S100A9 expressed in ER(-)PgR(-) breast cancers induces inflammatory cytokines and is associated with an impaired overall survival.
Breast cancer is the most common cancer form among women today. Depending on hormone receptor status, breast cancers are divided into different subtypes with vastly varying prognosis. S100A9 is a calcium-binding protein that is associated with inflammation and expressed not only in myeloid cells but also in some tumours. The role for S100A9 in the malignant cells is not well characterised; however, previous studies have shown that the protein could have important immune-modulating properties. Using a human breast cancer cohort consisting of 144 tumour samples and in vitro analysis of human breast cancer cell lines, we investigated the expression and function of S100A9 in human breast cancer. We show that S100A9 expression in breast cancer correlated with the ER(-)PgR(-) breast tumour subtype (P<0.001) and with Ki67 (P=0.024) and was expressed both in the malignant cells and in the tumour-infiltrating anti-inflammatory CD163(+) myeloid cells (P<0.001). Stromal expression of S100A9 also correlated to nodal stage, tumour size and Her2 positivity. Within the ER(-)PgR(-) subgroup, all Her2(+) and EGFR(+) tumours expressed S100A9 in the cytoplasm. Both cytoplasmic staining in the malignant cells as well as stromal S100A9 expression in myeloid cells correlated with a decreased overall survival in breast cancer patients. Furthermore, rS100A9 homodimers induced expression of pro-inflammatory cytokines (IL-6, IL-8 and IL-1β) in a TLR4- and EGFR-dependent manner in human breast cancer cells in vitro. We suggest that S100A9 could be viewed as a novel therapeutic target for patients with ER(-)PgR(-) breast cancers.


Ultrastructure of hair follicle afferent fibre terminations in the spinal cord of the cat.
In acute electrophysiological experiments on anaesthetized cats, single identified hair follicle afferent fibres were injected with horseradish peroxidase (HRP). The HRP was injected from an intra-axonal microelectrode in the lumbosacral spinal cord. One to six hours after injection the animals were perfused and the tissue prepared for light and electron microscopy (EM). Axon collateral arborizations containing HRP reaction product were identified in thick sections under the light microscope and the same tissue then cut on the ultramicrotome for EM study. The terminal branches of the collaterals kept their myelin sheaths until they were 0.45-1.0 micron in diameter, just before they formed synaptic boutons. Synaptic boutons (1.0-4.0 microns in diameter) were usually of the en passant variety and made contact with dendrites. The contacts were asymmetrical (Type I) and contained round, clear synaptic vesicles of 35-60 nm diameter. Both the non-myelinated portion of the terminal axon and the synaptic boutons received axo-axonic contacts. These axo-axonic boutons contained clear (agranular) vesicles irregular in profile.


Improvement of the spectroscopic parameters of the air- and self-broadened N2O and CO lines for the HITRAN2020 database applications.
This paper outlines the major updates of the line-shape parameters that were performed for the nitrous oxide (N2O) and carbon monoxide (CO) molecules listed in the HITRAN2020 database. We reviewed the collected measurements for the air- and self-broadened N2O and CO spectra to determine proper values for the spectroscopic parameters. Careful comparisons of broadening parameters using the Voigt and speed-dependent Voigt line-shape profiles were performed among various published results for both N2O and CO. Selected data allowed for developing semi-empirical models, which were used to extrapolate/interpolate existing data to update broadening parameters of all the lines of these molecules in the HITRAN database. In addition to the line broadening parameters (and their temperature dependences), the pressure shift values were revised for N2O and CO broadened by air and self for all the bands. The air and self speed-dependence of the broadening parameter for these two molecules were added for every transition as well. Furthermore, we determined the first-order line-mixing parameters using the Exponential Power Gap (EPG) scaling law. These new parameters are now available at HITRAN online.


Public purchasers contracting external primary care providers in Central America for better responsiveness, efficiency of health care and public governance: issues and challenges.
Several national health systems in Latin America initiated health reforms to counter widespread criticisms of low equity and efficiency. For public purchasing agencies, these reforms often consisted in contracting external providers for primary care provision. This paper intends to clarify both the complex and intertwined issues characterizing such contracting as well as health system performances within the context of four Central American countries. It results from a European Commission financed project lead between 2002 and 2005, involving participants from Costa Rica, Guatemala, Nicaragua, Salvador, United Kingdom, Netherlands and Belgium, whose aim was to promote exchanges between these participants. The findings presented in this paper are the results of a two stage process: (a) the design of an initial analytical framework, built upon findings from the literature, interlinking characteristics of contractual relation with health systems performances criteria and (b) the use of that framework in four case studies to identify cross-cutting issues. This paper reinforces two pivotal findings: (a) contracting requires not only technical, but also political choices and (b) it cannot be considered as a mechanical process. The unpredictability of its evolution requires a flexible and reactive approach. This should be better assimilated by national and international organizations involved in health services provision, so as to progressively come out of dogmatic approaches in deciding to initiate contractual relation with external providers for primary care provision.


The cleavage specificity of the aspartic protease of cocoa beans involved in the generation of the cocoa-specific aroma precursors.
Particular peptides generated from the vicilin-class(7S) globulin of the cocoa beans by acid-induced proteolysis during cocoa fermentation are essential precursors of the cocoa-specific aroma notes. As revealed by in vitro studies, the formation of the cocoa-specific aroma precursors depends on the particular cleavage specificity of the cocoa aspartic protease, which cannot be substituted by pepsin. Therefore, we have investigated the effects of aspartic protease inhibitors on both enzymes and comparatively studied their cleavage specificities using different protein substrates and MALDI-TOF mass spectrometric analyses of the generated oligopeptides. Three classes of cleavage sites have been identified and characterized: (I) sequences exclusively cleaved by the cocoa enzyme, (II) sequences cleaved by both pepsin and the cocoa enzyme, and (III) those cleaved exclusively by pepsin. In contrast to most aspartic proteases from other origins, basic amino acid residues, particularly lysine, were found to be abundant in the specific cleavage sites of the cocoa enzyme.


An overview of the extra-lipid effects of rosuvastatin.
Statins, in addition to their beneficial lipid modulation effects, exert a variety of several so-called "pleiotropic" actions that may result in clinical benefits. Rosuvastatin, the last agent of the class to be introduced, has proved remarkably potent in reducing low-density lipoprotein cholesterol levels. At present, no large-scale primary or secondary prevention clinical trials document either its long-term safety or its effectiveness in preventing cardiovascular events. A substantial number of experimental and clinical studies have indicate favorable effects of rosuvastatin on endothelial function, oxidized low-density lipoprotein, inflammation, plaque stability, vascular remodeling, hemostasis, cardiac muscle, and components of the nervous system. Available data regarding the effects of rosuvastatin on renal function and urine protein excretion do not seem to raise any safety concerns. Whether the established "pleiotropy" and/or lipid-lowering efficacy of rosuvastatin may translate into reduced morbidity and mortality remains to be shown in ongoing clinical outcome trials.


Search for chiral molecules and optical activity in extraterrestrial systems. Example of Titan.
One of the main characteristics of terrestrial life is the role of optically active organic substances. Thus a search for chiral compounds and optical activity on an extraterrestrial body may give an indication of the presence of life, either fossilized or still in existence. If only abiotic conditions are prevailing the same search may still provide interesting information on the possible origins of homochiral families of biomolecules on Earth (e.g. the amino acids). In this respect, Saturn's satellite Titan is exemplary. A list of some of the most simple chiral derivatives devoid of oxygen atoms possibly present on Titan is presented. The interest of an investigation of optical activity is discussed taking into account some significant parameters. This raises numerous difficult technical problems which once solved may be helpful for further exploration of other planets.


Wrist circumduction reduced by finger constraints.
Assessment of wrist motion is important in diagnosing and treating motion impairment after injuries to the wrist. Little is known of how finger posture influences the movement of the wrist. The aim of this study was to investigate the effect of finger constraints on the maximum circumduction movement of the wrist. Fifteen male right-handed subjects performed maximal circumferential wrist movements under 4 finger conditions: unconstrained fingers, holding a large cylinder (50 mm diameter), holding a small cylinder (25 mm diameter), and closed fist position. The wrist motion was captured by a surface marker-based motion analysis system. To quantify wrist motion capability, we constructed the maximal boundaries of wrist motion (circumduction envelope) from angular plots in flexion-extension (FE) and radial-ulnar deviation (RUD). The ranges of motion in FE and RUD and the envelope area were calculated. Finger constraints significantly reduced motion ranges in flexion and ulnar deviation, but not in extension and radial deviation. In comparison to the unconstrained finger condition, the motion ranges in flexion decreased by 13%, 16%, and 27% for the large cylinder, small cylinder, and fist conditions, respectively. The range of ulnar deviation was reduced by 10% for the large and small cylinder conditions and by 11% for the fist condition. The overall mobility in FE and RUD, as quantified by the area of circumduction envelope, decreased by 15%, 15%, and 23% for the large cylinder, small cylinder, and fist conditions, respectively. Wrist mobility is facilitated by the synergistic motion of finger joints. Constraining fingers in static flexion posture reduces wrist flexion and ulnar deviation without decreasing extension and radial deviation. A clinical implication is that wrist motion should be assessed under standardized finger joint configuration.


Nano-Crystalline Li1.2Mn0.6Ni0.2O₂ Prepared via Amorphous Complex Precursor and Its Electrochemical Performances as Cathode Material for Lithium-Ion Batteries.
An amorphous complex precursor with uniform Mn/Ni cation distribution is attempted for preparing a nano-structured layered Li-rich oxide (Li1.2Mn0.6Ni0.2O₂)cathode material, using diethylenetriaminepentaacetic acid (DTPA) as a chelating agent. The materials are characterized by powder X-ray diffraction (XRD), scanning electron microscopy (SEM), transmission electron microscopy (TEM), and electrochemical tests. The crystal structure of Li-rich materials is found to be closely related to synthesis temperature. As-obtained nano materials sintered at 850 °C for 10 h show an average size of 200 nm with a single crystal phase and good crystallinity. At a current density of 20 mA·g-1, the specific discharge capacity reaches 221 mAh·g-1 for the first cycle and the capacity retention is 81% over 50 cycles. Even at a current density of 1000 mA·g-1, the capacity is as high as 118 mAh·g-1. The enhanced rate capability can be ascribed to the nano-sized morphology and good crystal structure.


