Culling intervals and culling risks in four stages of the reproductive life of first service and reserviced female pigs in commercial herds.
The objectives of this study were to measure culling intervals and culling risks in the four stages of the reproductive life of female pigs and to compare culling intervals between the number of services and between herd groups, based on herd productivity. We also compared survival patterns of females pigs between these herd groups. Our data set included lifetime records of 52,792 females born between 2001 and 2004 in 101 commercial herds. Two herd groups were selected on the basis of the upper 25th percentile of pigs weaned per mated female per 5 yr between 2002 and 2006, namely the high-performing herds, and ordinary herds. Culled females were also allocated into four groups based on the stages of their reproductive life when culled: unmated gilts, mated gilts, unmated sows, and mated sows. Culling intervals in unmated gilts and mated gilts were defined as the number of days from birth to culling and from first mating to culling, respectively. Culling intervals in unmated sows and mated sows were the number of days from weaning to culling. The number of services was categorized into two groups: first service and reservice groups. Multilevel linear mixed-effects models and survival analysis were performed. Culling intervals (+/-SEM) in unmated gilts, mated gilts, unmated sows, and mated sows were 302.9+/-1.16, 98.4+/-0.92, 14.3+/-0.12, and 89.6+/-0.42 d, respectively. Culling risks in the four groups were 5.6%, 7.1%, 58.0%, and 29.3%, respectively. In unmated gilts, mated gilts, and mated sows, the culling intervals in the high-performing herds were 43.0, 18.9, and 16.0 d shorter than those in ordinary herds, respectively (P<0.05), but no difference was found between the herd groups for the culling interval of unmated sows. For mated sows in the reservice group, culling intervals of high-performing herds were >or=13.7 d shorter than those of the ordinary herds (P<0.05), but for mated sows in the first service group, there was no difference in the culling interval between the herd groups. The culling hazard from 8 wk postweaning for mated sows in high-performing herds increased more rapidly than that in ordinary herds. In conclusion, to reduce culling intervals and improve herd productivity, we recommend implementing a strict culling policy for mated gilts and mated sows, especially reserviced females.


Enterococcus spp. in endodontically treated teeth with and without periradicular lesions.
The purpose of this study was to determine if Enterococcus spp. are more prevalent in endodontically treated teeth with periradicular lesions compared with teeth that require retreatment but have no periradicular rarefaction. Fifty-eight teeth that had received root canal therapy more than 1 yr previously and required retreatment were included. Designation of lesion versus no lesion was determined by two experienced endodontists. DNA extraction and PCR amplification were performed using ubiquitous 16S rDNA bacterial primers, as well as Enterococcus spp.-specific primers. The results showed that the overall prevalence of bacteria was 90% and Enterococcus spp. was 12%. chi analysis revealed a statistically significant relationship between the presence of a lesion and the presence of bacteria, as detected by the universal primers (p = 0.032). Using logistic regression, a statistically significant relationship was found between teeth with normal periapex and the presence of Enterococcus spp. (p = 0.023). This study revealed that bacteria are significantly associated with endodontic treatment failure but enterococci are not associated with disease.


Active Ageing in Italy: An Evidence-Based Model to Provide Recommendations for Policy Making and Policy Implementation.
In ageing societies, active ageing (AA) has been recognized as a useful conceptual tool due to its holistic approach to social issues and recognised benefits from it at multiple levels (micro, meso and macro) for addressing demographic challenges. However, one of the main problems identified in relation to AA, is to turn into practice, at the policy making level, the many positive aspects that it promises at the conceptual level, as is demonstrated by the available evidence based on experiences carried out in some European countries. As an advancement in this field, through an original research experience carried out in Italy between 2019 and 2021, this study for the first time provides a model for producing recommendations for policy making and policy implementation in the field of AA, by managing the main problematic aspects related to the operationalization, at the policy making level, of the AA concept, with the potential for replication in other countries. The main challenges were identified, as well as the way to deal with them through a model, for a proper operationalization of the AA concept, based, among other aspects, on a solid international framework concerning this matter, on a mainstreaming ageing approach (at the public policy level) and on a wide stakeholder participation through co-decisional tools. A multi-level (national-regional-local) perspective was adopted to consider cultural and geographical diversity, among other challenges.


Cytotoxic activity of phytochemicals from Garcinia mangostana L. and G. benthamiana (Planch. & Triana) Pipoly against breast cancer cells.
Four xanthones, α-mangostin (1), β-mangostin (2), mangostenol (3), mangaxanthone B (4), three benzophenones, mangaphenone (5), benthamianone (6), congestiflorone (7) and one sterol, stigmasterol (8) were isolated from the stem barks of Garcinia mangostana L. and G. benthamiana (Planch. & Triana) Pipoly. Compounds 1, 2, 4 and 5 exhibited significant cytotoxicity through MTT assay towards MCF-7 and MDA-MB-231 cells with the IC50 values range from 4.4 to 12.0 µM. Remarkably, mangaphenone (5) showed non-cytotoxicity against normal Vero cells, revealing its potential as lead compound for anti-breast cancer drug. Structure-activity relationship postulated that the prenyl and hydroxyl groups present in xanthones are important in promoting anti-proliferative effects. Molecular docking simulation study of 1, 2, 4 and 5 with 2OCF and 4PIV implied that the induction of apoptosis for both cancer cells involve ER and FAS signaling pathways. Future study on the lead optimization of 5 is highly recommended.


Efficient downregulation of VEGF in retinal pigment epithelial cells by integrin ligand-labeled liposome-mediated siRNA delivery.
The purpose of this study was to demonstrate the effectiveness of an integrin peptide ligand-labeled liposomal delivery system loaded with vascular endothelial growth factor (VEGF)-siRNA in a model study of gene therapy for retinopathy using human retinal pigment epithelial cells. Arg(R)-Gly(G)-Asp(D) motif peptide conjugating polyethylene glycol modified (RGD-PEGylated) liposomes were prepared using a thin-film hydration method and optimized for surface charge, particle size, small interfering RNA (siRNA) load, and entrapment efficiency. Reverse transcriptase-polymerase chain reaction and enzyme-linked immunosorbent assays were used to determine VEGF levels in retinal pigment epithelial cells. Cytotoxicity was determined using the 3-[4, 5-dimethylthiazol-2-yl]-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTS) assay and flow cytometry. Physicochemical properties, including particle size, zeta potential, and siRNA load, of the prepared RGD-PEGylated liposomes and their entrapment efficiency were determined to be within the following ranges: 123.8-234.1 nm, 17.31-40.09 m V, 5.27%-6.33%, and >97%, respectively. RGD-PEGylated liposome-mediated fluorescent-labeled siRNA delivery demonstrated significantly enhanced cellular uptake, and 3 mol% RGD-PEGylated liposomes (having 3β-[N-(N', N'-dimethylaminoethane) carbamoyl] cholesterol (DC-cholesterol) DSPE and DSPE-PEG(2000)-RGD with molar ratio of 50/47/3) were shown to have better efficacy with regard to specificity for retinal pigment epithelial cells, reduced cytotoxicity, and knockdown of the target molecule. By integrin receptor-mediated endocytosis, 3 mol% RGD-PEGylated liposomes were shown to be a suitable vector when loaded with VEGF-siRNA for efficient downregulation of VEGF in retinal pigment epithelial cells at both the protein and gene levels. This integrin ligand-modified liposomal delivery system has therapeutic potential for ocular gene therapy.


Antimicrobial susceptibilities of four species of Mycoplasma isolated in 2008 and 2009 from cattle in Japan.
We examined the susceptibility in vitro of 99 isolates of Mycoplasma (M. bovirhinis, M. bovis, M. alkalescens and M. bovoculi) and one of Acholeplasma laidlawii collected in 2008 and 2009 from Japanese homebred cattle and cows imported from Australia to 13 antimicrobial agents. Mycoplasma species isolated from homebred cattle were susceptible to spectinomycin and fluoroquinolones but resistant to macrolides, tetracyclines, thiamphenicol and flumequine. Kanamycin, lincomycin and chloramphenicol maintained intermediate effectiveness against these species. The isolates from the imported cows maintained sufficient susceptibility to all antimicrobials examined except for erythromycin and thiamphenicol. These results showed that four species of Mycoplasma isolated from Japanese homebred cattle had obviously higher drug resistance compared with previous reports, even within this decade.


Enhanced colloidal stability of nanoscale zero valent iron particles in the presence of sodium silicate water glass.
A method for the stabilization of nanoscale zero valent iron (nZVI) particles using silica was developed. Stabilization can significantly improve the performance characteristics of currently available nZVI products containing agglomerated particles. In the first step of the method, the agglomerates were broken using a sonication. A subsequent stabilizing effect was brought about by the deposition of silica onto the surface of the nZVI particles. The method was tested on three commercially available nZVI suspensions which formed agglomerates with mean sizes ranging from 1000 to 5000 nm. The application of the method resulted in a significant reduction of the mean size of the agglomerates to the values from 100 to 200 nm. The stabilizing effect of silica was also evidenced using scanning electron microscopy, zeta potential measurements and sedimentation analysis. The introduction of typical groundwater ions did not significantly affect the colloidal stability of the treated nZVI suspensions. The results of this study indicate that the silica coating have the potential to protect nZVI against agglomeration.


Efficiency of cycling exercise: Quantification, mechanisms, and misunderstandings.
The energetics of cycling represents a well-studied area of exercise science, yet there are still many questions that remain. Efficiency, broadly defined as the ratio of energy output to energy input, is one key metric that, despite its importance from both a scientific as well as performance perspective, is commonly misunderstood. There are many factors that may affect cycling efficiency, both intrinsic (e.g., muscle fiber type composition) and extrinsic (e.g., cycling cadence, prior exercise, and training), creating a complex interplay of many components. Due to its relative simplicity, the measurement of oxygen uptake continues to be the most common means of measuring the energy cost of exercise (and thus efficiency); however, it is limited to only a small proportion of the range of outputs humans are capable of, further limiting our understanding of the energetics of high-intensity exercise and any mechanistic bases therein. This review presents evidence that delta efficiency does not represent muscular efficiency and challenges the notion that the slow component of oxygen uptake represents decreasing efficiency. It is noted that gross efficiency increases as intensity of exercise increases in spite of the fact that fast-twitch fibers are recruited to achieve this high power output. Understanding the energetics of high-intensity exercise will require critical evaluation of the available data.


