites strongly correlated with the biomechanical strength for the L4 vertebra (r=0.77) and proximal femur (r=0.89). The radius correlated significantly with biomechanical strength of the L4 vertebra for vBMD (r=0.85) and FE-derived strength (r=0.72), but not with femur strength. vBMD at the tibia correlated significantly with femoral biomechanical strength (r=0.74) and FE-estimated strength (r=0.83), and vertebral biomechanical strength for vBMD (r=0.97) and FE-estimated strength (r=0.91). The higher correlations at the tibia compared to radius are likely due to the tibia's weight-bearing function.


Pharmacological treatment for Type 2 diabetes integrating findings from cardiovascular outcome trials: an expert consensus in the UK.
In people with Type 2 diabetes, cardiovascular disease is a leading cause of morbidity and mortality. Thus, as well as controlling glucose, reducing the risk of cardiovascular events is a key goal. The results of cardiovascular outcome trials have led to updates for many national and international guidelines. England, Wales and Northern Ireland remain exceptions, with the most recent update to the National Institute for Health and Care Excellence (NICE) guidelines published in 2015. We reviewed current national and international guidelines and recommendations on the management of people with Type 2 diabetes. This article shares our consensus on clinical recommendations for the use of sodium-glucose co-transporter 2 inhibitors (SGLT-2is) and glucagon-like peptide 1 receptor agonists (GLP-1RAs) in people with Type 2 diabetes and established or at very high risk of cardiovascular disease in the UK. We also consider cost-effectiveness for these therapies. We recommend considering each person's cardiovascular risk and using diabetes therapies with proven cardiovascular benefits when appropriate to improve long-term outcomes and cost-effectiveness.


Social bonds and posttraumatic stress disorder.
Retrospective and prospective studies consistently show that individuals exposed to human-generated traumatic events carry a higher risk of developing Posttraumatic Stress Disorder (PTSD) than those exposed to other kinds of events. These studies also consistently identify perceptions of social support both before and after a traumatic event as an important factor in the determining vulnerability to the development of PTSD. We review the literature on interpersonal traumas, social support and risk for PTSD and integrate findings with recent advances in developmental psychopathology, attachment theory and social neuroscience. We propose and gather evidence for what we term the social ecology of PTSD, a conceptual framework for understanding how both PTSD risk and recovery are highly dependent on social phenomena. We explore clinical implications of this conceptual framework.


Lack of PTEN expression in non-small cell lung cancer could be related to promoter methylation.
The PTEN gene at chromosome 10q23.3 is a tumor-suppressor genethat is inactivated in several types of human tumors. Althoughmutation and homozygous deletion are the most commonmechanisms of PTEN inactivation, promoter methylation and translational modification can also account for PTEN silencing. The aim of this study was to investigate the expression of PTEN protein in primary non-small cell lung cancer (NSCLC) samples and to investigate the promoter methylation status of the gene in a panel of NSCLC cell lines as well as primary tumors. We analyzed PTEN expression by immunohistochemistry in tissue samples from 125 patients with early-stage NSCLC. We also evaluated PTEN promoter methylation status by methylation-specific PCR in 20 microdissected PTEN-negative primary tumors from among the last specimens as well as in a panel of 16 NSCLC cell lines. Western and Northern blotting were performed in the same panel of NSCLC cell lines. Thirty (24%) of the 125 specimens showed a lack of staining for PTEN. PTEN methylation was detected in 7 (35%) of the 20 PTEN-negative NSCLC samples and in none of the 10 PTEN-positive NSCLC samples that were microdissected. Furthermore, PTEN methylation was observed in 11 (69%) of the 16 NSCLC cell lines tested. PTEN mRNA expression was increased in the NCI-H1299 cell line by in vitro treatment with the demethylating agent 5-aza-2'-deoxycytidine. PTEN methylation was well correlated with PTEN expression in NSCLC cell lines by Western and Northern blot (P = 0.025). Although genetic alterations of the PTEN gene are rare in NSCLC, loss of PTEN protein is not an uncommon event in early-stage NSCLC. Lack of PTEN expression may be partially explained by promoter methylation.