Production of alcohol from raw wheat flour by Amyloglucosidase and Saccharomyces cerevisiae.
Ethanol production, by a simultaneous saccharification and fermentation process from raw wheat flour, has been performed by Saccharomyces cerevisiae and a low level of amyloglucosidase enzyme. The fermentation time was about 60 h after a 6 h pre-saccharification, with an amyloglucosidase (AMG) level of 270 AGU. kg(-1) starch, but only 31 h with a simultaneous saccharification fermentation process (SSF). When an AMG level of 540 AGU. kg(-1) starch was used, the time decreased to 21 h, giving an ethanol concentration of 67 g. l(-1). Sugar composition of the wort after the liquefaction may be responsible of the difference between these two process. Maltose, a fermentable sugar, was produced in high concentration during the liquefaction, allowing a shorter process period, counteracting the effect of the slow starch hydrolysis at 35 degrees C (SSF temperature).


Effects of fluorescent derivatives of TPA on HL60 cells: dissociation between the differentiation-induced and protein kinase C activity.
The four fluorescent derivatives of TPA--dansylaza-TPA, NBDaza-TPA, and (N)- and (P)-dansylamino-TPA--were synthesized and examined for their ability to induce differentiation in human promyelocytic leukemic HL60 cells. At a concentration of 20 nM, all the derivatives inhibited proliferation and induced 60-80% of the cells to differentiate into macrophage-like cells. Removal of dansylaza-TPA from the medium after 5 h did not arrest adherence or the expression of nonspecific esterase activity. However, upon removal of any of the other three compounds after 5 h, HL60 cells became nonadherent and expressed low nonspecific esterase activity after additional culture. To investigate the relationship between protein kinase C (PKC) activation and cell maturation, PKC activity and translocation were measured after 0.5, 5, 24, and 48 h of treatment with each compound. Cells induced to differentiate by dansylaza-TPA or (N)- or (P)-dansylamino-TPA exhibited enhanced PKC activity, 50-80% of which was located in the particulate fraction. In cells that differentiated with NBDaza-TPA, 65-70% of PKC activity remained in the cytosol. After removal of the TPA derivatives, all cells exhibited PKC activity in the cytosol. These results indicate that the fluorescent derivatives are as potent as TPA in inducing HL60 cell differentiation. However, in the case of NBDaza-TPA and (N)- or (P)-dansylamino-TPA, their continuous presence in the culture medium was required for the recruitment of cells to differentiate. Consequently, it is suggested that activation and translocation of PKC are among the early biochemical events that trigger HL60 cell differentiation. Nevertheless, these two events alone are not sufficient to induce differentiation.


Pedometer-determined physical activity is linked to low systemic inflammation and low arterial stiffness in Type 2 diabetes.
The aim of this study was to explore the association between pedometer-determined physical activity versus measures of obesity, inflammatory markers and arterial stiffness in people with Type 2 diabetes. We analysed data from 224 men and 103 women with Type 2 diabetes, aged 54-66 years. Physical activity was measured with waist-mounted pedometers during three consecutive days and the number of steps/day were calculated and classified in four groups: < 5000 steps/day, 5000-7499 steps/day, 7500-9999 steps/day and ≥ 10000 steps/day. Blood samples were analysed for lipids, HbA(1c), inflammatory markers including C-reactive protein and interleukin-6. Nurses measured blood pressure and anthropometrics. Aortic pulse wave velocity was measured with applanation tonometry over the carotid and femoral arteries. Mean steps/day was 7683 ± 3883 (median 7222, interquartile range 4869-10,343). There were no differences in age, diabetes duration, blood pressure, lipids or glycaemic control between the four groups of pedometer-determined physical activity. Subjects with higher steps/day had lower BMI (28.8 vs. 31.5 kg/m(2), P < 0.001), waist circumference (101.7 vs. 108.0 cm, P < 0.001), lower levels of C-reactive protein (1.6 vs. 2.6 mg/l, P = 0.007), lower levels of interleukin-6 (1.9 vs. 3.8 pg ml, P < 0.001) and lower pulse wave velocity (10.2 vs. 11.0 m/s, P = 0.009) compared with less physically active people. We conclude that physical activity measured with pedometer was associated not only with less abdominal obesity, but also with decreased systemic low-grade inflammation as well as with low arterial stiffness, in people with Type 2 diabetes.


Enhancement of Dissolution Rate and Intestinal Stability of Clopidogrel Hydrogen Sulfate.
Clopidogrel is an antiplatelet and antithrombotic prodrug. It has poor oral bioavailability due to poor dissolution and possible premature degradation in the intestine. Accordingly, the objective of this study was to enhance clopidogrel dissolution rate and to reduce its premature degradation in rabbit intestine. Solid dispersion (SD) systems of clopidogrel with gelucire 50/13 and/or cremophor RH40 were prepared using fusion technique. The SD systems were characterized with respect to drug dissolution. The characterization included thermal analysis and infrared investigations. The stability of clopidogrel in the fluid extracted from small intestinal and colonic mucosal surfaces was monitored both in absence and presence of cremophor or gelucire. SD formation enhanced drug dissolution with the enhancement increasing at higher concentrations of either cremophor or gelucire. The ternary SD of clopidogrel with cremophor and gelucire reflected synergism between them. This synergism was manifested by enhanced dissolution efficiency of drug to reach 85 % at pH 6.8 and 89 % at pH 7.4 compared to unprocessed drug which liberated 16.2 and 15.2 % at the same pH values, respectively. Enhanced dissolution from SD was mainly due to micellar solubilization for cremophor and was due to change in the crystalline nature of drug with a contribution to self-emulsification in case of gelucire. Clopidogrel showed premature degradation in the intestinal fluid. Cremophor RH 40 reduced this degradation but gelucire failed in this respect. The study introduced SD system for enhanced dissolution rate of clopidogrel with a potential of reduced premature degradation in the intestine.


A cost minimisation analysis comparing oral linezolid and intravenous daptomycin administered via an outpatient parenteral antibiotic therapy programme in patients requiring prolonged antibiotic courses.
This study is an economic analysis seeking to examine cost savings that may be accrued from usage of oral linezolid in place of OPAT IV daptomycin in patients requiring prolonged courses of IV or highly bioavailable oral antibiotic therapy. In order to do so we conducted a literature review to establish the scenarios in which the agents could be considered equivalent. We then, using a decision-tree model, conducted a cost analysis to establish differences in cost between the approaches. Under the model base-case, the total cost of treatment with OPAT daptomycin was €3,496.84 and the total cost of treatment with oral linezolid was €772.01. Therefore the oral linezolid strategy would be projected to save the Irish health service €2,724.83 per patient. These results were robust to one-way deterministic sensitivity analyses and probabilistic sensitivity analysis. Our study suggests that significant savings could be safely accrued in the management of these patients.


In-situ and quantitative imaging of evaporation-induced stratification in binary suspensions.
The drying of a multi-component dispersion, such as water-based paint, ink and sunscreen to form a solid film, is a widespread process. Binary colloidal suspensions have proven capable of spontaneous layer formation through size segregation during drying. To design bespoke stratification patterns, a deeper understanding of how these emerge is crucial. Here, we visualize and quantify the spatiotemporally evolving concentration profiles in situ and with high resolution using confocal fluorescence microscopy of custom-designed binary dispersions in a well-defined geometry. Our results conclusively establish two distinct stratification routes, which give rise to three layered structures. A first thin layer develops directly underneath the evaporation front in which large particles are kinetically trapped. At later times, asymmetrical particle interactions lead to the formation of two subsequent layers enriched in small and large particles, respectively. The spatial extent and magnitude of demixing strongly depend on the initial volume fraction. We explain and reproduce the experimental concentration profiles using a theoretical model based on dynamic arrest and higher-order thermodynamic and hydrodynamic interactions. These insights unravel the key mechanisms underlying colloidal auto-stratification in multi-component suspensions, and allow preprogramming of stratification patterns in single-deposition formulations for future applications.


The nude rat as an orthotopic model for cervical cancer.
The purposes of this study were to establish intracervical tumors of the nude rat as an orthotopic experimental model for human cervical cancer and to preliminary evaluate the effects of the adenoviral vector, Ad5CMV-p53, on orthotopic cervical tumor size. Human cervical cancer SiHa and ME-180 cells were injected into the cervix of the nude rat. Four days later, 1 x 10(9) plaque forming units (PFU) of Ad5CMV-p53 were injected into the cervix. The rats were later sacrificed to determine cervical tumor size. Eight of ten nude rats developed SiHa cell tumors; all ten nude rats developed ME-180 cell tumors. Four of ten SiHa cell tumors metastasized to the pelvic cavity; no ME-180 cell tumors did. The growth of Ad5CMV-p53-infected cells was greatly suppressed. The ad5CMV-p53 treatment significantly reduced both cell tumor volumes in nude rat cervixes. The nude rat cervix grows tumors similar to human cervical cancer tumors and makes an excellent experimental model. Transfection of cervical cancer cells with the wild-type p53 gene via Ad5CMV-p53 is a potential therapeutic approach to cervical cancer.


Induction of systemic and mucosal immunity and maintenance of its memory against influenza A virus by nasal vaccination using a new mucosal adjuvant SF-10 derived from pulmonary surfactant in young cynomolgus monkeys.
Induction of systemic and mucosal immunity and maintenance of its memory was investigated in 12 young male cynomolgus monkeys after intranasal instillation of flu vaccine using a new mucosal adjuvant SF-10 derived from pulmonary surfactant constituents. Split-product of influenza virus A/California/7/2009(H1N1)pdm hemagglutinin vaccine (HAv) at 15 μg with or without SF-10 and the adjuvant alone were instilled intranasally three times every 2 weeks. SF-10-adjuvanted HAv (SF-10-HAv) elicited significantly higher HAv-specific IgG and hemagglutinin inhibition (HI) titers in serum and HAv-specific secretory IgA and its neutralizing activities in nasal washes compared with HAv antigen and SF-10 alone. Significant cross-neutralizing activities of nasal washes after the third vaccination to several other H1N1 and H3N2 strains were observed. HI titers in serum and neutralizing activities in nasal washes reached peak levels at 6 weeks after initial vaccination, then gradually decreased after 10 weeks and returned to the baseline levels at 36 weeks. A single intranasal revaccination of SF-10-HAv at 36 weeks rapidly and significantly increased both immunity in serum and nasal washes compared with naïve monkeys. Revaccination by one or two doses achieved almost maximal immunity at 2 or 4 weeks after instillation. Statistically significant adverse effects (e.g., body weight loss, elevated body temperature, nasal discharge, change in peripheral blood leukocyte and platelet counts) were not observed for 2 weeks after vaccination of SF-10-HAv, HAv or SF-10 and also during the experimental period. These results in young monkey model suggest the potential of clinical use SF-10 for intranasal flu vaccine.


