Anti-red blood cell autoantibodies induced in rat by Plasmodium berghei infection bind to a cross-reacting determinant present also in other cell types.
It has been previously shown that rats injected with red blood cells (RBC) infected with Plasmodium berghei make autoantibodies reacting with normal rat RBC as demonstrated by a staphylococcal protein A binding assay. In this study, the serum of the infected rats was found to contain also antibodies to several other types of rat cells (lymphoid, liver, and kidney cells and brain tissue) as well as against RBC and lymphoid cells of mice and sheep, but not of man. Rat, mouse, and sheep RBC and rat and mouse lymphocytes absorbed completely the antibodies to rat RBC, suggesting that a single, cross-reacting determinant might be involved in these reactions.


Neighborhood & Family Effects on Learning Motivation among Urban African American Middle School Youth.
Motivational theorists in psychology have moved away from individual-based approaches to socio-cognitive and socio-ecological models to explain student engagement and motivation for learning. Such approaches consider, for example, the influence of family and neighborhood environments as important constructs in youth behavior. In this study, links between neighborhood condition (e.g. external appearance of the blocks nearest to the respondents' home), family dysfunction, and motivation for learning are investigated. Data were obtained from two hundred and sixteen (216) urban African American middle school children enrolled in a substance use prevention intervention. Analytic models show associations between poor neighborhood condition and both family dysfunction and lower learning motivation. Family dysfunction was also found to mediate the effect of neighborhood condition on motivated learning. Neighborhood and family characteristics are important determinants of urban schoolchildren's motivation for learning.


Impact of dialyzer membrane flux on metal clearance in hemodialysis patients.
Deficiency of essential trace elements (such as Cu or Zn) and accumulation of potentially toxic trace elements (as Cd or Pb) are both known to have adverse effects in hemodialysis (HD) patients. Up to our knowledge, no studies about the permeability of low and high flux polysulfone membranes on metal ions during hemodialysis are available. Therefore, the aim of the present study was to address this issue. Forty one hemodialysis patients (19 were using high flux polysulfone membrane while the remaining were using low flux one) participated in the study. Blood levels of Cu, Zn, Cd and Pb were determined by graphite furnace atomic absorption spectrometry among HD patients, before and after dialysis session, as well as among matched 40 healthy persons. Blood concentrations of Cu and Zn in the whole hemodialysis group was significantly lower than those of the healthy control group, on the other hand the toxic metals (Cd and Pb) levels were observed to be significantly higher among HD patients compared to the normal persons. Among the hemodialysis group, there were no significant differences between the low and high flux dialyzer groups in terms of pre-dialysis blood levels of Cu, Zn, Cd and Pb. In addition, significantly decreased levels of all metal ions were observed after dialysis sessions using either low or high flux membranes. An exception was Pb which did not show any difference between pre-dialysis and post-dialysis values in the low flux groupIn conclusion Zn and Cu deficiencies should be considered in the treatment of these patients. High flux membranes are more efficient than low flux ones in removing excess Cd and Pb. Therefore, when high flux membranes are used, chelation therapy might not be required for Cd and Pb overload.


Simultaneously improving the mechanical properties, dissolution performance, and hygroscopicity of ibuprofen and flurbiprofen by cocrystallization with nicotinamide.
To be fully exploitable in both formulation and manufacturing, a drug cocrystal needs to demonstrate simultaneous improvement of multiple key pharmaceutical properties over the pure drug crystal. The present work was aimed at investigating such feasibility with two model profen-nicotinamide cocrystals. Phase pure 1:1 ibuprofen-nicotinamide and flurbiprofen-nicotinamide cocrystals were prepared from solutions through rapid solvent removal using rotary evaporation,and characterized by DSC, PXRD, FTIR, phase solubility measurements, equilibrium moisture sorption analysis, dissolution testing and tabletability analysis. Temperature-composition phase diagrams constructed from DSC data for each profen and nicotinamide crystal revealed the characteristic melting point of the 1:1 cocrystal as well as the eutectic temperatures and compositions. Both cocrystals exhibited higher intrinsic dissolution rates than the corresponding profens. The cocrystals also sorbed less moisture and displayed considerably better tabletability than the individual profens and nicotinamide. Phase behaviors of 1:1 profen-nicotinamide cocrystal systems were delineated by constructing their temperature-composition phase diagrams. Cocrystallization with nicotinamide can simultaneously improve tableting behavior, hygroscopicity, and dissolution performance of ibuprofen and flurbiprofen. This could pave the way for further development of such cocrystal systems into consistent, stable, efficacious and readily manufacturable drug products.


Imperceptible, designable, and scalable braided electronic cord.
Flexible sensors, friendly interfaces, and intelligent recognition are important in the research of novel human-computer interaction and the development of smart devices. However, major challenges are still encountered in designing user-centered smart devices with natural, convenient, and efficient interfaces. Inspired by the characteristics of textile-based flexible electronic sensors, in this article, we report a braided electronic cord with a low-cost, and automated fabrication to realize imperceptible, designable, and scalable user interfaces. The braided electronic cord is in a miniaturized form, which is suitable for being integrated with various occasions in life. To achieve high-precision interaction, a multi-feature fusion algorithm is designed to recognize gestures of different positions, different contact areas, and different movements performed on a single braided electronic cord. The recognized action results are fed back to varieties of interactive terminals, which show the diversity of cord forms and applications. Our braided electronic cord with the features of user friendliness, excellent durability and rich interaction mode will greatly promote the development of human-machine integration in the future.


The Prevalence of Clinically Significant Prostate Cancer According to Commonly Used Histological Thresholds in Men Undergoing Template Prostate Mapping Biopsies.
Transrectal prostate biopsies are inaccurate and, thus, the prevalence of clinically significant prostate cancer in men undergoing biopsy is unknown. We determined the ability of different histological thresholds to denote clinically significant cancer in men undergoing a more accurate biopsy, that of transperineal template prostate mapping. In this multicenter, cross-sectional cohort of men who underwent template prostate mapping biopsies between May 2006 and January 2012, 4 different thresholds of significance combining tumor grade and burden were used to measure the consequent variation with respect to the prevalence of clinically significant disease. Of 1,203 men 17% (199) had no previous biopsy, 38% (455) had a prior negative transrectal ultrasound biopsy, 24% (289) were on active surveillance and 21% (260) were seeking risk stratification. Mean patient age was 63.5 years (SD 7.6) and median prostate specific antigen was 7.4 ng/ml (IQR 5.3-10.5). Overall 35% of the patients (424) had no cancer detected. The prevalence of clinically significant cancer varied between 14% and 83% according to the histological threshold used, in particular between 30% and 51% among men who had no previous biopsy, between 14% and 27% among men who had a prior negative biopsy, between 36% and 74% among men on active surveillance, and between 47% and 83% among men seeking risk stratification. According to template prostate mapping biopsy between 1 in 2 and 1 in 3 men have prostate cancer that is histologically defined as clinically significant. This suggests that the commonly used thresholds may be set too low.


Canadian stem cell scientists take the prize.
This year, the recipients of the Lasker Award for Basic Medical Research are Ernest A. McCulloch and James E. Till. The landmark studies of McCulloch and Till in the 1960s defined the hallmark properties of stem cells: the ability to self-renew and differentiate.


Fate of Cd during mineral transformation by sulfate-reducing bacteria in clay-size fractions from soils with high geochemical background.
Sulfate-reducing bacteria (SRB) can immobilize heavy metals in soils through biomineralization, and the parent rock and minerals in the soil are critical to the immobilization efficiency of SRB. To date, there is little knowledge about the fate of Cd associated with the parent rocks and minerals of soil during Cd immobilized by SRB. In this study, we created a model system using clay-size fraction of soil and SRB to explore the role of SRB in immobilizing Cd in soils from stratigraphic successions with high geochemical background. In the system, clay-size fractions (particle size < 2 µm) with concentration of Cd (0.24-2.84 mg/kg) were extracted from soils for bacteria inoculation. After SRB reaction for 10 days, the Cd fraction tended to transform into iron-manganese bound. Further, two clay-size fractions, i.e., the non-crystalline iron oxide (Fe-OX) and the crystalline iron oxide (Fe-CBD), were separated by extraction. The reaction of SRB with them verified the transformation of primary iron-bearing minerals into secondary iron-bearing minerals, which contributed to Cd redistribution. This study shows that SRB could exploit the composition and structure of minerals to induce mineral recrystallization, thereby aggravating Cd redistribution and immobilization in clay-size fractions from stratigraphic successions with high geochemical background.


Characterization of compound mechanisms and secondary activities by BioMAP analysis.
Unexpected drug activities account for many of the failures of new chemical entities in clinical trials. These activities can be target-dependent, resulting from feedback mechanisms downstream of the primary target, or they can occur as a result of unanticipated secondary target(s). Methods that would provide rapid and efficient characterization of compounds with respect to a broad range of biological pathways and mechanisms relevant to human disease have the potential to improve preclinical and clinical success rates. BioMAP assays containing primary human cells (endothelial cells and co-cultures with peripheral blood leukocytes) were stimulated in complex formats (specific combinations of inflammatory mediators) for 24 h in the presence or absence of test agents (drugs, experimental compounds, etc.). The levels of selected protein readouts (adhesion receptors, cytokines, enzymes, etc.) were measured and activity profiles (normalized data sets comprising BioMAP profiles) were generated for each test agent. The resulting profiles were compared by statistical methods to identify similarities and mechanistic insights. Compounds with known mechanisms including inhibitors of histamine H1 receptor, angiotensin converting enzyme, IkappaB kinase-2, beta2 adrenergic receptor and others were shown to generate reproducible and distinguishable BioMAP activity profiles. Similarities were observed between compounds targeting components within the same signal transduction pathway (e.g. NFkappaB), and also between compounds that share secondary targets (e.g. ibuprofen and FMOC-L-leucine, a PPARgamma agonist). Complex primary cell-based assays can be applied for detecting and distinguishing unexpected activities that may be of relevance to drug action in vivo. The ability to rapidly test compounds prior to animal or clinical studies may reduce the number of compounds that unexpectedly fail in preclinical or clinical studies.


Maternal phenylketonuria--the Irish experience.
The outcome of 48 pregnancies in 18 women with elevated phenylalanine was studied. The women were divided into two groups, diet and non-diet. All the women on diet had severe hyperphenylalaninaemia and only six were on the amino acid mixture at conception. In the non-diet group four women had concentrations of less than 1000 mumols/L and five had severe hyperphenylalaninaemia. Three deaths and one stillbirth occurred in the non-diet group. Microcephaly occurred in 24 children; 11 in the diet group and 13 in the non-diet group. There were five miscarriages in the non-diet group and one in the diet group. Diet is beneficial when started early but planned pregnancies, good motivation and compliance were difficult to achieve in this group of women despite every reasonable effort by the PKU clinic.