Nanoparticle-electrode collisions as a dynamic seeding route for the growth of metallic nanostructures.
The collisions between colloidal metal nanoparticles and a carbon electrode were explored as a dynamic method for the electrodeposition of a diverse range of electrocatalytically active Ag and Au nanostructures whose morphology is dominated by the electrostatic interaction between the charge of the nanoparticle and metal salt.


Prognostic and predictive significance of GITR in metastatic renal cell carcinoma.
Renal cell carcinoma (RCC) has gradually increased in recent years. There have been significant developments in metastatic RCC in recent years with the introduction of immune control point inhibitors. Glucocorticoid-induced tumor necrosis factor (TNF) receptor-related protein (GITR) is a co-stimulatory molecule and is seen in the highest amounts in activated CD4+ T lymphocytes and CD8+ T lymphocytes, forkhead box protein 3 (FOXP3) positive regulatory T cells (Treg). GITR leads to an increase in interleukin (IL)-2 and CD25 and Interferon Gamma. It shows an anti-tumoural effect by inhibiting the suppressive functions of FOXP3+ regulatory cells (Treg). Therefore, we aimed to evaluate the prognostic and predictive effect of GITR, tumor-infiltrating lymphocytes (CD4+CD8) (TIL), and FOXP3 in patients with metastatic RCC. Patients diagnosed with pathologically confirmed metastatic renal cancer between 2016 and 2021 were included in our study. Clinicopathological features and some laboratory tests were recorded. GITR, CD4, CD8, and FOXP3 were evaluated by immunohistochemistry (IHC) from biopsies or nephrectomy material and recorded. The study included 41 patients. The median progression-free survival (PFS) was 10.5 months, and the median overall survival (OS) was 13.9 months. Median PFS was 7.9 months for the GITR-low group and 18.9 months for the GITR-high group. Median PFS was statistically significant and longer for the GITR-high group than the GITR-low group (p=0.003). When patients who received nivolumab in the 2nd line were evaluated, median PFS was found to be 5.7 months in the GITR-low group and 15.7 months in the GITR-high group. Median PFS was statistically significantly higher in the GITR-high group than in the GITR-low group (p=0.026). In patients with metastatic RCC, higher GITR was associated with better PFS. At the same time, in patients using nivolumab, better PFS was seen in the GITR high group. If supported by prospective studies, GITR can be used as both a prognostic and predictive marker.


[Evaluation of Matas test by CBF studies, angiographic cross-filling and stump pressure in CEA patients].
Temporary and/or permanent occlusion of carotid artery is one of the most important strategy in the surgical treatment for cervical vascular and paraclinoid lesions. Cerebral ischemia produced by this procedure should be preoperatively grasped for the safety of the surgeries. Matas test has been widely applied to estimate the collateral blood flow at the main artery occlusion by observing only clinical features. In this study, we measured CBF and common carotid artery stump pressure (CCA stump P) in contrast to angiographic cross-filling in 43 carotid endarterectomy (CEA) patients to quantitatively evaluate Matas test. The patients were manually occluded the CCA on the affected side and clinically observed for at least 10 minutes. CCA stump P and cross-filling were evaluated in all patients and CBF measurements were carried out in the patients who clinically tolerated Matas test. CBF studies were performed by single photon emission CT with Xe-133 inhalation method and CBF values were obtained in the middle cerebral artery territory before and during Matas test. CCA stump P was measured during carotid angiography by manual occlusion of the proximal site of the CCA. Cross-filling was evaluated with the contralateral carotid angiography during the manual occlusion of the affected CCA. CBF changes in Matas test were classified into the following four patterns; bilateral mild decreases in CBF, unilateral severe decrease in CBF, unilateral mild decrease in CBF and no decrease in CBF. The frequency of bilateral mild decreases in CBF, unilateral severe decrease in CBF, unilateral mild decrease in CBF and no decrease in CBF were 8, 10, 14, 7 of 43 patients, respectively.(ABSTRACT TRUNCATED AT 250 WORDS)