Preoperative informed consent for mandibular third molar surgeries: A survey analysis in a subset of dentists and oral surgeons in Saudi Arabia.
The objective of this study was to identify patterns of obtaining preoperative informed consent from patients undergoing mandibular third molar surgeries, in a subset of general dentists and oral surgeons in Saudi Arabia, and to compare the consenting patterns based on the clinician's rank, years of experience, place of work and gender. A prospective questionnaire based study was designed and data was obtained through an online survey from 102 participants who were selected for the study. Demographic information, clinician experience, type of informed consent obtained and information related to discussion of legal implications and complications were collected. Descriptive analysis of the obtained data and statistical comparisons using cross tabulation and Pearson Chi-Square test with a 95% significance level (P < 0.05) were done between the independents demographic variables and dependent variables pertaining to patterns of preoperative consenting. The survey response rate was 81.3% (n = 83), with 59.04% general dentists and 40.96% oral surgeons. The ratio of male to female respondents was approximately 3:1. Majority of the respondents reported a clinical experience of less than 10 years (77.11%) and were reportedly working in the private sector (73.49%). Nearly 80% of the respondents (79.52%) mentioned obtaining preoperative consent for mandibular third molar surgeries and was significantly higher (p-value - 0.018) among clinicians with more than 5 years of experience (90%). While 38.5% of the respondents indicated obtaining both a written and verbal consent, 53.01% obtained only a verbal consent. Majority of the respondents were aware of the legal implications of obtaining informed consent (81.93%) and disclosed incidental complications to their patients (91.57%). However, differences in the perceived post-operative complications associated with mandibular third molar surgeries were observed between general dentists and oral surgeons. The results of this study indicate a good level of knowledge about informed consent for mandibular third molar surgery and its legal implications among the dentists and oral surgeons who were surveyed. However, to avoid painful medico-legal disputes, a written informed consent signed by patients along with a witness should be considered mandatory.


Molecular aspects of bone healing and remodeling.
The process of fracture healing involves a complex series of coordinated cellular and molecular processes leading to removal of material contaminating the wound, angiogenesis validating restoration of disrupted microcirculation, and restoration of bone continuity by activation, proliferation and chemotaxis of bone progenitors from surrounding periosteum and endosteum. The regulation of various cell populations that orchestrate in this process depends on the biological effect evoked by cytokines and growth factors. The paper presents the state-of-the-art knowledge of the influence of growth factors and cytokines, namely TGF-beta, BMP, IGF, PDGF, FGF, NO, IL-1, IL-6, IL-11 on the process of fracture healing.


Drug treatments for polycystic ovary syndrome.
Polycystic ovary syndrome is a condition present in approximately 5 to 10 percent of women of childbearing age. Diagnosis can be difficult because the signs and symptoms can be subtle and varied. These may include hirsutism, infertility, menstrual irregularities, and biochemical abnormalities, most notably insulin resistance. Treatment should target specific manifestations and individualized patient goals. When choosing a treatment regimen, physicians must take into account comorbidities and the patient's desire for pregnancy. Lifestyle modifications should be used in addition to medical treatments for optimal results. Few agents have been approved by the U.S. Food and Drug Administration specifically for use in polycystic ovary syndrome, and several agents are contraindicated in pregnancy. Insulin-sensitizing agents are indicated for most women with polycystic ovary syndrome because they have positive effects on insulin resistance, menstrual irregularities, anovulation, hirsutism, and obesity. Metformin has the most data supporting its effectiveness. Rosiglitazone and pioglitazone are also effective for ameliorating hirsutism and insulin resistance. Metformin and clomiphene, alone or in combination, are first-line agents for ovulation induction. Insulin-sensitizing agents, oral contraceptives, spironolactone, and topical eflornithine can be used in patients with hirsutism.


Electroencephalographic activity before and after cognitive effort in children with attention deficit/hyperactivity disorder.
Numerous studies have detected elevated electroencephalographic (EEG) theta/beta ratio (TBR) or theta power in children with attention deficit/hyperactivity disorder (ADHD) and therefore TBR has been suggested to be a promising biomarker of ADHD. At the same time, recent theoretical models have emphasized the heterogeneity of ADHD and the notion that cognitive deficits in ADHD are not fixed but fluctuate according to contextual and state factors. Surprisingly, so far the context- or state-dependency of EEG abnormalities in ADHD has hardly been addressed. Therefore, in the current study, 3 minutes eyes closed resting EEG before and after execution of 3 n-back tasks were compared between 21 children with ADHD and 22 typically developing children. No difference between groups was found for TBR or theta power (or other frequency bands), neither before nor after task execution, indicating that enhanced TBR or theta power is not to be considered universal for the disorder. Hence, cautiousness is warranted in using these indices for diagnostic purposes in ADHD. Across groups, posterior theta power, as well as central and posterior beta power was attenuated after task execution, which was interpreted as the children experiencing a more alert state after cognitive effort. Yet, this EEG modulation was similar in both groups, providing no support for a context-or state-dependency of EEG abnormalities in ADHD. However, in light of the absence of any group differences in EEG parameters, further research is warranted.


Erectile dysfunction in premature ejaculation.
Between October 1993 and December 1995, 45 patients with premature ejaculation were included in this study. The patients were divided into two groups etiologically, as having primary or secondary ejaculation. After a complete laboratory evaluation all patients underwent papaverine tests color Doppler ultrasonography pharmacocavernosometry-cavernosography and consequently organic etiology were investigated. Venous leakage was found in 5 (22,7%), arterial or mix insufficiency in 2 (9%) of the patients with primary premature ejaculation. In secondary premature ejaculation group, venous leakage was encountered in 9 patients (39,1%), arterial insufficiency in 2 patients (8,6%). In conclusion, investigation of organic etiology for the patients with premature ejaculation, particularly with secondary premature ejaculation, seems to be beneficial for correct diagnosis.


Cell surface expression of fusogenic vesicular stomatitis virus G protein from cloned cDNA.
Vesicular stomatitis virus (VSV) enters the host cell by the receptor-mediated endocytotic pathway. This brings the virus particle into acidic vesicles inside the cell where infection occurs through a fusion event between the viral and the host vesicle membrane. In this work we have shown that the VSV glycoprotein (G) carries the fusion activity of this virus. The G protein was expressed on the surface of baby hamster kidney 21 cells from cloned cDNA which had been engineered into an expression vector and introduced into cell nuclei with the aid of a glass microneedle. A short (60 s) treatment with acid (pH less than or equal to 6.0) medium induced fusion of cells having G protein on their surface. For efficient G protein expression and cell-cell fusion we had to trim the 5' end of the G cDNA and to use as promoter the long terminal repeat of the mouse Moloney sarcoma virus.


Use of ICP/MS with ultrasonic nebulizer for routine determination of uranium activity ratios in natural water.
A method is described that allows precise determination of 234U/238U activity ratios (UAR) in most natural waters using commonly available inductively coupled plasma/mass spectrometry (ICP/MS) instrumentation and accessories. The precision achieved by this technique (+/- 0.5% RSD, 1 sigma) is intermediate between thermal ionization mass spectrometry (+/- 0.25% RSD, 1 sigma) and alpha particle spectrometry (+/- 5% RSD, 1 sigma). It is precise and rapid enough to allow analysis of a large number of samples in a short period of time at low cost using standard, commercially available quadrupole instrumentation with ultrasonic nebulizer and desolvator accessories. UARs have been analyzed successfully in fresh to moderately saline waters with U concentrations of from less than 1 microg/L to nearly 100 microg/L. An example of the uses of these data is shown for a study of surface-water mixing in the North Platte River in western Nebraska. This rapid and easy technique should encourage the wider use of uranium isotopes in surface-water and groundwater investigations, both for qualitative (e.g. identifying sources of water) and quantitative (e.g. determining end-member mixing ratios purposes.


A robust weighted Kaplan-Meier approach for data with dependent censoring using linear combinations of prognostic covariates.
The weighted Kaplan-Meier (WKM) estimator is often used to incorporate prognostic covariates into survival analysis to improve efficiency and correct for potential bias. In this paper, we generalize the WKM estimator to handle a situation with multiple prognostic covariates and potential-dependent censoring through the use of prognostic covariates. We propose to combine multiple prognostic covariates into two risk scores derived from two working proportional hazards models. One model is for the event times. The other model is for the censoring times. These two risk scores are then categorized to define the risk groups needed for the WKM estimator. A method of defining categories based on principal components is proposed. We show that the WKM estimator is robust to misspecification of either one of the two working models. In simulation studies, we show that the robust WKM approach can reduce bias due to dependent censoring and improve efficiency. We apply the robust WKM approach to a prostate cancer data set.


A singly bonded derivative of endohedral metallofullerene: La@C82CBr(COOC2H5).
A novel Bingel monoadduct of La@C82 (mono-A) has been synthesized by the reaction with diethyl bromomalonate in the presence of DBU (Bingel-Hirsch reaction). Its structure has been fully determined by NMR spectroscopic and X-ray crystallographic analyses. The most distinct feature of mono-A is the single bond moiety between the functional group and fullerene cage, which is very different from the cyclopropane moiety in a conventional Bingel adduct of empty fullerenes. Further spectroscopic characterizations and calculations revealed the closed-shell structure of mono-A. Its formation mechanism was discussed according to calculation results.


A time-domain Kirchhoff formula for the convective acoustic wave equation.
Kirchhoff's integral method allows propagated sound to be predicted, based on the pressure and its derivatives in time and space obtained on a data surface located in the linear flow region. Kirchhoff's formula for noise prediction from high-speed rotors and propellers suffers from the limitation of the observer located in uniform flow, thus requiring an extension to arbitrarily moving media. This paper presents a Kirchhoff formulation for moving surfaces in a uniform moving medium of arbitrary configuration. First, the convective wave equation is derived in a moving frame, based on the generalized functions theory. The Kirchhoff formula is then obtained for moving surfaces in the time domain. The formula has a similar form to the Kirchhoff formulation for moving surfaces of Farassat and Myers, with the presence of additional terms owing to the moving medium effect. The equation explicitly accounts for the influence of mean flow and angle of attack on the radiated noise. The formula is verified by analytical cases of a monopole source located in a moving medium.


Composition of Stallion Seminal Plasma and Its Impact on Oxidative Stress Markers and Spermatozoa Quality.
The composition of seminal plasma of individual sires varies and so does the fertilizing ability. Micro and macro elements along with seminal enzymes, hormones, proteins, and lipids contained in seminal plasma are essential for the proper physiological function of spermatozoa. However, elevated levels against the normal physiological values, especially in the case of trace metals, result in the production of reactive oxygen species. The deficiency of antioxidants in the seminal plasma that could scavenge free radicals causes an impairment of spermatozoa quality. Ejaculates were obtained from 19 stallions. The fresh semen was analyzed to evaluate qualitative parameters of spermatozoa in terms of the motility, viability, and integrity of DNA. Separated seminal plasma underwent the assessment of the chemical and biochemical composition and RedOx markers. Based on the obtained concentrations of individual chemical elements, the correlation analysis suggested a negative impact of Cu in seminal plasma on the SOD, GPx, and LPO. Contrary, positive correlation was detected between FRAP and motility features. While Cu negatively correlated with sperm motion parameters, the adverse effect on viability was suggested for Cd. Our data suggest that seminal plasma has a potential due to its availability to become the potential biomarker of the reproductive health of farm animals.