Soft Tissue Trauma: Critical Recognition and Timing of Intervention in Emergency Presentations.
The facial trauma surgeon will see a variety of facial injuries. Recognition of emergency cases and proper intervention is and this article aims to highlight those cases and the respective proper interventions.


An integrated electrochemical fructose biosensor based on tetrathiafulvalene-modified self-assembled monolayers on gold electrodes.
An integrated amperometric fructose biosensor based on a gold electrode (AuE) modified with a self-assembled monolayer (SAM) of 3-mercaptopropionic acid (MPA) on which fructose dehydrogenase (FDH) and the mediator tetrathiafulvalene (TTF) are co-immobilized by cross-linking with glutaraldehyde is reported. Variables concerning the behavior of the biosensor were optimized by taking the slope value obtained for the fructose calibration plot in the 0.1-1.0 mM concentration range as a criterion of selection. At an applied potential of +0.20 V, a good repeatability of such slope values (RSD=6.7%, n=10) was obtained with no need to apply a cleaning or pretreatment procedure to the modified electrode. Moreover, results from five different TTF-FDH-MPA-AuEs yielded a RSD of 5.8%. The useful lifetime of one single biosensor was approximately 30 days, exhibiting a 93% of the original response on the 33rd day. A linear calibration graph was obtained for fructose over the 1.0 x 10(-5)-1.0 x 10(-3) M range, with a limit of detection of 2.4 x 10(-6) M. The effect of potential interferents was evaluated. The TTF-FDH-MPA-AuE also performed well in the flow-injection mode. The biosensor was used for the determination of fructose in real samples, and the results compared with those provided by using a commercial enzyme test kit.


The follow-up CT scan in childhood meningitis.
CT in childhood bacterial meningitis has indicated that bacterial meningitis is more than an inflammation of the subarachnoid space; it is a disease process which often exerts a profound effect on the brain parenchyma. In some cases ventricular widening may be a reflection of loss of brain substance rather than increased intracranial pressure. A vasculitis may occur in cerebral vessels with consequent ischemic infarct. The abnormalities found on CT during the acute illness are frequently persistent.


Ultrasound mediated delivery of oxygen and LLL12 loaded stimuli responsive microdroplets for the treatment of hypoxic cancer cells.
LLL12 exhibits high specificity for inhibiting STAT3 phosphorylation and dimerization, and inducing apoptosis to constitutively activated STAT3 cancer cells without cytotoxicity to normal cells with dormant STAT3. However, clinical deployment of LLL12 in cancer treatment is hindered by its low bioavailability and hypoxia-induced resistance. To overcome these limitations, we encapsulate both oxygen and LLL12 in stimuli responsive microdroplets (SRMs) by a gas-driven coaxial flow focusing (CFF) process for ultrasound mediated treatment of hypoxic cancer cells. Our benchtop experiments demonstrate that the CFF process is able to produce SRMs with uniform size distribution, large oxygen loading capacity, high LLL12 encapsulation efficiency, well protection of bioactivity, and steadily long shelf time. The in vitro therapeutic studies in pancreatic cancer cells (PANC-1 and CAPAN-1) demonstrate the immediate release of oxygen and LLL12 in exposure to therapeutic ultrasound pulses as well as the improved anticancer effects under hypoxic conditions. The findings suggest that the proposed oxygen and LLL12 loaded SRMs provide a promising drug delivery strategy for more effective treatment of hypoxic cancer cells.


Clinical applications of the p53 tumor suppressor gene.
The interest on the p53 gene has grown enormously in the last 4 years. It is now known that p53 is directly involved in important cellular functions including regulation of the cell cycle, and that its alteration may be one of the important steps in the initiation of cancer. In this review I will cover briefly basic and clinical aspects related to the p53 gene and protein and explore ways of using the accumulated knowledge for patient diagnosis and monitoring. The literature suggests that it is now appropriate to start assessing the p53 gene status of breast tumors for prognosis. Therapeutic options are at an infancy stage. A new diagnostic approach based on the immune response of cancer patients against mutant proteins is discussed, by using the p53 mutant protein as a model. Although the cancer patient has not as yet benefited directly from the enormous number of investigations on the p53 gene and protein, there is hope that in the long-term these studies will promote the understanding of cancer initiation and progression at the molecular level with a practical return at a later phase.


An update: the role of Nephrin inside and outside the kidney.
Nephrin is a key molecule in podocytes to maintain normal slit diaphragm structure. Nephin interacts with many other podocyte and slit diaphragm protein and also mediates important cell signaling pathways in podocytes. Loss of nephrin during the development leads to the congenital nephrotic syndrome in children. Reduction of nephrin expression is often observed in adult kidney diseases including diabetic nephropathy and HIV-associated nephropathy. The critical role of nephrin has been confirmed by different animal models with nephrin knockout and knockdown. Recent studies demonstrate that knockdown of nephrin expression in adult mice aggravates the progression of unilateral nephrectomy and Adriamycin-induced kidney disease. In addition to its critical role in maintaining normal glomerular filtration unit in the kidney, nephrin is also expressed in other organs. However, the exact role of nephrin in kidney and extra-renal organs has not been well characterized. Future studies are required to determine whether nephrin could be developed as a drug target to treat patients with kidney disease.


[Expression of neuronal nitric oxide synthase in brain tissue of hypoxic-ischemic neonatal rat and the cerebral protective effect of neuronal nitric oxide synthase inhibitor 7-nitroindazole].
To explore the effect of nNOS and its selective inhibitor 7-NI in neonatal rats with hypoxic-ischemic brain damage (HIBD). The model of HIBD in neonatal rat was made and the expression of nNOS mRNA was measured at different time points and compared with the control group. The levels of NOS, NO, MDA, SOD were also assayed at different time points after aderministering 7-NI, and compared with the HIBD group and the control group. In addition, we observed the degree of apoptosis after 24 h of HIBD. Expression of nNOS mRNA was higher at 2 h and 6 h after hypoxia than those in the control group (P < 0.05) and reached more at 6 h (P < 0.05). Concentrations of NOS, NO, MDA increased after HIBD, but concentration of SOD decreased (P < 0.01); 7-NI reduced the levels of NOS, NO, MDA, and level of SOD was higher than those in the HIBD group (P < 0.05). There was no apoptosis in the control group, 7-NI could effectively inhibit the degree of apoptosis after HIBD (P < 0.001). nNOS has an important role in the pathogenesis of neonatal rats with HIBD, its specific inhibitor 7-NI can protect brains from hypoxic-ischemic damage through anti-oxidation and prohibiting apoptosis.


Intermittent exposure to traces of green leaf volatiles triggers a plant response.
Plants are known to mount a defensive response when exposed to volatile chemicals from other plants, but the critical concentration required for this response is not known. We showed that intermittent exposure over a period of 3 weeks to trace amounts (less than 140 pptV) of green leaf volatiles emitted by a freshly damaged Arabidopsis plant induced physiological (defensive) responses in undamaged neighbouring plants. These results demonstrated that plants can respond to long-term repeated exposures to subcritical amounts of chemical signals.


Photosynthetic plasticity of populations of Heliotropium curassavicum L. originating from differing thermal regimes.
Plants of the widely distributed species Heliotropium curassavicum L. have a large photosynthetic acclimation potential to temperature. There are, however, some differences among the acclimation potentials of populations occupying dissimilar thermal regimes. Plants of populations originating from a cool maritime climate have a greater acclimation potential than plants of populations originating from a desert habitat, which is characterized by large seasonal changes in temperature.


Cu Nanoclusters/FeN4 Amorphous Composites with Dual Active Sites in N-Doped Graphene for High-Performance Zn-Air Batteries.
Exploring inexpensive and earth-abundant transition metal-nitrogen-based carbon (MNC) catalysts to substitute the scarce and costly Pt-based electrocatalysts for the oxygen reduction reaction (ORR) is quite anticipated in metal-air batteries (MABs). Here, we demonstrate a facile vacuum-annealing method to synthesize Cu nanoclusters/FeN4 amorphous composites embedded in N-doped graphene (Cu/Fe-NG). This approach avoids the long-term pyrolysis procedure and the use of an inert atmosphere in the conventional procedure for fabricating MNC catalysts. Interestingly, we discovered that the amorphous structure of Cu/FeN4 composites can provide high-activity bimetallic M-Nx sites (M = Cu, Fe), because of which the Cu/FeN4 composites exhibit boosted electrocatalytic activity with a positive half-wave potential of 0.88 V (vs RHE), long-term durability, and low hydrogen peroxide for the ORR. The origin of this enhancement was assigned to the concomitance of Fe-N4 and Cu-Nx moieties in Cu/Fe-NG, favoring adsorption and activation of the O2 molecule as suggested by X-ray absorption fine structure (XAFS) analyses and density functional theory (DFT) calculations. Moreover, examinations of Cu/Fe-NG in both liquid and quasi-solid-state Zn-air batteries (ZABs) can exhibit remarkable performances. This work may offer facile fabrication of enhanced performance MNC catalysts as well as a profound insight into the use of amorphous materials in the ORR and ZABs.


Improvement in thermal stability and substrate binding of pig kidney D-amino acid oxidase by chemical modification.
Chemical modification was evaluated to stabilize pig kidney D-amino acid oxidase (pkDAAO), which is required for analytical determination of D-amino acids. Optimization of modification conditions was performed to obtain high recovery yield and stability, and chemical modification at 30 degrees C for 12 h with a highly concentrated enzyme solution gave dextran-conjugated pkDAAO with a 70% yield of activity. pkDAAO was stable at less than 55 degrees C at pH 6.0, while the conjugated enzyme was stable even at 70 degrees C. In addition, the conjugated enzyme showed decreased Km values for D-amino acids. Because of these outstanding characteristics, this new material is expected to be available for use as a liquid assay reagent.


Central role of dendritic cells in the regulation and deregulation of immune responses.
Dendritic cells (DCs) play a critical role in orchestrating the innate and adaptive components of the immune system so that appropriate, coordinated responses are mounted against infectious agents. Tissue-resident DCs interact with microbes through germline-encoded pattern-recognition receptors (PRRs), which recognize molecular patterns expressed by various microorganisms. Antigens use PRR activation to instruct DCs for the appropriate priming of natural killer (NK) cells, followed by specific T-cell responses. Due to the central role of DCs in regulating the activation and progression of immune responses, minor imbalances in the feedback control of Toll-like receptor (TLR)-activated cells have been associated with autoimmunity in genetically prone individuals. We review here recent findings on the role of DCs in the priming of innate and adaptive immune responses and the possible involvement of DCs in inducing and maintaining autoimmune reactions.