Effect of Atorvastatin on Single Oral Pharmacokinetics and Safety of Daclatasvir in Rats: Emphasis on P-glycoprotein and Cytochrome P450.
This study aimed to investigate the effect of atorvastatin on daclatasvir oral pharmacokinetics and safety and assess the possible underlining mechanisms by targeting P-glycoprotein (P-gp) and cytochrome P450 (CYP3A4). The transport of daclatasvir, as well as the standard rhodamine 123 by P-gp across the rat intestine, was studied in vitro using the non-everted sac method. To assess the pharmacokinetic profile of daclatasvir in vivo, rats were divided into three groups receiving either saline, standard P-gp inhibitor verapamil (25 mg/kg), or atorvastatin (10 mg/kg), 2 hrs prior to a single dose of daclatasvir (7 mg/kg). In addition, the markers of liver and kidney functions and muscle rhabdomyolysis were assessed. Further, histopathological examination of liver and kidney tissue and assessment of CYP3A4 level was done. The inhibitory effect of atorvastatin on Pgp activity and expression was manifested by increased serosal transport of the standard rhodamine 123, as well as daclatasvir. In vivo, Cmax (peak plasma concentration) and area under the curve (AUC (0-t)) of daclatasvir after atorvastatin treatment increased compared to the vehicle group but not in a significant manner. On the other hand, atorvastatin caused a significant increase in the clearance of daclatasvir. Concomitant administration of atorvastatin with daclatasvir significantly decreased CYP3A4 content compared to the control group. The combination also showed increased liver enzymes and some pathological alterations in the liver. Atorvastatin has a significant effect on P-gp mediated intestinal transport of daclatasvir; however, it did not affect the systemic bioavailability of a single oral dose of daclatasvir.


MRI of double-bundle ACL reconstruction: evaluation of graft findings.
To demonstrate the magnetic resonance imaging (MRI) findings of double-bundle (DB) anterior cruciate ligament (ACL) reconstruction grafts. Sixty-six patients with DB ACL reconstruction were evaluated with MRI 2 years postoperatively. Graft thickness was measured separately by two musculoskeletal radiologists. The MRI findings of graft disruption, signal intensity (SI) changes, cystic degeneration, arthrofibrosis, and impingement were analyzed. The statistical significance of the association between MRI findings was calculated. The mean anteromedial (AM) graft thickness was reduced 9% and the mean posterolateral (PL) graft thickness was reduced 18% from the original graft thickness. Disruption was seen in 3% of AM grafts and 6% of PL grafts and a partial tear in 8 and 23%, respectively. Both grafts were disrupted in 3% of patients. Increased SI was seen in 14% of intact AM grafts and in 60% of partially torn AM grafts (p = 0.032). In PL grafts the increased SI was seen in 51% of the intact grafts and in 93% of the partially torn grafts (p = 0.005). Cystic degeneration was seen in 8% of AM grafts and in 5% of PL grafts. Diffuse arthrofibrosis was seen in 5% of patients and a localized cyclops lesion in 3% of patients. Impingement of the AM graft was seen in 8% of patients. Both grafts were disrupted in 3% of patients. Also, the frequencies of other complications were low. The use of orthogonal sequences in the evaluation of the PL graft SI seems to cause volume-averaging artefacts.


A scale for rating the quality of psychological trials for pain.
This paper reports the development of a scale for assessing the quality of reports of randomised controlled trials for psychological treatments. The Delphi method was used in which a panel of 15-12 experts generated statements relating to treatment and design components of trials. After three rounds, statements with high consensus agreement were reviewed by a second expert panel and rewritten as a scale. Evidence to support the reliability and validity of the scale is reported. Three expert and five novice raters assessed sets of 31 and 25 published trials to establish scale reliability (ICC ranges from 0.91 to 0.41 for experts and novices, respectively) and item reliability (Kappa and inter-rater agreement). The total scale score discriminated between trials globally judged as good and poor by experts, and trial quality was shown to be a function of year of publication. Uses for the scale are suggested.


The effect of a subsidy policy on the utilization of community care services under a public long-term care insurance program in rural Japan.
This cross-sectional study investigated the effect of an interim subsidy policy for low-income beneficiaries to buy home help services under Japanese long-term care insurance (LTCI) for the service utilization of home help and other community care services. Claims records during the period from October to December 2001 were reviewed to compare the monthly amounts for each service and out-of-pocket copayment for subsidized (n = 137) and non-subsidized (n = 124) beneficiaries. A multivariable linear regression analysis was conducted to control for age, gender, household size, length of care, and care eligibility level. Compared with the non-subsidized beneficiaries, the subsidized beneficiaries used 1.53 times as much home help service (p < 0.001), and spent 0.70 times as much on out-of-pocket copayments (p < 0.001), while they used fewer adult daycare services (0.71 times as much as the non-subsidized did, p = 0.063). The results strongly indicated that the interim subsidy for low-income beneficiaries alleviated the economic burden, but also resulted in service overuse and the substitution of home help services for other community care services. The policy implications of these findings were discussed.


Aesthetic discomfort in hand osteoarthritis: results from the LIège Hand Osteoarthritis Cohort (LIHOC).
The primary complaint of patients with hand osteoarthritis (OA) is frequently the inelegant appearance of their hands. Only one study has been conducted to assess the magnitude of and identify the determinants of aesthetic discomfort in hand OA. The LIège Hand Osteoarthritis Cohort is a prospective cohort of 203 patients diagnosed with hand OA. At baseline, these patients rated their aesthetic discomfort on a 100-mm visual analogue scale (VAS) and used a Likert scale (range 0-7) to quantify the magnitude of their aesthetic damage. The median value of the aesthetic discomfort VAS was 35.0 [interquartile range (Q1-Q3) 6.0-59.0]. The median damage was rated 3.0 (Q1-Q3 1.0-4.0), corresponding to a moderate level. Both were significantly (p < 0.02) associated with the female gender, the duration of hand OA, the radiological severity of OA (Verbruggen-Veys and Kellgren-Lawrence scales) and pain, disability, or stiffness [Australian Canadian Osteoarthritis Hand Index (AUSCAN) and Functional Index for Hand Osteoarthritis ]. After a stepwise analysis, the parameters correlated to the aesthetic discomfort were the presence of erosive joints (p = 0.0048), the AUSCAN score (p < 0.0001), the number of joints with severe radiological damage (p = 0.023), and gender (p = 0.0009). For aesthetic damage, the parameters associated were AUSCAN score (p < 0.0001), duration of hand OA >10 years (p = 0.001), and presence of erosive joints (p < 0.0001). Compared with patients with low aesthetic discomfort (VAS ≤33 mm), those with the highest discomfort (VAS ≥66 mm) had more erosive OA (p = 0.014), a higher Verbruggen and Veys score (p = 0.0039), and a higher AUSCAN score (p < 0.001). Aesthetic discomfort and damage are significant complaints in patients with hand OA. The determinants of the magnitude of these are gender, radiological severity, duration of hand OA, presence of erosive joints, and impact on pain, function, and stiffness as assessed with the AUSCAN.


Complications of intravitreal steroid injections.
Intravitreal corticosteroid injections are a new therapeutic procedure used to treat various retinal edematous and neovascular conditions. They have been used in the treatment of diabetic macular edema, exudative macular degeneration, pseudophakic cystoid macular edema, macular edema associated with retinal vein occlusion, and chronic uveitis as well as other conditions. Because the use of this therapeutic technique is becoming increasingly more common, adverse effects are now being seen. The most common adverse effects associated with intravitreal steroid injection are elevation of intraocular pressure and progression of cataract. Endophthalmitis, pseudoendophthalmitis, and retinal detachment have also been reported. This report describes 2 patients who were followed up at the VA Connecticut Healthcare System Newington Campus Optometry Clinic for steroid-induced elevation of intraocular pressure after intravitreal corticosteroid injection. One patient exhibited elevation of intraocular pressure after his first intravitreal steroid injection for treatment of clinically significant macular edema secondary to diabetes. The second patient did not exhibit a steroid response to the first intravitreal steroid injection utilized as treatment for choroidal neovascularization from age-related macular degeneration. However, he did show a rise in intraocular pressure after a second intravitreal corticosteroid injection. Intraocular pressures, treatment, and frequency of follow-up in both patients pre- and postinjection are discussed. Elevation of intraocular pressure after intravitreal steroid injection can commonly be controlled with topical glaucoma medications. Cataract progression is common in patients after intravitreal injection of corticosteroid; however, findings show these patients are at no additional risk for cataract surgery complications. Therefore, these do not appear to be major contraindications. However, because 30% to 50% of patients experience intraocular pressure rise up to a few months postinjection, and patients are at higher risk for complications such as endophthalmitis, optometrists should be aware of appropriate management after this increasingly utilized therapeutic procedure.


A Uridine Glucuronosyltransferase 2B7 Polymorphism Predicts Epirubicin Clearance and Outcomes in Early-Stage Breast Cancer.
Epirubicin is metabolized by uridine glucuronosyltransferase 2B7 (UGT2B7), an enzyme rich in single nucleotide polymorphisms (SNPs). We studied whether the -161 C > T germline SNP in UGT2B7 was related to epirubicin metabolism and whether differences exist in the toxicity and efficacy of epirubicin-based chemotherapy among patients who were TT homozygotes, CT heterozygotes, and CC homozygotes. A total of 132 women with non-metastatic breast cancer receiving FEC (5-fluorouracil 500 mg/m(2), epirubicin 100 mg/m(2), cyclophosphamide 500 mg/m(2)) were prospectively enrolled. Toxicity was assessed in cycle 1 using the National Cancer Institute Common Toxicity Criteria, version 2.0. The sequence at -161 was studied in 132 subjects; 37 were TT homozygotes, 63 were CT heterozygotes, 26 were CC homozygotes, and 6 could not be genotyped. The CC genotype patients had decreased epirubicin clearance (median, 103.3 L/hr) compared with the CT/TT genotype patients (median, 134.0 L/hr; P = .002). The CC homozygous patients had an increased risk of grade 3 to 4 leukopenia compared with the TT homozygotes or heterozygotes (P = .038 and P = .032, respectively). TT homozygotes or heterozygotes had an increased risk of early recurrence (P = .039; χ(2) test). The results of the present prospective pharmacogenetic study suggest that the UGT2B7 -161 C > T SNP correlate with drug metabolism, toxicity, and efficacy in patients receiving epirubicin chemotherapy. Further studies of this UGT2B7 SNP as a predictor of epirubicin toxicity and efficacy are warranted.


