One step purification of flavanone glycosides from Poncirus trifoliata by centrifugal partition chromatography.
Flavanone glycosides were successfully separated from the crude extract of Poncirus trifoliata by preparative centrifugal partition chromatography with a two-phase solvent system composed of ethyl acetate-acetonitrile-water (3:2:5, v/v/v). Naringin (50.0 mg), neoponcirin (16.8 mg), and poncirin (71.9 mg) were purified from the 524 mg crude extract in only one step. The purities of the isolated compounds were determined to be over 90% by HPLC analysis and their structures were elucidated by (1)H NMR, (13)C NMR, and ESI-MS.


[The electrodynamics of biological processes].
The relations between field-physics and biology are systematically and continuously being inspected. In this paper, the electromagnetic field is taken into consideration, too. An objective point for a biological system of equations corresponding with MAXWELL-equations is given. Therefore processes of the growth and procreation are to be regarded as a unit. A new natural constant called b can be predetermined from field-biology. A triplet of natural constants--c, h, and b--enables a solid connection between physics and biology.


Assessment of potential impacts associated with gene flow from transgenic hybrids to Mexican maize landraces.
Genetically modified (GM) maize has been grown and safely consumed on a global scale since its commercialization in 1996. However, questions have been raised about the potential impact that GM maize could have on native maize landraces in Mexico, which is the center of origin and diversity of maize. This research was conducted to evaluate potential changes to maize landraces in an unlikely event of transgene introgression. For this study, two GM traits that confer insect protection and herbicide tolerance in maize (MON 89034 and MON 88017), designated as VT3Pro, were introgressed into two Mexican landraces, Tuxpeño and Tabloncillo. Field trials were conducted across four environments to assess phenotypic characteristics, plant response to stressors, and kernel composition of landraces with and without VT3Pro traits. Furthermore, materials from four backcrossing generations were analyzed for segregation of these GM traits. Generally, no significant differences were observed between landraces with and without VT3Pro traits for the evaluated characteristics and the segregation analysis showed that GM traits, when introgressed into landraces, followed Mendelian principles. These results support the conclusion that, if inadvertently introgressed into landraces, VT3Pro traits are not expected to alter phenotypic or kernel characteristics, plant response to stressors (except for targeted insect protection and herbicide tolerance traits) and would segregate like any endogenous gene. These results should be taken into consideration when discussing benefits and risks associated with commercial production of GM maize hybrids in the centers of origin and diversity of maize.


Work-parenting linkages among dual-earner couples at the transition to parenthood.
This study evaluates the impact of work experiences on parenting quality during the transition to parenthood. Dual-earner parents (n = 83) completed measures of work experiences (autonomy and interpersonal atmosphere). Parenting was observed twice, 3 months apart. Hierarchical regression analyses showed that mothers who reported a more negative interpersonal atmosphere at work showed decreases in positive parenting and increases in negative parenting over time, after controlling for their occupational status and general well-being. Mothers' work experiences also predicted changes in fathers' observed parenting. In contrast, fathers' work experiences were largely unrelated to fathers' or mothers' parenting. With one exception, parents' occupational status did not moderate work-parenting relationships, and feelings of role overload did not mediate these relationships.


Chaos in the three-site Bose-Hubbard model: Classical versus quantum.
We consider a quantum many-body system-the Bose-Hubbard system on three sites-which has a classical limit, and which is neither strongly chaotic nor integrable but rather shows a mixture of the two types of behavior. We compare quantum measures of chaos (eigenvalue statistics and eigenvector structure) in the quantum system, with classical measures of chaos (Lyapunov exponents) in the corresponding classical system. As a function of energy and interaction strength, we demonstrate a strong overall correspondence between the two cases. In contrast to both strongly chaotic and integrable systems, the largest Lyapunov exponent is shown to be a multivalued function of energy.


Mitogen-activated protein kinase and phosphatidylinositol 3-kinase pathways are not sufficient for insulin-like growth factor I-induced mitogenesis and tumorigenesis.
Insulin-like growth factor-I (IGF-I) and insulin are known to activate a signaling cascade involving ras --> kappa raf-1 --> mitogen-activated protein (MAP) kinase kinase (MEK) --> p42/p44 MAP kinase (Erk-1 and -2). Recent reports suggest that activation of this ras/MAP kinase pathway is involved in mitogenesis and c-fos transcription but is not required for insulin action on metabolic processes such as glycogen synthesis, lipogenesis, and GLUT-4-mediated glucose transport. Previously we and others have demonstrated that substitution of both tyrosines at positions 1250 and 1251 in the carboxy-terminal region of the human IGF-I receptor has relatively small effects on receptor and endogenous substrate phosphorylation but completely abrogated the ability of these cells to form tumors in nude mice or proliferate in response to IGF-I in culture. Replacement of the tyrosine at position 1316 also did not affect the kinase activity of the receptor with respect to autophosphorylation or phosphorylation of endogenous substrates but did reduce the ability of the receptor to mediate mitogenic or tumorigenic signals. To further characterize the role of these tyrosines in IGF-I receptor function, we have used three distinct approaches to examine the ras/MAP kinase pathway in IGF-I-induced mitogenesis and tumorigenesis in NIH-3T3 cells overexpressing wild-type and mutated IGF-I receptors: 1) tyrosine phosphorylation of the MAP kinases Erk-1 and -2; 2), mobility shifts indicative of MAP kinase phosphorylation; and 3) in vitro MAP kinase activation. We have also examined IGF-I-induced phosphatidylinositol (PI) 3-kinase activation in the same cell lines. By each method we show that the IGF-I-induced MAP kinase phosphorylation/activation and PI 3-kinase activation, are not different between cells overexpressing wild-type IGF-I receptors and cells carrying IGF-I receptors having tyrosine motifs replaced at positions 1250 and 1251. We conclude that mitogenic and tumorigenic signals involving tyrosine residues in the C-terminal domain of the IGF-I-receptor include pathways other than the MAP kinase and PI 3-kinase pathways.


[Learning of flexible endoscopy, particularly endoscopic vacuum therapy (EVT)].
Anastomotic insufficiency and perforations in the gastrointestinal tract are severe complications associated with a high complication rate and mortality. Conventional treatment options (particularly re-operations) are often unsatisfactory. Endoscopic vacuum therapy (EVT) is increasingly being used as a promising alternative. The use of EVT requires a high level of competence in interventional flexible endoscopy, which is primarily not available to every surgeon. Special training programs are required here. Based on this need the long-proven Tübingen training system for flexible endoscopy was modified to meet the special requirements of surgeons and is currently being extended by a special training module for EVT. In addition to the theoretical principles, the training is focused on learning the manual skills for flexible endoscopy. A 2-stage process was developed for this purpose: 1) to become familiar with handling of the flexible endoscope and to learn spatial orientation by means of a didactically optimized abstract phantom (Tübingen Orientophant) and 2) learning and training of EVT using a newly developed patient-analogous training model with various insufficiencies and abscess cavities in the upper and lower gastrointestinal tract. The procedure can be trained hands-on step by step exactly as with the patient, whereby the sponge can be applied using different methods, such as overtube and dragging procedures. The consequences of mistakes and complication management can also ideally be trained hands-on using the phantom. Evaluations of the first course series show that surgeons achieve endoscopic competence very quickly and learn to master the new procedure. The structure of such a course must, however, be designed according to long-term experience in an optimal didactic manner. Decision-makers in healthcare policy should give much more support to such courses in order to improve patient care and to increase patient safety.


Cytidine Triphosphate Synthase Four From Arabidopsis thaliana Attenuates Drought Stress Effects.
Cytidine triphosphate synthase (CTPS) catalyzes the final step in pyrimidine de novo synthesis. In Arabidopsis, this protein family consists of five members (CTPS1-5), and all of them localize to the cytosol. Specifically, CTPS4 showed a massive upregulation of transcript levels during abiotic stress, in line with increased staining of CTPS4 promoter:GUS lines in hypocotyl, root and to lesser extend leaf tissues. In a setup to study progressive drought stress, CTPS4 knockout mutants accumulated less fresh and dry weight at days 5-7 and showed impaired ability to recover from this stress after 3 days of rewatering. Surprisingly, a thorough physiological characterization of corresponding plants only revealed alterations in assimilation and accumulation of soluble sugars including those related to drought stress in the mutant. Bimolecular fluorescence complementation (BiFC) studies indicated the interaction of CTPS4 with other isoforms, possibly affecting cytoophidia (filaments formed by CTPS formation. Although the function of these structures has not been thoroughly investigated in plants, altered enzyme activity and effects on cell structure are reported in other organisms. CTPS activity is required for cell cycle progression and growth. Furthermore, drought can lead to the accumulation of reactive oxygen species (ROS) and by this, to DNA damage. We hypothesize that effects on the cell cycle or DNA repair might be relevant for the observed impaired reduced drought stress tolerance of CTPS4 mutants.


Dipeptide Surrogates Containing Asparagine-Derived Tetrahydropyrimidinones: Structure and Amide Rotomer Stereochemistry of a Proline Mimic.
Novel dipeptide surrogates Fmoc-Xaa-(cyclo)Asn-OBut are prepared from asparagine, aromatic aldehydes, and Fmoc-protected amino acid chlorides. Previous studies have focused on basic approaches to key synthetic intermediates containing the (cyclo)Asn fragment for polypeptide preparation. The synthesis of these systems has now been extended to electron-rich aromatic aldehydes, exemplified by p-anisaldehyde. One-dimensional nuclear Overhauser effect experiments confirm the trans configuration around the central secondary amide bond and the boat conformation of the pyrimidinone ring. Variable-temperature NMR at 500 MHz was employed both to obtain thermodynamic data on the cis-trans amide bond interconversion barrier and to probe for the presence of hydrogen bonding. Single-crystal X-ray analysis of Ac-d-Ala-(cyclo)Asn-NHMe shows a high degree of structural similarity to a known proline-containing dipeptide.


Seroconversion after hepatitis B vaccination in healthy young adults, and the effect of a booster dose.
Previous studies have shown that 5% to 15% of healthy people do not show a protective antibody response following hepatitis B vaccination. The study was done to determine the protective efficacy of vaccination in healthy young adults 1 to 4 years after the three dose vaccination series and to study the effect of a booster dose on non-responders and hypo-responders. Prospective intervention study. From January to June 2000, Faculty of Medicine, University of Colombo. 258 volunteers from five batches of medical students vaccinated with three doses of the recombinant vaccine at 0, 1 and 6 months. 9.5% were non-responders. Duration of vaccination, sex and body mass index were not significantly associated with anti-HBs levels. 28.6% had potential risk factors for acquiring HBV infection. 86.3% of non-responders developed protective anti-HBs titres after a booster dose. The persistent non-responders did not have a chronic illness or past HBV infection. A substantial number do not seroconvert after hepatitis B vaccination. Testing of blood for anti-HBs one month after vaccination is recommended to recognise non-responders as a booster dose will be beneficial in the majority of them.