Perceived barriers to immunizations as identified by Latino mothers.
Identify low-income Latino mothers' perceived barriers to immunizations. A 1-month survey was conducted in a Southern California, low-income primary care clinic. Mothers of Latino children aged 2 months to 18 years were surveyed in Spanish using a 52-question immunization survey. Latino mothers' perception of immunizations and knowledge of up-to-date status greatly influenced their children's immunization status. In addition, Latino children had lower immunization rates and encountered more barriers to immunizations than children in other population groups despite government efforts to provide vaccines to healthcare providers at no cost to assist low-income children. Most mothers (92%) believed their children were up-to-date on immunizations; however, immunization records documented that 42% of the children were current based on age. The most common barriers reported were that the child was sick at the time of the visit (51%) and/or transportation problems (39%). When working with low-income, uninsured Latinos, nurse practitioners and other healthcare professionals with low immunization rates in their clinic must assess the barriers to immunization and implement an action plan.


Predicting exercise capacity after lobectomy by single photon emission computed tomography and computed tomography.
This study compared the prediction of postoperative exercise capacity by employing lung perfusion scintigraphy images obtained with single photon emission computed tomography together with computed tomography (SPECT/CT) versus the common method of counting subsegments (SC method). In 18 patients scheduled for lobectomy, predicted postoperative maximum oxygen uptake per kilogram body weight ([Formula: see text]) was calculated by the SPECT/CT and SC methods. Correlations were examined between the [Formula: see text] predicted by SPECT/CT or the SC method, and the actual [Formula: see text] measured at 2 weeks (mean 15.4 ± 1.5 days) and 1 month (mean 29.1 ± 0.75 days) after surgery to determine whether SPECT/CT was more accurate than SC for predicting postoperative exercise capacity. There was a significant positive correlation between the [Formula: see text] predicted by SPECT/CT and the actual value at 2 weeks (r = 0.802, p < 0.0001) or 1 month (r = 0.770, p < 0.0001). There was also a significant positive correlation between the [Formula: see text] predicted by SC and the actual value at 2 weeks (r = 0.785, p < 0.0001) or 1 month (r = 0.784, p < 0.0001). This study showed that both SPECT/CT and the SC method were useful for predicting postoperative [Formula: see text] in the clinical setting.


Timing of aspirin administration as a determinant of survival of patients with acute myocardial infarction treated with thrombolysis.
Unlike thrombolytic agents, there are conflicting data regarding the time-dependent effect of aspirin treatment on outcome in acute myocardial infarction (AMI). We sought to evaluate the impact of timing of aspirin administration (before vs after thrombolysis) on mortality of patients with AMI. Our study included 1,200 patients with ST elevation AMI treated with thrombolysis. Early (n = 364) versus late (n = 836) users were defined as those receiving emergency aspirin before versus after initiation of thrombolysis, respectively. Time (median) from symptom onset to initiation of aspirin treatment was significantly shorter in early versus late users (1.6 vs 3.5 hours; p <0.001). There were no significant differences between the 2 groups with respect to baseline clinical characteristics. Early aspirin users were more likely to develop reischemia, to be treated with beta blockers, to be referred to coronary angiography, percutaneous transluminal coronary angioplasty, or coronary artery bypass graft surgery. Early users experienced lower mortality at 7 days (2.5% vs 6.0%, p = 0.01), 30 days (3.3% vs 7.3%, p = 0.008), and 1 year (5.0% vs 10.6%, p = 0.002) than late users. This survival benefit persisted for patients with and without previous aspirin therapy or revascularization and after adjustment for baseline characteristics and therapies at 7 days (odds ratio 0.36, 95% confidence interval 0.15 to 0.79), at 30 days (odds ratio 0.39, 95% confidence interval 0.17 to 0.82), and at 1 year (odds ratio 0.41, 95% confidence interval 0.21 to 0.74). Our study proposes a time-dependent benefit from aspirin in patients with AMI treated with thrombolysis.


Assessment of possible transneuronal changes in the retina of rats with inherited retinal dystrophy: cell size, number, synapses, and axonal transport by retinal ganglion cells.
In pigmented RCS rats with inherited retinal dystrophy, most photoreceptor cells disappear between postnatal days 20 and 100. We have examined the time course of the degeneration of photoreceptor nuclei and synapses and determined whether transneuronal changes occur in the inner nuclear layer (INL), inner plexiform layer (IPL), and retinal ganglion cells following loss of photoreceptor cells in these animals. Electron microscopic photomontages of the entire thickness of the IPL of dystrophic (RCS-p+) and control (RCS-rdy+ p+) rats 334 to 515 days old were prepared, and synapses were counted and identified as either conventional (amacrine) or ribbon (bipolar) types. Neither the incidence of synapses in the IPL nor the ratio of conventional to ribbon synapses differed in the dystrophic and control retinas. Ganglion cell diameter, perimeter, area, and density were measured from drawings of wholemount preparations of dystrophic and control rats 105 days and older. Diameter, perimeter, area and number of ganglion cells were not significantly different in the two genotypes. Anterograde axonal transport was measured by studying the displacement of labeled material as it traveled along ganglion cell axons and accumulated in the superior colliculus. The normal and dystrophic rats showed no significant difference in (1) the rates of rapidly moving components (approximately 110-180 mm/day) and slowly moving components (1.7-2.5 mm/day) or (2) the amount of radioactive material transported to the superior colliculus. The absence of transneuronal changes in retinal ganglion cells of RCS rats contrasts with results obtained earlier in rd mice (Graftstein et al., '72). Unlike the RCS rat, retinal degeneration in rd mice occurs before the maturation of the retina. We hypothesize that the ganglion cells may be more affected by loss of input early in development, and, therefore, ganglion cells of retinal dystrophic rats are less affected despite little or no synaptic input for several months. Furthermore, any reduction in the electrical activity of retinal ganglion cells that might follow loss of photoreceptor cells does not result in a significantly decreased rate of axonal transport.


Nicotinamide ameliorates palmitate-induced ER stress in hepatocytes via cAMP/PKA/CREB pathway-dependent Sirt1 upregulation.
Nicotinamide (NAM) is the amide of nicotinic acid and a predominant precursor for NAD(+) biosynthesis via the salvage pathway. Sirt1 is a NAD(+)-dependent deacetylase, playing an important role in regulating cellular functions. Although hepatoprotective effect of NAM has been reported, the underlying mechanism remains elusive. ER stress, induced by saturated fatty acids, in specific palmitate, plays a pathological role in the development of nonalcoholic fatty liver disease. This study aims to determine the effect of NAM on palmitate-induced ER stress in hepatocytes and to elucidate molecular mechanisms behind. Both HepG2 cells and primary mouse hepatocytes were exposed to palmitate (conjugated to BSA at a 2:1 M ratio), NAM, or their combination for different durations. Cellular NAD(+) level, Sirt1 expression/activity, ER stress, as well as cAMP/PKA/CREB pathway activation were determined. NAM increased Sirt1 expression and enzymatic activity, which contributes to the ameliorative effect of NAM on palmitate-triggered ER stress. NAM increased intracellular NAD(+) level in hepatocytes, however, blocking the salvage pathway, a pathway for NAD(+) synthesis from NAM, only partially prevented NAM-induced Sirt1 upregulation while completely prevented NAD+ increase in response to NAM. Further mechanistic investigations revealed that NAM elevated intracellular cAMP level via suppressing PDE activity, leading to downstream PKA and CREB activation. Importantly, cAMP/PKA/CREB pathway blockade abolished not only NAM-induced Sirt1 upregulation, but also its protective effect against ER stress. Our results demonstrate that NAM protects hepatocytes against palmitate-induced ER stress in hepatocytes via upregulating Sirt1. Activation of the cAMP/PKA/CREB pathway plays a key role in NAM-induced Sirt1 upregulation.


Actions of general anaesthetics on a neuronal nicotinic acetylcholine receptor in isolated identified neurones of Lymnaea stagnalis.
1. Completely isolated identified neurones from the right parietal ganglion of the pond snail Lymnaea stagnalis were studied under two-electrode voltage-clamp. Neuronal nicotinic acetylcholine receptor currents were studied at low acetylcholine (ACh) concentrations (< or = 200 nM). At these levels, control currents were non-desensitizing and proportional to the square of the ACh concentration. 2. IC50 concentrations were determined for the steady-state inhibition of the ACh-activated current by 31 general anaesthetics plus the non-anaesthetic alcohol n-tridecanol. The general anaesthetics included inhalational agents, n-alcohols, n-alkane-(alpha,omega)-diols, cycloalcohols and an n-alkane. 3. Anaesthetic inhibition was independent of voltage and consistent with two anaesthetic-binding sites on the receptor. 4. IC50 concentrations for inhibiting the neuronal nicotinic ACh receptor correlated well (r = 0.97) with EC50 concentrations for general anaesthesia. The maximum deviation from the line of identity was less than fourfold. The inhalational agents tended to be more potent as inhibitors of the ACh receptor than as general anaesthetics, while the alcohols and diols were less potent. 5. The inhibition of the ACh-induced current by the homologous series of n-alcohols exhibited a cutoff at the same position (just after dodecanol) as found for the induction of general anaesthesia in tadpoles. 6. Polarity profile maps of the anaesthetic-binding sites on the neuronal nicotinic ACh receptor were calculated from IC50 concentrations for the homologous series of n-alcohols and n-alkane-(alpha,omega)-diols. They reveal amphiphilic sites with apolar regions capable of accommodating the hydrocarbon chains of n-alcohols as large as decanol. A striking resemblance was found to profiles previously calculated from data for tadpole general anaesthesia.


Dissecting the mode of action of various HIV-inhibitor classes in a stable cellular system.
We describe a stable and sensitive HIV evaluation system, which discriminates HIV-specific membrane fusion and early transcription events and is suitable for high-throughput inhibitor screening. A human lymphocytic line, constitutively producing infectious HIV-1, serves as Env-positive donor. A second indicator cell line carries a silent HIV-1 LTR lacZ reporter plasmid. A bicellular cocultivation setup allows titration and standardization of "fusion-induced gene stimulation (FIGS)" events. With few manipulations aspects of fusion and/or LTR induction can be distinguished and simultaneously assayed. Anti-Env-V3 antibodies prevent fusion and subsequent lacZ induction, and a Tat-specific inhibitor blocks only lacZ induction in a dose dependent manner without affecting membrane fusion. The LTR reporter is readily activated by Tat from HIV-1, HIV-2, or SIV and it responds to exogenous Tat protein. The reporter system is sensitive enough to detect single infection events on pre-seeded layers of indicator cells, which renders it potentially useful for direct virus quantification in patients' material. Moreover, our system allows to control and normalize DNA transfection efficiencies of HIV-derived plasmids. This aspect is particularly valuable for studies of RT- and protease-inhibitors and resistances, where p24 or supernatant reverse transcriptase, otherwise standard virus readouts, can be directly affected by inhibitors or mutations.