Extinction Risk Assessment and Chemical Composition of Aerial Parts Essential Oils from Two Endangered Endemic Malagasy Salvia Species.
Seven essential oil samples of two endemic species of Malagasy sage, Salvia sessilifolia Baker and Salvia leucodermis Baker, were investigated via GC(RI), GC-MS and 13C NMR spectrometry. In total, 81compounds were identified accounting for 93.5% to 98.7% of the total composition. The main constituents for the both species were (E)-β-caryophyllene (29.2% to 60.1%), myrcene (1.2% to 21.7%), α-humulene (5.2% to 19.7%), (E)-nerolidol (0.8% to 15.5%) and caryophyllene oxide (1.4% to 10.8%). Ethnobotanical survey of 46 informants revealed that decoctions of leafy twigs and chewed leaves were usually used. Due to the repeated fires, over-harvesting and grazing, the populations of S. sessilifolia and S. leucodermis are drastically fragmented. These risk factors led to threats to the habitats of the target species. Salvia sessilifolia Baker and Salvia leucodermis Baker are proposed to be classified as endangered species.


Broadband time-domain absorption spectroscopy with a ns-pulse supercontinuum source.
A Q-switched laser based system for broadband absorption spectroscopy in the range of 1390-1740 nm (7200-5750 cm(-1)) has been developed and tested. In the spectrometer the 1064 nm light of a 25 kHz repetition-rate micro-chip Nd:YAG laser is directed into a photonic crystal fiber to produce a short (about 2 ns) pulse of radiation in a wide spectral range. This radiation is passed through a 25 km long dispersive single-mode fiber in order to spread the respective wavelengths over a time interval of about 140 ns at the fiber output. This fast swept-wavelength light source allows to record gas absorption spectra by temporally-resolved detection of the transmitted light power. The realized spectral resolution is about 2 cm(-1). Examples of spectra recorded in a cell with CO(2):CH(4):N(2) gas mixtures are presented. An algorithm employed for the evaluation of molar concentrations of different species from the spectra with non-overlapping absorption bands of mixture components is described. The uncertainties of the concentration values retrieved at different acquisition times due to the required averaging are evaluated. As an example, spectra with a signal-to-noise ratio large enough to provide species concentrations with a relative error of 5% can be obtained in real time at a millisecond time scale. Potentials and limitations of this technique are discussed.


Chronic rhinosinusitis in children: race and socioeconomic status.
Although chronic rhinosinusitis (CRS) is common in children, the influence of race and socioeconomic status (SES) on the diagnosis/treatment of CRS has not been evaluated. We describe the epidemiology of children with CRS in a pediatric otolaryngology clinic and evaluate demographic differences when compared to a group of children referred for a general pediatric otolaryngology evaluation. Historical cohort study. Tertiary academic care center. All new/consult patients (March 1, 2008-July 1, 2011) in a tertiary pediatric otolaryngology clinic with primary diagnosis of CRS were compared to a control group that consisted of all new/consult patients seen in the same clinic over 3 months. Records were evaluated for variables including age, sex, race, and insurance. Characteristics were compared between groups using Mann-Whitney and Fisher's exact tests. One hundred and seventy-four children with CRS were compared to 430 controls. When compared to the general pediatric otolaryngology population, children with CRS were older (8.2 ± 4.4 years vs 5.9 ± 4.8 years; P < .0001) and more commonly male (63% vs 52%; P = .018). When compared to controls, children with CRS were more likely to be white (CRS 77% white, 10% black, 13% other vs control 47% white, 33% black, 20% other; P < .0001, risk ratio [RR] = 2.7; 95% confidence interval [CI], 2.0-3.7). Likewise, children with CRS were less commonly insured with medical assistance (CRS 14% vs control 44%; P < .0001; RR = 0.3; 95% CI, 0.21-0.45). When compared to the general population of children seen in this academic urban pediatric otolaryngology setting, children with CRS were more likely to be white and privately insured. This study is the first to evaluate race and SES in relationship to pediatric CRS. Future research should employ nationally representative data to assess the true demographic variation in children with CRS.