Identifying Electroencephalography Biomarkers in Individuals at Clinical High Risk for Psychosis in an International Multi-Site Study.
The clinical high-risk for psychosis (CHR-P) paradigm was introduced to detect individuals at risk of developing psychosis and to establish preventive strategies. While current prediction of outcomes in the CHR-P state is based mostly on the clinical assessment of presenting features, several emerging biomarkers have been investigated in an attempt to stratify CHR-P individuals according to their individual trajectories and refine the diagnostic process. However, heterogeneity across subgroups is a key challenge that has limited the impact of the CHR-P prediction strategies, as the clinical validity of the current research is limited by a lack of external validation across sites and modalities. Despite these challenges, electroencephalography (EEG) biomarkers have been studied in this field and evidence suggests that EEG used in combination with clinical assessments may be a key measure for improving diagnostic and prognostic accuracy in the CHR-P state. The PSYSCAN EEG study is an international, multi-site, multimodal longitudinal project that aims to advance knowledge in this field. Participants at 6 international sites take part in an EEG protocol including EEG recording, cognitive and clinical assessments. CHR-P participants will be followed up after 2 years and subcategorised depending on their illness progression regarding transition to psychosis. Differences will be sought between CHR-P individuals and healthy controls and between CHR-P individuals who transition and those who do not transition to psychosis using data driven computational analyses. This protocol addresses the challenges faced by previous studies of this kind to enable valid identification of predictive EEG biomarkers which will be combined with other biomarkers across sites to develop a prognostic tool in CHR-P. The PSYSCAN EEG study aims to pave the way for incorporating EEG biomarkers in the assessment of CHR-P individuals, to refine the diagnostic process and help to stratify CHR-P subjects according to risk of transition. This may improve our understanding of the CHR-P state and therefore aid the development of more personalized treatment strategies.


Modulation of Mg(2+)-dependent [3H]TCP binding by L-glutamate, glycine, and guanine nucleotides in rat cerebral cortex.
Biochemical and electrophysiological studies have demonstrated that phencyclidine (PCP) recognition site exists in the ion channel of the N-methyl-D-aspartate (NMDA) receptor ion channel complex. Using an extensively washed rat cortical membrane preparation, the effects of Mg2+ and guanylylimidodiphosphate (GppNHp) were examined on the binding of [3H]-N-[1-(2-thienyl)cyclohexyl]-3,4-piperidine ([3H]TCP). Low concentrations of Mg2+ (EC50 = 11 microM) stimulated [3H]TCP binding under the basal condition and high concentrations of Mg2+ (IC50 = 1 mM) inhibited it. In the presence of 10 microM L-glutamate and 10 microM glycine, their EC50 values for Mg2+ enhancement of [3H]TCP binding were markedly reduced (to 1.9 microM or 8.4 microM), respectively. By contrast, the IC50 values for Mg2+ inhibition of [3H]TCP binding were reduced in the presence of L-glutamate, but not glycine. Furthermore, a stimulatory effect of Mg2+ on [3H]TCP binding was additional to the [3H]TCP binding stimulated by a maximally effective concentration of L-glutamate (10 microM) or glycine (10 microM). In the kinetic study, 300 microM Mg2+ produced an increase in the rates of both association and dissociation of [3H]TCP. Similar results were obtained with L-glutamate (10 microM) and glycine (10 microM); 10 mM Mg2+ also caused an acceleration of the association rate but strongly decreased [3H]TCP binding at equilibrium. Compared with [3H]TCP binding under the basal condition, K+ (10 mM) alone decreased the maximal binding without producing any change in the association rate; 10 mM K+ also significantly decreased Mg(2+)-stimulated [3H]TCP binding but caused no change in the acceleration of the association rate caused by Mg2+.(ABSTRACT TRUNCATED AT 250 WORDS)


Clinical trials of immunogene therapy for spontaneous tumors in companion animals.
Despite the important progress obtained in the treatment of some pets' malignancies, new treatments need to be developed. Being critical in cancer control and progression, the immune system's appropriate modulation may provide effective therapeutic options. In this review we summarize the outcomes of published immunogene therapy veterinary clinical trials reported by many research centers. A variety of tumors such as canine melanoma, soft tissue sarcomas, osteosarcoma and lymphoma, feline fibrosarcoma, and equine melanoma were subjected to different treatment approaches. Both viral and mainly nonviral vectors were used to deliver gene products as cytokines, xenogeneic tumor associated antigens, specific ligands, and proapoptotic regulatory factors. In some cases autologous, allogenic, or xenogeneic transgenic cytokine producing cells were assayed. In general terms, minor or no adverse collateral effects appeared during this kind of therapies and treated patients usually displayed a better course of the disease (longer survival, delayed or suppressed recurrence or metastatic spread, and improvement of the quality of life). This suggests the utility of these methodologies as standard adjuvant treatments. The encouraging outcomes obtained in companion animals support their ready application in veterinary clinical oncology and serve as preclinical proof of concept and safety assay for future human gene therapy trials.


Risk-adjusted strategies in the prevention of early arterial thrombosis following lower extremity arterial reconstruction: a comparison of unfractionated versus low molecular weight heparin.
In vascular surgery postoperative thrombosis prophylaxis must sufficiently prevent arterial thrombosis. This cohort study examines different therapeutic approaches of unfractionated heparin (UFH) or low molecular weight heparin (LMWH) after vascular reconstruction. Four hundred seventy-five patients entered the study between 2005 and 2008. Our clinical routine made a differentiation between low-risk patients (N.=375) and patients with peripheral bypass, which were grouped as high-risk (N.=148). We changed our postoperative anticoagulation management after 24 months in the low-risk and after each 16 months in the high-risk group. The anticoagulation of low-risk patients consisted of either two applications of 7.500 IU UFH subcutaneously (N.=158) or one daily application of 40 mg LMWH each up to discharge (N.=169). High-risk patients received either 25.000 IU UFH i.v. over 24 hours and 4 days (N.=48), 2-times (N.=51) or one-time weight-adjusted LMWH (N.=49) up to discharge (1 mg/kg body weight). Minor complications (bleedings) were differentiated from major early graft occlusion during the postoperative course. Further follow-up was not done for this study. Low risk: under LMWH, complications could be significantly reduced (P=0.001). Under LMWH significantly fewer occlusion complications occurred (P=0.01) and operation-induced hemorrhages were less frequently observed (P=0.05), this was significant in the complete low-risk group. High-risk: the one-time weight-adjusted LMWH group similarly exhibited many occlusions, like the unfractionated group (NS). The two-time LMWH treatment was significantly superior to the one-time application with respect to occlusion followed by amputations (P=0.03). Minor complications could be minimized overall by administration of LMWH and its dose reduction (NS). The differentiation between patients with high and low risk seems reasonable. An improvement could be achieved by differentiated LMWH application. Synthetic specific antifactor Xa substances (fondaparinux) or other medications could lead in future to other changes in the management of vascular surgery patients and should be further evaluated.


Large Volume Paracentesis in Patients with Liver Cirrhosis Temporarily Diminishes Blood Cell Count.
Large-volume paracentesis is the preferred treatment for patients with severe and refractory ascites. Several complications were reported during therapeutical paracentesis. However, there are very few published studies on the change in blood cell count after paracentesis. This study aimed to evaluate any changes in blood cell counts after ascites fluid drainage. This study was conducted on patients with severe ascites and chronic liver disease who underwent large-volume paracentesis at Namazi Hospital, in Shiraz, Iran, between March 2021 and February 2022. A data gathering form containing the patient's medical history, cause of cirrhosis, ascites fluid volume, as well as routine tests including primarily sodium, potassium, and basal creatinine, was filled out. Before and after the surgery, the total blood cell count was measured. Before the procedure, adjustment was made in the case of coagulopathy and albumin deficiency. The effect of factors such as the volume of drained fluid, splenomegaly, antibiotics, and steroid use was assessed on the changes in the number of blood cells. Using the JAMOVI 2.3.9 software, a paired t test and multiple regression were applied for statistical analysis (P<0.001). The study included 37 patients. After the paracentesis procedure, the number of blood cells significantly decreased in all groups (P<0.001). The followings are the amounts of each type of blood cells before and after the procedure: Platelet=153837±91862 and 115648±69136, red blood cells=3.53±0.784 and 3.22±0.705, white blood cells=12.3±7.78 and 8.6±5.5. None of the study variables, including drained volume, splenomegaly, antibiotics, and steroid use, were significant predictors of the changes in the blood cell count after paracentesis (P>0.001). The findings of the present study showed that children with tense ascites who had large-volume paracentesis might experience a sharp drop in blood cell count after the procedure, which was a transient physiological condition.


Composition of cytochrome P-450 isozymes from hepatic microsomes of C57BL/6 and DBA/2 mice assessed by warfarin metabolism, immunoinhibition, and immunoelectrophoresis with anti-(rat cytochrome P-450).
The composition of isozymes of hepatic microsomal cytochrome P-450 (P-450) from C57BL/6 (B6) and DBA/2 (D2) inbred strains of mice, uninduced and treated with phenobarbital (PB), pregnenolone-16 alpha-carbonitrile (PCN), and beta-naphthoflavone (BNF), were assessed. The assessment was based on comparisons of the regioselectivity and stereoselectivity of warfarin metabolism, inhibition of metabolism by antibodies raised against highly purified rat P-450 isozymes (the specificity of these antibodies in inhibiting rat microsomal metabolism of warfarin was also determined), and by immunoelectrophoresis using the anti-(rat P-450). Isozymes were considered to be the same or very similar if they exhibited the same substrate specificity, antigenicity, and molecular weight. Untreated D2 and B6 mice had similar compositions of P-450 isozymes which were independent of the Ah receptor. Both strains contained an isozyme very similar to rat P- 450UT -A. PB or PCN induction of the two mouse strains proceeded independently of the Ah receptor and yielded isozymes which are the same as or very similar to the rat P- 450UT -A, P- 450PB -B, P- 450PB -C, and P- 450PB /PCN-E isozymes. D2 and B6 mice had similar isozyme compositions when treated with PB or PCN, but isozyme compositions varied in differently induced mice. BNF treatment of the D2 mice was without effect, but the B6 mice produced an isozyme(s) which was different from rat P- 450BNF -B, the major BNF-induced isozyme in rat liver.