[Is necessary to perform a transthoracic echocardiogram in all the patients with cryptogenic stroke during hospitalization?].
From 15 to 30% of all ischemic strokes are cardioembolic. Transthoracic echocardiography plays a key role in the evaluation, diagnosis and management of the embolic source. The absence of official recommendations for the use of echocardiography in patients with ischemic stroke leads to a universal application showing low diagnostic efficiency. To analyze the diagnostic accuracy of echocardiograpm in patients with ischemic stroke in two situations: with universal indication and after the application of risk clinical criteria. Analysis of the echocardiograms performed on patients with acute ischemic stroke from the stroke unit during the years 2009-2011. We study the diagnostic and etiological contribution to the etiological study. Apply a selection criteria: 'high risk patient with need of test performance during admission' (age < 60 years, abnormal baseline electrocardiogram, cardiomegaly on chest radiograph or baseline history of heart disease, suspected endocarditis and/or active neoplasia) and analyzed their validity. From 930 inpatients, 201 (21.6%), underwent echocardiogram. Cardioembolic source was detected in 9.95%. After application of selection criteria, only 97 patients (10.4%) should have undergone it. The proposed criteria have a sensitivity 95%, specificity 56.9%, positive predictive value 19.6% and negative predictive value of 99%. The application of our criteria in undetermined stroke patients help us to identify with high efficiency cardioembolic sources postponing the test to an ambulatory scenario in the rest of the patients.


Selection of donors for living donor liver transplantation in a single center of a developing country: lessons learned from the first 100 cases.
The selection of donors for living donor liver transplantation (LDLT) is one of the most important features in this kind of surgery. The aim of this study is to describe our initial experience in the donor evaluation process. From December 2001 to January 2005, 104 donors were evaluated for 70 recipients (65 potential donors were evaluated for 39 adult recipients, and 39 donors for 31 pediatric recipients). Only 30 donors were able to donate: 13 for the adult group, and 17 for the pediatric one. In general, the utilization rate of potential donors was 28.8% (30/104). For the adult patients, 65 potential donors were seen to perform 13 LDLT, which represents a utilization rate of potential donors of 20%. For the pediatric patients, this rate was 43.6%. The exclusion criteria were clinical in 22 cases (21%), anatomical in 13 cases (13%), psychosocial in nine cases (9%), and others in 12 (12%). Death of recipients led to exclusion 18 of donors (17%). Thirty-three percent of adults and 55% of pediatric recipients who had at least one potential donor to start the evaluation process were able to identify a living donor. In conclusion, the first limit for LDLT is the rigorous donor evaluation.


Influence of Psychological Need-Based Teachers' Autonomy Support on Effectiveness and Engagement in English Learning.
It is to explore the effects of psychological needs and teachers' autonomy support on students' English learning, hoping to provide a realistic basis for the innovation of English teaching. Four hypotheses are proposed first, and 420 students are then randomly selected from seven classes in Xi'an Ai Zhi Zhong Xue in Shaanxi Province as the research subjects, with 420 questionnaires distributed. After invalid questionnaires are excluded, a total of 400 valid questionnaires are collected. Then, SPSS26.0 is employed to analyze the collected scale data. The average value of emotional exhaustion and the average value of low sense of achievement are relatively close in the direction of English learning burnout, which are larger than that in the other two directions. The three directions in English learning burnout are positively correlated with each other. The three directions in English learning engagement are positively correlated with those of psychological needs. Besides, there is an obvious positive correlation between the three directions of English learning engagement with teachers' autonomy support (p < 0.01). However, the three directions in teachers' autonomy support and the three directions in psychological needs are all negatively correlated with English learning burnout (p < 0.01). To sum up, the psychological needs of middle school students are effectively met, and teachers' autonomy support can obviously promote the effectiveness and engagement of middle school students' English learning.


Long non-coding RNA tumor protein 53 target gene 1 promotes cervical cancer development via regulating microRNA-33a-5p to target forkhead box K2.
Long non-coding RNA tumor protein 53 target gene 1 (TP53TG1) has been unraveled to exert regulatory effects on cancer progression, while the regulatory function of TP53TG1 on cervical cancer (CC) via regulating microRNA (miR)-33a-5p/Forkhead box K2 (FOXK2) axis remains rarely explored. This study aims to unearth the regulatory mechanism of TP53TG1/miR-33a-5p/FOXK2 axis in CC. The CC clinical samples were collected, and CC cells were cultured. TP53TG1, miR-33a-5p and FOXK2 levels were examined in CC tissues and cells. The CC cells were transfected with high- or low-expressed TP53TG1, FOXK2 or miR-33a-5p to determine the changes of CC cell biological activities and the status of phosphatidylinositol 3-kinase/protein kinase B/mammalian target of rapamycin (PI3K/AKT/mTOR) pathway. The tumorigenesis in nude mice was conducted. The relationship among TP53TG1, miR-33a-5p and FOXK2 was validated. TP53TG1 and FOXK2 expression levels were increased and miR-33a-5p expression level was reduced in CC cells and tissues. The silenced TP53TG1 or FOXK2, or elevated miR-33a-5p decelerated the CC cell development and restrained the activation of PI3K/AKT/mTOR signaling pathway. The depleted FOXK2 or elevated miR-33a-5p reversed the effects of decreased TP53TG1 on CC cell progression. TP53TG1 sponged miR-33a-5p, which targeted FOXK2. The experiment in vivo validated the outcomes of the experiment in vitro. TP53TG1 accelerates the CC development via regulating miR-33a-5p to target FOXK2 with the involvement of PI3K/AKT/mTOR signaling pathway. This study provides novel theory basis and distinct therapeutic targets for CC treatment.


The development potential of ethanol-induced monosomic and trisomic conceptuses in the mouse.
The development potential of fertilized embryos isolated from female mice previously given a single dose of either a dilute solution of ethanol or distilled water (controls) by mouth was studied. Exposure to ethanol occurred at various times during the cycle leading to ovulation and shortly after fertilization. The chromosome constitution of all preimplantation embryos isolated from these females was determined either at the first cleavage mitosis or at the morula stage. The incidence of aneuploidy in the ethanol-exposed groups at these times was approximately 19% and 13.5%, respectively, with a similar number of monosomic and trisomic conceptuses observed at these times. In addition, about 2% of all conceptuses examined were triploid. Further females were autopsied on the 10th or 11th day of gestation, though the chromosome constitution of only the morphologically abnormal or developmentally retarded embryos was determined. Eight embryos out of a total of 16 studied in the ethanol-exposed group were either aneuploid or triploid, whereas in the control group only one out of 11 examined proved to be aneuploid. The triploids and ethanol-induced aneuploid conceptuses appeared to be capable of surviving to the morula stage but generally failed to survive to the 10th/11th day. No monosomics were in fact observed in the postimplantation series. The present findings are briefly discussed with reference to the possible pathogenesis of spontaneous abortions in man, which often possess similar types of chromosomal anomalies.


The Effect of Increasing Volume of Exercise on Activation Pattern of Vastus Medialis and Lateralis and its Correlation With Anterior Knee Pain in Karate Elites.
The effects of exercise volume on the pattern of muscle activity is one of the most important factors in training management and injury risk reduction. In the lower limb, the quadriceps muscle which plays a determining role in performing the stance and other karate techniques could be injured in intensive exercise and may induce anterior knee pain in athletes. The aim of this study was to determine the relationship between training volume and muscle activity of vastus medialis and vastus lateralis and its association with anterior knee pain in karate elites. Male and female athletes from national junior and cadet karate team (14 to 18 years) were invited to participate in the study at the beginning and the end of the training camps. Studies involved measurement of electromyographic muscle activity of vastus medialis and vastus lateralis in both lower extremities with surface electromyography device and assessment of movement by electrogoniometery. Muscle activity was recorded in three tests of dachi, walking up and walking down stairs. Simultaneously, anterior knee pain was evaluated using visual analogue scale and anterior knee pain scale questionnaire. Eight athletes of a total number of 23 reported increased ratings of pain in their right knees. No differences in muscle activity were observed in tests of Dachi and stairs between the groups with and without pain. Comparing Dachi task pattern at the beginning and end of training camps, there was no significant difference in pattern of biomechanical movement; however, reducing the amount of muscle activity in early and late phases of tasks was observed in electromyographic assessment. The results showed that performing the same task after a six-week training period, less muscle activity was required in all phases in two groups of tasks, including karate-specific movement (dachi) and activities of daily living (up or down stairs).


An invertible element of DNA controls phase variation of type 1 fimbriae of Escherichia coli.
The expression of type 1 fimbriae (pili) of Escherichia coli is turned on and off at the transcriptional level at a high frequency (10(-3) per cell per generation) in a process termed phase variation. Using Southern blot and DNA sequence analysis, we have detected a genomic rearrangement in the switch region immediately upstream of the fimbrial structural gene. This rearrangement involves an invertible 314-base-pair segment of DNA whose alternating orientation apparently results in the on-and-off activation of a promoter that determines the state of fimbrial expression.


Clinical Study of Oral Mucosal Lesions in the Elderly-Prevalence and Distribution.
The aim of the study was to determine the prevalence, pattern, and distribution of oral mucosa lesions in elderly patients attending an outpatient oral pathology clinic. A retrospective analysis of 2747 patients' medical files was performed. Of these, 1398 (50.6%) belonged to seniors. The mean age was 69.8 ± 7.1, and women were in the majority. Among the entire group of patients, the three most common mucosal lesions were: oral lichenoid diseases (OLDs), lingual changes, and small nodules. In the female group, the most common diagnoses were: OLDs, lingual changes, and oral candidiasis; in the male population, the most common diagnoses were: lingual changes, small nodules, and OLDs. Those suffering from OLDs were the youngest patients, and those with focal oral mucosa erosions and ulcerations were the oldest patients. In the groups aged 60-70 and 71-80 years old, the most common diagnoses were OLDs, and in the oldest group (+80 years old), they were lingual lesions. Oral health has an obvious impact on the functional, psychological, and behavioral quality of life. There is a small number of publications concerning the prevalence and distribution of oral mucosa lesions in the senior population of European countries. Our study is filling in that void.