Transcript Profiling and Gene Identification Involved in the Ethylene Signal Transduction Pathways of Creeping Bentgrass (Agrostis stolonifera) during ISR Response Induced by Butanediol.
Creeping bentgrass (Agrostis stolonifera) is the preferred green lawn grass, with excellent turf characteristics but poor disease resistance. At present, the mechanisms of disease resistance in creeping bentgrass are poorly understood, especially the ethylene signal transduction pathway under the induced systemic resistance (ISR) response. In this study, butanediol (BDO), as a new type of disease-resistance compound, was applied to creeping bentgrass seedlings to induce the ISR response. Then, we measured ethylene production and related enzyme activities. Additionally, transcript profiling and gene identification were performed in association to ethylene signal transduction pathways. The changes of ethylene production and related enzyme 1-aminocyclopropane-1-carboxylic acid oxidase (ACO) and 1-aminocyclopropane-1-carboxylic acid synthases (ACS) activities showed significant difference at 24 h after Rhizoctonia solani inoculation among five treatments of various BDO concentrations. After 100 µmol L-1 BDO treatment, ethylene production and related enzyme activities reached their peak levels. Additionally, 208,672 unigenes of creeping bentgrass were obtained by de novo assembly. In total, 15,903 annotated unigenes were grouped into 33 canonical pathways in the KEGG (Kyoto Encyclopedia of Genes and Genomes) analysis. Among those, 1803 unigenes were classified as 'signal transduction'. There were 6766 differentially expressed genes (DEGs) among B24 (inoculated-rhizobacteria in MS medium with 100 µmol L-1 BDO for 24 h), NB24, B72 and NB24 (no rhizobacteria in MS medium with 100 µmol L-1 BDO for 24 h) libraries, and 4,639 DEGs between B24 and B72 (inoculated-rhizobacteria in MS medium with 100 µmol L-1 BDO for 72 h) libraries, with 4489 DEGs in all three libraries. As suggested by the RT-PCR assay, the expression levels of ethylene-responsive and defense-related genes were variable among treated samples during the BDO-induced ISR responses. The expression levels of EIN, ERF, NPR1, PR3 and PR4 genes increased and reached their peaks in the first 24 h after R. solani infection in the BDO-induced ISR reaction compared with NB24 treatments. This results is consistent with the changes of important ethylene biosynthetic enzymes and ethylene concentrations during the BDO-induced ISR responses. We further found the intermediate substances for the signaling pathway, and the relationships between the expression levels of BDO-induced ISR disease-resistance genes and those of the response genes for ethylene signal pathway. Our findings present a genetic basis for systemic resistance of creeping bentgrass through transcriptomic analysis and our study provides a theoretical and practical basis for the improvement of turfgrass disease resistance and quality.


Findings on brain MRI from research studies of occupational exposure to known neurotoxicants.
The expanding use of MRI in large-scale epidemiologic studies of CNS outcomes has led to increasing concern for the consistent handling of incidental findings. Our purpose is to identify the prevalence of incidental neuroradiologic abnormalities in an adult population with past occupational exposure to lead who underwent brain MRI as part of a large, longitudinal cohort study. Structural MR images obtained for the research study were reviewed by a neuroradiologist on an ongoing basis for findings of clinical concern. The subjects were recruited as part of a longitudinal research study designed to examine the effects of past lead exposure on CNS structures. The cohort examined consisted of 589 men who previously worked at an organolead manufacturing facility and had a wide range of lead exposure durations and intensities. MR images were also reviewed from 67 population-based control subjects from the same study who had no history of occupational lead exposure. Incidental findings were detected in 84% (548/656) of research subjects. Of the 548 subjects with abnormalities, 30% (167) required no referral, 51% (280) required routine referral, 17% (93) required urgent referral, and 1.5% (8) required immediate referral. Incidental findings of all categories were observed in a larger percentage of older subjects, aged 60 years and above. Using multivariable logistic regression analysis, we found that age (p < 0.0001), but not lead history, was associated with an increased risk of incidental neuroradiologic abnormalities. In the population examined, there was a high prevalence of incidental brain and head and neck abnormality. Of particular concern were the serious health problems observed requiring urgent or immediate referral in 18.5% of the subjects. These findings underscore the need for radiologists to evaluate the anatomic images generated by research studies, particularly those with an older population base.