Responsiveness of five outcome measurement instruments in total elbow arthroplasty.
To quantify and compare the sensitivity to change of 5 outcome instruments for the elbow joint. In a prospective cohort study (n = 65), outcome was measured by the Short Form 36 (SF-36), the Disabilities of the Arm, Shoulder, and Hand questionnaire (DASH), the modified American Shoulder and Elbow Surgeons questionnaire for the elbow, patient part (pmASES) and examiner/clinical part (cmASES), and the Patient-Rated Elbow Evaluation form (PREE). Responsiveness was quantified by the effect size (ES) and standardized response mean (SRM) before and 6 months after total elbow arthroplasty. Receiver operating characteristic (ROC) curves were used to determine the instruments' ability to classify effects into global health change assessment categories. For the total scores, the ES were 1.50 for the PREE, 1.32 for the pmASES, 0.86 for the cmASES, 0.56 for the DASH, and 0.11 for the SF-36 (P ≤ 0.002 for all differences, except the cmASES and DASH). The same order was found within the subdomains of pain/symptoms and function and remained consistent when using the SRM and in ROC curve analysis. PREE total (area under the ROC curve 0.68), DASH function, and pmASES total and pain (area under the ROC curve range 0.64-0.67) discriminated best between "much better" and the other categories. The PREE was the most responsive instrument and can be recommended for every set of measures for elbow joint disorders. The pmASES was slightly less responsive but is a valid alternative. The examiner-assessed cmASES is affected by concerns regarding validity and was relatively less responsive. The DASH for comprehensive measurement of the entire upper extremity and the SF-36 for chronic pain conditions complete the assessment set.


Dietary supplementation with black raspberries prolongs survival in ApcMin/+ mice.
A diet supplemented with freeze-dried black raspberries (BRBs) has been demonstrated to modulate various biochemical and physiological pathways in both colorectal cancer (CRC) patients and ApcMin/+ mice, which are utilized to model CRC. These changes have been previously shown to exert beneficial chemopreventive effects against CRC, with outcomes such as reduction of adenoma development and inflammation. This study aimed to assess whether these effects manifest in a meaningful change in survival rates by comparing these rates between ApcMin/+ mice administered a 5% BRB-supplemented diet or a control AIN-76A diet. Percent survival over days elapsed was assessed in order to determine a median length of survival for each group of mice. Significant increases in survival rates with consumption of the BRB diet versus the control diet were demonstrated in both male and female mouse study groups. Male and female control groups were also compared in order to reduce confounding due to the sex of the mice; the difference in survival rates between male and female mice was not significant (p = 0.07, *p < 0.05), as male mice lived for a median of 143 days and females for 194 days. The results of this study suggest that administration of a BRB-supplemented diet may potentially prolong the lifespan and increase survival rates of colorectal cancer patients.


Quantitative evaluation of CXCL8 and its receptors (CXCR1 and CXCR2) gene expression in Iranian patients with multiple sclerosis.
CXCL8 and its receptors (CXCR1 and CXCR2) play important roles in CNS development, neuronal survival, modulation of excitability, and neuroimmune response. The aim of this study is to evaluate gene expression of CXCL8 and CXCR1/CXCR2 in peripheral blood cells (PBCs) of Iranian patients with relapsing remitting (RR) form of Multiple sclerosis (MS). We explored the mRNA expression of CXCL8 and its receptors in PBCs of 49 RR-MS patients in remitting status and 60 healthy controls by quantitative Real-Time PCR. Median expression of CXCL8 mRNA in peripheral blood of MS patients decreased more than 3-fold compared to control group (p < 0.001), while there were not significant differences in CXCR1 and CXCR2 gene expression between MS patients and healthy subjects (p = 0.159 and p = 0.248, respectively). There was a significant negative correlation of CXCR2 expression with EDSS (rs = -0.432, p = 0.004). It appears that decreased expression of CXCL8 may lead to a raised risk of MS.


Polymorphism of short tandem repeat (STR) loci PLA2, D3S2459, D8S315 (kw38), CYP19, D3S1359, FGA, and D8S1132 in the Japanese and Chinese populations.
We report a study of polymorphism for seven short tandem repeat (STR) loci in Japanese and Chinese populations. Among 104 to 134 individuals in the both population samples, eight alleles were revealed for locus PLA2, thirteen for D3S1359, eleven for FGA, eight for D8S315 (kw38), ten for D8S1132, five for CYP19, and seven for D3S2459. They correspondingly constituted 10 to 39 genotypes therein. For most of the STRs, there was only a single allele active as the most frequent one among the others, except locus D3S1359 in Chinese samples (two alleles, 206 bp and 210 bp, frequency = 0.273 each). Also, the population genotype configurations were locus specific, varying in the patterns of commonest genotypes on each locus, e.g., one pattern for loci CYP19, D3S1359, and D8S315, one and two for loci PLA2 and D3S2459, two for locus D8S1132, and one and four for locus FGA. The distributions of observed genotypes were in Hardy-Weinberg Equilibrium. Furthermore, the seven STRs were exhibited highly polymorphic and informative for the both populations, and the alleles could be easily separated in electrophoresis and correctly interpreted with side-to-side allelic ladders. Together, the results suggest that the tri- and tetra-meric STRs are useful genetic markers for forensic practice.


Purification and structure of exendin-3, a new pancreatic secretagogue isolated from Heloderma horridum venom.
An amino-terminal histidyl structure (His1) is characteristic of most peptides in the glucagon superfamily. An assay for His1 peptides performed by amino-terminal amino acid sequencing was used to screen venom from the Gila monster lizard, Heloderma horridum. Two His1 peptides were identified: helospectin and a new His1 peptide that has been named exendin-3 to indicate that it is the third peptide to be found in an exocrine secretion of Heloderma lizards which has endocrine activity, the first two being helospectin (exendin-1) and helodermin (exendin-2). In the lot of H. horridum venom tested, exendin-3 was 5-10-fold more abundant in molar concentration than helospectin. The structure of exendin-3 was analyzed by amino acid sequencing and mass spectrometry. Exendin-3 is a 39-amino acid peptide with a mass of 4200. It contains a carboxyl-terminal amide and has a strong homology with secretin at its amino-terminal 12 amino acids. The complete structure of exendin-3 is His-Ser-Asp-Gly-Thr-Phe-Thr-Ser-Asp-Leu-Ser-Lys-Gln-Met-Glu-Glu-Glu-Ala- Val-Arg - Leu-Phe-Ile-Glu-Trp-Leu-Lys-Asn-Gly-Gly-Pro-Ser-Ser-Gly-Ala-Pro-Pro-Pro- Ser- amide. It is 32 and 26% homologous with helospectin and helodermin, respectively. It has greatest homology with glucagon (48%) and human glucagon-like peptide-1 (50%). Exendin-3 (3 microM) stimulated increases in cellular cAMP and amylase release from dispersed guinea pig pancreatic acini.


First Report of Botryosphaeria dothidea Causing Stem Canker and Dieback of Apple Trees in Ontario, Canada.
Apple trees (Malus domestica L. Borkh.) exhibiting extensive and unknown tree fruit decline symptoms were observed in commercial orchards over the past 5 years in Ontario, Canada. The trees exhibited shoot dieback, attached wilted leaves and cankers on the main trunk. Trees with rapid development of cankers upward from the graft union developed extensive vascular discoloration that resulted in sudden collapse of the entire tree. In 2018-19, up to 42% mortality was observed on 2- to 8-year old apple trees. Nine symptomatic trees were collected from two orchards located in southwestern and one in southcentral Ontario. Samples (1 cm length) were collected from symptomatic trunk and shoot tissue, surface sterilized with 70% ethanol for 30 sec, followed by 1% NaClO for 20 min and three rinses in sterile water. The samples were air-dried and placed on a 2% potato dextrose agar (PDA, Difco) culture media supplemented with kanamycin (50 mg L-1). The PDA plates were incubated at 22°C for 5 days in the dark. All fungal colony-forming units that developed were hyphal-tip transferred to individual PDA plates and incubated at 22°C for 7 days in the dark. Purified mycelial isolates were classified into morphotypes prior to molecular identification. One morphotype showed white to olive-green colonies that developed on a moderately dense mycelial mat with aerial hyphae. Several solitary and globose black pycnidia that contained a single ostiole were produced on pine needles on PDA after incubation at 22°C for 14 days in the dark. Conidia were hyaline, fusiform, aseptate with an average size of 3.9 - 5.2 x 21.4 - 26.6 μm (N=50). Genomic DNA was extracted from 5-day old culture of a representative isolate, #M68-17, grown on PDA using the Plant/Fungi DNA Isolation Kit (Norgen Biotech Corp., Thorold, ON, Canada). The rDNA internal transcribed spacer (ITS), translation elongation factor 1- alpha gene (TEF-1α), and β-tubulin gene (TUB2) were amplified using the primer pairs ITS1/ITS4 (White et al. 1990), EF1-728F/EF1-986R (Carbone and Kohn 1999), and Bt2a/Bt2b (O'Donnell et al. 1998), respectively, and sequenced. The nucleotide sequences obtained (GenBank # MZ926850, MZ934654, MZ934655) were 100.00% similar to those of Botyrosphaeria dothidea (Moug. ex Fr.) Ces. & De Not. isolates from other hosts in several countries in the NCBI database (MT111097, MT309401, MN515421, respectively). Randomized Accelerated Maximum Likelihood (RAxML) analysis using the three gene sequence data was completed (Stamatakis et al. 2008). The isolate #M68-17 was clustered with high bootstrap support values with the B. dothidea isolates from the fungal biodiversity centre (CBS) collection, including the ex-epitype (CBS 115476, CBS 110302) (Fig. 1). A living culture of the representative isolate, #M68-17, was deposited in the Canadian Collection of Fungal Cultures (DAOMC 252246). Pathogenicity tests were conducted in the lab on wood cuttings and in planta. Ten 20 cm apple cuttings and five potted one year old apple seedlings were surface sterilized, wounded and inoculated with 4 mm mycelium agar plug from a 5-day old culture of isolate #M68-17 and wrapped with Parafilm. Three control apple cuttings and seedlings were inoculated with PDA plugs and incubated in the same environment. Cuttings were placed inside a clear plastic container with moist paper towels and incubated at room temperature in the dark. Seedlings were placed in an open-air area between two greenhouses and watered as needed. Twelve days post-inoculation, the average length of the developed necrotic lesions on cuttings was 8.8 ± 0.4 cm. Necrotic and sunken canker symptoms appeared at 10 days, spread upward from the inoculation point and by 6 weeks the upper portion of the seedling was dead (Fig. 2). B. dothidea was isolated from all the inoculated cuttings and seedlings, thus fulfilling Koch's postulates. Control cuttings and seedlings showed no symptoms, and the fungus was not isolated from the wood. B. dothidea was reported as a pathogen causing cankers on a wide range of woody crop plants including apples, almonds, pistachios, hazelnut, walnut, olive and grapes in the United States, China, Uruguay, Spain, Tunisia and Turkey (Chebil et al. 2014; Delgado-Cerrone et al. 2016; Moral et al. 2019; Tang et al. 2012; Türkölmez et al. 2016). However, this is the first report of B. dothidea causing stem canker and death of young apple seedlings in Ontario, Canada. The findings suggest that B. dothidea has the potential to severely affect apple production in Ontario. Accurate identification of pathogen(s) associated with apple decline will support management of the disease.