Role of cholinergic, vagal reflexes on the bronchoconstrictor responses to histamine during carbon dioxide inhalation in conscious guinea-pigs.
Histamine-induced bronchoconstriction in conscious guinea-pigs involves a cholinergic, bronchoconstrictor reflex, but the role of this reflex during elevated levels of inspired carbon dioxide is unknown. In this study we examined the role of cholinergic, vagal reflexes on the bronchoconstrictor responses to histamine during CO2 inhalation. Guinea-pigs were placed inside a whole body plethysmograph for measurement of tidal volume (VT), respiratory rate (f) and minute volume (V) and a head chamber was used to deliver a hypercapnic gas mixture (10% CO2, 21% O2, 69% N2) and for inhalation of aerosolized drugs. Inhalation of CO2 caused an increase in VT, f and V and these effects were reduced by exposure to aerosolized histamine (0.01-0.05% for 30 s). The histamine-induced reduction of VT was significantly (P less than 0.05) attenuated following a 60 s exposure to inhaled atropine (0.03 and 0.1%) as was the reduction of VT due to inhaled methacholine. Intravenous atropine (1 mg/kg) also blocked the VT reduction due to aerosolized histamine. Intravenous administration of the ganglionic blockers hexamethonium (1 mg/kg) and mecamylamine (10 mg/kg) did not inhibit the histamine-induced reduction of VT at doses of these drugs that revealed systemic evidence of ganglionic blockade, i.e. inhibition of vagally stimulated bronchoconstriction and bradycardia. The results demonstrate that the bronchoconstrictor responses to histamine during CO2 inhalation in guinea-pigs involves stimulation of airway cholinergic receptors, but this response does not involve ganglionic neurotransmission. It is speculated that histamine's bronchoconstrictor effects during CO2 breathing involves stimulation of postganglionic, parasympathetic nerves innervating airway smooth muscle.


'Living in a shrinking world'-The experience of loneliness among community-dwelling older people with reduced mobility: a qualitative grounded theory approach.
Loneliness is associated with severe consequences for both the physical and mental health of older people. Research has shown that mobility limitations are an important risk factor for the emergence and maintenance of feelings of loneliness. The aim of this study was to explore the experiences of loneliness among community-dwelling older people with reduced mobility and its underlying dynamics. This study employed a qualitative, descriptive design, inspired by a grounded theory approach. Participants were purposively recruited in collaboration with home health care providers. The main inclusion criteria were as follows: aged 75 years or older, being mobile impaired, experiencing feelings of loneliness, and living at home and being cognitively able to be interviewed. Data analysis of 15 semi-structured, in-depth interviews was conducted based on the Qualitative Analysis Guide of Leuven (QUAGOL). Loneliness was expressed through all the stories, but it appeared as an elusive, intangible phenomenon. Both indicating the presence of loneliness and describing what this phenomenon means were difficult to express for most participants. Loneliness was experienced as an inherent part of the ageing process characterised by losses, limitations and meaninglessness. Participants described how they have experienced losing grip on the world in which they live and feel isolated in a literally and figuratively shrinking world. Loneliness is described as the feeling that one is unable to address the situation that results in deep sadness and the feeling of no longer being of value to their environment. This study shows that loneliness among community-dwelling older persons with reduced mobility is embedded in experiences of loss related to ageing, among which reduced mobility plays a significant role. The results suggest the existence of a more profound experience of loneliness than might appear at first glance. How to recognise experiences of loneliness and how to support a meaningful existence for community-dwelling older persons should be given priority in health care. The findings of this study can increase professional caregivers' sensitivity to implicit signals of loneliness. Further research is necessary to refine the outcomes and to further explore the role of reduced mobility in the experience of loneliness.


Plasma endothelin in patients with acute aortic disease.
We investigated the plasma levels of endothelin 1/2 in patients with acute symptoms relating to a known or newly diagnosed aortic aneurysm in order to investigate the possible role of peptides in the development of the disease. Endothelin 1/2 plasma levels were determined in patients admitted to the emergency unit with suspected acute aortic disease. The history, type of aneurysm, outcome and laboratory findings were determined and compared to endothelin 1/2 levels collected on admission. In patients with ruptured aneurysm (n=27) or acute aortic dissection (n=18) the endothelin 1/2 median levels were higher 1.1 (25th and 75th quartile 0.7, 1.7) fmol/ml than in patients (n=20) with pre-existing aneurysm 0.7 (0.4, 1.1) fmol/ml (P=0.013). Patients who died had significantly higher endothelin levels 1.3 (0.8, 1.9) fmol/ml than the survivors 0.8 (0.5, 1.4) fmol/ml (P=0.04). In a logistic regression analysis, only a higher blood pressure on admission was an independent predictor of survival. Endothelin 1/2 levels are elevated in patients with acute dissection or ruptured aneurysm, but they are not an independent predictor of survival.


Reduction of voluntary ethanol consumption in alcohol-preferring Alko alcohol (AA) rats by desoxypeganine and galanthamine.
The effects of desoxypeganine, an alkaloid from Peganum harmala L., and of galanthamine, an alkaloid from Galanthus nivalis L., on voluntary ethanol consumption were investigated in female Alko alcohol (AA) rats. Desoxypeganine-HCl reduced ethanol intake and ethanol preference dose-dependently at a dose range between 10 and 30 mg/kg body weight when given by gavage. Subcutaneous and intraperitoneal applications of desoxypeganine lead to even more pronounced decreases of ethanol intake and ethanol preference. The effects of desoxypeganine and galanthamine seem to be additive. A combination of both substances in doses, which were ineffective when administered alone, caused a significant decrease of ethanol preference. To exclude habituation to desoxypeganine treatment, the substance was given once daily over a period of 16 days. No decreases of the desoxypeganine effects on ethanol intake, total fluid intake, and ethanol preference were observed. This attenuation of ethanol preference combined with unchanged total fluid intake and food consumption represents a promising activity especially because no acquirement of tolerance after repeated administration was observed.


Occurrence of illicit drugs in surface waters in China.
Illicit drugs have been recognized as a group of emerging contaminants. In this work, occurrence of common illicit drugs and their metabolites in Chinese surface waters was examined by collecting samples from 49 lakes and 4 major rivers across the country. Among the drugs examined, methamphetamine and ketamine were detected with highest frequencies and concentration levels, consistent with the fact that these are primary drugs of abuse in China. Detection frequencies and concentrations of other drugs were much lower than in European lakes and rivers reported in the literature. In most Chinese surface waters methamphetamine and ketamine were detected at concentrations of several ng L(-1) or less, but in some southern lakes and rivers, these two drugs were detected at much higher concentrations (up to several tens ng L(-1)). Greater occurrence of methamphetamine and ketamine in southern surface waters was attributed to greater abuse and more clandestine production of the two drugs in southern China.


Chemical properties of lipopolysaccharide-like substance (LLS) extracted from Leptospira interrogans serovar canicola strain Moulton.
The aqueous layer was isolated from Leptospira interrogans serovar canicola strain Moulton by the hot phenol-water method. After ultracentrifugation, the precipitate was designated as lipopolysaccharide-like substance (LLS) fraction and the chemical composition was compared with that of bacterial LPS. The LLS fraction consists of 35.2% carbohydrate, 3.8% amino sugar, 36.4% lipid, 15.2% protein, and 0.3% phosphorus. Neutral sugars were detected as rhamnose, arabinose, xylose, 4-O-methylmannose, mannose, galactose, and a small amount of erythrose, fucose and glucose by gas-liquid chromatography (GLC), but 2-keto-3-deoxyoctonic acid was not detected in the LLS by thiobarbituric acid test and high voltage paper electrophoresis. Fatty acids detected by GLC were decanoic acid (C10: 0), dodecanoic acid (C12: 0), dodecenoic acid (C12: 1), tridecenoic acid (C13: 1), tetradecanoic acid (C14: 0), hexadecanoic acid (C16: 0), hexadecenoic acid (C16: 1), and octadecenoic acid (C18: 1). With SDS-polyacrylamide gel electrophoresis, bacterial LPS showed many orderly bands, while the banding pattern of the leptospiral LLS was very simple. These findings demonstrate that the physicochemical properties and chemical composition of LLS fraction from Leptospira are different from those of LPS extracted from gram-negative bacteria such as Enterobacteriaceae, and suggesting that Leptospira has no typical LPS.


Correlation of serum interleukin 17 and macrophage migration inhibitory factor levels with clinical response to intralesional Candida antigen and their potential use as predictors of clinical outcome in patients with multiple common warts.
Candida antigen injection is one of the most widely used intralesional immunotherapy in the treatment of warts. It acts through the induction of systemic immune response. The pattern of cytokines production may play an integral role in its mechanism of action. To investigate the possible relation between serum levels of IL17 and MIF, and the clinical response to intralesional Candida antigen in multiple common warts. A total of 90 patients with multiple common warts were divided into 2 groups. Sixty patients received intralesional Candida antigen injection into the largest wart, controlled against thirty patients who had intralesional saline, as placebo. The injection was done at a 2-week interval for 5 doses. Blood samples were obtained from both groups, and serum levels of IL17A and MIF were estimated at baseline and 2 weeks after the last session using ELISA kits. Complete clearance of warts was statistically higher in the Candida antigen group (40% of the patients) compared to the saline group (p < 0.05). The serum levels of IL17 had significantly declined from baseline, while the level of MIF had risen after intralesional Candida antigen injection, but not in the saline group. At a cutoff level of 316 pg/ml, IL17 had a sensitivity of 83.3% to predict therapeutic response. IL17A and MIF may have possible roles in the mechanism of action of Candida antigen in the treatment of common warts. At a certain level, serum IL17A may be a potential predictor of response to treatment.


Cross-desensitization of capsaicin-evoked oral irritation by high but not low concentrations of nicotine in human subjects.
We previously showed that sequential application of a relatively low nicotine concentration (7.4 mM) to the tongue, followed by a rest period, induced self-desensitization, i.e. a reduction of oral irritation elicited by subsequent nicotine, but not cross-desensitization of capsaicin-evoked irritation. We presently investigated if cross-desensitization of capsaicin-evoked irritation might be induced by higher concentrations of nicotine. Nicotine (74 or 300 mM) was applied once, unilaterally to the tongue of human subjects. When the irritant sensation had completely subsided, capsaicin (33 microM) was applied bilaterally. In a 2-alternative forced-choice (2-AFC) test, subjects indicated which side of the tongue had a stronger irritant sensation. They additionally rated the intensity of irritation on each side separately. Pretreatment with 74 mM nicotine did not induce cross-desensitization, since no significant difference was observed in intensity ratings or in choice of the treated vs. non-treated side in the 2-AFC. However, 300 mM nicotine did induce cross-desensitization, since a significant majority of subjects chose the non-treated side as having a stronger capsaicin-evoked irritant intensity and assigned significantly higher ratings to that side. These psychophysical findings are discussed in terms of possible neural mechanisms of desensitization.