Purification of a novel factor which binds to the mouse alpha 2 (I) collagen promoter.
We have identified and purified a DNA binding protein which specifically binds to a segment of the mouse alpha 2 (I) collagen promoter between -420 and -399 bp upstream of the start of transcription. Purification included heparin-agarose and sequence-specific DNA-affinity chromatography, followed by SDS-PAGE and renaturation of the DNA binding activity after elution from SDS-polyacrylamide gel. The DNA binding activity resides in two species of 42 kDa and 40 kDa, respectively. The levels of DNA binding activity of this factor, which has been tentatively designated as ColF1, are considerably higher in nuclear extracts of NIH-3T3 fibroblasts than in nuclear extracts from epidermal cells, lymphoid cells and transformed NIH-3T3 fibroblasts.


Experiments with very high energy synchrotron radiation.
The use of synchrotron radiation with very high photon energies has become possible only with the latest generation of storage rings. All high-electron-energy synchrotron sources will have a dedicated program for the use of very high photon energies. The high-energy beamline ID15 at the ESRF was the first beamline built and dedicated to this purpose, and it has now been in user operation for more than three years. The useful energy range of this beamline is 30-1000 keV and the superconducting insertion device for producing the highest attainable photon energies is described in detail. The techniques most often used today are diffraction and Compton scattering; an overview of the most important experiments is given. Both techniques have been used in the investigation of magnetic systems, and, additionally, the high resolution in reciprocal space, which can be achieved in diffraction, has led to a series of applications. Other fields of research are addressed, and attempts to indicate possible future research areas of high-energy synchrotron radiation are made.


CMUT Fabrication Based On A Thick Buried Oxide Layer.
We introduce a versatile fabrication process for direct wafer-bonded CMUTs. The objective is a flexible fabrication platform for single element transducers, 1D and 2D arrays, and reconfigurable arrays. The main process features are: A low number of litho masks (five for a fully populated 2D array); a simple fabrication sequence on standard MEMS tools without complicated wafer handling (carrier wafers); an improved device reliability; a wide design space in terms of operation frequency and geometric parameters (cell diameter, gap height, effective insulation layer thickness); and a continuous front face of the transducer (CMUT plate) that is connected to ground (shielding for good SNR and human safety in medical applications). All of this is achieved by connecting the hot electrodes individually through a thick buried oxide layer, i.e. from the handle layer of an SOI substrate to silicon electrodes located in each CMUT cell built in the device layer. Vertical insulation trenches are used to isolate these silicon electrodes from the rest of the substrate. Thus, the high electric field is only present where required - in the evacuated gap region of the device and not in the insulation layer of the post region. Array elements (1D and 2D) are simply defined be etching insulation trenches into the handle wafer of the SOI substrate.


The Revised Helping Alliance Questionnaire (HAq-II) : Psychometric Properties.
The concept of the therapeutic alliance and its operationalization have received much attention in recent years. One of the early self-report measures of the therapeutic alliance was the Helping Alliance questionnaire (HAq-I). This scale was recently revised to exclude the items that explicitly reflect improvement. Using the revised 19-item HAq-II on a sample of 246 patients diagnosed with DSM-III-R cocaine dependence, the authors found that the new scale had excellent internal consistency and test-retest reliability. Further, the HAq-II demonstrated good convergent validity with the California Psychotherapy Alliance Scale (CALPAS) total score. Alliance levels as measured by the CALPAS or the Helping Alliance questionnaire during early sessions were not associated with pretreatment psychiatric severity or level of depression.