The Course of Orofacial Pain and Jaw Disability After Whiplash Trauma: A 2-year Prospective Study.
Prospective cohort study. The aim of this study was to evaluate the course of orofacial pain and jaw disability in relation to neck pain, neck disability, and psychosocial factors at the acute stage and the chronic stage after whiplash trauma. Many individuals report chronic pain in the orofacial region after whiplash trauma. The possible association between whiplash trauma and orofacial pain is debated. Prospective studies are therefore needed to evaluate the development of orofacial pain after whiplash trauma. Within 1 month following a whiplash trauma, 176 cases were examined and compared to 116 controls with questionnaires concerning neck and jaw pain and related disability, nonspecific physical symptoms and depression. At the 2-year follow-up, 119 cases (68%) and 104 controls (90%) were re-examined. Compared to controls, cases reported more jaw and neck pain, both at baseline and follow-up. A majority (68%) of cases with pain in the jaw region in the acute stage also reported jaw pain at the follow-up. The intensity of jaw and neck pain was correlated both at baseline and follow-up. Both neck pain and jaw pain were correlated to nonspecific physical symptoms and to depression. Orofacial pain and jaw disability related to neck pain are often present already at the acute stage after whiplash trauma and persist into the chronic stage for most individuals. Assessment following whiplash trauma should therefore include both the neck and the orofacial regions. More studies are needed to further evaluate risk factors for development of orofacial pain after whiplash trauma. 3.


Vitamin D deficiency is associated with inflammatory cytokine concentrations in patients with diabetic foot infection.
Vitamin D has been recognised as a potent immunomodulator and its deficiency is common in different population groups including patients with diabetic foot infection. Diabetic foot infection reflects the altered immune status of the host. As cytokine regulation plays a significant role in infection and wound-healing processes, the present study aimed to evaluate the association between vitamin D status and inflammatory cytokine profiles in patients with diabetic foot infection. The serum concentrations of vitamin D (25-hydroxyvitamin D), IL-1β, IL-6, TNF-α and interferon-γ (IFN-γ) were measured in 112 diabetic foot infection cases and 109 diabetic controls. Severe vitamin D deficiency (25-hydroxyvitamin D concentration < 25 nmol/l) was more common in cases than in controls (48.2 v. 20.5%). Although age, duration of diabetes, HbA1C (glycosylated Hb) concentration and BMI were similar, cases had significantly higher concentrations of IL-6 (P≤ 0.001), IL-1β (P≤ 0.02) and TNF-α (P≤ 0.006) than controls. A significant negative correlation was also observed between 25-hydroxyvitamin D concentration and circulating concentrations of IL-1β (r -0.323; P≤ 0.001) as well as IL-6 (r -0.154; P≤ 0.04), but not between 25-hydroxyvitamin D and TNF-α and IFN-γ concentrations. Furthermore, a significant difference in IL-1β (P≤ 0.007) and IL-6 (P≤ 0.02) concentrations was observed in patients with severe 25-hydroxyvitamin D deficiency compared with patients with 25-hydroxyvitamin D concentration ≥ 25 nmol/l, and this difference was remarkable for TNF-α. In conclusion, severe vitamin D deficiency is associated with elevated inflammatory cytokine concentrations in diabetic patients, particularly in those with foot infection. A 25-hydroxyvitamin D concentration value < 25 nmol/l is suggested as the 'cut-off' for such immunological alterations in patients with diabetes mellitus.


Immunogenicity of epikeratophakia tissue lenses containing living donor keratocytes.
The purpose of this study was to compare the survival of epikeratophakia tissue lenses prepared with cryolathing and lyophilization (frozen lenses) and without (fresh lenses) in donor-sensitized recipients with vascularized corneas. Fresh lenses placed in vascularized corneas of immune recipients were subjected to immune attack. Frozen lenses placed in vascularized corneas of immunized recipients did not elicit an immune reaction. Neither fresh nor frozen lenses elicited immune reactions in nonvascularized corneas of immune recipients or in nonvascularized, nonimmune recipients. These results indicate that although the fresh lenses are more antigenic than the lenses in which the cells have been killed by freezing and lyophilization, the fresh lenses prepared using the BKS-1000 technique containing living stromal keratocytes are not likely to stimulate allograft immune reactions in unsensitized patients with avascular graft sites.


Light-driven eco-evolutionary dynamics in a synthetic replicator system.
Darwinian evolution involves the inheritance and selection of variations in reproducing entities. Selection can be based on, among others, interactions with the environment. Conversely, the replicating entities can also affect their environment generating a reciprocal feedback on evolutionary dynamics. The onset of such eco-evolutionary dynamics marks a stepping stone in the transition from chemistry to biology. Yet the bottom-up creation of a molecular system that exhibits eco-evolutionary dynamics has remained elusive. Here we describe the onset of such dynamics in a minimal system containing two synthetic self-replicators. The replicators are capable of binding and activating a co-factor, enabling them to change the oxidation state of their environment through photoredox catalysis. The replicator distribution adapts to this change and, depending on light intensity, one or the other replicator becomes dominant. This study shows how behaviour analogous to eco-evolutionary dynamics-which until now has been restricted to biology-can be created using an artificial minimal replicator system.


A redox reversible endoplasmic reticulum-targeted fluorescent probe for revealing the redox status of living cells.
The endoplasmic reticulum (ER) is one of the most important organelles in cells and is involved in protein synthesis, folding and orderly transport. Redox balance is the key to its normal function. In this work, we designed and synthesized an endoplasmic reticulum-targeted fluorescent probe N-Se with selenomorpholine as the redox reversible detection moiety. N-Se could selectively respond to ClO- within only 8 s with a LOD of 28.8 nM. Furthermore, such a response is reversible in the regulation of GSH. Confocal fluorescence imaging confirmed the excellent endoplasmic reticulum targeting ability of N-Se. Thus, it could real-time monitor the dynamic changes of the redox status in the endoplasmic reticulum through the variation of the fluorescence intensity.


Comparison of LightCycler PCR and culture for detection of group B streptococci from vaginal swabs.
Group B streptococci (GBS) are an important cause of neonatal sepsis and meningitis. New rapid, sensitive and specific methods for detection of GBS in pregnant women are needed in order to provide timely treatment of neonates. The sensitivity, specificity and cost of a LightCycler PCR method was compared with selective culture for the detection of GBS from 400 vaginal swabs. In addition, two DNA extraction methods (simple boiling and automated DNA extraction by Roche MagNA Pure LC) were compared for a subgroup of 100 clinical samples. The sensitivity of the LightCycler PCR assay for the detection of GBS from vaginal swabs was significantly higher than that of culture. There were no culture-positive, LightCycler PCR-negative cases. The efficiencies of the two DNA extraction procedures were not significantly different. The detection of GBS from vaginal swabs by the molecular method (including simple boiling extraction) required the same hands-on time, but the procedure was completed in 1.5 h, compared with c. 48 h for the culture-based approach. Disadvantages of the molecular method are the increased costs (45%) and the absence of antibiogram data. The LightCycler PCR is a promising tool for sensitive, specific and rapid detection of GBS directly from clinical specimens of pregnant women.


Multifunctional yolk-shell nanoparticles: a potential MRI contrast and anticancer agent.
We report a new type of multifunctional nanomaterials, FePt@Fe2O3 yolk-shell nanoparticles, that exhibit high cytotoxicity originated from the FePt yolks and strong MR contrast enhancement resulting from the Fe2O3 shells. Encouraged by the recently observed high cytotoxicity of FePt@CoS2 yolk-shell nanoparticles, we used Fe2O3 to replace CoS2 as the shells to further explore the applications of the yolk-shell nanostructures. The ultralow IC50 value (238 +/- 9 ng of Pt/mL) of FePt@Fe2O3 yolk-shell nanoparticles likely originates from the fact that the slow oxidation and release of FePt yolks increases the cytotoxicity. Moreover, compared with two commercial magnetic resonance imaging (MRI) contrast agents, MION and Sinerem, the FePt@Fe2O3 yolk-shell nanoparticle showed stronger contrast enhancement according to their apparent transverse relaxivity values (r2* = 3.462 (microg/mL)(-1) s(-1)). The bifunctional FePt@Fe2O3 yolk-shell nanoparticles may serve both as an MRI contrast agent and as a potent anticancer drug. This work indicates that these unique yolk-shell nanoparticles may ultimately lead to new designs of multifunctional nanostructures for nanomedicine.


QSAR for anti-HIV activity of HEPT derivatives.
QSARs were derived for 103 analogues of 1-[(2-hydroxyethoxy)methyl]-6-(phenylthio)thymine (HEPT), a potent inhibitor of the HIV-1 reverse transcriptase (RT). The activity of these compounds was investigated by means of multiple linear regression (MLR) and artificial neural network (ANN) techniques. Considering the relevant descriptors obtained from the MLR, a correlation coefficient of 0.92 (n = 95) was obtained with a 4-5-1 ANN model. The contribution of each descriptor to the structure-activity relationships was evaluated. The results showed that the anti-HIV activity of HEPT derivatives was strongly dependent on hydrophobic character and also steric factors of substituents.


Genes Involved in Maintaining Mitochondrial Membrane Potential Upon Electron Transport Chain Disruption.
Mitochondria are biosynthetic, bioenergetic, and signaling organelles with a critical role in cellular physiology. Dysfunctional mitochondria are associated with aging and underlie the cause of a wide range of diseases, from neurodegeneration to cancer. Through signaling, mitochondria regulate diverse biological outcomes. The maintenance of the mitochondrial membrane potential, for instance, is essential for proliferation, the release of mitochondrial reactive oxygen species, and oxygen sensing. The loss of mitochondrial membrane potential triggers pathways to clear damaged mitochondria and often results in cell death. In this study, we conducted a genome-wide positive selection CRISPR screen using a combination of mitochondrial inhibitors to uncover genes involved in sustaining a mitochondrial membrane potential, and therefore avoid cell death when the electron transport chain is impaired. Our screen identified genes involved in mitochondrial protein translation and ATP synthesis as essential for the induction of cell death when cells lose their mitochondrial membrane potential. This report intends to provide potential targets for the treatment of diseases associated with mitochondrial dysfunction.