Composite vascular grafts with high cell infiltration by co-electrospinning.
There is an increasing demand for functional small-diameter vascular grafts (diameter<6mm) to be used in clinical arterial replacement. An ideal vascular graft should have appropriate biomechanical properties and be biocompatible. Electrospinning has become a popular polymer processing technique for vascular tissue engineering, but the grafts fabricated by electrospinning often have relatively small pores and low porosity, which limit cell infiltration into scaffolds and hinder the regeneration and remodeling of grafts. In the present study, we aimed to develop an efficient method to prepare electrospun composite vascular grafts comprising natural and synthetic materials. We fabricated grafts made of polycaprolactone, gelatin, and polyvinyl alcohol (PVA) by co-electrospinning, and the scaffolds were further functionalized by immobilizing heparin on them. The PVA fibers degraded rapidly in vivo and generated electrospun scaffolds with high porosity, which significantly enhanced cell proliferation and infiltration. The mechanical properties of the grafts are suitable for use in artery replacement. Heparin functionalization of the grafts yielded a good antithrombogenic effect, which was demonstrated in platelet adhesion tests. Moreover, in vitro and in vivo results demonstrated that the heparin release from the grafts enhanced the growth of endothelial cells, which is important for the endothelium of implanted grafts. The results of this study indicate that our method is effective and controllable for the fabrication of vascular grafts that meet the clinical requirements for blood vessel transplantation.


The Associated Factors of Anxiety and Depressive Symptoms in COVID-19 Patients Hospitalized in Wuhan, China.
This study was aimed to investigate the prevalence and factors associated with anxiety and depressive symptoms among hospitalized patients with COVID-19 during the epidemic outbreak in Wuhan, China. A total of 99 COVID-19 patients were recruited and completed the Hospital Anxiety and Depression Scale (HADS) and the modified Medical Research Council (mMRC) Scale. Results showed there was no significant difference in anxiety or depressive symptoms between male and female. Patients aged 46-60 years old had a higher ratio of both anxiety and depressive symptoms. Besides, patients whose hospital stays was longer than 14 days had a higher risk of depressive symptoms than those stays was less than 7 days. There was no significant difference in the correlation between level of dyspnea and the levels of anxiety or depressive symptoms. In conclusion, COVID-19 patients might have anxiety and depressive symptoms during hospitalization. Clinicians should pay attention to the middle age group and patients with longer hospital stays.


Hormonal aspects of fluid and sodium balance in the elderly.
Normal aging is accompanied by many changes in the hormonal systems involved in control of sodium and water balance. These alterations, together with age-associated changes in renal function, diminish the capacity of the elderly person to withstand the challenges of illness, drugs, and physiologic stresses. Thus, the aging individual is at increased risk of developing clinically significant alterations in sodium and water balance, which are commonly expressed as hyperatremia and hypoatremia. Awareness of the limitations of homeostatic ability allow the physician to anticipate the impact of illness and drugs on volume and electrolyte status and lead to prevention or to a rational approach to therapeutic intervention and management.


Intercellular: local and systemic actions of skeletal muscle PGC-1s.
Physical exercise promotes complex adaptations in skeletal muscle that benefit various aspects of human health. Many of these adaptations are coordinated at the gene expression level by the concerted action of transcriptional regulators. Peroxisome proliferator-activated receptor gamma (PPARγ) coactivator-1 (PGC-1) proteins play a prominent role in skeletal muscle transcriptional reprogramming induced by numerous stimuli. PGC-1s are master coactivators that orchestrate broad gene programs to modulate fuel supply and mitochondrial function, thus improving cellular energy metabolism. Recent studies unveiled novel biological functions for PGC-1s that extend well beyond skeletal muscle bioenergetics. Here we review recent advances in our understanding of PGC-1 actions in skeletal muscle, with special focus on their systemic effects.


Cyclic nucleotides and protein kinase systems in the developing chick retina and pigment epithelium.
During embryonic development of the chick neural retina, cyclic nucleotide levels are relatively uniform but rise abruptly at the time of hatching. The rise is thus not temporally correlated with features of morphological development such as outer segment elongation but rather with the onset of actual visual function. In the pigment epithelium, the cyclic AMP level declines throughout the embryonic period studied and does not rise at hatching. Cyclic GMP levels are much lower in both retina and pigment epithelium but rise several-fold at hatching. A binding protein is observed for cyclic AMP in the retina prior to outer segment development; cyclic GMP binding is considerably lower. Retinal ATP-kinase activity is high throughout the embryonic period studied and is stimulated up to 6-fold by 1 muM cyclic AMP and by 100 muM cyclic GMP. The major rise in GTP-kinase activity correlates temporally with photoreceptor outer segment development and may be involved intimately in the visual process.


Evaluation of lime and hydrothermal pretreatments for efficient enzymatic hydrolysis of raw sugarcane bagasse.
Ethanol production from sugarcane bagasse requires a pretreatment step to disrupt the cellulose-hemicellulose-lignin complex and to increase biomass digestibility, thus allowing the obtaining of high yields of fermentable sugars for the subsequent fermentation. Hydrothermal and lime pretreatments have emerged as effective methods in preparing the lignocellulosic biomass for bioconversion. These pretreatments are advantageous because they can be performed under mild temperature and pressure conditions, resulting in less sugar degradation compared with other pretreatments, and also are cost-effective and environmentally sustainable. In this study, we evaluated the effect of these pretreatments on the efficiency of enzymatic hydrolysis of raw sugarcane bagasse obtained directly from mill without prior screening. In addition, we evaluated the structure and composition modifications of this bagasse after lime and hydrothermal pretreatments. The highest cellulose hydrolysis rate (70 % digestion) was obtained for raw sugarcane bagasse pretreated with lime [0.1 g Ca(OH)2/g raw] for 60 min at 120 °C compared with hydrothermally pretreated bagasse (21 % digestion) under the same time and temperature conditions. Chemical composition analyses showed that the lime pretreatment of bagasse promoted high solubilization of lignin (30 %) and hemicellulose (5 %) accompanied by a cellulose accumulation (11 %). Analysis of pretreated bagasse structure revealed that lime pretreatment caused considerable damage to the bagasse fibers, including rupture of the cell wall, exposing the cellulose-rich areas to enzymatic action. We showed that lime pretreatment is effective in improving enzymatic digestibility of raw sugarcane bagasse, even at low lime loading and over a short pretreatment period. It was also demonstrated that this pretreatment caused alterations in the structure and composition of raw bagasse, which had a pronounced effect on the enzymes accessibility to the substrate, resulting in an increase of cellulose hydrolysis rate. These results indicate that the use of raw sugarcane bagasse (without prior screening) pretreated with lime (cheaper and environmentally friendly reagent) may represent a cost reduction in the cellulosic ethanol production.


Immunogenicity and reactogenicity of a combined high dose hepatitis A and hepatitis B vaccine, compared to that of Twinrix in healthy Indian children.
Hepatitis A virus (HAV) and hepatitis B virus (HBV) are vaccine preventable important childhood acquired infectious diseases in developing countries. In the changing epidemiology of HAV, the utility of such a vaccine in India needs urgent attention. Further, the efficacy of two versus three dose schedule needs to be assessed to improve compliance. One hundred healthy school children, aged 1-15 years were recruited in a randomised open study to receive either vaccination schedule: Group I: combined high-dose hepatitis A and B vaccine to be administered on a 0, 6 month schedule intramuscularly; Group II: to be administered on 0, 1, 6 month Twinrix (GlaxoSmithKline Biologicals, Rixensart, Belgium) schedule intramuscularly. The seroconversion (> or =1MIU/ml for anti-HBs antibodies and > or =33MIU/ml for anti-HAV antibodies) and seroprotection (anti-HBs > or =10MIU/ml after the third dose of vaccine) rates were determined at months 1, 2, and 7. The mean age and gender was similar between groups: 7.9+/-2.6 years (range 3-15 years). At month 7 all subjects (100%) in both groups were seropositive for anti-HAV antibodies, Group I had higher anti-HAV titres at months 1 or 2 compared to Group II (P=0.025, P=0.040). Group II developed higher seroprotection rates (month 2, P=0.002, month 6, P=0.003) compared to Group I and higher titres (month 2, P=0.001, month 6 P=0.001) compared to Group I. At month 7, the geometric mean titres (GMTs) were comparable between groups and seroprotection reached 100% in both the groups. The incidence of any symptom per dose analysis reported during a 4-day follow-up period was significantly higher in Group I, 53% (52/98) of the documented doses compared to 37% (54/146) in Group II (P=0.018). Twinrix vaccine is safe and highly immunogenic in Indian children. Further study of the high dose vaccine would determine if its two dose regimen is a feasible advantage.


Atovaquone-proguanil exposure in pregnancy and risk for adverse fetal and infant outcomes: A retrospective analysis.
Malaria in pregnancy can cause severe maternal and fetal complications. Chloroquine (CQ) and mefloquine (MQ) are recommended for chemoprophylaxis in pregnancy, but are not always suitable. Atovaquone-proguanil (AP) might be a viable option for malaria prevention in pregnancy, but more safety data are needed. Data for pregnancies and live births among active duty military women, 2003-2014, from the Department of Defense Birth and Infant Health Research program were linked with pharmacy data to determine antimalarial exposure. Multivariable Cox and logistic regression models were used to assess the relationship of antimalarial exposure with fetal and infant outcomes, respectively. Among 198,164 pregnancies, 50 were exposed to AP, 156 to MQ, and 131 to CQ. Overall, 17.6% of unexposed pregnancies and 28.0%, 16.0%, and 6.1% of pregnancies exposed to AP, MQ, and CQ, respectively, ended in fetal loss (spontaneous abortion or stillbirth) (adjusted hazard ratios [aHR] = 1.46, 95% confidence interval [CI] 0.87-2.46; aHR = 1.06, 95% CI 0.72-1.57; and aHR = 0.47, 95% CI 0.24-0.94, respectively). The small number of AP exposed pregnancies highlights the difficulty in assessing safety. While definitive conclusions are not possible, these data suggest further research of AP exposure in pregnancy and fetal loss is warranted. More research on fetal loss following atovaquone-proguanil exposure in pregnancy is warranted.