Autologous platelet concentrate for post-extraction socket healing: a systematic review.
Autologous platelet concentrates are claimed to enhance hard and soft tissue healing due to the considerable amount of growth factors that are released after application in the surgical site. However, their actual efficacy for improving tissue healing and regeneration in oral surgery applications is controversial. Tooth extraction socket healing represents a proper model to study the effect of autologous platelet-enriched preparations due to the concomitant occurrence of different processes of both hard and soft tissue healing. To evaluate the efficacy of platelet concentrates for alveolar socket healing after tooth extraction, by conducting a systematic review. Medline, Embase and Cochrane Central Register of Controlled Trials were searched using a combination of specific search terms. The last electronic search was performed on 15 June, 2014. Manual searching of the relevant journals and of the reference lists of reviews and all identified randomised controlled trials was also performed. Randomised controlled trials evaluating the effect of a platelet concentrate on fresh extraction sockets were included. Further inclusion criteria were that at least 10 patients were treated (at least 5 per group) and there was a minimum follow-up duration of 3 months. Primary outcomes were postoperative complications, patient satisfaction and postoperative discomfort. Secondary outcomes were any clinical, radiographic, histological and histomorphometric variables used to assess hard and soft tissue healing. Assessment of the methodological quality of the trials was made. RESULTS were expressed as fixed-effects models using mean differences for continuous outcomes and risk ratios for dichotomous outcomes, with 95% confidence intervals (CI). The initial search yielded 476 articles. After the screening process, six articles met the inclusion criteria (199 teeth in 156 patients). Three studies were considered at high risk of bias, two at medium risk and one at low risk. A large heterogeneity in study characteristics and outcome variables used to assess hard tissue healing was observed. A meta-analyses of two studies reporting histomorphometric evaluation of bone biopsies at 3 months' follow-up showed greater bone formation when platelet concentrates were used, as compared to control cases (P <0.001; mean difference 20.41%, 95% C.I. 13.29%, 27.52%). Beneficial effects of platelet concentrates were generally but not systematically reported in most studies, in particular when considering the effects on soft tissue healing and the patient's reported postoperative symptoms like pain and swelling, although no meta-analysis could be done for such parameters. Although the results of the meta-analysis of the present review are suggestive for a positive effect of platelet concentrates on bone formation in post-extraction sockets, due to the limited amount and quality of the available evidence, they need to be cautiously interpreted. A standardisation of the experimental design is necessary for a better understanding of the true effects of the use of platelet concentrates for enhancing post-extraction socket healing.


Recombination of the porcine X chromosome: a high density linkage map.
Linkage maps are essential tools for the study of several topics in genome biology. High density linkage maps for the porcine autosomes have been constructed exploiting the high density data provided by the PorcineSNP60 BeadChip. However, a high density SSCX linkage map has not been reported up to date. The aim of the current study was to build an accurate linkage map of SSCX to provide precise estimates of recombination rates along this chromosome and creating a new tool for QTL fine mapping. A female-specific high density linkage map was built for SSCX using Sscrofa10.2 annotation. The total length of this chromosome was 84.61 cM; although the average recombination rate was 0.60 cM/Mb, both cold and hot recombination regions were identified. A Bayesian probabilistic to genetic groups and revealed that the animals used in the current study for linkage map construction were likely to be carriers of X chromosomes of European origin. Finally, the newly generated linkage map was used to fine-map a QTL at 16 cM for intramuscular fat content (IMF) measured on longissimus dorsi. The sulfatase isozyme S gene constitutes a functional and positional candidate gene underlying the QTL effect. The current study presents for the first time a high density linkage map for SSCX and supports the presence of cold and hot recombination intervals along this chromosome. The large cold recombination region in the central segment of the chromosome is not likely to be due to structural differences between X chromosomes of European and Asian origin. In addition, the newly generated linkage map has allowed us to fine-map a QTL on SSCX for fat deposition.