Design and methods of the Hospital Elder Life Program (HELP), a multicomponent targeted intervention to prevent delirium in hospitalized older patients: efficacy and cost-effectiveness in Dutch health care.
The Hospital Elder Life Program (HELP) has been shown to be highly efficient and (cost-)effective in reducing delirium incidence in the USA. HELP provides multicomponent protocols targeted at specific risk factors for delirium and introduces a different view on care organization, with trained volunteers playing a pivotal role. The primary aim of this study is the quantification of the (cost-)effectiveness of HELP in the Dutch health care system. The second aim is to investigate the experiences of patients, families, professionals and trained volunteers participating in HELP. A multiple baseline approach (also known as a stepped-wedge design) will be used to evaluate the (cost-) effectiveness of HELP in a cluster randomized controlled study. All patients aged 70 years and older who are at risk for delirium and are admitted to cardiology, internal medicine, geriatrics, orthopedics and surgery at two participating community hospitals will be included. These eight units are implementing the intervention in a successive order that will be determined at random. The incidence of delirium, the primary outcome, will be measured with the Confusion Assessment Method (CAM). Secondary outcomes include the duration and severity of delirium, quality of life, length of stay and the use of care services up to three months after hospital discharge. The experiences of patients, families, professionals and volunteers will be investigated using a qualitative design based on the grounded theory approach. Professionals and volunteers will be invited to participate in focus group interviews. Additionally, a random sample of ten patients and their families from each hospital unit will be interviewed at home after discharge. We hypothesize that HELP will reduce delirium incidence during hospital admission and decrease the duration and severity of delirium and length of hospital stays among these older patients, which will lead to reduced health care costs. The results of this study may fundamentally change our views on care organization for older patients at risk for delirium. The stepped-wedge design was chosen for ethical, practical and statistical reasons. The study results will be generalizable to the Dutch hospital care system, and the proven cost-effectiveness of HELP will encourage the spread and implementation of this program. Netherlands Trial register: NTR3842.


Structural-functional changes in detached cucumber leaves, and modelling these by hormone-treated leaf discs.
Senescence and rejuvenation were investigated in detached cucumber (Cucumis sativus L.) leaves after cultivation in nutrient solution for one week or four weeks. Rooting of the petiole (visible generally from the 7th day) elicited a combination of different morphological, anatomical, and physiological changes in the lamina. Extensive growth in area and thickness, extreme regreening, changes of chloroplast structure and activity, as well as the pattern of Chl-protein complexes were observed and compared either to the corresponding parameters of young detached leaves or mature attached leaves. These responses could be provoked separately by treating excised leaf discs with kinetin, benzyladenine, or indolylacetic acid. The hormones showed mutuality in their effects, benzyladenine being responsible for the growth of cells, while indolylacetic acid and kinetin promoted an increase in chlorophyll content. However, none of the treatments resulted in the growth of the chloroplasts in the leaf discs, which was only prominent in the rooting leaves.


Recumbent Total Skin Electron Beam Therapy.
Our purpose was to describe preliminary dosimetric and clinical results of a recumbent total skin electron beam therapy (TSEBT) technique and compare this to a conventional standing TSEBT technique. A customized treatment platform with recessed side wheels was constructed and commissioned for patients to be treated in a recumbent position. Dosimetric and clinical information was collected for patients treated with this new recumbent technique in addition to that of a cohort of patients treated contemporaneously using the conventional standing method. Dose delivery and clinical outcomes were compared for patients treated with the recumbent and standing techniques. Between 2017 and 2019, 27 patients were treated with TSEBT with the recumbent (n = 13) or conventional standing technique (n = 14) at our institution. Measured dose at 15 body sites could be directly compared. Of these, 10 showed no significant difference between the two techniques while five sites showed significant differences in median measured dose, including the top of left shoulder, right biceps, bend of left elbow, upper back, and medial right thigh (P < .003). Measured dose was significantly higher with the standing technique at these sites with the exception of the upper back. Rates of complete response (25% vs 23%), partial response (50% vs 69%), and stable disease (17% vs 8%) were similar between the standing and recumbent cohorts, respectively (P = .78). We have developed, commissioned, and implemented a floor-based, recumbent technique that allows for treatment of patients who would otherwise not be eligible for TSEBT. Dosimetric and clinical measurements suggest that this technique is a viable alternative to the standing method.


Structural variation in the brown alga Sargassum cymosum and its effects on associated amphipod assemblages.
The presence of diverse biological substrates adds complexity to coastal landscapes and increases the number of ecological niches that can be used by the mobile epifauna. Studies on the influence of structural complexity have focused mainly on algal host species, but there is little information about the influence of intraspecific structural variation on the associated mobile epifauna. In this work, we examined whether intraspecific variation in the brown alga Sargassum cymosum influenced the structure of amphipod assemblages on two shores with different wave exposure. At least 15 fronds were randomly sampled at Fortaleza and Perequê-Mirim beaches, on the Atlantic coast of São Paulo state, southeastern Brazil, and 12 variables were measured for each alga. The amphipods were identified and counted. The greatest structural variation in S. cymosum occurred within shores, whereas the differences between shores were mainly related to algal size. These characteristics influenced amphipod assemblages differently on each shore, with the greatest effects being associated with variables related to morphological complexity, such as holdfast size, the number and size of branches, and the extent of cover by sessile colonial animals. These findings show that monospecific algal banks are not homogeneous, and that morphological differences and interactions with other biological substrates can influence the mobile epifaunal assemblages.


Fall prevention among children in the presence of caregivers in a paediatric ward: a best practice implementation.
This study aims to reduce the incidence of falls in paediatric inpatients aged 3 and below by implementing fall prevention strategies. The Joanna Briggs Institute Practical Application of Clinical Evidence System and Getting Research into Practice programmes were used for this project. The project was carried out in three phases over a 4-month period from March to June 2011. A fall prevention poster was introduced during the implementation phase, and pre- and post-implementation audits were carried out in a 43-bed acute care paediatric ward in Singapore, with a sample size of 30. The audit result of Criterion 1, evaluating the effectiveness of the fall prevention measures, improved by 13%, to 93%. Criterion 2, measuring nurses' compliance in the regular reinforcement of safety, improved significantly by 27%, to 40%. However, Criterion 3, which measured nurses' compliance in identifying patients at high risk of falls by placing a green identification wrist tag on such patients, decreased by 23%, to 50%. A multi-language poster on fall prevention was strategically positioned at the foot of all the cots. The poster served as an effective reminder and communication method between nurses and caregivers and also among caregivers of the child. Caregivers' increased awareness and knowledge of fall prevention contributed to a 50% decrease in fall incidence of patients aged 3 and below in the presence of a caregiver from January to June 2011, as compared with the incidence rate in 2010. This project has shown that fall incidents can be reduced when caregivers' awareness of fall prevention measures in the hospital setting were to be improved. The poster on fall prevention has increased caregivers' awareness and reduced inpatient falls in the acute care setting. The pre- and post-implementation audits showed that the presence of a poster on fall prevention to remind parents/caregivers to raise and securely lock the cot rails at all times was effective in reducing the number of falls for children aged 3 years and below. The experience gained from this project was that communication to every staff member and caregivers is essential in implementing practice change. As a result of the study, the hospital plans to implement constant monitoring and reminders to nurses and caregivers to improve compliance to the recommended measures on fall prevention in the near future.


Olefinic peptide nucleic acid (OPA).
The olefinic peptide nucleic acid analogues (OPA) monomers containing the bases thymine and adenine were synthesised in 11 steps. Fully modified oligomers containing these units were prepared and their pairing properties assessed by means of UV-melting experiments.


Awareness of malalignment and malocclusion in children and their guardians.
We conducted a survey on the awareness of abnormalities of dentition and occlusion in 1,904 children (0-15 years old) and their guardians (parents, grandmothers, grandfathers and siblings) on their initial visit to the Department of Pediatric Dentistry, Chiba Hospital, Tokyo Dental College. The location and type of abnormality for which the children and their guardians most commonly sought treatment were crowding of the upper and lower anterior teeth and inverted occlusion. The most common initial triggers for concern were "guardians noticed abnormalities themselves". It seems logical that where malocclusions that children and guardians can easily notice for themselves are present, they are likely to make an early visit to a clinic in the hope of improving the abnormality. We suggest that further effect is made to educate guardians of children with abnormalities of dentition and occlusion on the importance of obtaining treatment.


Endoscopic characteristics and high-risk background mucosa factors of early gastric cancer after helicobacter pylori eradication: a single-center retrospective study.
Gastric cancer still develops after successful Helicobacter pylori(Hp)eradication. In this study, we aimed to explore the characteristics and risks of mucosal factors. A total of 139 early gastric cancers (EGC) diagnosed in 133 patients after successful eradication from January 2016 to December 2021 were retrospectively included in the Hp-eradication EGC group and 170 EGCs diagnosed in 158 patients were included in the Hp-positive EGC group. We analyzed the clinical, pathological, and endoscopic characteristics between the two groups to identify the features of EGC after Hp eradication. Another 107 patients with no EGC after Hp eradication were enrolled in a Hp-eradication non-EGC group. The background mucosal factors between the Hp-eradication EGC group and the Hp-eradication non-EGC group were compared to analyze the high-risk background mucosal factors of EGC after eradication. In addition, we divided the EGC group after Hp eradication into IIc type and non-IIc type according to endoscopic gross classification to assess the high-risk background factors of IIc-type EGC after Hp eradication. The endoscopic features of EGC after Hp eradication included location in the lower part of the stomach (p=0.001), yellowish color (p= 0.031), and smaller size (p=0.001). The moderate/severe gastric atrophy (GA), intestinal metaplasia (IM) in the corpus, severe diffuse redness, and map-like redness were risk factors for EGC after eradication (p=0.001, p=0.001, p=0.001, and p= 0.005, respectively). The Kyoto classification total score in the EGC group was higher than the non-EGC group (4 vs.3 p<0.001). A multivariate analysis revealed that depressed erosion (OR=3.42, 95% CI 1.35-8.65, p= 0.009) was an independent risk factor for IIc-type EGC after Hp eradication. EGC after eradication are smaller and yellowish lesions located in the lower part of the stomach. The risk background mucosal factors include moderate/severe GA, IM in the corpus, severe diffuse redness, and map-like redness. The Kyoto classification total score of 4 or more after successful eradication treatment might indicate EGC risk. In addition, the IIc-type EGC should be cautioned in the presence of depressed erosion after Hp eradication.