Relation between HLA and copy number variation of steroid 21-hydroxylase in a Swedish cohort of patients with autoimmune Addison's disease.
Autoantibodies against the adrenal enzyme 21-hydroxylase is a hallmark manifestation in autoimmune Addison's disease (AAD). Steroid 21-hydroxylase is encoded by CYP21A2, which is located in the human leucocyte antigen (HLA) region together with the highly similar pseudogene CYP21A1P. A high level of copy number variation is seen for the 2 genes, and therefore, we asked whether genetic variation of the CYP21 genes is associated with AAD. Case-control study on patients with AAD and healthy controls. Using next-generation DNA sequencing, we estimated the copy number of CYP21A2 and CYP21A1P, together with HLA alleles, in 479 Swedish patients with AAD and autoantibodies against 21-hydroxylase and in 1393 healthy controls. With 95% of individuals carrying 2 functional 21-hydroxylase genes, no difference in CYP21A2 copy number was found when comparing patients and controls. In contrast, we discovered a lower copy number of the pseudogene CYP21A1P among AAD patients (P = 5 × 10-44), together with associations of additional nucleotide variants, in the CYP21 region. However, the strongest association was found for HLA-DQB1*02:01 (P = 9 × 10-63), which, in combination with the DRB1*04:04-DQB1*03:02 haplotype, imposed the greatest risk of AAD. We identified strong associations between copy number variants in the CYP21 region and risk of AAD, although these associations most likely are due to linkage disequilibrium with disease-associated HLA class II alleles.


Synthesis and characterization of 8-ethynyl-1,3-dihydro-benzo[b][1,4]diazepin-2-one derivatives: new potent non-competitive metabotropic glutamate receptor 2/3 antagonists. Part 1.
A series of 1,3-dihydrobenzo[b][1,4]diazepin-2-one derivatives was evaluated as non-competitive mGluR2/3 antagonists. Attachment of an 8-(2-aryl)-ethynyl-moiety produced compounds inhibiting the binding of [(3)H]-LY354740 to rat mGluR2 with low nanomolar affinity and consistent functional effect at both mGluR2 and mGluR3.


β-Arrestin 2 acts an adaptor protein that facilitates viral replication in silkworm.
β-Arrestin 2 is known to be a widely distributed adaptor protein in mammals but its function has never been reported in Lepidoptera insects. Herein, the β-Arrestin 2 (BmArrestin 2) gene from silkworm was cloned and characterized. The spatiotemporal expression level of BmArrestin 2 was highest in the gonads at the 3rd day of 5th instar, whereas the highest and lowest abundance of BmArrestin 2 were identified in the tracheal and testis, respectively. BmArrestin 2 is mainly distributed in the cytoplasm. Furthermore, in BmN cells,overexpression of BmArrestin 2 promoted Bombyx mori nucleopolyhedrovirus (BmNPV) and B. mori cytoplasmic polyhedrosis virus (BmCPV) replication as the increment of the concentration of plasmid transfection, whereas silencing the gene with specific siRNA inhibited viral replication. Replication of BmNPV and BmCPV also was weakened using BmArrestin 2 antiserum as the increment of the concentration. Immunofluorescent staining revealed the invasion of recombinant BmNPV or BmCPV was decreased after blocking endogenous BmArrestin 2. On the other hand, BmArrestin 2 co-localizes with recombinant BmNPV and BmCPV virions in BmN cells. These results suggest that BmArrestin 2 may represent a novel target for antiviral strategies, as it is an adaptor protein that plays a key role in virus replication.


Frailty still matters to health and survival in centenarians: the case of China.
Frailty indicates accumulated vulnerability of adverse health outcomes in later life. Its robustness in predicting dependent living, falls, comorbidity, disability, health change, mortality, and health care utilization at older ages is well-documented. However, almost no studies have ever attempted to examine its robustness in centenarians, mainly due to data unavailability. This study examines prevalence of frailty in centenarians and its predictive powers on subsequent mortality and health conditions. We use a sample of 4434 centenarians from the 2002, 2005, 2008, and 2011 waves of the Chinese Longitudinal Healthy Longevity Survey (CLHLS), with elders in three younger age groups 65-79, 80-89, and 90-99 as comparisons. Frailty is measured by a cumulative deficit index (DI) that is constructed from 39 variables covering physical and cognitive function, disease conditions, psychological well-being, and other health dimensions. Survival analysis is conducted to examine how frailty is associated with subsequent mortality at an average follow-up length of 3.7 years (2.6 years for deceased persons died in 2002-2011 and 7.6 years for survived persons at the 2011 wave). Logistic regressions are applied to examine how frailty is associated with subsequent physical and cognitive functions, disease conditions, and self-rated health with an average follow-up length of 3.0 years. The study reveals that centenarians are frailer than younger elders. The DI scores increase from less than 0.1 at ages 65-79 to over 0.30 in centenarians. Women are frailer than men at all ages. However, there is a great variation in frailty among all age groups. We also find that each additional increase of 0.01 score of the DI is associated with 1.6 % higher mortality risk (95 % CI: 1.014-1.018) in female centenarians and 1.4 % higher mortality risk (95 % CI: 1.010-1.018) in male centenarians, although these associations are weaker than those in other three younger age groups. Frailty still plays an important role in determining subsequent health outcomes and mortality in centenarians.


Pleiotropic effects of NOACs with focus on edoxaban: scientific findings and potential clinical implications.
Non-vitamin K antagonist oral anticoagulants (NOACs) are well-established as inhibitors of factor Xa (FXa) and thrombin in the treatment and prevention of thrombosis. However, there is growing evidence that beneficial outcomes might be based on additional pleiotropic effects beyond anticoagulation. FXa and thrombin are also known to activate protease-activated receptors (PARs), which can mediate pro-inflammatory and pro-fibrotic effects. Since PAR‑1 and PAR‑2 play an important role in the development of atherosclerosis, the inhibition of this pathway represents an interesting potential target for preventing the progression of atherosclerosis and fibrosis. This review focuses on potential pleiotropic effects of FXa inhibition with edoxaban seen in a variety of studies in different in vitro and in vivo test systems. As common findings from these experiments, edoxaban was able to attenuate FXa- and thrombin-induced pro-inflammatory and pro-fibrotic effects and decrease pro-inflammatory cytokine expression. In some, but not all experiments edoxaban was also shown to decrease the levels of PAR‑1 and PAR‑2 expression. Further studies are required to clarify the clinical implications of the pleiotropic effects mediated by NOACs.


Lipoprotein(a) and family history for cardiovascular disease in paediatric patients: A new frontier in cardiovascular risk stratification. Data from the LIPIGEN paediatric group.
Little is known about the role of Lp(a) in the assessment of cardiovascular risk in the paediatric population. Trying to clarify the clinical relevance of Lp(a) in risk stratification, the aim of the study is to evaluate the association between Lp(a) plasma levels in children with familial hypercholesterolaemia (FH) and positive family history for premature cardiovascular disease (pCVD) in first- and second-degree relatives. 653 Caucasian children and adolescents (334 females and 319 males), aged 2-17 years, with diagnosis of FH from a paediatric cohort included in the LIPIGEN Network, were selected. We compared family history of pCVD, lipid and genetic profile in two groups based on Lp(a) levels below or above 30 mg/dL. To determine the independent predictors of pCVD, a multivariate logistic regression was used, with all clinical characteristics and blood measurements as predictors. Subjects with Lp(a) > 30 mg/dl more frequently reported positive family history of pCVD compared to subjects with Lp(a)≤30 mg/dl (69.90% vs 36.66%, p < 0.0001), while did not show differences in terms of median [interquartile range] LDL-cholesterol level (153.00 [88.00 vs 164.50 [90.25] mg/dL, p = 0.3105). In the regression analysis, Lp(a) > 30 mg/dl was an independent predictor of family history of pCVD. Comparing subjects with or without family history of pCVD, we reported significant differences for Lp(a) > 30 mg/dl (46.25% vs 17.65%, p < 0.0001), FH genetic mutation (50.48% vs 40.75%, p = 0,0157), as well as for LDL-cholesterol (p = 0.0013) and total cholesterol (p = 0.0101). Children/adolescents with FH and Lp(a) > 30 mg/dl where more likely to have a positive family history of pCVD. Lp(a) screening in children and adolescents with FH may enhance risk assessment and help identify those subjects, children and relatives, at increased pCVD risk.


The clinical spectrum of anomalous pancreatobiliary junction.
Anomalous junction of pancreatobiliary ducts (AJPB) is known to be associated with choledochal cyst and gallbladder cancer. This report describes our experience with eight patients with AJPB, six of whom were diagnosed at endoscopic retrograde cholangiography and two at transhepatic cholangiography. Four of the patients with AJPB had an associated congenital choledochal cyst, two had a normal biliary tree and one each had inspissated bile syndrome and cholangiocarcinoma. The last two associations have not been previously described. It is felt that the clinical spectrum of AJPB may unfold further with the widespread use of the cholangiographic techniques.


miR-216b enhances the efficacy of vemurafenib by targeting Beclin-1, UVRAG and ATG5 in melanoma.
Autophagy maintains cells survival in many stressful conditions including starvation, growth factor deprivation and misfolded protein accumulation. Additionally, autophagic survival mechanisms are used by transformed tumor cells to inhibit cell death, limit drug effectiveness and possibly generate drug resistance. However, the mechanism of how cells utilize autophagy during drug resistance is not fully understood. Here, we demonstrate that miR-216b plays an important role in alleviating drug resistance by regulating autophagy in melanoma. We show that miR-216b attenuates autophagy by directly targeting three key autophagy genes Beclin-1, UVRAG and ATG5. Overexpression of these genes from miRNA immune cDNA constructs rescue autophagic activity in the presence of miR-216b. Antagomir-mediated inactivation of endogenous miR-216b led to an increase of Beclin-1, UVRAG, ATG5, and subsequent autophagic activity. More importantly, we have discovered that BRAF(V600E) inhibitor vemurafenib suppresses miR-216b activity, which in turn activates autophagy to generate drug resistance in both BRAFi-sensitive and -resistant cells. Strikingly, ectopic expression of miR-216b increases the efficacy of vemurafenib both in vitro and in vivo. Taken together, these data indicate that miR-216b regulates autophagy by suppressing three key autophagy genes, and enhances the antitumor activity of vemurafenib in BRAF(V600E) melanoma cells.


Epistemic Justification and Methodological Luck in Inflationary Cosmology.
I present a recent historical case from cosmology-the story of inflationary cosmology-and on its basis argue that solving explanatory problems is a reliable method for making progress in science. In particular, I claim that the success of inflationary theory at solving its predecessor's explanatory problems justified the theory epistemically, even in advance of the development of novel predictions from the theory and the later confirmation of those predictions. 1Introduction2A Portrait of the Argument as a Short Précis3A Brief History of Inflationary Theory4Empiricist Interpretations of Inflationary Case5A Confutation of Empiricist Interpretations6Explanationist Interpretation of Inflationary Case7Concluding Remarks.