Mahidol Scoring for Assessing Various Grades of β Thalassemia Intermedia.
To determine the effectiveness of Mahidol Scoring for assessing various grades of β thalassemia intermedia. Cross-sectional study. Fatima Memorial Hospital, Shadman, Lahore, from August 2016 to August 2017. A total of 150 patients, both inpatient and outpatient diagnosed as thalassaemia intermedia fulfilling inclusion criteria, were enrolled, interviewed and examined after an informed consent. All patients were assessed by using Mahidol Scoring system. Their scores were documented on a predesigned evaluation porforma. Patients were labelled as mild, moderate or severe diseased. In a total of 150 patients, 88 (58.7%) were males and 62 (41.3%) were females. Using Mahidol Scoring, 88 (58.7%) were labelled as mild, 53 (35.3%) as moderate and 9 (6%) as severe diseased. Patient who were labelled as mild diseased according to Mahidol Scoring, had no blood transfusions and showed good response to hydroxyurea. Patients with moderate score, were infrequently transfused and showed variable response to hydroxyurea. Patients in severe group received first transfusion at an early age, maintained lower Hb levels, showed growth retardation, splenomegaly, and had poor response to hydroxyurea. Mahidol Scoring system is an easy, safe and effective way for classification of thalassaemia Intermedia severity. The grades according to Mahidol Scoring system will aid in the management of patients as the score can be quickly calculated, and can assist the clinician in an initial evaluation for disease severity in patients of thalassaemia intermedia.


Effects of dietary restriction on cellular immunity in rats.
Phagocytosis of opsonized sheep red blood cells (SRBC) by alveolar macrophages (AM) was measured in rats fasted for 1 to 9 days or fed on diets restricted 20 to 95% compared to control group for 2 and 8 weeks. In rats fasted for 1 to 6 days, AM showed an increased phagocytosis at 2 days after fasting, but their phagocytic activity remarkably decreased afterwards. Furthermore, phagocytic activity of AM per rat revealed much more decrease at 3 to 6 days after fasting. Then the production of interleukin-1 (IL-1) by AM increased with prolonged fasting, but the production of prostaglandin E2 (PGE2) by AM cultured with lipopolysaccharide (LPS) conversely decreased in rats fasted for 2 days or longer. The proliferation of splenocytes increased with prolonged fasting. On the other hand, 20 to 95% restricted diets induced the increased phagocytosis of AM with prolonged experimental period. However, phagocytic activity of AM per rat showed significant increase only in rats on a 40% restricted diet. The findings suggest that differences in both duration and degree of dietary restriction modulate phagocytic function of AM, and may contribute to explaining, in part, conflicting observations which have been obtained on the immunologic state in malnourished animals.


Identifying actives from HTS data sets: practical approaches for the selection of an appropriate HTS data-processing method and quality control review.
High-throughput screening (HTS) has achieved a dominant role in drug discovery over the past 2 decades. The goal of HTS is to identify active compounds (hits) by screening large numbers of diverse chemical compounds against selected targets and/or cellular phenotypes. The HTS process consists of multiple automated steps involving compound handling, liquid transfers, and assay signal capture, all of which unavoidably contribute to systematic variation in the screening data. The challenge is to distinguish biologically active compounds from assay variability. Traditional plate controls-based and non-controls-based statistical methods have been widely used for HTS data processing and active identification by both the pharmaceutical industry and academic sectors. More recently, improved robust statistical methods have been introduced, reducing the impact of systematic row/column effects in HTS data. To apply such robust methods effectively and properly, we need to understand their necessity and functionality. Data from 6 HTS case histories are presented to illustrate that robust statistical methods may sometimes be misleading and can result in more, rather than less, false positives or false negatives. In practice, no single method is the best hit detection method for every HTS data set. However, to aid the selection of the most appropriate HTS data-processing and active identification methods, the authors developed a 3-step statistical decision methodology. Step 1 is to determine the most appropriate HTS data-processing method and establish criteria for quality control review and active identification from 3-day assay signal window and DMSO validation tests. Step 2 is to perform a multilevel statistical and graphical review of the screening data to exclude data that fall outside the quality control criteria. Step 3 is to apply the established active criterion to the quality-assured data to identify the active compounds.


[New developments in cancer chemotherapy (author's transl)].
Based on new drugs, new combinations, earlier onset of treatment and aggressive doses in the induction phase chemotherapeutic results in advanced cancer have been improved within the last decade. The present review presents the currently available realistic results obtained with both curative and palliative chemotherapy. In addition, the concepts of preoperative ("debulking") chemotherapy and of adjuvant chemotherapy are described for the adequate clinical indications. Finally, guidelines for chemotherapeutic measures in elderly patients are presented.