Assessment of stability of sulphated lactosyl archaeol archaeosomes for use as a vaccine adjuvant.
Archaeosomes, composed of sulphated lactosyl archaeol (SLA) glycolipids, have been proven to be an effective vaccine adjuvant in multiple preclinical models of infectious disease or cancer. In addition to efficacy, the stability of vaccine components including the adjuvant is an important parameter to consider when developing novel vaccine formulations. To properly evaluate the potential of SLA glycolipids to be used as vaccine adjuvants in a clinical setting, a comprehensive evaluation of their stability is required. Herein, we evaluated the long term stability of preformed empty SLA archaeosomes prior to admixing with antigen at 4 °C or 37 °C for up to 6 months. In addition, the stability of adjuvant and antigen was evaluated for up to 1 month following admixing. Multiple analytical parameters evaluating the molecular integrity of SLA and the liposomal profile were assessed. Following incubation at 4 °C or 37 °C, the SLA glycolipid did not show any pattern of degradation as determined by mass spectroscopy, nuclear magnetic resonance (NMR) and thin layer chromatography (TLC). In addition, SLA archaeosome vesicle characteristics, such as size, zeta potential, membrane fluidity and vesicular morphology, were largely consistent throughout the course of the study. Importantly, following storage for 6 months at both 4 °C and 37 °C, the adjuvant properties of empty SLA archaeosomes were unchanged, and following admixing with antigen, the immunogenicity of the vaccine formulations was also unchanged when stored at both 4 °C and 37 °C for up to 1 month. Overall this indicates that SLA archaeosomes are highly stable adjuvants that retain their activity over an extended period of time even when stored at high temperatures.


Optical single sideband modulation based on a high-order birefringent filter using cascaded Solc-Sagnac and Lyot-Sagnac loops.
We propose and experimentally demonstrate a simple and flexible photonic approach to implementing single sideband (SSB) modulation based on optical spectral filtering. The high-order birefringent filter is realized through the cascaded Solc-Sagnac and Lyot-Sagnac loops. By adjusting the rotation angle of the polarization controller (PC), the notch position to remove undesired sidebands changes. The frequency for SSB modulation varies accordingly. The periodical response of the filter spectrum allows both the carrier wavelength and the optical carrier to sideband ratio (OCSR) to be tunable. SSB modulation over a frequency range from 5 to 40 GHz and tunable OCSR ranging from -9.174 to 34.408 dB are obtained. The significant merits of the proposed approach are the simple structure, easy operation, large frequency range, tunable OCSR, and wavelength independence. The approach has potential applications in optimizing the transmission performance of photonic microwave signal processing systems.


Electronic Structure Calculations of Static Hyper(Polarizabilities) of Substrate-Supported Group-IV and -V Elemental Monolayers.
The substrate-induced effects on the polarizability (α) and first dipole hyperpolarizability (β) of group-IV (i.e., graphene, silicene, germanene, stanene) and group-V (i.e., phosphorene, arsenene, antimonene, and bismuthene) elemental monolayer nanoflakes are investigated. Density functional theory calculations show that these monolayers are bound with varying degrees of interaction strength with the Ag(111) substrate surface. Calculated dipole moment and β values are zero for the centrosymmetric configurations of the pristine elemental monolayers. On the other hand, substrate-induced changes in the electronic densities at the interface lead to substantially enhanced values of β, making these materials attractive for applications in the next-generation photonic technologies at the nanoscale.


Fatigue in knee and hip osteoarthritis: the role of pain and physical function.
It is suggested that serious levels of fatigue are present in nearly half of patients with OA. However, it is unclear which dimensions of fatigue are involved, if fatigue is related to pain and physical function, and if fatigue is influenced by therapy. The aims of this study were to measure levels of different dimensions of fatigue before and after evidenced-based conservative treatment and to investigate the association between fatigue and pain and physical function in patients with knee or hip OA. In this observational cohort study, levels of different dimensions of fatigue were measured in knee and/or hip OA patients before and after 12 weeks of conservative treatment. Cross-sectional and longitudinal relations between (change in) fatigue dimensions and (change in) pain or physical function were studied using association models, controlling for predefined possible confounders. A total of 231 patients was included, with 47% experiencing severe fatigue. A small decrease in levels of fatigue was seen after standardized treatment. The level of fatigue severity was cross-sectionally and longitudinally associated with physical function, whereas the level of physical fatigue was cross-sectionally and longitudinally associated with pain and physical function. No confounders were identified. Important levels of fatigue are common in knee and hip OA patients. After evidence-based tailored conservative treatment targeted to improve pain and physical function, a small decrease in fatigue levels was found. Reduction in levels of different fatigue dimensions were related to the change in physical function and pain.


Association Between Maternal Glucose Levels in Gestational Diabetes Screening and Subsequent Hypertension.
We assessed the association between maternal glucose levels in pregnancy and subsequent hypertension. This population-level, retrospective cohort study examined women aged 12 to 54 years with singleton pregnancies completed at ≥29 weeks of gestation from October 1, 2008 to December 1, 2018 followed until March 31, 2019 in Alberta, Canada. Women were stratified by results in the 50-gram glucose challenge test and by 75-gram oral glucose tolerance test subtypes (normal oral glucose tolerance test, elevated fasting plasma glucose only [elevated fasting], elevated postload glucose only, or both elevated fasting and postload glucose [combined]. Time to development of hypertension was modeled using Cox proportional hazards models. Of 313 361 women, 231 008 (79.1%) underwent a glucose challenge test only while 60 909 (20.9%) underwent either an oral glucose tolerance test only or both. Nine thousand five hundred eighty (3.1%) developed hypertension, and 2824 (0.9%) developed cardiovascular disease over a median follow-up of 5.7 years. Every 1-mmol/L increase in glucose in the glucose challenge test increased the risk of subsequent hypertension by 15% (adjusted hazard ratio and 95% CI, 1.15 [1.14-1.16]). Among those who underwent the oral glucose tolerance test, the combined group conferred the highest risk of subsequent hypertension, followed by elevated fasting, then elevated postload glucose only (reference: glucose challenge test ≤7.1 mmol/L, adjusted hazard ratio [95% CI]: elevated postload glucose only, 1.83 [1.68-2.00]; elevated fasting 2.02 [1.70-2.40]; combined, 2.65 [2.33-3.01]). No significant associations between maternal glucose levels and cardiovascular disease were observed. Increasing maternal glucose levels in pregnancy were associated with increasing risk of subsequent hypertension. These findings may help identify higher-risk women who should be targeted for earlier postpartum cardiovascular risk reduction.


High-k polymeric gate insulators for organic field-effect transistors.
Gate insulators play a role as important as that of the semiconductor in high performance OFETs, with a high on/off current ratio, low hysteresis, and device stability. The essential requirements for gate dielectrics include high capacitance, high dielectric breakdown strength, solution-processibility, and flexibility. In this paper we review progress in recent years in developing high-k gate polymeric insulators for modern organic electronic applications. After a general introduction to OFETs, three types of high-k polymeric gate insulating materials are enumerated in achieving high-quality OFETs, including polymer gate insulators, polymer-inorganic gate composites or bilayers, and ion gel electrolytes. Especially, we emphasize the significance, implementation and development of high-k polymeric gate insulators used in OFETs for future low voltage operated and flexible electronics. Finally, a brief summary and outlook are presented.


Transcriptome signature of cellular senescence.
Cellular senescence, an integral component of aging and cancer, arises in response to diverse triggers, including telomere attrition, macromolecular damage and signaling from activated oncogenes. At present, senescent cells are identified by the combined presence of multiple traits, such as senescence-associated protein expression and secretion, DNA damage and β-galactosidase activity; unfortunately, these traits are neither exclusively nor universally present in senescent cells. To identify robust shared markers of senescence, we have performed RNA-sequencing analysis across eight diverse models of senescence triggered in human diploid fibroblasts (WI-38, IMR-90) and endothelial cells (HUVEC, HAEC) by replicative exhaustion, exposure to ionizing radiation or doxorubicin, and expression of the oncogene HRASG12V. The intersection of the altered transcriptomes revealed 50 RNAs consistently elevated and 18 RNAs consistently reduced across all senescence models, including many protein-coding mRNAs and some non-coding RNAs. We propose that these shared transcriptome profiles will enable the identification of senescent cells in vivo, the investigation of their roles in aging and malignancy and the development of strategies to target senescent cells therapeutically.


Expression of tumour necrosis factor-alpha is associated with apoptosis in lungs of pigs experimentally infected with porcine reproductive and respiratory syndrome virus.
Expression of tumour necrosis factor (TNF)-alpha and its association with apoptosis was examined in lungs from pig experimentally inoculated with porcine reproductive and respiratory syndrome virus (PRRSV). The TNF-alpha nucleic acid was detected in PRRSV-infected alveolar macrophages by reverse transcription-polymerase chain reaction (RT-PCR). The TNF-alpha antigen was detected in alveolar macrophages by immunohistochemisty. DNA fragmentation in apoptotic cells was detected by in situ terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labelling (TUNEL). A double-labelling procedure using immunohistochemisty for the detection of PRRSV and the in situ TUNEL assay for the detection of apoptosis demonstrated that a majority of labelled cells were infected with PRRSV or apoptotic, but not both. Apoptotic cells were more abundant than PRRSV-infected cells in all lungs examined. A double-labelling procedure using immunohistochemisty for the detection of TNF-alpha and the in situ TUNEL assay for the detection of apoptosis demonstrated that a majority of labelled cells were apoptotic or for TNF-alpha, but not both. These data suggested that TNF-alpha released from macrophages after infection by PRRSV may induce apoptosis in uninfected bystander cells.


Influence of Fiber Orientation on the Water and Ions Transportation of Engineered Cementitious Composite (ECC).
An engineered cementitious composite (ECC) belongs to a type of high-performance fiber-reinforced materials. Fiber alignment causes the anisotropy of such materials. Herein, the influence of the fiber orientation on water and ion penetration into an ECC was studied. Fiber alignment was achieved using an extrusion approach. Water absorption, sorptivity, chloride penetration resistance, sulfate attack resistance, and freezing-thawing resistance of specimens with fiber aligned horizontally (AH), vertically (AV), and randomly (R), corresponding to the direction of the exposure surface that was studied. The results showed that fibers oriented perpendicular to the water path delayed water migration into the ECC matrix. The sorptivity was significantly affected by the fiber direction. The sorptivity of the AH specimens was 35% and 13% lower than that of the AV and R specimens, respectively. After 180 days of exposure, the chloride penetration depth of the AH specimens was 5.7 mm, which is 13.6% and 20.8% lower than that of the AV and R specimens, respectively. The sulfate ingress profile indicates that the fiber-matrix interface oriented perpendicular to the penetration path can effectively delay sulfate migration. The fiber orientation also influences the compressive strength gain under immersion conditions (Na2SO4 solution, Na2SO4 + NaCl solution, and water). Compared with the AH and R specimens, the AV specimens are more sensitive to the immersion condition. In contrast, the fiber orientation has no significant effect on ECC specimens under freeze-thaw cycles. These findings indicate that controlling the fiber alignment and orientation in an ECC can improve its durability under certain exposure conditions.