Constraints on integration of orthographic information across multiple stimuli: effects of contiguity, eccentricity, and attentional span.
Five flanked lexical decision experiments investigated the integration of information across spatially distinct letter strings. Experiment 1 found no significant difference between quadrigram flankers (e.g., CKRO ROCK CKRO) and double bigram flankers (e.g., CK RO ROCK CK RO). Experiment 2 varied the eccentricity of single bigram flankers and found that closer flankers generated greater effects. A combined analysis of these experiments revealed that the double bigram condition (Experiment 1) was less effective than the close single bigram condition (Experiment 2). Experiment 3 tested one explanation for this pattern - that the outer bigrams in the double bigram condition interfered with processing the inner bigrams, and that spatial integration only operates across adjacent stimuli. In Experiment 3, outer bigrams were now a repeat of the inner bigram (e.g., RO RO ROCK CK CK), and this repeated bigram condition was still found to be significantly less effective than single bigrams. Experiments 4 and 5 tested an alternative explanation whereby the addition of spatially distinct flanking stimuli increases the spread of spatial attention, hence reducing the impact of proximal flankers. In line with this explanation, we found no significant difference between repeated bigram flankers and a condition where only the inner bigram was related to the target (e.g., CA RO ROCK CK SH). We conclude that spatial integration processes only operate across the central target and proximal flankers, and that these effects are diluted by the increased spread of spatial attention caused by additional spatially distinct flankers.


Relationships between self-perceived and clinical expression of pain and function differ based on the underlying pathology of the human hip.
Patient-reported outcomes are commonly used to assess patient symptoms. The effect of specific hip pathology on relationships between perceived and objectively measured symptoms remains unclear. The purpose of this study was to evaluate differences of function and pain in patients with FAIS and DDH, to assess the correlation between perceived and objective function, and to determine the influence of pain on measures of function. This prospective cross-sectional study included 35 pre-operative patients (60% female) with femoroacetabular impingement syndrome (FAIS) and 37 pre-operative patients (92% female) with developmental dysplasia of the hip (DDH). Objectively measured function (6-min walk [6MWT], single leg hop [SLHT], Biodex sway [BST], hip abduction strength [HABST], and STAR excursion balance reach [STAR] tests), patient-reported function (UCLA Activity, Hip Outcome Score [HOS], Short Form 12 [SF-12], and Hip Disability and Osteoarthritis Outcome Score [HOOS]), and patient-reported pain (HOOS Pain, visual analogue scale (VAS), and a pain location scale) were collected during a pre-surgical clinic visit. Between-group comparisons of patient scores were performed using Wilcoxon Rank-Sum tests. Within-group correlations were analyzed using Spearman's rank correlation coefficients. Statistical correlation strength was defined as low (r = ± 0.1-0.3), moderate (r = ± 0.3-0.5) and strong (r > ± 0.5). Patients with DDH reported greater pain and lower function compared to patients with FAIS. 6MWT distance was moderately-to-strongly correlated with a number of patient-reported measures of function (FAIS: r = 0.37 to 0.62, DDH: r = 0.36 to 0.55). Additionally, in patients with DDH, SLHT distance was well correlated with patient reported function (r = 0.37 to 0.60). Correlations between patient-reported pain and objectively measured function were sparse in both patient groups. In patients with FAIS, only 6MWT distance and HOOS Pain (r = -0.53) were significantly correlated. In patients with DDH, 6MWT distance was significantly correlated with VAS Average (r = -0.52) and Best (r = -0.53) pain. Pain is greater and function is lower in patients with DDH compared to patients with FAIS. Moreover, the relationship between pain and function differs between patient groups. Understanding these differences is valuable for informing treatment decisions. We recommend these insights be incorporated within the clinical continuum of care, particularly during evaluation and selection of surgical and therapeutic interventions.


Oocyte cytoplasmic diameter of ≥130 μm can be used to determine human giant oocytes.
To determine if a cytoplasmic diameter of ≥130 μm can help identify human giant oocytes (GOs) in clinical practice and confirm the presence of genetic abnormalities in GOs by assessing the spindle length and centromere numbers. Case-control study. Private in vitro fertilization clinic. The subjects were women aged 20-49 years who underwent oocyte retrieval after ovarian stimulation from January 2014 to December 2020. None. The oocyte diameter was measured; immunofluorescent staining was performed to assess the spindle diameter and centromere numbers. Among the 254,337 oocytes examined, 561 (0.22%) had a diameter of ≥130 μm. The mean diameter ranges in the normal-sized metaphase II (MII) oocytes (MII group) and GO group were 103.0-119.0 and 132.3-175.9 μm. Spindle size could be measured in 6 GOs with 1 spindle (GO1), 10 GOs with 2 spindles (GO2), and 16 MII groups. The equatorial plane and pole-to-pole distance in the GO1 were significantly longer than in the GO2 and MII groups. The median numbers of centromeres were 86 in GOs with 1 spindle and 42 in each spindle for GOs with 2 spindles among 11 GO1s and 5 GO2s. This study is the first to define GOs as oocytes with a diameter of ≥130 μm and is a large-scale study surveying the incidence of GO. It is also the first study to analyze and elucidate the relationship between spindle numbers within the cytoplasm of GOs and spindle size and centromeres.


Hydrothermal hydrolysis of algal biomass for biofuels production: A review.
Hydrothermal hydrolysis is an energy-efficient and economical pretreatment technology to disrupt the algal cells and hydrolyze the intracellular compounds, thereby promoting the biofuels production of fermentation. However, complex reaction mechanisms, unpredictable rheological properties of algal slurry, and immature continuous reactors still constrain the commercialization of such a process. To systematically understand the existing status and lay a foundation for promoting the technology, the chemical mechanism of hydrothermal hydrolysis of algal biomass is elaborated in this paper, and the influences of temperature, residence time, total solid content, and pH, on the biomethane production of hydrolyzed algal biomass are summarized. Besides, a comprehensive overview of the rheological behavior of algal slurries is discussed at various operational factors. The recent advances in flow, heat and mass transfer model coupling with the generic kinetics model in continuous reactors and the application of energy-saving strategies for efficient algal biomass pretreatment are detailed reviewed.


Identification of volatile active components in Acori Tatarinowii Rhizome essential oil from different regions in China by C6 glioma cells.
Acori Tatarinowii Rhizome (ATR) is a well-recognized Chinese herbal medicine prescribed to treat neurological disorders. The essential oil (ATEO) is considered as the active fraction of ATR and the content of ATEO is used as the only indicator for ATR content determination. The quality of ATEO varies widely due to region difference; however, little is known about how to study ATEO quality chemically and biologically in response to region difference. Thus, it is of great importance to identify volatile active components in ATEO to conduct quality study. In this study, we analyzed ATEO from different regions in China using chemical component analysis combined with biological activity evaluation. GC-MS was used to obtain different volatile component profiles of ATEO and significantly changed volatile components were screened out. The neuroprotective activities of ATEO, including anti-oxidation, anti-inflammation and neurotrophic functions, were revealed in C6 glioma cells. The correlation study between the bioactivities and the components was performed. 57 volatile components, including terpenoids, phenylpropanoids, aromatic compounds, and other aliphatic compounds, were identified. 8 volatile components (β-asarone, cis-methyl isoeugenol, γ-asarone, methyleugenol, calarene, longifolene, β-caryophyllene and caryophyllene oxide) from ATEO were significantly changed due to region difference and 2 of them (β-asarone and γ-asarone) showed strong correlation with neuroprotective activities. Our results reveal that ATEO from different regions in China show great changes in chemical composition and biological activity. Moreover, phenylpropanoids (β-asarone and γ-asarone) present strong correlation with the bioactivities, which are considered as volatile active components in ATEO. The findings will be useful for the development of quality study of ATEO.


The prevalence of comorbid symptoms of central sensitization syndrome among three different groups of temporomandibular disorder patients.
Symptoms of central sensitization syndrome (CSS) were evaluated among three different groups of temporomandibular disorder (TMD) patients. Additionally, TMD group differences in pain and pain-related disability were assessed, as well as emotional distress. Participants were 250 patients with symptoms of acute TMD, recruited from dental clinics within a major metropolitan area. Sequential regressions and multivariate analyses of covariance were conducted in order to make group comparisons. Those with a TMD Muscle Disorder (ie, myofacial TMD [m-TMD]) and those with more than one TMD diagnosis had the most symptoms of CSS and higher reports of pain and pain-related disability. Moreover, emotional distress accounted for a substantial amount of the variance for physical symptoms and mediated all TMD comparisons. Myofacial TMD is characterized by a high degree of comorbidity of symptoms of CSS and associated emotional distress.


Post-surgical hallux varus. A case report.
Sometimes a hallux varus occurs as a consequence of the treatment for a hallux valgus, determined by an alteration of the balance between the muscolo-ligamentous structure which crosses the first metatarso-phalangeal joint. This complication is poorly tolerated by patients. Various operations have been proposed to correct the varus, which includes the bone and the soft tissue. A clinical case has been presented which uses a split extensor hallucis longus transfer, distally sectioned and made to pass under the deep transverse intermetatarsal ligament, used as a pulley, and through a tunnel made in the bone at the proximal-lateral base of the proximal phalanx, and then tensioned and sutured to its medial side.


Identification of conserved regions from 230,163 SARS-CoV-2 genomes and their use in diagnostic PCR primer design.
As the rapidly evolving characteristic of SARS-CoV-2 could result in false negative diagnosis, the use of as much sequence data as possible is key to the identification of conserved viral sequences. However, multiple alignment of massive genome sequences is computationally intensive. To extract conserved sequences from SARS-CoV-2 genomes for the design of diagnostic PCR primers using a bioinformatics approach that can handle massive genomic sequences efficiently. A total of 230,163 full-length viral genomes were retrieved from the NCBI SARS-CoV-2 Resources and GISAID EpiCoV database. This number was reduced to 14.11% following removal of 5'-/3'-untranslated regions and sequence dereplication. Fast, reference-based, multiple sequence alignments identified conserved sequences and specific primer sets were designed against these regions using a conventional tool. Primer sets chosen among the candidates were evaluated by in silico PCR and RT-qPCR. Out of 17 conserved sequences (totaling 4.3 kb), two primer sets targeting the nsp2 and ORF3a genes were picked that exhibited > 99.9% in silico amplification coverage against the original dataset (230,163 genomes) when a 5% mismatch between the primers and target was allowed. In addition, the primer sets successfully detected nine SARS-CoV-2 variant RNA samples (Alpha, Beta, Gamma, Delta, Epsilon, Zeta, Eta, Iota, and Kappa) in experimental RT-qPCR validations. In addition to the RdRp, E, N, and S genes that are targeted commonly, our approach can be used to identify novel primer targets in SARS-CoV-2 and should be a priority strategy in the event of novel SARS-CoV-2 variants or other pandemic outbreaks.