Droplet size prediction in ultrasonic nebulization for non-oxide ceramic powder synthesis.
Spray pyrolysis process has been used for the synthesis of non-oxide ceramic powders from liquid precursors in the Si/C/N system. Particles with a high thermal stability and with variable composition and size distribution have been obtained. In this process, the mechanisms involved in precursor decomposition and gas phase recombination of species are still unknown. The final aim of this work consists in improving the whole process comprehension by an experimental/modelling approach that helps to connect the synthesized particles characteristics to the precursor properties and process operating parameters. It includes the following steps: aerosol formation by a piezoelectric nebulizer, its transport and the chemical-physical phenomena involved in the reaction processes. This paper focuses on the aerosol characterization to understand the relationship between the liquid precursor properties and the liquid droplet diameter distribution. Liquids with properties close to the precursor of interest (hexamethyldisilazane) have been used. Experiments have been performed using a shadowgraphy technique to determine the drop size distribution of the aerosol. For all operating parameters of the nebulizer device and liquids used, bimodal droplet size distributions have been obtained. Correlations proposed in the literature for the droplet size prediction by ultrasonic nebulization were used and adapted to the specific nebulizer device used in this study, showing rather good agreement with experimental values.


Adsorption and degradation of thiazopyr in compost-amended and non-amended soils.
Adsorption and degradation of thiazopyr on two unamended soils and a soil amended annually during 8 years with compost were studied under laboratory conditions and compared with the results obtained on soils amended with fresh sewage sludge compost. The adsorption isotherms fitted the Freundlich equation well and a marked sorption increase was found in amended soils. Degradation data followed first-order kinetics and thiazopyr had a half-life of about 75 days at 25 degrees C and 60% water-holding capacity of soil. The addition of fresh compost markedly decreased the rate of thiazopyr degradation, whereas the compost mineralised in the field after annual additions had only a small influence. Incubation studies with sterile soils showed a very significant decrease of the degradation rate, indicating that degradation by micro-organisms was the main pathway of thiazopyr degradation in the soils studied.


Plasma levels of soluble programmed death ligand-1 may be associated with overall survival in nonsmall cell lung cancer patients receiving thoracic radiotherapy.
Immune-checkpoint signaling plays an important role in immunosuppression of tumors. We aimed to investigate the association of soluble programmed death-ligand 1 (sPD-L1) level in plasma with overall survival (OS) in locally advanced or inoperable nonsmall-cell lung cancer (NSCLC) patients treated with thoracic radiotherapy (TRT). We used ELISA to evaluate the sPD-L1 levels at diagnosis and during TRT in 126 clinically inoperable NSCLC patients. OS rates were followed up and recorded. SPSS software and GraphPad Prism 5 were used for statistics. In this study, the average sPD-L1 levels at baseline, week 2, and week 4 during TRT and post-TRT were 107.2, 51.3, 65.4, and 111.1 pg/mL, respectively. Levels of sPD-L1 at week 2 and week 4 were significantly less than at baseline, with both P values < 0.001. Using 96.5 pg/mL as the cutoff, patients with lower baseline sPD-L1 level had longer OS than those with higher sPD-L1 level (27.8 months vs 15.5 months, P = 0.005). Using multivariate analysis, the following factors were significantly associated with longer OS: female, adenocarcinoma, higher TRT dose, and lower baseline sPD-L1 level. Patients with both characteristics of lower baseline sPD-L1 level and higher TRT dose (BED10 ≥84 Gy) had the longest OS. To conclude, the lower baseline sPD-L1 level was significantly associated with longer OS in NSCLC patients treated with TRT, which may serve as an independent biomarker and needs further clinical study.


Development of an Experimental Drug Eluting Suprachoroidal Microstent as Glaucoma Drainage Device.
A novel glaucoma drainage device (GDD) with local drug delivery (LDD) system was created and characterized for safety and effectiveness after implantation into the suprachoroidal space (SCS) of rabbit eyes. Thin films of two different polymers, Poly(3-hydroxybutyrate) (P(3HB)) and Poly(4-hydroxybutyrate) (P(4HB)), containing the drugs mitomycin C (MitC) or paclitaxel (PTX) were attached to silicone-tubes to create LDD devices. The release kinetics of these drugs were explored in vitro using high performance liquid chromatography (HPLC). Twenty-four New Zealand white rabbits, randomly divided into eight groups, were implanted with different kinds of microstents into SCS. The intraocular pressure (IOP) was monitored noninvasively. After 6 weeks, rabbits were sacrificed and enucleated eyes were used for anterior segment optical coherence tomography (OCT), micro magnetic resonance imaging (MRI), and histology. In vitro, faster drug release from both polymers was observed for MitC compared to PTX. Comparing polymers, the release from P(3HB) matrix was slower for both drugs. MRI and OCT showed all implants maintained a proper location. An effective IOP reduction was observed for up to 6 weeks in eyes with microstents combined with a drug-releasing LDD system. Overall, the surrounding tissue revealed mild-to-moderate inflammation. No pronounced fibrosis was observed in any of the groups. However, both drugs caused damage to the neighboring retina. The suprachoroidal microstent reduced IOP with mild inflammation in rabbit eyes. To avoid negative effects on the retina, it is necessary to identify novel drugs with less cytotoxicity. Future studies are needed to explore the fibrotic process over the long-term. The presented data serve as a proof of principle study for the concept of a suprachoroidal drug eluting microstent. Future device improvements will be focused on the design of LDD systems and the use of specific anti-inflammatory or antifibrotic agents with less cytotoxicity compared to MitC or PTX. Long-term animal studies using a reliable glaucoma model will be a further step towards clinical application and improvement of surgical glaucoma therapy.


The development of a thermostable CiP (Coprinus cinereus peroxidase) through in silico design.
Protein thermostability is a crucial issue in the practical application of enzymes, such as inorganic synthesis and enzymatic polymerization of phenol derivatives. Much attention has been focused on the enhancement and numerous successes have been achieved through protein engineering methods. Despite fruitful results based on random mutagenesis, it was still necessary to develop a novel strategy that can reduce the time and effort involved in this process. In this study, a rapid and effective strategy is described for increasing the thermal stability of a protein. Instead of random mutagenesis, a rational strategy was adopted to theoretically stabilize the thermo labile residues of a protein using computational methods. Protein residues with high flexibility can be thermo labile due to their large range of movement. Here, residue B factor values were used to identify putatively thermo labile residues and the RosettaDesign program was applied to search for stable sequences. Coprinus cinereus (CiP) heme peroxidase was selected as a model protein for its importance in commercial applications, such as the polymerization of phenolic compounds. Eleven CiP residues with the highest B factor values were chosen as target mutation sites for thermostabilization, and then redesigned using RosettaDesign to identify sequences. Eight mutants based on the redesigns, were produced as functional enzymes and two of these (S323Y and E328D) showed increased thermal stability over the wild-type in addition to conserved catalytic activity. Thus, this strategy can be used as a rapid and effective in silico design tool for obtaining thermostable proteins.


Blast cell proliferative activity and sensitivity to GM-CSF in vitro are associated with early response to TAD-9 induction therapy in acute myeloid leukemia.
The current study was undertaken to determine the relevance of leukemic blast cell proliferative activity, cellular parameters of Ara-C metabolism and the in vitro sensitivity to GM-CSF in association with the clinical response to TAD-9 induction therapy in 66 patients with de novo acute myeloid leukemia (AML). Proliferative activity was assessed by 3H-thymidine (3H-TdR) incorporation and thymidine kinase (TK) activity, parameters of Ara-C metabolism comprised the activities of deoxycytidine kinase (DCK) and DNA polymerase alpha (poly alpha) as well as Ara-CTP concentrations and 3H-Ara-C uptake into DNA. GM-CSF sensitivity was determined by in vitro incubation of blasts for 48 h with or without GM-CSF (100 U/ml) followed by an additional 4 h concurrent exposure to GM-CSF and 3H-TdR (0.5 microCi/ml). The following results were obtained as expressed by median values and ranges: 3H-TdR incorporation: 1.07 pmol/10(5) cells (0.0-10.1), TK: 7.3 pmol/min/mg protein (1.3-56.0), DCK: 9.3 pmol/min/mg protein (0.77-47.1), poly alpha: 1.7 pmol/min/mg protein (0.00-28.9), Ara-CTP: 53.3 ng/10(7) cells (13.3-211.0), 3H-Ara-C uptake: 0.06 pmol/10(5) cells (0.0-0.57). 3H-Ara-C uptake was correlated with 3H-TdR incorporation (r = 0.74) and with the (S-phase dependent) activities of TK (r = 0.73) and poly alpha (r = 0.71, but not with DCK activity or intracellular Ara-CTP content. Blast cells of 37 from 55 analyzed patients were found to be sensitive to GM-CSF stimulation as defined by an increase in 3H-TdR incorporation > or = 1.5-fold over control values after the 48 h GM-CSF exposure. In vitro data were related with clinical response to TAD-9 induction therapy in 43 patients with newly diagnosed AML, taking the blast cell reduction at day 10 or 16 to < 5% or > or = 5% residual blasts as early parameter for adequate or inadequate response, respectively. While neither 3H-Ara-C uptake, nor intracellular Ara-CTP concentration, TK nor DCK activity were predictive for response, a high 3H-TdR incorporation and a high poly alpha activity were associated with adequate blast cell reduction. Median values of 3H-TdR incorporation were 2.26 pmol/10(5) cells for patients with adequate blast cell clearance and 0.80 pmol/10(5) cells for patients with inadequate blast cell clearance (P = 0.11), the respective values for poly alpha were 3.22 pmol/min/mg protein for responders and 1.1 pmol/min/mg protein for non-responders (P = 0.0085).(ABSTRACT TRUNCATED AT 400 WORDS)


Metformin effects on clinical features, endocrine and metabolic profiles, and insulin sensitivity in polycystic ovary syndrome: a randomized, double-blind, placebo-controlled 6-month trial, followed by open, long-term clinical evaluation.
In the last few years some studies assessed the effects of attenuation of hyperinsulinemia and insulin resistance, obtained by insulin sensitizing agents, in women with polycystic ovary syndrome (PCOS), suggesting potential scope for these drugs in treating the whole spectrum of reproductive, endocrine, and metabolic abnormalities found in such subjects. However, the results of these studies, mostly uncontrolled and short-term, are still inconclusive, and there is no long-term follow-up. In the present study, 23 PCOS subjects [mean (+/- SE) body mass index 30.0+/-1.1 kg/m2] were randomly assigned to double-blind treatment with metformin (500 mg tid) or placebo for 6 months, while maintaining their usual eating habits. Before and after treatment, menstrual history, endocrine and metabolic profiles, serum 17-hydroxyprogesterone response to GnRH-agonist testing, and insulin sensitivity measured by the glucose clamp technique were assessed. Eighteen of these women, as well as 14 additional PCOS patients, were subsequently given metformin in an open trial for 11.0+/-1.3 months (range 4-26), to assess long-term effects of treatment and baseline predictors of metformin efficacy on reproductive abnormalities. After metformin treatment, mean frequency of menstruation improved (P = 0.002), due to striking amelioration of menstrual abnormalities in about 50% of subjects. Women given metformin showed reduced plasma insulin (at fasting: P = 0.057; during the clamp studies: P<0.01) and increased insulin sensitivity (P<0.05). Concurrently, ovarian hyperandrogenism was attenuated, as indicated by significant reductions in serum free testosterone (P<0.05) and in the 17-hydroxyprogesterone response to GnRH-agonist testing (P<0.05). No changes were found in the placebo group. Only comparable minor changes in body mass index were found both in the metformin group and in the placebo group. In the open, long-term trial 17 women (54.8%) showed striking improvements of their menstrual abnormalities and were considered as responders. Logistic regression analysis of baseline characteristics in responders and nonresponders showed that plasma insulin, serum androstenedione, and menstrual history were independent predictors of the treatment's clinical efficacy. In 10 subjects whose menses proved regular after treatment, the great majority of cycles became ovulatory (32 out of 39 assessed, 79%). In conclusion, in women with PCOS metformin treatment reduced hyperinsulinemia and hyperandrogenemia, independently of changes in body weight. In a large number of subjects these changes were associated with striking, sustained improvements in menstrual abnormalities and resumption of ovulation. Higher plasma insulin, lower serum androstenedione, and less severe menstrual abnormalities are baseline predictors of clinical response to metformin.