Horizontal Extraocular Muscle and Scleral Anatomy in Children: A Swept-Source Anterior Segment Optical Coherence Tomography Study.
To evaluate the efficacy of anterior segment swept-source optical coherence tomography (SS-OCT) for examining horizontal extraocular muscle thickness, distance from the corneal limbus to the insertion of the horizontal extraocular muscle (limbus to insertion distance), and scleral thickness in Korean pediatric strabismus patients. This study included pediatric strabismus patients between 5 and 10 years of age. Children with any ocular disease other than strabismus or a history of ocular surgery were excluded. SS-OCT was used to measure horizontal extraocular muscle thickness, limbus to insertion distance, and scleral thickness. Eyes were classified into subgroups by sex, spherical equivalent of the refractive error (measured with cycloplegic refraction), and fixating/deviating eye. One patient initially included in this study was excluded due to poor cooperation. Of the remaining 35 eyes of 20 patients, 19 eyes (54.3%) were from male patients and 16 eyes (45.7%) were from female patients. The mean patient age was 7.86 ± 1.38 years. Lateral scleral thickness was greater in male eyes than in female eyes (p = 0.048). No other differences were noted between male and female children. Additionally, there were no statistically significant differences between fixating and deviating eyes or among spherical equivalent groups for any parameter examined. Anterior segment SS-OCT can successfully and comfortably measure horizontal extraocular muscle and scleral anatomy in children. These measurements may be helpful for treatment and follow-up of pediatric strabismus patients.


Synthetic Guaiacol Derivatives as Promising Myeloperoxidase Inhibitors Targeting Atherosclerotic Cardiovascular Disease.
Myeloperoxidase (MPO) is known to cause oxidative stress and inflammation leading to cardiovascular disease (CVD) complications. MPO-mediated oxidation of lipoproteins leads to dysfunctional entities altering the landscape of lipoprotein functionality. The specificity of guaiacol derivatives toward preventing MPO-mediated oxidation to limit MPO's harmful effects is unknown. Diligent in silico studies were accomplished for a portfolio of compounds with guaiacol as a building block. The compounds' activity toward MPO inhibition was also validated. The role of these chemical entities in controlling MPO-mediated oxidation of lipoproteins (LDL and HDL) was shown to agree with our approach of developing powerful MPO inhibitors. The mechanism of MPO inhibition was demonstrated to be reversible in nature. This study reveals that there is great potential for guaiacol derivatives as therapeutics for CVD by modulating lipid profiles, reducing atherosclerotic plaque burden, and subsequently optimizing cardiovascular functions.


Influence of feeding thermally peroxidized soybean oil on growth performance, digestibility, and gut integrity in growing pigs.
Consumption of highly peroxidized oils has been shown to affect pig performance and oxidative status through the development of compounds which differ according to how oils are thermally processed. The objective of this study was to evaluate the effect of feeding varying degrees of peroxidized soybean oil (SO) on parameters of growth performance; lipid, N, and GE digestibility, gut integrity in growing pigs, and plasma Trp. Fifty-six barrows (25.3 ± 3.3 kg initial BW) were randomly assigned to one of four diets containing either 10% fresh SO (22.5 °C) or thermally processed SO (45 °C for 288 h, 90 °C for 72 h, or 180 °C for 6 h), each with an air infusion of 15 L/min. Peroxide values for the 22.5, 45, 90, and 180 °C processed SO were 2.0, 96, 145, and 4.0 mEq/kg, respectively; 2,4-decadienal values for 22.5, 45, 90, and 180 °C processed SO were 2.11,5.05, 547.62, and 323.57 mg/kg, respectively; and 4-hydroxynonenal concentrations of 0.05, 1.05, 39.46, and 25.71 mg/kg with increasing SO processing temperature. Pigs were individually housed and fed ad libitum for a 49 d period to determine the effects of SO peroxidation status on growth performance, including a metabolism period for assessing GE and N digestibility, and N retention. In vivo urinary lactulose to mannitol ratio was also assessed to evaluate potential changes in small intestinal integrity. Although there were no differences observed in ADFI (P = 0.19), ADG was decreased in pigs fed 90 °C SO diet (P = 0.01), while G:F was increased (P = 0.02) in pigs fed 45 °C SO diet compared to the other SO diets. Pigs fed the 90 °C processed SO had the lowest (P = 0.01) DE as a percentage of GE, whereas ME as a percentage of DE was lowest (P = 0.05) in pigs fed the 180 °C SO and 90 °C SO followed by 45 °C SO and fresh SO. Ether extract (EE) digestibility was lowest (P = 0.01) in pigs fed 90 °C SO followed by pigs fed 180 °C SO, 45 °C SO, and fresh SO. The percent of N retained was greatest (P = 0.01) in pigs fed fresh SO followed by pigs fed 45 °C SO, 180 °C SO, and 90 °C, respectively. There were no differences observed among SO treatments for urinary lactulose to mannitol ratio (P = 0.60). Pigs fed SO processed at 90 °C and 180 °C had lower concentrations (P < 0.01) of serum Trp compared to pigs fed the 22.5 °C and 45 °C SO treatments. The presence of lipid peroxidation products, namely several aldehydes, contained in the 90 °C SO diet reduced ADG, GE and EE digestibility, and N balance, but had no impact on gut permeability.


Interpreting γ-hydroxybutyrate concentrations for clinical and forensic purposes.
γ-Hydroxybutyric acid is an endogenous substance, a therapeutic agent, and a recreational drug of abuse. This psychoactive substance acts as a depressant of the central nervous system and is commonly encountered in clinical and forensic practice, including impaired drivers, poisoned patients, and drug-related intoxication deaths. The aim of this review is to assist clinical and forensic practitioners with the interpretation of γ-hydroxybutyric acid concentrations in blood, urine, and alternative biological specimens from living and deceased persons. The information sources used to prepare this review were PubMed, Scopus, and Web-of-Science. These databases were searched using keywords γ-hydroxybutyrate (GHB), blood, urine, alternative specimens, non-conventional biological matrices, saliva, oral fluid, sweat, hair, vitreous humor (VH), brain, cerebrospinal fluid (CSF), dried blood spots (DBS), breast milk, and various combinations thereof. The resulting 4228 references were screened to exclude duplicates, which left 1980 articles for further consideration. These publications were carefully evaluated by taking into account the main aims of the review and 143 scientific papers were considered relevant. Analytical methods: The analytical methods used to determine γ-hydroxybutyric acid in blood and other biological specimens make use of gas- or liquid-chromatography coupled to mass spectrometry. These hyphenated techniques are accurate, precise, and specific for their intended purposes and the lower limit of quantitation in blood and other specimens is 0.5 mg/L or less. Human pharmacokinetics: GHB is rapidly absorbed from the gut and distributes into the total body water compartment. Only a small fraction of the dose (1-2%) is excreted unchanged in the urine. The plasma elimination half-life of γ-hydroxybutyric acid is short, being only about 0.5-0.9 h, which requires timely sampling of blood and other biological specimens for clinical and forensic analysis. Endogenous concentrations of GHB in blood: GHB is both an endogenous metabolite and a drug of abuse, which complicates interpretation of the laboratory results of analysis. Moreover, the concentrations of GHB in blood and other specimens tend to increase after sampling, especially in autopsy cases. This requires the use of practical "cut-off" concentrations to avoid reporting false positive results. These cut-offs are different for different biological specimen types. Concentrations of GHB in clinical and forensic practice: As a recreational drug GHB is predominantly used by young males (94%) with a mean age of 27.1 years. The mean (median) and range of concentrations in blood from apprehended drivers was 90 mg/L (82 mg/L) and 8-600 mg/L, respectively. The concentration distributions in blood taken from living and deceased persons overlapped, although the mean (median) and range of concentrations were higher in intoxication deaths; 640 mg/L (280 mg/L) and 30-9200 mg/L, respectively. Analysis of GHB in alternative specimens: All biological fluids and tissue containing water are suitable for the analysis of GHB. Examples of alternative specimens discussed in this review are CSF, saliva, hair strands, breast milk, DBS, VH, and brain tissue. Body fluids for the analysis of GHB must be obtained as quickly as possible after a poisoned patient is admitted to hospital or after a person is arrested for a drug-related crime to enhance chances of detecting the drug. The sampling of urine lengthens the window of detection by 3-4 h compared with blood samples, but with longer delays between last intake of GHB and obtaining specimens, hair strands, and/or nails might be the only option. In postmortem toxicology, the concentrations of drugs tend to be more stable in bladder urine, VH, and CSF compared with blood, because these sampling sites are protected from the spread of bacteria from the gut. Accordingly, the relationship between blood and urine concentrations of GHB furnishes useful information when drug intoxication deaths are investigated.


Investigation of interfacial behavior of glycyrrhizin with a lipid raft model via a Langmuir monolayer study.
An interaction of glycyrrhizin (GC) with a lipid raft biomembrane model that consisted of N-palmitoyl-d-erythro-sphingosylphosphorylcholine (PSM), 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC), and cholesterol (CHOL) was systematically studied using the Langmuir monolayer technique. To construct the lipid raft model, the surface pressure (π)-molecular area (A) and surface potential (ΔV)-A isotherms for three-component (PSM/DOPC/CHOL) systems on 0.02M Tris buffer with 0.13M NaCl (pH7.4) were primarily measured by changing their compositions. Thermodynamic and interaction parameters for binary PSM/DOPC and PSM/CHOL systems revealed that PSM interacts more strongly with CHOL than with DOPC. In addition, a morphological analysis performed with Brewster angle microscopy (BAM) and fluorescence microscopy (FM) revealed an optimal ratio of PSM/DOPC/CHOL (1/1/1, by mole) as a model of lipid rafts. Second, the interaction of GC with the ternary PSM/DOPC/CHOL monolayers was investigated on Tris buffer solutions containing different GC concentrations (1, 5, 10, 25, and 50μM). In BAM and FM images, microdomains were found to become smaller by increasing the GC concentration in the subphase, suggesting that GC regulates the size of raft domains, which provide dynamic scaffolding for numerous cellular processes. More interestingly, the distinctive GC striped regions were formed at the interface at 50μM, which shows that GC divides the ternary monolayer into pieces. This phenomenon was observed only in the presence of CHOL in the monolayer. These results suggest that CHOL plays an essential role in the interaction with GC, which results in one of the major activities associated with saponins' membrane disruption.