Omics analyses of a somatic Trp53R245W/+ breast cancer model identify cooperating driver events activating PI3K/AKT/mTOR signaling.
Alterations of the tumor suppressor TP53, one of the most common events in cancer, alone are insufficient for tumor development but serve as drivers of transformation. We sought to identify cooperating events through genomic analyses of a somatic Trp53R245W mouse model (equivalent to the TP53R248W hot spot mutation in human cancers) that recapitulates metastatic breast-cancer development. We identified cooperating lesions similar to those found in human breast cancers. Moreover, we identified activation of the Pi3k/Akt/mTOR pathway in most tumors via mutations in Pten, Erbb2, Kras, and/or a recurrent Pip5k1c mutation that stabilizes the Pip5k1c protein and activates Pi3k/Akt/mTOR signaling. Another PIP5K1C family member, PIP5K1A, is coamplified with PI4KB in 18% of human breast cancer patients; both encode kinases that are responsible for production of the PI3K substrate, phosphatidylinositol 4,5-bisphosphate. Thus, the TP53R248W mutation and PI3K/AKT/mTOR signaling are major cooperative events driving breast-cancer development. Additionally, a combination of two US Food and Drug Administration (FDA)-approved drugs, tigecycline and metformin, which target oxidative phosphorylation downstream of PI3K signaling, inhibited tumor cell growth and may be repurposed for breast-cancer treatment. These findings advance our understanding of how mutant p53 drives breast-tumor development and pinpoint the importance of PI3K/AKT/mTOR signaling, expanding combination therapies for breast-cancer treatment.


Extending the conditions of application of an inversion of the Hodgkin-Huxley gating model.
We present an inversion of the Hodgkin-Huxley formalism to estimate initial conditions and model parameters, including functions of voltage, from the solutions of the underlying ordinary differential equation (ODE) subjected to multiple voltage step stimulations. As such, the procedure constitutes a means to estimate the parameters including functions of voltage of an Hodgkin-Huxley formalism from experimental data.The basic idea was developed in a previous communication (SIAM J. Appl. Math. 64:1264-1274, 2009). The inversion in question applies to currents exhibiting activation and inactivation, but the version, as published previously, cannot estimate the unknowns for channels that rapidly inactivate just after a brief opening. In such cases, the amplitude of the current, in a given voltage range, is too small to be detectable by the instrumentation using previously applied experimental protocols. This is, for example, the case for the sodium channels in a number of excitable tissue for potential in the vicinity of the cell resting potential. The current communication extends the inversion procedure in a manner to overcome this limitation.Furthermore, within the inversion framework, we can determine whether the data at the basis of the estimation sufficiently constrains the estimation problem, i.e., whether it is complete. We exploit this element of our method to document a set of stimulation protocols that constitute a complete data set for the purpose of inverting the Hodgkin-Huxley formalism.


Determination of 4-amino-1-hydroxybutane-1,1-bisphosphonic acid in urine by automated pre-column derivatization with 2,3-naphthalene dicarboxyaldehyde and high-performance liquid chromatography with fluorescence detection.
A sensitive (5 ng/ml) method for the determination of 4-amino-1-hydroxybutane-1,1-bisphosphonic acid in human urine is described. The procedure includes (1) the isolation of the drug from urine by co-precipitation of its calcium salt with endogenous phosphates in the presence of base, (2) a solid-phase anion-exchange sample clean-up and (3) automated pre-column derivatization of the primary amino group with 2,3-naphthalene dicarboxyaldehyde-cyanide reagent followed by fluorescence detection of the N-substituted cyanobenz[f]isoindole derivative. The derivative of the drug was synthesized and its spectral and fluorescence properties were evaluated. The fluorescence quantum efficiency was determined to be 0.82 in the mobile phase used for the assay. The derivative is also capable of accepting energy in an oxalate ester-hydrogen peroxide chemiluminescence system.


Regulatory implications of protein assemblies at the gamma origin of plasmid R6K - a review.
Recognition of the replication origin (ori) by initiator protein is a recurring theme for the regulated initiation of DNA replication in diverse biological systems. The objective of the work reviewed here is to understand the initiation process focusing specifically on the gamma-ori of the antibiotic-resistance plasmid R6K. The control of gamma-ori copy number is determined by both plasmid-encoded and host-encoded factors. The two central regulatory elements of the plasmid are a multifunctional initiator protein pi, and sequence-related DNA target sites, the inverted half-repeats (IRs) and the direct repeats (DRs). The replication activator and inhibitor activities of pi seem to be at least partially distributed between two naturally occurring pi polypeptides (designated by their molecular weights pi35.0 and pi30.5). Regulatory variants of pi with altered states of oligomerization in nucleoprotein complexes with DRs and IRs have been isolated. The properties of these mutants laid the foundation for our model of pi protein activity which proposes that different protein surfaces are required for the formation of functionally distinct complexes of pi with DRs and IRs. These mutants also suggest that pi polypeptides have a modular structure; the C-terminus contains the DNA-binding domain while the N-terminus controls protein oligomerization. Additionally, pi35.0 binds to a novel DNA sequence in the A+T-rich segment of gamma-ori. This binding site is at or near the site from which synthesis of the leading strand begins.


In vitro activities against mycobacteria of two long-acting rifamycins, FCE22807 and CGP40/469A (SPA-S-565).
The in vitro activities of two new long-acting rifamycins, FCE22807 a derivative of FCE22250, and CGP40/469A (SPA-S-565) were studied. When compared with rifampicin, the minimal inhibitory concentrations (MIC) against Mycobacterium tuberculosis of both were 4 times lower but neither was particularly active against rifampicin-resistant strains of M. tuberculosis nor against M. avium-intracellulare-scrofulaceum complex strains. A drug is likely to be particularly effective in widely spaced intermittent dosage if it has a long half-life and high bactericidal activity. When tested against M. tuberculosis in the logarithmic and in the stationary phase of growth, FCE22807 was amongst the most bactericidal of the rifamycins while CGP40/469A had little bactericidal activity.


Removal of heavy metal from sludge by the combined application of a biodegradable biosurfactant and complexing agent in enhanced electrokinetic treatment.
In this work, the heavy metal removal potentiality of an electrokinetic (EK) decontamination treatment enhanced by a biodegradable complexing agent Tetrasodium of N, N-bis (carboxymethyl) glutamic acid (GLDA) also in combination with a biodegradable biosurfactant (rhamnolipid) was investigated to decontaminate heavy metals from the sludge. The main results explored that the nature of sludge and their interactions with different improving agents significantly influenced the electrokinetic removal processes. A general increase of pH values from anode to cathode in the sludge-cell was observed due to the strong buffering capacity of carbonates. Compared with the deionized water, the use of GLDA as an electrolyte, Cu, Zn, Cr, Pb, Ni and Mn removal efficiencies were 53.2 ± 3.12%, 67.4 ± 3.45%, 59.2 ± 4.78%, 45.4 ± 4.15%, 72.8 ± 3.68% and 45.0 ± 4.85%, respectively, whereas a further improvement heavy metals removal efficiencies (Cu, Zn, Cr, Pb, Ni and Mn removal efficiencies were 64.8 ± 2.34%, 56.8 ± 4.12%, 49.4 ± 4.45%, 46.6 ± 2.35%, 60.4 ± 3.45% and 69.6 ± 3.54%, respectively) were achieved by repalcing rhamnolipid as the electrolyte. Significantly higher removal efficiencies (Cu, Zn, Cr, Pb, Ni and Mn removal efficiencies were 70.6 ± 3.41%, 82.2 ± 5.21%, 89.0 ± 3.34%, 60.0 ± 4.67%, 88.4 ± 4.43% and 70.0 ± 3.51%, respectively) were obtained by the simultaneous use of GLDA and rhamnolipid due to their synergic action in electrokinetic process.


Spontaneous myocardial calcium oscillations: are they linked to ventricular fibrillation?
The physiological oscillation of cytosolic [Ca2+] that underlies each heart beat is generated by the sarcoplasmic reticulum (SR) in response to an action potential (AP) and occurs relatively synchronously within and among cells. When the myocardial cell and SR Ca2+ loading become sufficiently high, the SR can also generate spontaneous, i.e., not triggered by sarcolemmal depolarization, Ca2+ oscillations (S-CaOs). The purpose of this review is to describe properties of S-CaOs in individual cells, myocardial tissue, and the intact heart, and to examine the evidence that may link S-CaOs to the initiation or maintenance of ventricular fibrillation (VF). The SR Ca2+ release that generates S-CaOs occurs locally within cells and spreads within the cell via Ca(2+)-induced Ca2+ release. The localized increase in cytosolic [Ca2+] due to S-CaOs may equal that induced by an AP and causes oscillatory sarcolemmal depolarizations of cells in which it occurs. These oscillatory depolarizations are due to Ca2+ activation of the Na/Ca exchanger and of nonspecific cation channels. Asynchronous occurrence of diastolic S-CaOs among cells within the myocardium causes inhomogeneity of diastolic SR Ca2+ loading; this leads to inhomogeneity of the systolic cytosolic [Ca2+] transient levels in response to a subsequent AP, which leads to heterogeneity of AP repolarization, due to heterogeneous Ca2+ modulation of the Na/Ca exchanger, nonspecific cation channels, and of the L-type Ca2+ channel. In a tissue in which asynchronous S-CaOs are occurring in diastole, the subsequent AP temporarily synchronizes SR Ca2+ loading and release within and among cells. Varying extents of synchronized S-CaOs then begin to occur during the subsequent diastole. The partial synchronization of this diastolic S-CaOs among cells within myocardial tissue produces aftercontractions and diastolic depolarizations. When S-CaOs are sufficiently synchronized, the resultant depolarizations summate and can be sufficient to trigger a spontaneous AP.S-CaOs occurrence within some cells during a long AP plateau also modulates the removal of voltage inactivation of L-type Ca2+ channels and increases the likelihood for "early afterdepolarizations" to occur in myocardial tissue. S-CaOs have an apparent modulatory role in the initiation of VF in the Ca2+ overload model and in the reflow period following ischemia. Likewise, in non-a priori Ca2+ overloaded hearts, S-CaOs modulate the threshold for VF induction (induced typically by alternating current) but may not be essential for VF induction.(ABSTRACT TRUNCATED AT 400 WORDS)