CXCL12 rs266085 and TNF-α rs1799724 polymorphisms and susceptibility to cervical cancer in a Chinese population.
Further research is required to identify single nucleotide polymorphisms (SNPs) associated with cervical cancer. The aim of this study was to assess the association of TNF-α/rs1799724 and CXCL12/rs266085 polymorphisms with susceptibility to cervical cancer in Han Chinese population in Shandong Province. 348 patients with cervical squamous cell carcinoma, including CIS (121) and invasive carcinoma (227), and 351 healthy controls. Genomic DNA was isolated from peripheral blood and genotyping for TNF-α/rs1799724 and CXCL12/rs266085 was carried out using TaqMan SNP Genotyping Assays. TNF-α/rs1799724 polymorphism showed the C-allele was less prevalent among cases as compared to controls (74.3% vs. 92.0%), while the T-allele was more prevalent among cases (P=0.000, OR=3.99, 95% C.I.: 2.90-5.51). CXCL12/rs266085 polymorphism showed the C-allele was less prevalent among cases as compared to controls (41.2% vs. 49.7%), while the T-allele was more prevalent among cases (P=0.001, OR=1.41, 95% C.I.: 1.14-1.74). The genotype and allele frequencies of these two SNPs did not differ between CIS and invasive squamous cell carcinoma (P>0.05). Moreover, the allele frequencies of rs1799724 were significantly different between controls without or with HPV infection (P<0.05). Neither the genotype nor allele frequencies of rs266085 were statistically different between HPV-negative and positive controls. TNF-α/rs1799724 and CXCL12/rs266085 polymorphisms are associated with cervical cancer. C->T polymorphism of these two SNPs and HPV infection are linked to high risk for cervical cancer.


Psychoneuroendocrinology of stress and emotions: issues for future research.
It is stated that much psychoneuroendocrinological research is carried out that lacks a theoretical framework and therefore adds little to our current knowledge. Therefore, five research issues have been presented which need further exploration: (1) the temporal relation between subjective and biological stress responses; (2) the question of the psychobiological specificity; (3) the relationship between endocrine responses to acute and to chronic stressors; (4) the relationship between endocrine activity and susceptibility to disease, and (5) the mediating and moderating effects of personal factors (coping, personality, and task performance) on psychobiological responses. It is argued that the study of these issues will increase our understanding of the stress phenomenon.


TEMPO-Mediated Cross-Dehydrogenative Coupling of Indoles and Imidazo[1,2-a]pyridines with Fluorinated Alcohols.
A simple and highly efficient metal-free method has been developed for hydroxyfluoroalkylation of indoles and imidazo[1,2-a]pyridines via TEMPO-mediated C(sp3)-H and C(sp2)-H bond cross-dehydrogenative coupling of fluorinated alcohols and indoles. The protocol showed broad substrate scope, afforded good yields of hydroxyfluoroalkylated products, and was amenable for scale-up. Mechanistic investigation indicated involvement of the radical pathway.


Accurate control of hyperbolic trajectories in any dimension.
The unsteady (nonautonomous) analog of a hyperbolic fixed point is a hyperbolic trajectory, whose importance is underscored by its attached stable and unstable manifolds, which have relevance in fluid flow barriers, chaotic basin boundaries, and the long-term behavior of the system. We develop a method for obtaining the unsteady control velocity which forces a hyperbolic trajectory to follow a user-prescribed variation with time. Our method is applicable in any dimension, and accuracy to any order is achievable. We demonstrate and validate our method by (1) controlling the fixed point at the origin of the Lorenz system, for example, obtaining a user-defined nonautonomous attractor, and (2) the saddle points in a droplet flow, using localized control which generates global transport.


Effect of high-pressure homogenization on different matrices of food supplements.
There is a growing demand for food supplements containing high amounts of vitamins, phenolic compounds and mineral content that provide health benefits. Those functional compounds have different solubility properties, and the maintenance of their compounds and the guarantee of their homogenic properties need the application of novel technologies. The quality of different drinkable functional foods after thermal processing (0.1 MPa) or high-pressure homogenization under two different conditions (80 MPa, 33 ℃ and 120 MPa, 43 ℃) was studied. Physicochemical characteristics and sensory qualities were evaluated throughout the six months of accelerated storage at 40 ℃ and 75% relative humidity (RH). Aroma and color were better maintained in high-pressure homogenization-treated samples than the thermally treated ones, which contributed significantly to extending their shelf life. The small particle size obtained after high-pressure homogenization treatments caused differences in turbidity and viscosity with respect to heat-treated samples. The use of high-pressure homogenization, more specifically, 120 MPa, provided active ingredient homogeneity to ensure uniform content in functional food supplements. Although the effect of high-pressure homogenization can be affected by the food matrix, high-pressure homogenization can be implemented as an alternative to conventional heat treatments in a commercial setting within the functional food supplement or pharmaceutical industry.


In vitro simulated semi-dynamic gastrointestinal digestion: evaluation of the effects of processing on whey proteins digestibility and allergenicity.
The effect of thermal processing on digestibility of milk proteins should be better understood as this can greatly affect their immunoreactivity. The aim of this study was to evaluate the effects of thermal processing and lactosylation on digestibility and allergenicity, by comparing non heat-treated with industrially processed whey proteins. A semi-dynamic model was used to mimic the kinetics of digestion, and ELISA inhibition tests against human specific serum IgE were performed on the mass-spectrometry characterized products. A quicker gastric digestion of the industrially treated sample produced a lower immunogenic response in comparison with the raw sample, where intact conformational epitopes remained. In later digests, greater IgE reactivity was shown in the heat treated product, probably due to the release of linear epitopes, while at intestinal level the immunogenic response was negligible. Moreover, transepithelial transport of a reported β-lactoglobulin-derived allergen, KIDALNENVLVL, produced during digestion was assayed. It was found that the epitope-belonging peptide could be transported through the cell monolayer, both in the native and mono-lactosylated forms, with a favored passage of the native peptide.


Meteorological factors-based spatio-temporal mapping and predicting malaria in central China.
Despite significant reductions in the overall burden of malaria in the 20th century, this disease still represents a significant public health problem in China, especially in central areas. Understanding the spatio-temporal distribution of malaria is essential in the planning and implementing of effective control measures. In this study, normalized meteorological factors were incorporated in spatio-temporal models. Seven models were established in WinBUGS software by using Bayesian hierarchical models and Markov Chain Monte Carlo methods. M₁, M₂, and M₃ modeled separate meteorological factors, and M₃, which modeled rainfall performed better than M₁ and M₂, which modeled average temperature and relative humidity, respectively. M₇ was the best fitting models on the basis of based on deviance information criterion and predicting errors. The results showed that the way rainfall influencing malaria incidence was different from other factors, which could be interpreted as rainfall having a greater influence than other factors.


CdSe/ZnS core/shell quantum dot sensitization of low index TiO(2) single crystal surfaces.
Quantum dots (QDs) are actively explored as alternative sensitizers to inorganic complexes in sensitized solar cells (SSC) due to their interesting physical, optical, and electronic properties. It is thought that the inorganic nature of QDs should provide enhanced stability over the entirely organic or inorganic complex dyes, yet the long-term stability of laboratory QD-SSC devices has not been investigated in detail. A general approach to synthesize high stability QDs involves coating the core material with a wide band gap inorganic shell material (type-I CS QD). However, the electronic structure of the resulting core/shell (CS) structure has potential barriers for both electron and hole transfer, suggesting inefficient charge carrier separation for type-I CS QDs. Herein we demonstrate that type-I CdSe/ZnS CS QDs can effectively sensitize single crystal TiO(2) electrodes and continue to operate in a regenerative mode in an aerated iodide electrolyte for more than 20 h. Core CdSe QDs degrade rapidly in the same electrolyte presumably due to CdI(2) formation. The possibility of exploring new core/shell nanomaterials in a variety of electrolyte/mediator combinations may result in more efficient and stable QD-SSCs.


Experimental verification of dynamic soaring in albatrosses.
Dynamic soaring is a small-scale flight manoeuvre which is the basis for the extreme flight performance of albatrosses and other large seabirds to travel huge distances in sustained non-flapping flight. As experimental data with sufficient resolution of these small-scale movements are not available, knowledge is lacking about dynamic soaring and the physical mechanism of the energy gain of the bird from the wind. With new in-house developments of GPS logging units for recording raw phase observations and of a dedicated mathematical method for postprocessing these measurements, it was possible to determine the small-scale flight manoeuvre with the required high precision. Experimental results from tracking 16 wandering albatrosses (Diomedea exulans) in the southern Indian Ocean show the characteristic pattern of dynamic soaring. This pattern consists of four flight phases comprising a windward climb, an upper curve, a leeward descent and a lower curve, which are continually repeated. It is shown that the primary energy gain from the shear wind is attained in the upper curve where the bird changes the flight direction from windward to leeward. As a result, the upper curve is the characteristic flight phase of dynamic soaring for achieving the energy gain necessary for sustained non-flapping flight.