Surgical Informed Consent Process in Neurosurgery.
The doctrine of informed consent, as opposed to medical paternalism, is intended to facilitate patient autonomy by allowing patient participation in the medical decision-making process. However, regrettably, the surgical informed consent (SIC) process is invariably underestimated and reduced to a documentary procedure to protect physicians from legal liability. Moreover, residents are rarely trained in the clinical and communicative skills required for the SIC process. Accordingly, to increase professional awareness of the SIC process, a brief history and introduction to the current elements of SIC, the obstacles to patient autonomy and SIC, benefits and drawbacks of SIC, planning of an optimal SIC process, and its application to cases of an unruptured intracranial aneurysm are all presented. Optimal informed consent process can provide patients with a good comprehension of their disease and treatment, augmented autonomy, a strong therapeutic alliance with their doctors, and psychological defenses for coping with stressful surgical circumstances.


Statistical Analysis of the Impact of Molecular Descriptors on Cytotoxicity of Thiourea Derivatives Incorporating 2-Aminothiazole Scaffold.
Chemical reactivity descriptors and lipophilicyty (log P) were evaluated via semi-empirical method for the quantum calculation of molecular electronic structure (PM3) in order to clarify the structure-cytotoxic activity relationships of disubstutited thioureas. Analysed compounds were obtained by the linkage of 2-aminothiazole ring, thiourea and substituted phenyl ring. The detailed examination was carried out to establish correlation between descriptors and cytotoxic activity against the MT-4 cells for 11 compounds. For the most active compounds (6 compounds) cytotoxic activity against three cancer cell lines (CCRF-CEM, WIL-2NS, CCRF-SB) and normal human cell (HaCaT) was determined. 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) reduction and lactate dehydrogenase (LDH) release were assessed. Regression analysis revealed that electrophilicity index and chemical potential significantly contributed to expain the thioureas cytotoxic potential.


Index distribution of gaussian random matrices.
We compute analytically, for large N, the probability distribution of the number of positive eigenvalues (the index N+) of a random N x N matrix belonging to Gaussian orthogonal (beta=1), unitary (beta=2) or symplectic (beta=4) ensembles. The distribution of the fraction of positive eigenvalues c=N+/N scales, for large N, as P(c,N) approximately = exp[-betaN(2)Phi(c)] where the rate function Phi(c), symmetric around c=1/2 and universal (independent of beta), is calculated exactly. The distribution has non-Gaussian tails, but even near its peak at c=1/2 it is not strictly Gaussian due to an unusual logarithmic singularity in the rate function.


Gout: state of the art after a decade of developments.
This review article summarizes the relevant English literature on gout from 2010 through April 2017. It emphasizes that the current epidemiology of gout indicates a rising prevalence worldwide, not only in Western countries but also in Southeast Asia, in close relationship with the obesity and metabolic syndrome epidemics. New pathogenic mechanisms of chronic hyperuricaemia focus on the gut (microbiota, ABCG2 expression) after the kidney. Cardiovascular and renal comorbidities are the key points to consider in terms of management. New imaging tools are available, including US with key features and dual-energy CT rendering it able to reveal deposits of urate crystals. These deposits are now included in new diagnostic and classification criteria. Overall, half of the patients with gout are readily treated with allopurinol, the recommended xanthine oxidase inhibitor (XOI), with prophylaxis for flares with low-dose daily colchicine. The main management issues are related to patient adherence, because gout patients have the lowest rate of medication possession ratio at 1 year, but they also include clinical inertia by physicians, meaning XOI dosage is not titrated according to regular serum uric acid level measurements for targeting serum uric acid levels for uncomplicated (6.0 mg/dl) and complicated gout, or the British Society for Rheumatology recommended target (5.0 mg/dl). Difficult-to-treat gout encompasses polyarticular flares, and mostly patients with comorbidities, renal or heart failure, leading to contraindications or side effects of standard-of-care drugs (colchicine, NSAIDs, oral steroids) for flares; and tophaceous and/or destructive arthropathies, leading to switching between XOIs (febuxostat) or to combining XOI and uricosurics.


Parallax diagnostics of radiation source geometric dilution for iron opacity experiments.
Experimental tests are in progress to evaluate the accuracy of the modeled iron opacity at solar interior conditions [J. E. Bailey et al., Phys. Plasmas 16, 058101 (2009)]. The iron sample is placed on top of the Sandia National Laboratories z-pinch dynamic hohlraum (ZPDH) radiation source. The samples are heated to 150-200 eV electron temperatures and 7× 10(21)-4× 10(22) cm(-3) electron densities by the ZPDH radiation and backlit at its stagnation [T. Nagayama et al., Phys. Plasmas 21, 056502 (2014)]. The backlighter attenuated by the heated sample plasma is measured by four spectrometers along ±9° with respect to the z-pinch axis to infer the sample iron opacity. Here, we describe measurements of the source-to-sample distance that exploit the parallax of spectrometers that view the half-moon-shaped sample from ±9°. The measured sample temperature decreases with increased source-to-sample distance. This distance must be taken into account for understanding the sample heating.


Three new dicyemids from Octopus sasakii (Mollusca: Cephalopoda: Octopoda).
Three new species of dicyemid mesozoan are described from Octopus sasakii Taki, 1942, collected from Tosa Bay and Kii Strait in Japan. Dicyema shimantoense n. sp. is a medium-size species that reaches about 3,000 microm in length, and lives in folds of the renal appendages. The vermiform stages are characterized by having 22 peripheral cells, a conical calotte, and an axial cell that extends to the base of the propolar cells. Infusoriform embryos consist of 37 cells; a single nucleus is present in each urn cell, and the refringent bodies are solid. Dicyema codonocephalum n. sp. is also a medium-size species that reaches about 2,000 microm in length. It too lives in folds of the renal appendages. The vermiform stages are characterized by having 17-22 peripheral cells, an elongated calotte, and an axial cell that extends to the middle of propolar cells. Infusoriform embryos consist of 37 cells; a single nucleus is present in each urn cell, and the refringent bodies are solid. Dicyemennea pileum n. sp. is a medium species that reaches about 2,000 microm in length. It attaches to the surface of the renal appendages. The vermiform stages are characterized by having 23 peripheral cells, a disc-shaped calotte, and an axial cell that extends to the propolar cells. An anterior abortive axial cell is absent in vermiform embryos. Infusoriform embryos consist of 38 cells; 2 nuclei are present in each urn cell, and the refringent bodies are liquid. These are the first dicyemids to be described from Octopus sasakii.


Diagnostic Accuracy of Monocyte Chemotactic Protein (MCP)-2 as Biomarker in Response to PE35/PPE68 Proteins: A Promising Diagnostic Method for the Discrimination of Active and Latent Tuberculosis.
Several studies have been conducted to find new biomarkers for the discrimination of Latent Tuberculosis Infection (LTBI) from active TB (ATB); however, their findings are inconsistent. The aim of the current study was to evaluate the potential of in vitro antigenspecific expression of Monocyte Chemotactic Protein (MCP)-2 for discrimination of ATB and LTBI after stimulation of whole blood with PE35 and PPE68 recombinant proteins. The recombinant PE35 and PPE68 proteins were evaluated at a final concentration of 5 µg/ml by a 3-day whole blood assay. Secreted MCP-2 from the culture supernatants were measured by commercially available Human MCP2 ELISA Kit. The diagnostic performance of MCP-2 was ascertained by Receiver Operator Characteristic (ROC) curve and measuring the Area Under the Curve (AUC) and their 95% Confidence Intervals (CI). Cut-offs was estimated at various sensitivities and specificities and at the maximum Youden's index (YI), i.e. sensitivity specificity-1. The median MCP-2 response to both PE35 and PPE68 in those with LTBI was significantly higher than patients with ATB. The discrimination performance of MCP-2 response following stimulation of PE35 (assessed by AUC) between LTBI and patients with ATB was 0.98 (95%CI: 0.94-1.00). Maximum discrimination was reached at a cut-off of 86pg/mL with 100% sensitivity and 97% specificity. The highest sensitivity and specificity was obtained using cut off 58 pg/mL following stimulation with PPE68 (100% and 90%, respectively; AUC: 0.94, 95%CI: 0.85- 1.00). MCP-2 induced by PE35 and PPE68 shows good discriminatory power for discrimination of ATB and LTBI. Additional studies with a larger sample size are needed to confirm the advantage of this marker, alone or combined with other markers; however, these findings present a promising method, which can discriminate between ATB and LTBI.


[Effects of insulin on proteolysis of cultured rabbit skeletal muscular myotubes in vitro].
To study the effects of insulin on the proteolysis of cultured rabbit skeletal muscular myotubes in vitro, and their possible mechanisms. Muscles of lower limbs of juvenile rabbits were isolated for tissue-block culture. After passage, myoblasts were formed and fused into myotubes. Then the protein in myotubes was radiolabelled with L-[ 3,5-3H] tyrosine. The myotubes were cultured in DMEM medium containing 100 nmol/L insulin (n = 24, group B) , 100 nmol/L dexamethasone (n = 24, group C) , 100 nmol/L insulin and 100 nmol/L dexamethasone (n = 24, group D) , no insulin or dexamethasone (n =24, group A), respectively. Twenty-four hours after culture, the L-[3,5-3H] tyrosine content in culture medium and cells were determined, and the degradation rates of protein were calculated. The mRNA expressions of ubiquitin and protease C2 subunit were determined by Northern blot. The degradation rates of myotube protein in group A(0. 38+/-0.04) was obviously lower than that in group C (0.50+/-0.03, P <0.01), but it was obviously higher than that in group B(0. 35+/-0.03, P <0.05). Though the degradation rates of myotube protein in group D (0.41+/-0. 03) was evidently lower than that in group C ( P < 0.01) , it was still higher than that in group A( P < 0.05 ). The mRNA expressions of ubiquitin and protease C2 subunit in group A ( the scale: 2. 4 kb ubiquitin was 0. 82+/-0. 15, 1. 2 kb ubiquitin was 0. 60+/-0. 10, C2 subunit was 0. 75+/-0. 16) was obviously lower than that in group C ( the scale: 2.4 kb ubiquitin was 2. 15+/-0. 23, 1.2 kb ubiquitin was 1.50+/-0. 14,C2 subunit was 1.50+/-0. 13 , P <0. 01) , but it in group D was lower than that in group C (the scale: 2. 4 kb ubiquitin was 1. 25+/-0. 17, 1. 2 kb ubiquitin was 0. 85+/-0. 09, C2 subunit was 0. 90+/-0. 15, P <0. 01) , and it was similar to that in group B (the scale: 2.4 kb ubiquitin was 0. 85+/-0.07, 1.2 kb ubiquitin was 0. 65+/- 0. 12, C2 subunit was 0. 76 +/-0. 09, P > 0. 05). The effects of insulin on the activity of ubiquitin-proteasome pathway and the proteolytic rate in normal myotubes were relatively weak. However, insulin can significantly inhibit the effects of dexamethasone on the gene expressions of ubiquitin system and the proteolytic rate in myotubes, but the mechanism needs further research.