Addressing buprenorphine bottlenecks in the context of MAT Act implementation: A shared responsibility.
Medications for opioid use disorder (OUD) such as buprenorphine reduce overdose mortality and other opioid related acute health events but have historically been highly regulated. The recent Mainstreaming Addiction Treatment (MAT) Act ended the requirement clinicians complete a specified training and apply for a DATA 2000 ("X") waiver on their Drug Enforcement Administration (DEA) number, to prescribe buprenorphine. With the MAT Act, any practitioner with Schedule III prescribing authority (a regular DEA number) can now prescribe buprenorphine for OUD. While this has potential to improve OUD treatment access, the impact will depend on implementation. Though the MAT Act may facilitate increased buprenorphine prescribing, ensuring adequate buprenorphine dispensing is also critical to improving Medications for opioid use disorder. Recognized buprenorphine bottlenecks arise from a complex convergence of factors in community pharmacies, threatening to undercut the benefits of the MAT Act. If prescribing increases but is not matched by increased dispensing, bottlenecks may worsen. Any worsening of buprenorphine bottlenecks could have a disproportionate impact in rural areas where residents may rely on fewer pharmacies to fill prescriptions for people in larger geographic area and where larger prescribing-dispensing gaps already exist such as in Southern states. Rigorous research will be needed to document the overall impact of the MAT Act on community pharmacists and their patients. At the federal level, pharmacists and their professional organizations should lobby the DEA to de-schedule or re-schedule buprenorphine. The DEA should announce a moratorium on enforcement actions against wholesalers and pharmacies related to buprenorphine distribution and dispensing. More supports should be offered to community pharmacies by state pharmacy boards and associations including continuing pharmacy education and technical assistance for advocating with wholesalers to increase buprenorphine order sizes, and to more effectively communicate with prescribers. Pharmacies should not have to face these challenges alone. Regulators, wholesalers, and researchers must join together with community pharmacies to further reduce regulatory barriers to dispensing, provide evidence-based interventions where needed to support pharmacy dispensing efforts, conduct rigorous implementation research, and be constantly vigilant in identifying and addressing multi-level buprenorphine bottlenecks in the wake of the MAT Act.


Inadequate follow-up controller medications among patients with asthma who visit the emergency department.
The purpose of this study is to determine the frequency with which primary care physicians add inhaled corticosteroids to the regimen of asthmatic patients after a visit to the emergency department (ED) among patients not previously prescribed inhaled corticosteroids and to determine the rate at which inhaled corticosteroids prescribed in the ED were continued by primary care physicians. We conducted a structured retrospective cohort study using electronic medical record review of consecutive patients aged 6 to 45 years, treated for acute asthma exacerbation (International Classification of Diseases, Ninth Revision code 493.00 through 493.99) in the ED during a specified 6-month period, and followed up for 1 year. The patients' first ED visit for asthma exacerbation during the study period was considered the index visit for purposes of this study. Six hundred twenty-nine patients met study inclusion criteria, 414 of whom were not previously receiving inhaled corticosteroid therapy. On ED or hospital discharge, 99 (24%) of these 414 patients were prescribed an inhaled corticosteroid. Of these 99 patients, 37 patients had a primary care follow-up visit within 6 months, with 4 receiving an inhaled corticosteroid dose change and no patients having the inhaled corticosteroids discontinued. Of the 315 patients not prescribed an inhaled corticosteroid on ED or hospital discharge, 128 had a primary care follow-up visit within 6 months, with 32 (25%) patients having an inhaled corticosteroid added to their therapeutic regimen. After primary care follow-up, only 69 (42%) of the 165 patients treated in clinic were receiving an inhaled corticosteroid for control of their asthma. Patients without insurance (odds ratio 0.14; 95% confidence interval 0.027 to 0.71) and patients initially discharged home from the ED (odds ratio 0.17; 95% confidence interval 0.05 to 0.53) were much less likely to receive inhaled corticosteroids at follow-up on multivariate logistic regression adjusting for race, sex, insurance status, and initial disposition. Primary care physicians infrequently add controller medications (inhaled corticosteroids) at follow-up to the regimen of asthmatic patients after a visit to the ED. Emergency physicians should be encouraged to evaluate chronic asthma burden among patients presenting with exacerbation, educate asthmatic patients, and prescribe controller medications, such as inhaled corticosteroids, for those with persistent symptoms.


Osteochondral Fracture Fixation With Fragment Preserving Suture Technique.
Osteochondral fractures are relatively uncommon injuries that typically present after an acute or subacute traumatic injury. Osteochondral fracture fixation is traditionally performed in the acute setting with internal fixation procedures using pins or compression screws through the fragment. Outcomes have generally been good, but cartilage thinning, subchondral remodeling, and tissue reactions can occur after internal fixation through the fragment. This article describes osteochondral fracture fragment fixation with a fragment-preserving technique that does not violate the articular cartilage of the fragment. This technique minimizes risk to articular cartilage that has already sustained injury and also provides superior fixation.


Self-template-assisted micro-phase segregation in blended liquid-crystalline block copolymers films toward three-dimensional structures.
In-plane mesopatterns derived from block-copolymer (BCP) micro-phase segregation in thin films have attracted much interest in practical applications as well as fundamental research programs. However, phase segregation along the film-normal direction has been less studied. Here, we describe a strategy to concurrently, yet independently, control in-plane micro-phase and out-of-plane macro-phase segregation in multiblended films composed of liquid-crystalline BCPs (LCBCPs), affording spontaneously layered three-dimensional (3D) mesostructures. This strategy relies on sequential liquid crystallization during the cooling process in thermal annealing as follows. The constituent LCBCP with the highest isotropic-transition temperature (Tiso) first liquid-crystallizes and segregates from the other LCBCP mixture remaining in isotropic states to form a noncontaminated layer at the top surface. This preformed LCBCP layer preserves its inherent in-plane pattern and acts as a template guiding the subsequent micro-phase segregations of the other low-Tiso LCBCPs underneath. This self-template-assisted micro-phase segregation (STAMPS) readily provides 3D mesostructures, the potential toward rational material design of which is also demonstrated in water-separation applications.


The inhibition of movement by morphine or haloperidol depends on an intact nucleus reticularis tegmenti pontis.
Electrolytic damage of the nucleus reticularis tegmenti pontis (NRTP) in rats produces a form of accelerating forward locomotion, indicating that this region is part of a system that inhibits locomotion and movement. In animals with such damage, 5 mg/kg haloperidol does not block forward locomotion although it produces complete akinesia in normal rats [2]. Our present results demonstrate that doses of morphine sulfate that render normal rats completely akinetic (40, 50, or 60 mg/kg) also fail to block forward locomotion. Furthermore, 200 micrograms gamma-aminobutyric acid applied intracranially in the region of the NRTP can reverse the akinesia produced by systemically administered haloperidol or morphine. We suggest that there are two types of akinesia--direct and indirect--and that morphine and haloperidol may produce akinesia indirectly via an inhibitory system which includes the NRTP.


Bipartite tight spectral clustering (BiTSC) algorithm for identifying conserved gene co-clusters in two species.
Gene clustering is a widely used technique that has enabled computational prediction of unknown gene functions within a species. However, it remains a challenge to refine gene function prediction by leveraging evolutionarily conserved genes in another species. This challenge calls for a new computational algorithm to identify gene co-clusters in two species, so that genes in each co-cluster exhibit similar expression levels in each species and strong conservation between the species. Here, we develop the bipartite tight spectral clustering (BiTSC) algorithm, which identifies gene co-clusters in two species based on gene orthology information and gene expression data. BiTSC novelly implements a formulation that encodes gene orthology as a bipartite network and gene expression data as node covariates. This formulation allows BiTSC to adopt and combine the advantages of multiple unsupervised learning techniques: kernel enhancement, bipartite spectral clustering, consensus clustering, tight clustering and hierarchical clustering. As a result, BiTSC is a flexible and robust algorithm capable of identifying informative gene co-clusters without forcing all genes into co-clusters. Another advantage of BiTSC is that it does not rely on any distributional assumptions. Beyond cross-species gene co-clustering, BiTSC also has wide applications as a general algorithm for identifying tight node co-clusters in any bipartite network with node covariates. We demonstrate the accuracy and robustness of BiTSC through comprehensive simulation studies. In a real data example, we use BiTSC to identify conserved gene co-clusters of Drosophila melanogaster and Caenorhabditis elegans, and we perform a series of downstream analysis to both validate BiTSC and verify the biological significance of the identified co-clusters. The Python package BiTSC is open-access and available at https://github.com/edensunyidan/BiTSC. Supplementary data are available at Bioinformatics online.


Cardiovascular autonomic neuropathy and proliferative diabetic retinopathy.
To determine prospectively whether cardiovascular autonomic neuropathy is a risk factor for proliferative diabetic retinopathy. A five-year follow-up study of 88 diabetic persons was performed at a center providing primary and specialized care for diabetes. Participants were white, insulin-dependent patients with diabetes of 15 to 21 years' duration. The primary end point was the presence of proliferative diabetic retinopathy, seen either on fundus photography or ophthalmologic examination. Cardiovascular autonomic neuropathy was measured at baseline by using a standard protocol. Fourteen patients developed proliferative diabetic retinopathy during follow-up. One measure of cardiovascular autonomic neuropathy, the 30:15 ratio, the heart rate variation at the 30th beat compared with that at the 15th beat, was lower among patients with proliferative diabetic retinopathy (P = .0049) The risk of proliferative diabetic retinopathy in patients with an abnormal cardiovascular autonomic neuropathy index was 2.59, although the estimate was not statistically significant because of the small number of patients who developed proliferative diabetic retinopathy. This study provides prospective evidence that cardiovascular autonomic neuropathy is associated with proliferative diabetic retinopathy. In addition to ocular determinants of proliferative diabetic retinopathy, systemic risk factors also should be considered when examining patients with diabetes mellitus.


Juxtaposed half-cystines as disulphide bridged partners in protein tertiary structure.
Disulphide bridges involving juxtaposed half-cystines are observed in a number of protein three-dimensional structures analyzed from the Protein Data Bank. These disulphide bridges comprise a 'ring of 8-atoms' corresponding to Calpha1-C'-N-Calpha2-Cbeta2-Sgamma2-Sgamma1-Cbeta1-Calpha1 in the two half-cystines. The presence of such disulphide bridges introduces a 'bend' or 'kink' in the protein polypeptide chain.


Adaptive polarization photoacoustic computed tomography for biological anisotropic tissue imaging.
Most photoacoustic computed tomography (PACT) systems usually ignore the anisotropy of the tissue absorption coefficient, which will lead to the lack of information in reconstructed images. In this work, the effect is addressed of the possible optical absorption anisotropy of tissue on PACT images. The functional relationship is derived between the photoacoustic response and the polarization angle of the excitation light. An adaptive polarized light photoacoustic imaging (AP-PACT) approach is proposed and shown to make up for the lack of imaging information and achieve optimal image contrast when imaging samples with anisotropic optical absorption, by utilizing the standard deviation of photoacoustic response as the feedback signal in an adaptive data acquisition process. The method is implemented both on phantom and in vitro experiments, which show that AP-PACT can recover anisotropic absorption-related information from reconstructed images and thus significantly improve their quality.