Investigation of Bar-seq as a method to study population dynamics of Saccharomyces cerevisiae deletion library during bioreactor cultivation.
Despite the latest advancements in metabolic engineering for genome editing and characterization of host performance, the successful development of robust cell factories used for industrial bioprocesses and accurate prediction of the behavior of microbial systems, especially when shifting from laboratory-scale to industrial conditions, remains challenging. To increase the probability of success of a scale-up process, data obtained from thoroughly performed studies mirroring cellular responses to typical large-scale stimuli may be used to derive crucial information to better understand potential implications of large-scale cultivation on strain performance. This study assesses the feasibility to employ a barcoded yeast deletion library to assess genome-wide strain fitness across a simulated industrial fermentation regime and aims to understand the genetic basis of changes in strain physiology during industrial fermentation, and the corresponding roles these genes play in strain performance. We find that mutant population diversity is maintained through multiple seed trains, enabling large scale fermentation selective pressures to act upon the community. We identify specific deletion mutants that were enriched in all processes tested in this study, independent of the cultivation conditions, which include MCK1, RIM11, MRK1, and YGK3 that all encode homologues of mammalian glycogen synthase kinase 3 (GSK-3). Ecological analysis of beta diversity between all samples revealed significant population divergence over time and showed feed specific consequences of population structure. Further, we show that significant changes in the population diversity during fed-batch cultivations reflect the presence of significant stresses. Our observations indicate that, for this yeast deletion collection, the selection of the feeding scheme which affects the accumulation of the fermentative by-product ethanol impacts the diversity of the mutant pool to a higher degree as compared to the pH of the culture broth. The mutants that were lost during the time of most extreme population selection suggest that specific biological processes may be required to cope with these specific stresses. Our results demonstrate the feasibility of Bar-seq to assess fermentation associated stresses in yeast populations under industrial conditions and to understand critical stages of a scale-up process where variability emerges, and selection pressure gets imposed. Overall our work highlights a promising avenue to identify genetic loci and biological stress responses required for fitness under industrial conditions.


Toll-like receptor 9 promoter polymorphism is associated with decreased risk of Alzheimer's disease in Han Chinese.
Toll-like receptors (TLRs), as major innate immune mediators, may be involved in clearance of cerebral amyloid-β (Aβ) deposits. Recently, a novel TLR9 signaling pathway has been uncovered, which is functionally associated with the immune inflammatory response and reducing Aβ burden in Alzheimer's disease (AD) mice. Therefore, TLR9 might represent a reasonable functional candidate gene for AD. Our study investigated 1,133 sporadic late-onset AD (LOAD) and 1,159 healthy controls matched for sex and age in a large Han Chinese population. One selected functional rs187084 polymorphism within the TLR9 gene was genotyped by polymerase chain reaction-ligase detection reaction in a case-control associated study. The TLR9 rs187084 variant homozygote GG was significantly associated with a decreased LOAD risk after adjusting for age, gender, and ApoE ε4 status by logistic regression analysis (P = 0.035). Our result showed significant evidence of the interaction of ApoE ε4 with rs187084. When we further stratified our data by the ApoE ε4 status, we detected significant differences in the genotype and allele distributions of rs187084 between LOAD patients and controls in ApoE ε4 carriers (P < 0.001, P = 0.003, respectively). Moreover, we examined TLR9 expression in peripheral blood monocytes by flow cytometry, and the GG genotype of the TLR9 rs187084 polymorphism was associated with a higher TLR9 expression than two other genotypes in LOAD patients. Our findings support the hypothesis that the TLR9 polymorphism may modify LOAD risk in the Han Chinese population.


Models for the active transport of cations...the steady-state analysis.
We summarise the progress that has been made in the analysis of active transport models, at the steady-state level. The two general classes of such model, counter-and co-transport, can be treated by a kinetic analysis which makes no assumptions as to the symmetry or asymmetry of the systems nor as to the presence of any particular rate-limiting steps. Precisely the same formalism is obeyed for primary active transport as for secondary active transport. Both are merely a generalisation of facilitated diffusion, in that they follow directly from accepted properties of carrier models. How affinities of such carriers for their substrates affect the efficiency of active transport is discussed and it is shown that in a number of cases, the affinity changes that the carrier demonstrates arise from inherent properties of the free carrier and not from any "high energy" properties of the chemical reactants. Methods of obtaining the kinetic parameters of the system from experimental data are reviewed, together with methods for testing and characterising the different transport models.


pCLE highlights distinctive vascular patterns in early gastric cancer and in gastric diseases with high risk of malignant complications.
Endoscopy is widely used to detect and diagnose precancerous lesions and gastric cancer (GC). The probe-based Confocal Laser Endomicroscopy (pCLE) is an endoscopic technique suitable for subcellular resolution and for microvasculature analyses. The aim of this study was to use pCLE to identify specific vascular patterns in high-risk and early stage GC. Mucosal architecture, vessel tortuosity, enlargements and leakage were assessed in patients with autoimmune gastritis and early gastric cancer (EGC). We were able to stratify gastritis patients by identifying distinct vascular profiles: gastritis was usually associated with increased vascularization characterized by a high number of tortuous vessels, which were also found in atrophic autoimmune disease. Leaky and tortuous vessels, distributed in a spatially irregular network, characterized the atrophic metaplastic mucosa. The mucosal vasculature of EGC patients displayed tortuous vessels, but unlike what detected in atrophic gastritis, they appeared patchy, as is in neoplastic gastric tissue. Very importantly, we detected vascular changes even in areas without lesions, supporting the contention that vascular alterations may provide a favorable microenvironment for carcinogenesis. This report confirms that pCLE is a valid endoscopic approach to improve the definition of patients with malignant lesions or at increased risk for GC by assessing vascular changes.


Dose-response association between adult height and all-cause mortality: a systematic review and meta-analysis of cohort studies.
We conducted a systematic review and meta-analysis from published cohort studies to examine the association of adult height and all-cause mortality and to further explore the dose-response association. PubMed, The Cochrane Library, The Ovid, CNKI, CQVIP and Wanfang databases were searched for articles published from database inception to 6 February 2018. We used the DerSimonian-Laird random-effects model to estimate the quantitative association between adult height and all-cause mortality and the restricted cubic splines to model the dose-response association. We included 15 articles, with 1 533 438 death events and 2 854 543 study participants. For each 5-cm height increase below the average, the risk of all-cause mortality was reduced by 7% [relative risk (RR) = 0.93, 95% confidence interval (CI), 0.89-0.97] for men and 5% (RR = 0.95, 95% CI, 0.90-0.99) for women. All-cause mortality had a U-shaped association with adult height, the lowest risk occurring at 174 cm for men and 158 cm for women (both Pnonlinearity < 0.001). Relative to the shortest adult height (147 cm for men and 137 cm for women), men at 174 cm had a 47% lower likelihood of all-cause mortality and women at 158 cm a 33% lower risk of all-cause mortality. Our study suggests that the relation between adult height and all-cause mortality is approximately U-shaped in both men and women.


The importance of hyperkalemia in a cold perfusion solution: a correlative study examining myocardial function, metabolism, tissue gases, and substrates.
Twenty-four pigs were studied to assess the effect of potassium in a cardioplegic solution on the ability of the swine myocardium to maintain functional and metabolic integrity following induced ischemia. The pigs were evaluated on total and right heart bypass with measurement at normothermia and after a one-hour intervention of stroke volume (SV), coronary blood flow (CBF), myocardial oxygen consumption (MVO2), and lactate extraction. Myocardial tissue gases (PmO2 and PmCO2) were continuously monitored and, at the conclusion of the procedure tissues were analyzed for adenosine triphosphate (ATP). There were five interventions: (1) hypothermic perfusion (28 degrees C) (Group 1); (2) hypothermic ischemia (28 degrees C) (Group 2); and hypothermic ischemia with a cardioplegic solution (nonlactated Ringer's solution, pH 7.4, 4 degrees C) using (3) normokalemia (4 mEq of potassium chloride/L, 300 mOsm/L (Group 3), (4) hyperkalemia (43 mEq of KCl/L, 390 mOsm/L) (Group 4), and (5) normokalemia with increased osmolarity (3.6 mEq of KCl/L, 400 mOsm/L) (Groups 5). A significant decrease in SV and elevation in peak PmCO2 were seen in all groups subjected to ischemia except those protected with hyperkalemic solution. We conclude that the presence of hyperkalemia in a cold root perfusion solution provides better myocardial protection than cold root perfusion alone. Furthermore, potassium arrest appears to be more protective than coronary perfusion at 28 degrees C.


Does machine-vision-assisted dynamic navigation improve the accuracy of digitally planned prosthetically guided immediate implant placement? A randomized controlled trial.
This randomized controlled clinical trial was designed to compare the accuracy of machine-vision (MV)-based dynamic navigation (DN)-assisted immediate implant placement with the conventional freehand technique. A total of 24 subjects requiring immediate implant placement in maxillary anterior teeth were randomly assigned to either the control (freehand by an experienced surgeon, n = 12) or the test group (MV-DN, n = 12). Implant platform, implant apex, angular, and depth deviations with respect to prosthetically guided digital planning and differences in implant insertion torque (ITV) and implant stability quotient (ISQ) were compared between the groups. MV-DN resulted in more accurate immediate implant position: significantly smaller global platform deviation (1.01 ± 0.41 mm vs. 1.51 ± 0.67 mm, p = .038), platform depth deviation (0.44 ± 0.46 mm vs. 0.95 ± 0.68 mm, p = .045), global apex deviation (0.88 ± 0.43 mm vs. 1.94 ± 0.86 mm, p = .001), and lateral apex deviation (0.68 ± 0.30 mm vs. 1.61 ± 0.88 mm, p = .004) were found in MV-DN compared to controls. No significant intergroup differences were observed for ITV and ISQ. MV-DN achieved more precise immediate implant position and comparable primary stability. Further trials are necessary to assess the benefits in terms of esthetics and tissue health/stability.


Stem cell marker TRA-1-60 is expressed in foetal and adult kidney and upregulated in tubulo-interstitial disease.
The kidney has an intrinsic ability to repair itself when injured. Epithelial cells of distal tubules may participate in regeneration. Stem cell marker, TRA-1-60 is linked to pluripotency in human embryonic stem cells and is lost upon differentiation. TRA-1-60 expression was mapped and quantified in serial sections of human foetal, adult and diseased kidneys. In 8- to 10-week human foetal kidney, the epitope was abundantly expressed on ureteric bud and structures derived therefrom including collecting duct epithelium. In adult kidney inner medulla/papilla, comparisons with reactivity to epithelial membrane antigen, aquaporin-2 and Tamm-Horsfall protein, confirmed extensive expression of TRA-1-60 in cells lining collecting ducts and thin limb of the loop of Henle, which may be significant since the papillae were proposed to harbour slow cycling cells involved in kidney homeostasis and repair. In the outer medulla and cortex there was rare, sporadic expression in tubular cells of the collecting ducts and nephron, with positive cells confined to the thin limb and thick ascending limb and distal convoluted tubules. Remarkably, in cortex displaying tubulo-interstitial injury, there was a dramatic increase in number of TRA-1-60 expressing individual cells and in small groups of cells in distal tubules. Dual staining showed that TRA-1-60 positive cells co-expressed Pax-2 and Ki-67, markers of tubular regeneration. Given the localization in foetal kidney and the distribution patterns in adults, it is tempting to speculate that TRA-1-60 may identify a population of cells contributing to repair of distal tubules in adult kidney.