Prognostic factors affecting response to aminoglutethimide in advanced breast cancer.
An analysis of 122 patients with advanced breast cancer treated with aminoglutethimide was carried out. Forty-one patients (34%) achieved a complete response or partial response (CR + PR), while 81 patients (66%) remained stable or progressed (S + P). A number of variables were analyzed for their prognostic importance in terms of response to aminoglutethimide. These variables were dominant site of metastases, disease-free survival, estrogen receptor status, age, response to prior tamoxifen, response to chemotherapy administered for metastatic disease, and dose of aminoglutethimide. The most important factors in predicting response were found to be prior response to tamoxifen and a longer disease-free survival after diagnosis.


p16INK4A and CDH13 hypermethylation in tumor and serum of non-small cell lung cancer patients.
Aberrant promoter hypermethylation of several known or putative tumor suppressor genes occurs frequently during the etiopathogenesis of lung cancer and is a promising tool for cancer detection. In the present study, promoter hypermethylation of p16INK4A and CDH13 genes was investigated in tumor tissue and in matched serum from 61 patients with histologically confirmed non-small cell lung cancer. Using a fluorescence-based method of methylation-specific PCR (F-MSP), methylation of p16INK4A and CDH13 was detected in 79% and 66% of tumors, respectively, and was not significantly related to conventional clinicopathological characteristics of patients or tumors. Methylation of both genes was observed in 52% of tumors and of at least one gene in 92% of lesions. In matched serum, hypermethylation of p16INK4A and CDH13 was observed in 26% and 23% of patients, respectively, but as they were not associated, the methylation of at least one gene was detected in 39% of patients. In conclusion, the frequency of p16INK4A or CDH13 hypermethylation in patient serum, together with evidence of their early occurrence in lung cancerogenesis and the total lack of methylation in serum from healthy individuals, offer a promising tool for non invasive early detection of lung cancer.


Immunocytochemical correlates of an extrapituitary adrenocortical regulation in man.
Investigations reviewed in this article provide cytochemical and functional support for a significant involvement of extrapituitary factors in human adrenocortical functions. Among these factors neural messengers may play a crucial role in the adrenocortical regulation, arising from specifically coded postganglionic neurons with both, extrinsic and intrinsic locations, as well as from chemically characteristic afferent neurons. The close association of varicose transmitter segments with steroid hormone synthesizing cells and their occurrence at arteries and sinusoid capillaries are indicative for both direct and indirect regulatory mechanisms on cortical functions. The immunohistochemical presence of neuropeptides and cytokines in endocrine and/or immune cells of the human adrenal medulla and cortex as well as specific binding sites on steroidogenic cells indicate the modulatory implication of additional short-paracrine- and ultrashort-autocrine-feedback loops on cortical cell proliferation and steroid metabolism. The summarized data suggest that basal endocrine influence of the hypothalamo-pituitary axis on adrenocortical growth and functions in man is controlled by the nervous system that also regulates local fine-tuning of human cortical activity.


The insulin receptor of a human monocyte-like cell line: characterization and function.
Most clinical studies of the insulin receptor in man have been carried out in circulating cells, mainly monocytes. It is important to establish whether the function of the insulin receptor in blood cells reflects that of the major target tissues. Since peripheral monocytes do not divide, they cannot be studied under continuous cell culture conditions. We, therefore, studied the insulin receptor of a monocyte-like human cell line (U-937) to determine whether it could serve as a model for further investigation of the function of the insulin receptor. We found that the U-937 cells bind [125I]insulin specifically and in a time-, temperature-, concentration-, and pH-dependent fashion, similar to circulating monocytes. In addition, they internalize the hormone-receptor complex rapidly and extensively at 37 C in a fashion analogous to that of blood monocytes and hepatocytes. U-937 cells show a similar affinity for insulin as monocytes. Unlike hepatocytes, these cells do not appear to release extensive insulin-degrading activity and do not degrade cell surface-associated ligand during initial incubations. [125I] Insulin extracted or dissociated spontaneously from the cell surface shows full rebindability to fresh cells. The insulin receptor of the U-937 cells can be down-regulated as in the major target cells. As in cultured human lymphocytes, preincubation of U-937 cells with prednisolone results in significantly increased insulin binding. Incubation with a phorbol ester tumor promoter, in contrast, inhibits the extent of specific [125I]insulin binding to U-937 cells by altering the receptor affinity. The insulin receptor of the U-937 monocyte-like cell line mirrors the insulin receptor of blood monocytes and target cells; it is a useful tool for further in vitro studies of the insulin receptor.


Direct observation of temperature-driven magnetic symmetry transitions by vectorial resolved MOKE magnetometry.
Angle- and temperature-dependent vectorial magnetometry measurements are necessary to disentangle the effective magnetic symmetry in magnetic nanostructures. Here we present a detailed study on an Fe(1 0 0) thin film system with competing collinear biaxial (four-fold symmetry) and uniaxial (two-fold) magnetic anisotropies, carried out with our recently developed full angular/broad temperature range/vectorial-resolved magneto-optical Kerr effect magnetometer, named TRISTAN. The data give direct views on the angular and temperature dependence of the magnetization reversal pathways, from which characteristic axes, remanences, critical fields, domain wall types, and effective magnetic symmetry are obtained. In particular, although the remanence shows four-fold angular symmetry for all investigated temperatures (15 K-400 K), the critical fields show strong temperature and angular dependencies and the reversal mechanism changes for specific angles at a given (angle-dependent) critical temperature, showing signatures of an additional collinear two-fold symmetry. This symmetry-breaking is more relevant as temperature increases to room temperature. It originates from the competition between two anisotropy contributions with different symmetry and temperature evolution. The results highlight the importance of combining temperature and angular studies, and the need to look at different magnetic parameters to unravel the underlying magnetic symmetries and temperature evolutions of the symmetry-breaking effects in magnetic nanostructures.


Responses of the entomopathogenic nematode, Steinernema riobrave to its insect hosts, Galleria mellonella and Tenebrio molitor.
Potential hosts for infective juveniles of entomopathogenic nematodes can vary considerably in quality based on the characteristics of the host species/stage, physiological status (e.g. stress, feeding on toxins), and infection status (heterospecific or conspecific infection). In this study, we investigated responses of the entomopathogenic nematode Steinernema riobrave to hosts (Galleria mellonella or Tenebrio molitor) that were previously parasitized with conspecifics or injected with the nematode-symbiotic bacterium, Xenorhabdus sp., to determine if there is a preference for previously parasitized/injected hosts and when this preference might occur. In no-choice bioassays, the number of juveniles infecting both host species decreased with increasing time post-infection. However, infective juveniles continued to infect previously parasitized hosts up to 72 h. Significant preference was exhibited by S. riobrave for 24 h post-infection G. mellonella larvae over uninfected, and by 24 h post-injection G. mellonella larvae over 48 h post-injection larvae. No significant preference was exhibited by S. riobrave for T. molitor hosts previously parasitized with conspecifics or those injected with bacteria in any treatment combination. Such preference for, or continued infection of parasitized insects, has the potential to impact nematode efficacy.


Assembly of water-soluble chlorophyll-binding proteins with native hydrophobic chlorophylls in water-in-oil emulsions.
The challenges involved in studying cofactor binding and assembly, as well as energy- and electron transfer mechanisms in the large and elaborate transmembrane protein complexes of photosynthesis and respiration have prompted considerable interest in constructing simplified model systems based on their water-soluble protein analogs. Such analogs are also promising templates and building blocks for artificial bioinspired energy conversion systems. Yet, development is limited by the challenge of introducing the essential cofactors of natural proteins that are highly water-insoluble into the water-soluble protein analogs. Here we introduce a new efficient method based on water-in-oil emulsions for overcoming this challenge. We demonstrate the effectiveness of the method in the assembly of native chlorophylls with four recombinant variants of the water-soluble chlorophyll-binding protein of Brassicaceae plants. We use the method to gain new insights into the protein-chlorophyll assembly process, and demonstrate its potential as a fast screening system for developing novel chlorophyll-protein complexes.


Diffusion and perfusion MRI in patients with ruptured and unruptured intracranial aneurysms treated by endovascular coiling: complications, procedural results, MR findings and clinical outcome.
Our purpose was to evaluate treatment safety as well as complications frequency and management in endovascular coiling of intracerebral aneurysms using MR diffusion and perfusion imaging. In this prospective study, 77 MR examinations were performed in conjunction with 43 procedures in 40 patients, 14 patients presented with ruptured and 26 with unruptured aneurysms. Mean time interval between treatment and post-procedure MRI was 29 and 25 h for the ruptured and unruptured aneurysm group, respectively. Peri-procedural complications, including five major events and five minor transient events, occurred in 10/43 procedures (23%), necessitating thrombolytic therapy in two patients and angioplasty in one, all three within the unruptured aneurysm group. Fifty-one new lesions were found on post-treatment DWI and 47 of them were regarded as of ischemic origin. Most lesions were small (<3 mm), ipsilateral to the treated aneurysm and asymptomatic (37/40 patients). Sixty-seven percent of the lesions were found in the ruptured and 33% in the unruptured aneurysm group. The ischemic lesions did occur more frequently in patients treated for aneurysm of large neck size and according to the remodelling technique. The overall morbidity and mortality rates were 14.6 and 7.3% whereas morbidity and mortality rates related to the technique were only 2.6 and 0%, respectively. Silent embolism seems to be more common than clinically evident and partially related to patient presentation, heparinazation and treatment strategy. The capability to depict early complications and analyse their potential causes by using MR with DWI has been of great importance in our modification and improvement of therapeutic protocols, evaluations and strategies.


A tightly regulated molecular motor based upon T7 RNA polymerase.
Controlled movement of materials or molecules within the nanometer range is essential in many applications of nanotechnology. Here we report the capture, movement, and release of cargo molecules along DNA by a modified form of T7 RNA polymerase (RNAP) in a manner that is controlled by the sequence of the DNA. Using single-molecule methods, we visualize the assembly and manipulation of nanodevices and the ability to harness rotary and linear forces of the RNAP motor.