Study of platelet-rich fibrin promoting endothelial cell differentiation and angiogenesis induced by transplantation of adipose-derived stem cells.
Diabetic patients are characterized by long wound healing time, and adipose stem cells (ADSCs) can secrete growth factors to promote angiogenesis and improve diabetic wound healing. In this research, we attempted to interrogate the impact of platelet-rich fibrin (PRF) on ADSCs in diabetic wound healing. ADSCs were harvested from human adipose tissues and identified through flow cytometry. After pretreatment with cultured medium supplemented with different concentrations of PRF (2.5%, 5%, and 7.5%), proliferation and differentiation capacity of ADSCs were assessed by CCK-8 assay, qRT-PCR and immunofluorescence (IF), respectively. Tube formation assay measured angiogenesis. Western blot analysis analyzed expression of endothelial markers and the extracellular signal-regulated kinase (ERK) and serine/threonine kinase (Akt) pathways in PRF-induced ADSCs. The CCK-8 experiment indicated that PRF enhanced proliferation of ADSCs in dose-dependent manner, relative to normal control group. The expression of endothelial markers and the capacity of tube formation were significantly promoted by 7.5% PRF. The release of growth factors containing vascular endothelial grow factor (VEGF) and insulin-like growth factor-1 (IGF-1) from PRF was increased with the extension of detection time. When the receptors of VEGF or/and IGF-1 were neutralized, ADSCs differentiation into endothelial cells were obviously inhibited. Additionally, PRF stimulated ERK and Akt pathways, and the inhibitors of ERK and Akt attenuated PRF-induced differentiation of ADSCs into endothelial cells. In conclusion, PRF promoted endothelial cell differentiation and angiogenesis induced by ADSCs in diabetic wound healing, which appears to give guidance for treating patients.


"Ethics and clinical research" revisited. A tribute to Henry K. Beecher.
The doctrine of informed consent, borrowed from the law of torts, cannot be readily transplanted into therapeutic settings. The broader, as yet unrealized, idea of informed consent, which suggests that parties must make decisions jointly, should guide interactions between physicians and patients or investigators and subjects.


The prevention of injury from motorcycle use: epidemiologic success, legislative failure.
Following the repeal of the North Dakota mandatory motorcycle helmet law in 1977, the State Highway and Health Departments initiated a study to examine the impact of the change on driver safety. Motorcycle crash data gathered by the State Highway Patrol and other law enforcement agencies was augmented by death and injury reports submitted to the State Health Department by medical care providers. The combined data set produced 2934 crashes, 2162 traumatized victims, 3718 injuries and 53 deaths between January 1977 and December 1980. Reporting from medical providers increased the volume of crash reports, improved the reliability of the highway data and added an unrecognized population of victims to the data base. In spite of the clear indication that injuries and deaths had increased among motorcycle users who rode without helmet protection, legislative passage of a mandatory helmet law in either the 1979 or the 1981 North Dakota legislative session failed. The reasons behind the legislative rejection of efforts to improve highway safety are examined. The forces that affect politics are reviewed.


In vitro sonothrombolysis of human blood clots with BR38 microbubbles.
Microbubble-mediated sonothrombolysis is a promising approach for ischemic stroke treatment. The aim of this in vitro study was to evaluate a new microbubble (MB) formulation (BR38) for sonothrombolysis and to investigate the involved mechanisms. Human whole-blood clots were exposed to different combinations of recombinant tissue plasminogen activator (rtPA), ultrasound (US) and MB. Ultrasound at 1.6 MHz was used at 150, 300, 600 and 1000 kPa (peak-negative pressure). Thrombolysis efficacy was assessed by measuring clot diameter changes during 60-min US exposure. The rate of clot diameter loss (RDL) in μm/min was determined and clot lysis profiles were analyzed. The most efficient clot lysis (5.9 μm/min) was obtained at acoustic pressures of 600 and 1000 kPa in combination with MB and a low concentration of rtPA (0.3 μg/mL). This is comparable with the rate obtained with rtPA at 3 μg/mL alone (6.6 μm/min, p > 0.05). Clot lysis profiles were shown to be related to US beam profiles and microbubble cavitation.


X-ray scattering studies of a model complex of lipid and basic protein of myelin.
Low-angle and wide-angle X-ray scattering data from phosphatidylglycerol complexed with myelin basic protein, poly(L-lysine) and calcium ions are analyzed. The results confirm our earlier report (Brady, G.W., Murthy, N.S., Fein, D.B., Wood, D.D. and Moscarello, M.A. (1981) Biophys. J. 34, 345-350) that the basic protein interacts primarily with the polar headgroups of the lipid; and that at high protein concentrations (greater than 35%) the bilayers aggregate to form multilayers with a repeat period of 68 A, the single bilayer to multilayer transition being a cooperative process. Polylysine and Ca2+ produce multilayers with a smaller repeat of approx. 55 A. Basic protein and polylysine do not change the fluid-like arrangement of the hydrocarbon chains (diffuse 4.6 A peak in the wide-angle pattern), whereas Ca2+ probably induces a two-dimensional order (4.3 A and 3.9 A peak in the wide-angle pattern). Electron density profiles of the lipid and lipid-basic protein vesicles indicate that the basic protein penetrates into the bilayer.


Cell-specific localization of the sulphydryl oxidase QSOX in rat peripheral tissues.
Rat quiescin/sulphydryl oxidase (rQSOX) introduces disulphide bridges into peptides and proteins with the reduction of molecular oxygen to hydrogen peroxide. Its occurrence has been previously highlighted in a wide range of organs by reverse transcription-polymerase chain reaction (RT-PCR) and Northern blot analyses, methods that have provided information concerning its expression in whole organs but that do not reveal the cell types expressing this enzyme. In this report, in addition to RT-PCR and Western blot experiments, the cell-specific localization of rQSOX has been investigated in a wide range of male and female adult rat tissues by using in situ hybridization and immunohistochemistry. Labelling was detected in most organs and systems including the immune, endocrine and reproductive systems, the respiratory, digestive and urinary tracts and the skin. No labelling was observed in the heart, blood vessel endothelium, liver or smooth and skeletal muscles. rQSOX expression was mainly localized in epithelial cells specialized in secretion, strengthening the hypothesis that QSOX enzymes play an important role in the mechanism of secretion, notably in the folding of secreted proteins. The intracellular patterns of immunolabelling indicate that the protein usually follows the secretory pathway, which is in accordance with its secreted nature and its presumed involvement in the elaboration of the extracellular matrix. In seminiferous tubules, where a high level of expression was noticed, QSOX might play an important physiological role in sperm function and serve as a marker for the diagnosis of male infertility.


Likelihood ratios for fetal trisomy 21 based on nasal bone length in the second trimester: how best to define hypoplasia?
To determine the best measure of fetal nasal bone hypoplasia for trisomy 21 risk assessment in the second trimester. This was a prospective, observational study performed at a single institution between February 2003 and December 2005. Fetuses with nasal bone length recorded sonographically between 16 and 20.9 weeks and known karyotype were included. Definitions of nasal bone hypoplasia assessed included: non-visualized nasal bone, nasal bone < 10th percentile, nasal bone < 2.5th percentile, biparietal diameter/nasal bone ratio >or= 10 and >or= 11 and nasal bone multiples of the median (MoM) <or= 0.5, <or= 0.6, and <or= 0.7. A total of 371 chromosomally normal and 11 trisomy 21 fetuses were included. Nasal bone hypoplasia based on MoM was superior to the other measures of hypoplasia for trisomy 21 risk assessment as reflected by a higher positive likelihood ratio. The risk for fetal trisomy 21 was higher with greater degrees of nasal bone hypoplasia. Hypoplasia, as defined by MoM, is the superior approach when incorporating nasal bone evaluation into fetal trisomy 21 risk assessment in the second trimester.


Death due to non-traumatic hemoperitoneum in Milan 2002-2016, with focus on two cases of abdominal apoplexy (idiopathic spontaneous hemoperitoneum) and review of the literature.
Abdominal apoplexy, also known as idiopathic spontaneous hemoperitoneum, is a rare event related to sudden death, whose diagnosis and treatment is extremely challenging and whose pathologic mechanisms are still mainly unknown. It is frequently associated with vascular disorders (alterations in the vascular structures) and mainly linked to liver cirrhosis. However, the presence of hemoperitoneum may in such cases pose many challenges to forensic post-mortem examinations since the source of bleeding remains often unknown even after a careful and thorough dissection. The Authors present two cases of sudden death related to massive hemoperitoneum (4,650ml in case 1 and 5,100ml in case 2) occurred in two cirrhotic males aged 49 and 51: no traumatic injuries were detected and the source of bleeding was not identified although a careful dissection of organs and vascular structures was performed. Rare cases have been already described in the literature, only as case reports, and no systematic studies have been performed on this issue. Nevertheless, this event ought to be taken into account asa cause of sudden death in people with advanced liver disease. A general glimpse is provided among the different causal mechanisms and the challenges within forensic pathology.


Pharmacometrics of pterostilbene: preclinical pharmacokinetics and metabolism, anticancer, antiinflammatory, antioxidant and analgesic activity.
The present study evaluated the preclinical pharmacokinetics and pharmacodynamics of trans-pterostilbene, a constituent of some plants. Right jugular vein cannulated male Sprague-Dawley rats were dosed i.v. with 20 mg/kg of pterostilbene and samples were analysed by the reverse phase HPLC method. Serum AUC, serum t(1/2), urine t(1/2), Cl(total) and Vd(beta) were 17.5 +/- 6.6 microg/h/mL, 1.73 +/- 0.78 h, 17.3 +/- 5.6 h, 0.960 +/- 0.025 L/h/kg and 2.41 +/- 1.13 L/kg (mean +/- SEM), respectively. A pterostilbene glucuronidated metabolite was detected in both serum and urine. The in vitro metabolism in rat liver microsomes furthermore suggests phase II metabolism of pterostilbene. Pterostilbene demonstrated concentration-dependent anticancer activity in five cancer cell lines (1-100 microg/mL). An in vitro colitis model showed concentration-dependent suppression of PGE(2) production in the media of HT-29 cells. Antiinflammatory activity was examined by inducing inflammation in canine chondrocytes followed by treatment with pterostilbene (1-100 microg/mL). The results showed decreased levels of MMP-3, sGAG and TNF-alpha compared with control levels. Pterostilbene exhibited concentration-dependent antioxidant capacity measured by the ABTS method. Pterostilbene increased the latency period to response in both tail-flick and hot-plate analgesic tests.