Legume effects in a native community invaded by alien Asteraceae in a multi-species comparison.
Facilitation has been a long-neglected type of interaction but received more attention recently. Legumes are commonly involved in facilitative interactions due to their nitrogen fixation. Facilitative interactions are so far underappreciated yet potentially important for biological invasions, especially given increasing numbers of alien species. In a common garden experiment using 30 annual Asteraceae species (neophytes, archaeophytes, plus some natives), grown in communities with or without legume presence, we measured functional traits and fitness in focal Asteraceae, as well as nitrogen characteristics of Asteraceae and two native community phytometer species. We investigated how legume presence affects relationships between trait and nitrogen concentration and Asteraceae fitness; and whether mechanisms of facilitation in legume presence and its effects on aboveground performance differ among native phytometer, neophyte, and archaeophyte Asteraceae using the δ15N natural abundance method. Lower specific leaf area was associated with higher aboveground biomass and seed production, with a stronger effect in legume absence. Nitrogen concentration had a positive relationship with biomass, but did not generally increase seed production. Our results hint at N facilitation for the native grass phytometer Festuca rupicola when growing in legume presence, whereas the forb Potentilla argentea and 27 alien Asteraceae species did not indicate facilitative effects. Intriguingly, direct legume facilitation in native phytometer species was only detected when growing with archaeophytes neighbors, not with neophytes. This hints at varied mechanisms of competition for nitrogen between natives and alien species of different residence time and deepens the understanding of altered facilitative leguminous effects in alien species presence.


Knee joint kinematics and kinetics during the hop and cut after soft tissue artifact suppression: Time to reconsider ACL injury mechanisms?
The recent development of a soft tissue artifact (STA) suppression method allows us to re-evaluate the tibiofemoral kinematics currently linked to non-contact knee injuries. The purpose of this study was therefore to evaluate knee joint kinematics and kinetics in six degrees of freedom (DoF) during the loading phases of a jump lunge and side cut using this in silico method. Thirty-five healthy adults completed these movements and their surface marker trajectories were then scaled and processed with OpenSim's inverse kinematics (IK) and inverse dynamics tools. Knee flexion angle-dependent kinematic constraints defined based on previous bone pin (BP) marker trajectories were then applied to the OpenSim model during IK and these constrained results were then processed with the standard inverse dynamics tool. Significant differences for all hip, knee, and ankle DoF were observed after STA suppression for both the jump lunge and side cut. Using clinically relevant effect size estimates, we conclude that STA contamination had led to misclassifications in hip transverse plane angles, knee frontal and transverse plane angles, medial/lateral and distractive/compressive knee translations, and knee frontal plane moments between the NoBP and the BP IK solutions. Our results have substantial clinical implications since past research has used joint kinematics and kinetics contaminated by STA to identify risk factors for musculoskeletal injuries.


Time-course changes in the expression of heme oxygenase-1 in human subcutaneous hemorrhage.
To determine the time-course of human subcutaneous hemorrhage, heme oxygenase (HO)-1 expression and macrophage infiltration were observed using an immunohistochemical technique and semiquantitative analysis. The number of immunoreactive cells and the number of all infiltrating cells of each microscopic field were counted, and the ratio of the former to the latter was calculated as the positive cells ratio. An increase in the HO-1-positive cells ratio was observed starting at 3 h after injury, and the maximum ratio was observed 3 days after injury. The pattern of the increase in the macrophage ratio was similar to that of the HO-1-positive cells ratio in the early period after injury. Observation of serial sections revealed that the expression of HO-1 in the cells corresponded to the localization of macrophage. The present results suggest that the determination of HO-1 expression, as derived from macrophages, might be useful for the estimation of the time-course of subcutaneous hemorrhage.


Results of extraocular muscle surgery in WEBINO bilateral internuclear ophthalmoplegia patients.
To evaluate the results of extraocular surgery in patients with large exotropias from "wall-eyed" bilateral internuclear ophthalmoplegia (WEBINO). Eight patients with WEBINO evaluated between November 1994 and October 2006 underwent extraocular muscle surgery. Three patients required a second procedure. An 85% reduction in exodeviation occurred postoperatively. Fusion responses were demonstrable in 87.5% of patients. Other ocular signs of internuclear ophthalmoplegia (INO) such as adduction limitation, abducting nystagmus, and ocular dysmetria also improved. These results were obtained following two-staged unilateral recession-resection procedures in seven patients; the remaining patient underwent bilateral lateral rectus recessions. Binocular outcomes are achievable in patients with WEBINO following extraocular muscle surgery, and associated ocular signs may be reduced or eliminated.


Medical information networking and standardization.
There are few environments that generate and manage as much information as the hospital. Although computers are used extensively to manage this information, their application has been opportunistic rather than strategic. This has led to a multi-vendor information processing environment in many hospitals. Hospitals today have financial systems, order-entry communication systems, ADT systems, and clinical systems such as laboratory, pharmacy, etc. Communication between these islands of automation however are still, for the most part, primitive. Exceptions are in the large teaching hospitals where internal staff have been utilized to link systems together. The effort has traditionally been huge and fraught with difficulties related to the heterogeneous environment that is involved. In order to overcome these problems and enable us to truly bring the power of modern computing to bear on hospital-wide problems, as well as to make feasible, for example, the computerized medical record, we must develop an appropriate strategy. It seems unlikely that a single-vendor solution will entirely answer hospital information processing needs. Therefore, a perpetuation of the heterogeneous environment is inevitable. With this in mind, an alternative approach is proposed: to develop networking standards for the hospital, as has been done for the manufacturing and technical office environment, such that vendors can build systems that can be easily integrated.


Management of neonatal candidiasis. Neonatal Candidiasis Study Group.
To identify areas of consensus and controversy in the management of neonatal candidiasis. A questionnaire was distributed to US-based members of the Pediatric Infectious Diseases Society and a sampling of US neonatologists. Three hundred eighty evaluable questionnaires were returned (42% of those mailed). Ninety-five percent of respondents have cared for an infant with systemic candidiasis in the past 2 years. Fluconazole and liposomal amphotericin are used to some extent by 90 and 69% of respondents, respectively. A single blood culture positive for Candida led to a recommendation for immediate treatment by 99%; amphotericin B was the preferred therapy for candidemia (88%). More than 80% of respondents would request cerebrospinal fluid, urine and repeat blood cultures and ophthalmologic examination in the evaluation of candidemia. If a cerebrospinal fluid culture is positive, 25% would use amphotericin B alone whereas 62% would add flucytosine. For candiduria Society members chose fluconazole therapy more often than did neonatologists, 23% vs. 3.4% (P<0.001). There was no consensus concerning duration of therapy, use of an amphotericin B test dose or management of a central catheter in place during candidemia. Systemic candidiasis in neonates is a frequently encountered clinical problem. There is agreement that prompt therapy with amphotericin B is required if a blood culture is positive for Candida and that such infants require additional evaluations. Other antifungals (fluconazole, liposomal amphotericin B) are used to some extent in this population. Many issues in management have no clear consensus and warrant further research.


Association of physical activity with overall mortality among long-term testicular cancer survivors: A longitudinal study.
Physical activity (PA) has been associated with reduced mortality among cancer survivors, but no study has focused on testicular cancer survivors (TCSs). We aimed to investigate the association of PA measured twice during survivorship with overall mortality in TCSs. TCSs treated during 1980 to 1994 participated in a nationwide longitudinal survey between 1998 to 2002 (S1: n = 1392) and 2007 to 2009 (S2: n = 1011). PA was self-reported by asking for the average hours per week of leisure-time PA in the past year. Responses were converted into metabolic equivalent task hours/week (MET-h/wk) and participants were categorized into: Inactives (0 MET-h/wk), Low-Actives (2-6 MET-h/wk), Actives (10-18 MET-h/wk) and High-Actives (20-48 MET-h/wk). Mortality from S1 and S2, respectively, was analyzed using the Kaplan-Meier estimator and Cox proportional hazards models until the End of Study (December 31, 2020). Mean age at S1 was 45 years (SD 10.2). Nineteen percent (n = 268) of TCSs died between S1 and EoS, with 138 dying after S2. Compared to Inactives at S1, the mortality risk among Actives was 51% lower (HR 0.49, 95% CI: 0.29-0.84) with no further mortality reduction among High-Actives. At S2, the mortality risk was at least 60% lower among the Actives, High-Actives and even the Low-Actives compared to the Inactives. Persistent Actives (≥10 MET-h/wk at S1 and S2) had a 51% lower mortality risk compared to Persistent Inactives (<10 MET-h/wk at S1 and S2; HR 0.49, 95% CI: 0.30-0.82). During long-term survivorship after TC treatment, regular and maintained PA were associated with an overall mortality risk reduction of at least 50%.


How Brain Fat Conquers Stress.
A new paper by Bailey et al. reveals that lipid droplets, crucial organelles for energy storage, can also protect against oxidative stress. In Drosophila larvae, lipid droplets in glia allow neuronal stem cells to keep proliferating under hypoxic conditions. Protection likely involves sequestering vulnerable membrane lipids away from reactive oxygen species.


Risk for cardiovascular disease associated with metabolic syndrome and its components: a 13-year prospective study in the RIVANA cohort.
We aimed to investigate the association of metabolic syndrome (MetS) and its single components with cardiovascular risk and estimated their impact on the prematurity of occurrence of cardiovascular events using rate advancement periods (RAPs). We performed prospective analyses among 3976 participants (age range: 35-84, 55% female) in the Vascular Risk in Navarre (RIVANA) Study, a Mediterranean population-based cohort. MetS was defined based on the modified criteria of the American Heart Association/National Heart, Lung, and Blood Institute and the International Diabetes Federation. The primary endpoint was major cardiovascular event (a composite of myocardial infarction, stroke, or mortality from cardiovascular causes). Secondary endpoints were incidence of non-fatal myocardial infarction and non-fatal stroke, cardiovascular mortality, and all-cause mortality. Cox proportional hazards models, adjusted for potential confounders, were fitted to evaluate the association between MetS and its single components at baseline with primary and secondary endpoints. During a median follow-up of 12.8 years (interquartile range, 12.5-13.1), we identified 228 primary endpoint events. MetS was associated with higher risk of incidence of major cardiovascular event, cardiovascular and all-cause mortality, but was neither associated with higher risk of myocardial infarction nor stroke. Compared with participants without MetS, the multivariable hazard ratio (95% confidence interval [CI]) among participants with MetS was 1.32 (1.01-1.74) with RAP (95% CI) of 3.23 years (0.03, 6.42) for major cardiovascular event, 1.64 (1.03-2.60) with RAP of 3.73 years (0.02, 7.45) for cardiovascular mortality, and 1.45 (1.17-1.80) with RAP of 3.24 years (1.21, 5.27) for all-cause mortality. The magnitude of the associations of the single components of MetS was similar than the predicted by MetS. Additionally, for each additional trait of MetS, incidence of major cardiovascular event relatively increased by 22% (1.22, 95% CI 1.09-1.36) with RAP of 2.31 years (0.88, 3.74). MetS was independently associated with CVD risk, cardiovascular and all-cause mortality. Components of the MetS were associated with similar magnitude of increased CVD, which suggests that MetS was not in excess of the level explained by the presence of its single components. Further research should explore the association of different combinations of the components of MetS with CVD.