Transcription factor motif activity as a biomarker of muscle aging.
In prior work, we analyzed gene expression profiles of mouse, rat and human gastrocnemius muscles to identify conserved regulators of muscle aging processes. By further comparing data obtained from different muscles we found stronger conservation of aging-related factors at the level of molecular pathways than at the level of individual genes. Here we compared the predictive power of models based on gene expression levels and those based on transcription factor motif activities for an individual's age. Although somewhat less accurate than models based on gene expression, models based on motif activities achieve good prediction of muscle age, further providing insights into aging-related molecular pathways.


The emergence of zoonotic Fusarium oxysporum infection in captive-reared fingerlings of golden mahseer, Tor putitora (Hamilton, 1822) from the central Himalayan region of India.
Zoonotic Fusarium oxysporum infection was identified in captive-reared fingerlings of golden mahseer, Tor putitora (Hamilton, 1822) from the central Himalayan regions, India. Initially, fingerlings of T. putitora (mean length 10.8 ± 0.002 and weight 18.58 ± 0.054 g) were observed with cottony mass like growth completely covering the dorsal and caudal fins. The infected fingerlings were showing clinical signs such as sluggish, erratic movement, gasping, flared operculum and settling at one corner of the rearing tanks. The microscopic observation of 8-day old culture of cottony mass like growth showed the presence of septate macroconidia, randomly spread microconidia and chlamydospores in short-chain. From sequence analysis of ITS amplified fragment, the isolate was identified as Fusarium oxysporum, TPFCF 214 (MH464266.1) and clustered with F. oxysporum, strain NRRL 43504 (EF453107.1) and F. oxysporum, strain 20736 (JX 270150.1) isolated from the human in phylogenetic tree. An experimental infection of healthy golden mahseer fingerlings with 20 µl of F. oxysporum spore suspension (2.5 × 109 spore ml-1 ) showed the development of lesion 6-dpi at the site of injection. Experimental trial on EPC-2 cell culture recorded detachment in the monolayer, clumping and shrinking of the cell line 6-8 dpi with a spore suspension of F. oxysporum, TPFCF 214 (5.68 × 102 cell/ml). From the severity of its infection, there is a chance that F. oxysporum may emerge as pathogenically and pose a significant health risk on captive-reared golden mahseer in other Asian countries and world. As Fusarium solani and F. oxysporum are known to cause invasive fusariosis in human especially in immunocompromised patients, localized infection in immunocompetent individuals as well as osteomyelitis, arthritis, otitis, sinusitis and brain abscess, the global fish farmers, handlers and aquaculturist need to be aware of possible health hazards caused by Fusarium spp. and should adopt proper fish health management and animal husbandry practice to control the infection of Fusarium in culture environment.


Mast cell growth factor (c-kit ligand) enhances cytokine stimulation of proliferation of the human factor-dependent cell line, M07e.
Murine mast cell growth factor (muMGF), a c-kit ligand, has additive to greater-than-additive effects on in vitro colony formation of murine and human myeloid progenitor cells stimulated with erythropoietin, granulocyte-macrophage colony-stimulating factor (GM-CSF), and/or interleukin (IL)-3. To confirm direct-acting effects on responding cells, MGF was assessed alone and in combination with other cytokines for effects on the proliferation of the human factor-dependent cell line, M07e. Proliferation was assayed in liquid culture by [3H]thymidine uptake and in semisolid medium by colony formation. Purified recombinant (r) muMGF (25-50 ng/ml) by itself had proliferative activity but less than r human (hu) GM-CSF. In combination with rhuGM-CSF (250 U/ml) or IL-3 (500 U/ml), rmuMGF (25 ng/ml) enhanced [3H]thymidine uptake two- to sevenfold greater than the sum of the effects of each factor alone. Similar enhancement was seen in the number and size of colonies formed. When MGF was used in combination with rhuIL-4 (500-1000 U/ml), rhuIL-6 (5 ng/ml), rhuIL-9 (5-10 U/ml), or rhu interferon gamma (IFN-gamma; 250-500 U/ml) (factors that alone stimulate little proliferation), [3H]thymidine uptake and colony formation were respectively increased 2- to 11- and 3- to 55-fold over the sum of each of the effects of the factors alone. Exposure of 5 x 10(5) cells/ml to 50 ng/ml MGF for 24 h, a time during which synergism is noted with MGF plus either GM-CSF or IL-3, did not change GM-CSF or IL-3 receptor binding affinity or the number of binding sites. Exposure of cells to MGF for 48 h did not alter subsequent GM-CSF- or IL-3-stimulated proliferation. The results suggest that M07e cells will be useful as a model for the analysis of intracellular biochemical mechanisms of the direct-acting proliferative and synergistic effects of MGF.


Expression of vascular endothelial growth factor (VEGF) and its soluble receptor-1 in endometriosis.
The aim of this study is to evaluate the expression of vascular endothelial growth factor (VEGF) and its soluble receptor (sFlt-1) in peritoneal fluid (PF), peritoneal endometriotic lesions and eutopic endometrial tissues of patients with endometriosis. Peritoneal fluid, peritoneal endometriotic lesions and eutopic endometrial samples from patients with endometriosis, and peritoneal fluid, peritoneal tissue and endometrial samples from patients without endometriosis were obtained during an operative laparoscopy. The mean PF concentrations of VEGF and sFlt-1 were significantly higher in patients with endometriosis than in the controls. In the peritoneal tissue, the expressions of VEGF and sFlt-1 were significantly higher, where the expression of sFlt-1 in endometrium was significantly lower in patients with endometriosis. These findings indicate that not only abnormal expressions of angiogenic factors, but also aberrant expressions of antiangiogenic factors in the peritoneal and endometrial environment seem to be involved in the pathogenesis of endometriosis.


Unilateral intrauterine insemination with cryopreserved caudal epididymal sperm recovered from refrigerated canine epididymides.
Canine epididymides were excised and immediately stored at 4 degrees C for 48 hr, and the qualities of caudal epididymal sperm after recovery and cryopreservation were evaluated. To confirm the fertility of the cryopreserved caudal epididymal sperm, artificial intrauterine insemination was performed. The sperm motility (61.0%) immediately after recovery from caudal epididymis stored at 4 degrees C for 48 hr was significantly lower than those of sperm stored for 0 and 24 hr (88.6 and 80.7%, respectively), but there was no significant difference after freeze-thawing (0-, 24-, and 48-hr storage groups: 27.9, 24.3, and 28.3%, respectively). The incidence of abnormal sperm immediately after recovery was significantly higher in the 24-hr and 48-hr storage groups (19.3 and 27.7%, respectively) than in the 0-hr storage group (5.6%), and a significant difference was also observed after freeze-thawing. The incidence of immature sperm with cytoplasmic droplets was significantly higher in the 48-hr storage group (18.4%) than in the 0-hr storage group (4.7%), but there was no difference after freeze-thawing. By unilateral intrauterine insemination (2x10(8) sperm), 4 of 5 bitches (80%) conceived. The above findings demonstrated that sperm motility was good even enough the incidence of abnormal sperm was high in canine epididymal sperm that were recovered from the epididymis stored at 4 degrees C for 48 hr and cryopreseved, and that artificial intrauterine insemination resulted in a high conception rate.


Occupational therapists and patients' rights: their level of Clinical knowledge.
Addressing patients' rights issues brings occupational therapists ethical and political responsibilities that involve patients' privileges and new facilitating factors which influence their needs. The goal of this study was to determine the level of occupational therapists' knowledge about patients' rights. The present research was a cross-sectional study which involved 125 occupational therapists chosen by a convenience sampling strategy in Tehran during the year of 2012. A four-part questionnaire was used for data collection, and the degree of the subjects' self-assessment of their knowledge was measured based on the obtained numbers of correct answers in the third part. The validity and reliability of this questionnaire were assessed prior to its being distributed among participants. The results demonstrated no significant association between the level of occupational therapists' knowledge about patients' rights and their existing experiences within their areas of occupational therapy (P>0.05). Based on the result, 53.6% of the respondents had high level of knowledge about patients' rights. Facilitating factors which influence the attainment of patients' rights were classified into three groups: organizations, therapists and clients. The results of the present research demonstrated that the level of occupational therapists' knowledge about patients' rights were high. Furthermore, this study showed that for optimal result, there is a need to provide milieu for observing the patients' rights in clinical occupational therapy services.


YouTube™ as a Source of Information for Patients Regarding Dental Implant Failure: A Content Analysis.
The aim of this study is to investigate the quality of the information YouTube TM offers to patients concerning dental implant failure. YouTube TM was searched systematically using the keyword 'dental implant failure'. The first 100 videos were viewed by two independent researchers. For each video, its purpose, target audience and source were also recorded. A 10 point content scale (CS) was used to evaluate the video content. The Global Quality Scale (GQS) was also used to determine the quality of videos. Statistical analyses were performed using the Kruskal Wallis and Mann Whitney test and correlation coefficient analyses were performed using Spearman's Rho. While 92.2% of the videos included in the study were uploaded by dentists/health institutions, only 3.1% were uploaded by laypersons. Of the videos, 40.6% were aimed at informing laypersons and 56.3% targetted professionals. The content which was discussed most (71.9%) was 'definition of a dental implant' followed by 'reasons for failure' (65.6%). The mean CS and GQS score of the videos were 3.75 ± 2.35 (moderate) and 2.07 ± 1.05 (generally poor), respectively. There was a positive correlation between the CS and GQS score (r = 0.620). The quality of information on YouTube TM regarding dental implant failure was found inadequate. Thus, the information currently available online needs to be constantly checked and improved by professionals. In addition, clinicians should improve the ways in which they use YouTube TM to better inform patients about the causes, risks and treatment choices involved with failed dental implants.


The effect of aging on constitutive mRNA levels and lipopolysaccharide inducibility of acute phase genes.
Eukaryotic organisms possess natural defense processes associated with their response to injury, inflammation and pollutants. One of these, the acute phase (AP) host response, is characterized by a series of hepatic physiological reactions triggered by factors released as a result of bacterial infection, inflammation or tissue injury and is believed to be the mechanism by which cells and tissues are protected against further damage and injury. The capacity to respond to these physiological insults is known to be affected by aging. We propose that the AP response represents a series of intrinsic processes and interactions that may be affected by aging. Furthermore, we propose that this may be due to the progressive failure of the acute phase response. In this study we examine the relationship between aging and the expression of both positive and negative acute phase reactants, i.e., acute phase serum proteins whose levels are increased or decreased in response to systemic injury and infection. The mRNA levels of the positive acute phase reactants, alpha 1-acid glycoprotein (AGP), alpha 1-antitrypsin (AT), and the negative acute phase reactant, albumin were measured in both normal and inflammation-induced mice of ages 2, 7, 12, and 24 months. A significant decrease in the constitutive levels of AT and albumin mRNAs occurred as a function of increased age. Furthermore, aging decreased the ability of the AGP and albumin genes to respond to inflammation. Our studies indicate that aging may affect the transcription of these genes, processing of their mRNA or stability of the mRNA levels.