Possible role of dopamine in changes in LH and prolactin concentrations after experimentally induced hyperprolactinaemia in rats.
Nine-month-old female rats bearing an ectopic pituitary gland (from a litter-mate) under the right kidney capsule since day 30 of life and their sham-operated controls, were treated with a dopamine agonist (lysuride) or antagonist (metoclopramide). Plasma prolactin and LH levels were measured by double-antibody radioimmunoassays. Vaginal smears were taken before and during the treatment periods. Eight months after the operation, a significant (P less than 0.01) increase in basal prolactin levels together with a significant (P less than 0.05) reduction in LH values and permanent dioestrus occurred in the grafted animals when compared with controls. Lysuride treatment resulted in a marked reduction in plasma prolactin levels both in control and grafted rats over the whole 12 days of treatment, together with a partial restoration of plasma LH levels on day 1. From day 7 onwards a depression in LH values was again observed. Oestrous cycles were partially restored at the beginning of the treatment, but after 7 days dioestrus returned. Metoclopramide administration induced a significant (P less than 0.001) increase in basal prolactin levels in both grafted and control rats. Basal plasma LH values were unaffected in controls when compared with vehicle-treated animals. An increase could be seen in hyperprolactinaemic rats after 7 or 12 days of treatment however. The LH response to the administration of LH releasing hormone (LHRH) was greater in the experimental and control metoclopramide-treated rats when compared with vehicle-treated rats.(ABSTRACT TRUNCATED AT 250 WORDS)


CCL16/LEC powerfully triggers effector and antigen-presenting functions of macrophages and enhances T cell cytotoxicity.
The human CC chemokine CCL16, a liver-expressed chemokine, enhances the killing activity of mouse peritoneal macrophages by triggering their expression of tumor necrosis factor alpha (TNF-alpha) and Fas ligand. Macrophages also respond to CCL16 by enhancing their production of monocyte chemoattractant protein-1, regulated on activation, normal T cells expressed and secreted chemokines, and interleukin (IL)-1 beta, TNF-alpha, and IL-12. The effect of CCL16 is almost as strong as that of lipopolysaccharide and interferon-gamma, two of the best macrophage activators. Moreover, CCL16-activated macrophages overexpress membrane CD80, CD86, and CD40 costimulatory molecules and extensively phagocytose tumor cell debris. On exposure to such debris, they activate a strong, tumor-specific, cytolytic response in virgin T cells. Furthermore, cytolytic T cells generated in the presence of CCL16 display a higher cytotoxicity and activate caspase-8 in tumor target cells. This ability to activate caspase-8 depends on their overexpression of TNF-alpha and Fas ligand induced by CCL16. These data reveal a new function for CCL16 in the immune-response scenario. CCL16 significantly enhances the effector and the antigen-presenting function of macrophages and augments T cell lytic activity.


Ward staff perceptions of the role of the advanced nurse practitioner in a 'hospital at day' setting.
to examine ward staff perceptions on the role of the 'hospital at day' advanced nurse practitioner (ANP). This term is used locally to refer to a model first introduced into 'hospital at night' teams, in response to changes in working patterns of junior doctors, where an advanced nurse practitioner is based on the ward-the model was subsequently rolled out to daytime teams. a qualitative descriptive phenomenological approach was adopted to carry out this study with a purposive sample of ten participants. Participants were interviewed individually using semi-structured interviews to gain insights into their perceptions on the role of the ANP, following Colaizzi's analytical model of data analysis. four major themes were identified. The first overarching theme was effectiveness of the ANP role. There was evidence that the role was not only clinically effective in the ward but also in relation to organisational priorities such as impact on patient flow. Secondly it was acknowledged that having an ANP based on the ward and as a 'constant' in the ward area, when medical teams rotate, provides team stability and improves the functioning of the ward area. Despite the positivity recognised, the third theme identified that having a ward-based ANP may affect skill acquisition and development and may lead to the 'de-skilling' of staff. Lastly it was acknowledged that there remains an overall confusion about the role of the ANP. the themes identified reflect what is already known about the wider perceived role of the ANP. Moreover, this study adds to the limited literature available specifically on the benefits of the ANP within 'hospital at day' roles.