Expression of proinflammatory cytokine interleukin-6 in tissue samples of human prostate obtained by needle biopsy.
Interleukin-6 (IL-6) has been associated with the development of prostate cancer. The aim of the study was to clarify whether IL-6 expression in prostate tissue could be a useful marker in differentiation of prostate diseases in small foci by pathologist visual scoring. Archival paraffin-embedded specimens of benign prostate hyperplasia (BPH), high-grade prostatic intraepithelial neoplasia (PIN), prostatitis and prostate adenocarcinoma were studied by immunohistochemistry with a mouse monoclonal antibody IL-6 using the streptavidin-biotin method. Significantly, lower IL-6 immunoreactivity was observed in normal epithelial cells (p=0.000) and basal cells (p=0.000) in the samples of prostate adenocarcinoma in comparison to the samples with BPH, PIN and prostatitis. There was no significant difference in IL-6 expression in malignant and premalignant cells (p=0.814) as well as in stromal cells among the four diagnoses (p=0.22). IL-6 was expressed in normal epithelial cells, premalignant epithelial cells and malignant epithelial cells as well as in stromal cells. However, in our research IL-6 was of limited utility as a single marker for differential diagnosis of the prostate diseases in small foci needle biopsy by pathologist visual scoring. The standardization of immunohistochemical (IHC) staining protocol for IL-6 is required to determine IL-6 expression in order to avoid possible misinterpretation of the IHC results.


Measurement of corneal elasticity with an acoustic radiation force elasticity microscope.
To investigate the role of collagen structure in corneal biomechanics, measurement of localized corneal elasticity with minimal destruction to the tissue is necessary. We adopted the recently developed acoustic radiation force elastic microscopy (ARFEM) technique to measure localize biomechanical properties of the human cornea. In ARFEM, a low-frequency, high-intensity acoustic force is used to displace a femtosecond laser-generated microbubble, while high-frequency, low-intensity ultrasound is used to monitor the position of the microbubble within the cornea. Two ex vivo human corneas from a single donor were dehydrated to physiologic thickness, embedded in gelatin and then evaluated using the ARFEM technique. In the direction perpendicular to the corneal surface, ARFEM measurements provided elasticity values of E = 1.39 ± 0.28 kPa for the central anterior cornea and E = 0.71 ± 0.21 kPa for the central posterior cornea in pilot studies. The increased value of corneal elasticity in the anterior cornea correlates with the higher density of interweaving lamellae in this region.


Chemical shift assignments for the Ig2 domain of human obscurin A.
The giant sarcomeric protein obscurin (~720 kDa) is an essential contributor to myofibrillogenesis and acts as the only known tether between the contractile apparatus and the surrounding membrane structures in myofibrils. Genomic characterization of OBSCN suggests a modular architecture, consisting of dozens of individually-folded Ig-like and FnIII-like domains arranged in tandem. Here we describe the sequence-specific chemical shift assignments of the second putative obscurin Ig-like domain (Ig2). This domain specifically binds to MyBP-C slow variant-1 through an unknown mechanism. Ultimately, the assignments presented here will facilitate high-resolution structure determination of Ig2 and provide insight into the specificity of the obscurin-MyBP-C interaction.


Changes in pyridine and adenine nucleotide levels in Friend erythroleukaemia cells during growth and differentiation.
Pyridine and adenine nucleotide levels were measured in Friend erythroleukaemia cells (FELC) stimulated to growth and induced to differentiate by hexamethylene bisacetamide (HMBA) and N'-methylnicotinamide (N'-MNAM). A three- to fourfold increase in the NADP(H) was found to parallel cell growth stimulation in both the presence and absence of differentiation inducers. NAD(H) increased about twofold in control and to a minor extent in HMBA-treated FELC but did not vary significantly in N'-MNAM-treated cells. ATP was significantly higher in control cells stimulated to growth than in resting ones, but it did not vary in inducer-treated cells. These data confirm the relationship between high NADP(H) levels and cell resumption to growth; moreover they show that NAD(H) pool reduction and NAD/NADH ratio rise are associated with the process of FELC differentiation. The activities of NAD pyrophosphorylase and NAD kinase are much more enhanced in growth-stimulated FELC than in resting ones. On the other hand transition from the quiescent to the proliferative state was accompanied by a decrease in the activity of poly(ADP-ribose) polymerase. A decrease in poly(ADP-ribose) polymerase activity was also found in differentiated cells in contrast to controls.


Fabricating Dual-Atom Iron Catalysts for Efficient Oxygen Evolution Reaction: A Heteroatom Modulator Approach.
Understanding the thermal aggregation behavior of metal atoms is important for the synthesis of supported metal clusters. Here, derived from a metal-organic framework encapsulating a trinuclear FeIII 2 FeII complex (denoted as Fe3 ) within the channels, a well-defined nitrogen-doped carbon layer is fabricated as an ideal support for stabilizing the generated iron nanoclusters. Atomic replacement of FeII by other metal(II) ions (e.g., ZnII /CoII ) via synthesizing isostructural trinuclear-complex precursors (Fe2 Zn/Fe2 Co), namely the "heteroatom modulator approach", is inhibiting the aggregation of Fe atoms toward nanoclusters with formation of a stable iron dimer in an optimal metal-nitrogen moiety, clearly identified by direct transmission electron microscopy and X-ray absorption fine structure analysis. The supported iron dimer, serving as cooperative metal-metal site, acts as efficient oxygen evolution catalyst. Our findings offer an atomic insight to guide the future design of ultrasmall metal clusters bearing outstanding catalytic capabilities.


MiR-181a and -b expression in acute lymphoblastic leukemia and its correlation with acute graft-versus-host disease after hematopoietic stem cell transplantation, COVID-19 and torque teno viruses.
Acute lymphoblastic leukemia (ALL), a malignant transformation and proliferation of the lymphoid line of blood cells, is characterized by chromosomal abnormalities and genetic changes. The purpose of this research was the evaluation of expression level of miR-181a and -b in patients with ALL compared to the control group. Furthermore, we examined their expression level in hematopoietic stem-cell transplantation (HSCT) patients who developed acute graft-versus-host disease (aGVHD) in comparison with those without aGVHD and explore the relationship between their expression level and cytogenetic abnormalities. In this cross-sectional study, 76 newly diagnosed adult De novo ALL patients were enrolled who were admitted to our referral hospital. All patients received standard chemotherapy, consisting of daunorubicin. A total of 37 patients underwent HSCT from the related human leukocyte antigen-matched donors. ALL patients have been diagnosed with the coronavirus disease 2019 (COVID-19) and Torque teno viruses (TTVs). We assessed the expression levels of miR-181a and -b in the peripheral blood sample of ALL patients at the time of diagnosis prior to chemotherapy, and healthy matched individuals by RT-PCR. TTVs and COVID-19 load were also determined via RT-PCR. In conclusion, the expression level of miR-181a and -b were significantly higher in ALL patients than healthy controls and also increased in patients who developed aGVHD in comparison with those without aGVHD. MiR-181a and -b can be a useful biomarker in ALL and a useful indicator of aGVHD. The expression level of miR-181a in ALL patients with COVID-19 is significantly up-regulated, while it is reduced in these patients with TTV.


Genetic deletion of microsomal prostaglandin E synthase-1 suppresses mouse mammary tumor growth and angiogenesis.
The cyclooxygenase/prostaglandin (COX/PG) signaling pathway is of central importance in inflammation and neoplasia. COX inhibitors are widely used for analgesia and also have demonstrated activity for cancer prophylaxis. However, cardiovascular toxicity associated with this drug class diminishes their clinical utility and motivates the development of safer approaches both for pain relief and cancer prevention. The terminal synthase microsomal PGE synthase-1 (mPGES-1) has attracted considerable attention as a potential target. Overexpression of mPGES-1 has been observed in both colorectal and breast cancers, and gene knockout and overexpression approaches have established a role for mPGES-1 in gastrointestinal carcinogenesis. Here we evaluate the contribution of mPGES-1 to mammary tumorigenesis using a gene knockout approach. Mice deficient in mPGES-1 were crossed with a strain in which breast cancer is driven by overexpression of human epidermal growth factor receptor 2 (HER2/neu). Loss of mPGES-1 was associated with a substantial reduction in intramammary PGE2 levels, aromatase activity, and angiogenesis in mammary glands from HER2/neu transgenic mice. Consistent with these findings, we observed a significant reduction in multiplicity of tumors ≥1mm in diameter, suggesting that mPGES-1 contributes to mammary tumor growth. Our data identify mPGES-1 as a potential anti-breast cancer target.


EZH2-Myc driven glioblastoma elicited by cytomegalovirus infection of human astrocytes.
Mounting evidence is identifying human cytomegalovirus (HCMV) as a potential oncogenic virus. HCMV has been detected in malignant gliomas. EZH2 and Myc play a potential oncogenic role, correlating with the glioma grade. Herewith, we present the first experimental evidence for HCMV as a reprogramming vector, straight through the dedifferentiation of mature human astrocytes, and generation of CMV-Elicited Glioblastoma Cells (CEGBCs) possessing glioblastoma-like traits. HCMV counterparts the progression of the perceived cellular and molecular mechanisms succeeding the transformation and invasion processes with CEGBCs involved in spheroid formation and invasiveness. Glioblastoma multiforme (GBM) biopsies were characterized by an elevated EZH2 and Myc expression, possessing a strong positive correlation between the aforementioned markers in the presence of HCMV. From GBM tissues, we isolated HCMV clinical strains that transformed HAs toward CEGBCs exhibiting upregulated EZH2 and Myc. Spheroids generated from CEGBCs possessed invasion potential and were sensitive to EZH2 inhibitor, ganciclovir, and temozolomide triple therapy. HCMV clinical strains transform HAs and fit with an HCMV-induced glioblastoma model of oncogenesis, and supports the tumorigenic properties of Myc and EZH2 which might be highly pertinent in the pathophysiology of astrocytic brain tumors and thereby paving the way for new therapeutic strategies.