[A Research on Drug Discovery for Intracerebral Hemorrhage Focusing on Leukotriene B4 and Its Receptor].
Intracerebral hemorrhage (ICH) results from blood vessels rupture in the brain, forming a blood clot in the brain parenchyma. Leakage of blood constituents causes detrimental tissue damages, ensuing long-lasting neurological deficits; however, effective therapeutic approaches are not yet developed to date. In this study, leukotriene B4 (LTB4) and its receptor leukotriene B4 receptor 1 (BLT1) are proposed as novel therapeutic targets for ICH therapy. After the onset of ICH, the LTB4 content in the brain transiently elevated. Microglia are considered as the source of LTB4 production. Thrombin, a blood constituent, activated the BV-2 microglia and increased the LTB4 secretion from the BV-2 cells. Microglia-released LTB4 promoted its own microglial activation and neutrophil-like differentiated HL-60 cell migration activity. LTB4 receptors comprised of two types: BLT1 and BLT2, with BLT1 known to be a high-affinity receptor associated with chemotaxis. BLT1 knockout mice showed decreased neutrophil invasion, attenuating sensorimotor dysfunction after ICH. Furthermore, therapeutic administration of ONO-4057, an orally active LTB4 receptor antagonist, attenuated neutrophil invasion, microglial activation, axonal fragmentation, and sensorimotor deficits induced by ICH. These results suggest that LTB4 and its receptor BLT1 can be potential promising therapeutic targets that prevent tissue damages following ICH.


Efficacy and Limitations of Chemically Diverse Small-Molecule Enzyme-Inhibitors against the Synergistic Coagulotoxic Activities of Bitis Viper Venoms.
Snakebite remains a significant public health burden globally, disproportionately affecting low-income and impoverished regions of the world. Recently, researchers have begun to focus on the use of small-molecule inhibitors as potential candidates for the neutralisation of key snake venom toxins and as potential field therapies. Bitis vipers represent some of the most medically important as well as frequently encountered snake species in Africa, with a number of species possessing anticoagulant phospholipase A2 (PLA2) toxins that prevent the prothrombinase complex from inducing clot formation. Additionally, species within the genus are known to exert pseudo-procoagulant activity, whereby kallikrein enzymatic toxins cleave fibrinogen to form a weak fibrin clot that rapidly degrades, thereby depleting fibrinogen levels and contributing to the net anticoagulant state. Utilising well-validated coagulation assays measuring time until clot formation, this study addresses the in vitro efficacy of three small molecule enzyme inhibitors (marimastat, prinomastat and varespladib) in neutralising these aforementioned activities. The PLA2 inhibitor varespladib showed the greatest efficacy for the neutralisation of PLA2-driven anticoagulant venom activity, with the metalloproteinase inhibitors prinomastat and marimastat both showing low and highly variable degrees of cross-neutralisation with PLA2 anticoagulant toxicity. However, none of the inhibitors showed efficacy in neutralising the pseudo-procoagulant venom activity exerted by the venom of B. caudalis. Our results highlight the complex nature of snake venoms, for which single-compound treatments will not be universally effective, but combinations might prove highly effective. Despite the limitations of these inhibitors with regards to in vitro kallikrein enzyme pseudo-procoagulant venom activity, our results further support the growing body of literature indicating the potential use of small molecule inhibitors to enhance first-aid treatment of snakebite envenoming, particularly in cases where hospital and thus antivenom treatment is either unavailable or far away.


In vitro inhibition of MAP kinase (ERK1/ERK2) activity by phosphorylated glia maturation factor (GMF).
We report that recombinant glia maturation factor (GMF), a 17-kDa brain protein, inhibits the activity of mitogen-activated protein (MAP) kinase in the test tube assay, in particular the ERK1/ERK2 isoforms. A preliminary phosphorylation of GMF by protein kinase A (PKA) dramatically increases its inhibitory effect by over 600-fold (Ki approximately 3 nM), making it the most potent MAP kinase inhibitor ever reported. Immunoprecipitation of GMF from cell extracts using its specific antibody coprecipitates ERK (and vice versa), suggesting the association of the two proteins in the cell. The inhibitory effect of PKA-phosphorylated GMF is specific, as it does not suppress the activity of cdc2 kinase, another proline-directed kinase. Nor does it inhibit MAP kinase kinase (MEK) and MAP kinase-activated protein (MAPKAP) kinase-2, the two enzymes immediately upstream and downstream, respectively, of ERK. Of the other three enzymes that can phosphorylate GMF, only p90 ribosomal S6 kinase (RSK) enhances the inhibitory function of GMF on ERK; protein kinase C (PKC) and casein kinase II (CKII) are without effect. The inhibition of ERK by PKA-phosphorylated GMF suggests that GMF could be one of the mediators of the suppressive effect of the PKA pathway on the MAP kinase pathway. On the other hand, that RSK-phosphorylated GMF also inhibits ERK implies a negative feedback loop in the regulation of MAP kinase activity.


Preparation and Thermal Properties of Polycarbonates/esters Catalyzed by Using Dinuclear Salph-Al from Ring-Opening Polymerization of Epoxide Monomers.
A dinuclear Salph-Al complex/bis(triphenylphosphine)iminium chloride catalyst system was synthesized and employed for cyclohexene oxide (CHO) and CO2 copolymerization. The catalyst system had an excellent selectivity of 99 % for carbonate linkages and the resultant poly(cyclohexene carbonate) (PCHC) had a high glass transition temperature (Tg ) of 123.8 °C and a thermal decomposition temperature (5 % weight loss; Td 5 % ) of 265 °C. Furthermore, this catalyst system was active in the polymerization of phthalic anhydride (PA) and epoxides. Poly(CHO-alt-PA) was completely alternating, and had improved thermal properties (Tg =142.7 and Td 5 % =295 °C) compared with PCHC. The Tg values of the polyesters could be adjusted by addition of PO to the CHO/PA reaction system. For the CHO/PO/PA terpolymerization, CHO and PO participated concurrently and proportionally in the chain growth and the obtained terpolyesters had tunable Tg values from 62.8 to 142.7 °C depending on the CHO/PO feed ratio.


Dihydroartemisinin increases temozolomide efficacy in glioma cells by inducing autophagy.
Artemisinin, a powerful antimalarial medicine, is extracted from the Chinese herb, Artemisia annua L., and has the ability to inhibit the proliferation of cancer cells. Dihydroartemisinin (DHA), the major active metabolite of artemisinin, is able to inhibit the growth of a variety of types of human cancer. However, the effect of DHA on human glioma cells remains unclear. The aim of the present study was to investigate the effect of DHA on the proliferation of glioma cells, and whether DHA was able to enhance temozolomide (TMZ) sensitivity in vitro and in vivo. In total, 10 human glioma cell lines were used to analyze the growth inhibition ability of DHA by MTT assay. The typical autophagic vacuoles were monitored by the application of the autofluorescent agent, monodansylcadaverine. Western blotting was used to detect markers of apoptosis and autophagy, namely Caspase-3, Beclin-1 and LC3-B. The combination efficiency of DHA and TMZ was assessed in vitro and in vivo. The half maximal inhibitory concentration (IC50) of DHA differed among the ten human glioma cell lines. The number of autophagic vacuoles was higher in DHA-treated SKMG-4 cells; this was highest of all cell lines analyzed. The expression of autophagy molecular markers, Beclin-1 and LC3-B, was increased following DHA treatment, while no significant alteration was detected in the expression of apoptotic marker Caspase-3. When combined with DHA, the IC50 of TMZ decreased significantly in the four glioma cell lines analyzed. Furthermore, DHA enhanced the tumor inhibition ability of TMZ in tumor-burdened mice. The results of the present study demonstrated that DHA inhibited the proliferation of glioma cells and enhanced the tumor inhibition efficacy of TMZ in vitro and in vivo through the induction of autophagy.


Transforming growth factor beta 1 down-regulates vascular endothelial growth factor receptor 2/flk-1 expression in vascular endothelial cells.
Although the importance of the vascular endothelial growth factor (VEGF)/VEGF tyrosine kinase receptor (VEGFR) system in angiogenesis is well established, very little is known about the regulation of VEGFR expression in vascular endothelial cells. We have cloned partial cDNAs encoding bovine VEGFR-1 (flt) and -2 (flk-1) and used them to study VEGFR expression by bovine microvascular- and large vessel-derived endothelial cells. Both cell lines express flk-1, but not flt. Transforming growth factor beta 1 (TGF-beta 1) reduced the high affinity 125I-VEGF binding capacity of both cell types in a dose-dependent manner, with a 2.0-2.7-fold decrease at 1-10 ng/ml. Cross-linking experiments revealed a decrease in 125I-VEGF binding to a cell surface monomeric protein corresponding to Flk-1 on the basis of its affinity for VEGF, molecular mass (185-190 kDa), and apparent internalization after VEGF binding. Immunoprecipitation and Western blot experiments demonstrated a decrease in Flk-1 protein expression, and TGF-beta 1 reduced flk-1 mRNA levels in a dose-dependent manner. These results imply that TGF-beta 1 is a major regulator of the VEGF/Flk-1 signal transduction pathway in endothelial cells.


Nonalcoholic Fatty Liver Disease: Review of Management for Primary Care Providers.
Nonalcoholic fatty liver disease (NAFLD) is one of the most common causes of chronic liver disease in the United States and worldwide. The progressive form of NAFLD, nonalcoholic steatohepatitis (NASH), is a leading indication for liver transplant. Comorbidities associated with NAFLD development and NASH include type 2 diabetes, obesity, metabolic syndrome, and dyslipidemia. Extrahepatic morbidity and mortality are considerable as NAFLD is associated with an increased risk of cardiovascular disease and chronic kidney disease. Once NAFLD is diagnosed, the presence of liver fibrosis is the central determinant of hepatic prognosis. Severe liver fibrosis requires aggressive clinical management. No pharmacologic agents have regulatory approval in the United States for the treatment of NAFLD or NASH. Management is centered on efforts to reduce underlying obesity (lifestyle, medications, surgical or endoscopic interventions) and metabolic derangements (prediabetes, type 2 diabetes, hypertension, hyperlipidemia, and others). Current pharmacologic therapy for NAFLD is limited mainly to the use of vitamin E and pioglitazone, although other agents are being investigated in clinical trials. Cardiovascular and metabolic risk factors must also be assessed and managed. Here, NAFLD evaluation, diagnosis, and management are considered in the primary care setting and endocrinology clinics.