Comparison of ECG variables of dispersion of ventricular repolarization with direct myocardial repolarization measurements in the human heart.
QT dispersion (QTD) from the 12-lead ECG has been widely adopted as a noninvasive index of dispersion of ventricular repolarization (DVR). QTD, however, has never been validated by direct comparison with myocardial DVR in the human heart. Monophasic action potential (MAP) recordings obtained in an earlier study were retrospectively matched with 12-lead ECGs available from within 24 hours of the invasive procedure. MAPs were available from an average of 8+/-3 left endocardial sites in 4 patients with left ventricular hypertrophy (LVH) and 7 patients with normal ECGs, and 6+/-2 epicardial sites in 3 patients of each group during normal ventricular activation. Local repolarization time (RT) was determined as MAP duration at 90% repolarization plus the local activation time. Dispersion of RT was calculated as the difference between the earliest and latest RT. ECGs were digitized and analyzed with recently described interactive QTD analysis software. In addition to standard QTD (defined as QTmax-QTmin), all currently proposed ECG dispersion variables were compared and correlated with the invasive measurements of DVR. QTD exhibited a reasonable correlation with dispersion of RT (R = 0.67; P < 0.01). Several other variables designed to measure DVR exhibited a similar, but not better, correlation. Among them, the QT peak/QT end ratio in V3 (R = -0.72; P < 0.01) and averaged over all analyzable leads (R = -0.59; P < 0.01) exhibited a good correlation with dispersion of RT, which was further improved when endocardial measurements were considered alone. T area measures did not correlate with dispersion of RT, but discriminated LVH. DVR can be assessed by means of a 12-lead surface ECG. Several of the variables under study exhibit a similar accuracy in determination of true myocardial dispersion of repolarization. Variables involving the terminal part of repolarization, such as the QT peak/QT ratio, even from a single lead, may add to the determination of DVR from the human heart.


Incidence and risk factors of early deep venous thrombosis after varicose vein surgery with routine use of a tourniquet.
The incidence of early deep venous thrombosis (DVT) following varicose vein surgery (traditional open stripping) with routine use of a tourniquet remains unknown. A retrospective analysis of all patients who underwent varicose vein surgery with a tourniquet in the authors' unit between 1 January 2012 and 30 November 2013 was undertaken. Cases of postoperative DVT were identified from the unit database, and re-assessments conducted 1, 3 and 6 months after the initial diagnosis were recorded from the outpatient department. Out of 1461 patients, 113 (7.7%) developed postoperative DVT. Nineteen (1.3%) patients had proximal DVT, and 94 (6.4%) patients had isolated distal DVT. The risk factors for postoperative DVT included old age (≥65 years), female sex and gastrocnemius vein dilation (GVD). GVD was found to be a significant independent risk factor for the occurrence of DVT, with an odds ratio of 2.437 (95% confidence interval 1.644-3.611). Five patients with distal DVT (5.7%) and eight patients with proximal DVT (44.4%) still exhibited a thrombus at 6-month follow-up, but with decreased size and at various stages of resolution. This study found a higher incidence of postoperative DVT (7.7%) with routine use of a tourniquet during varicose vein surgery than has been reported previously. Among the factors examined, GVD had the highest predictive power for postoperative DVT. Both distal and proximal DVT were associated with acceptable outcomes.


The Shortened Dental Arch Concept: Awareness, Knowledge, and Practice of Dentists in Dubai and the Northern Emirates, United Arab Emirates.
This is a cross-sectional study utilizing an online questionnaire anonymously to investigate the awareness and views of dentists about SDA. The questionnaire was sent to all 901 dentists registered with the Emirates Medical Association (EMA). The questionnaire consists of 17 questions, which comprise demographics, awareness, and application in dental practice, preferred treatment modality, and risks and benefits associated with SDA. The data were analyzed using SPSS Statistics. The response rate reported was 40.3%. Two-thirds of the respondents (65.8%) were aware of the SDA concept; however, it was not usually applied in clinical practice (n = 196, 54.7%). Specialists were more aware of the concept (p ≤ 0.001) and applied it more frequently in their clinical practice (p=0.041) than general dental practitioners (GDPs). Respondents agreed that SDA was associated with the risks of teeth migration (n = 211, 59.9%), tooth wear (n = 196, 55.8%), and/or temporomandibular disorder (TMD) (n = 163, 45.3%). The implant was the treatment of choice for many of the participants (n = 169, 46.6%) to replace missing molars, followed by the acrylic removal partial denture (RPD) (n = 129, 35.5%). Most dentists who responded to this survey were aware of the SDA concept and had a positive attitude about it. However, they did not apply it frequently in their clinical practice.


Variance parameter estimation for age at puberty phenotypes under 2 levels of phenotype censorship.
Age at puberty (AGEP) is a moderately heritable trait in cattle that may be predictive of an animal's genetic merit for reproductive success later in life. In addition, under some mating strategies (for example, where mating begins before all animals have attained puberty) animals that attain puberty at a relatively young age will also likely conceive earlier than their herd mates, and thus begin their productive life earlier. Unfortunately, AGEP is challenging to measure because animals must be observed over a period of several months. Our objectives for this study were twofold. The first objective was to produce variance components for AGEP. The second objective was to investigate the implications of a simplified phenotyping strategy for AGEP, when the interval between repeated blood plasma progesterone measures was extended from weekly to monthly, increasing the extent of left, interval, and right censoring. We measured AGEP in a closely monitored population of around 500 Holstein-Friesian heifers, born in 2015 and managed under a seasonal, pasture-based dairy system. Animals were blood tested weekly from approximately 240 to 440 d of age and were deemed to have reached puberty when blood plasma progesterone elevation (>1 ng/mL) was detected in 2 of 3 consecutive blood tests (AGEP_Weekly). To simulate a simplified phenotyping strategy based on monthly herd visits (AGEP_Monthly), we selectively disregarded data from all but 3 blood test events, when animals were around 300, 330, and 360 d of age (standard deviation = 14.5 d). The posterior mean of estimated heritabilities for AGEP_Weekly was 0.54, with a 90% credibility interval (90% CRI) of 0.41 to 0.66, whereas it was 0.44 (90% CRI 0.32 to 0.57) for AGEP_Monthly. The correlation between EBVs for AGEP_Weekly and AGEP_Monthly was 0.87 (90% CRI, 0.84 to 0.89). We conclude that in this population, AGEP is a moderately heritable trait. Further, increasing phenotype censorship from weekly to monthly observations would not have altered the main conclusions of this analysis. Our results support the strategic use of censoring to reduce costs and animal ethics considerations associated with collection of puberty phenotypes.


Exact solutions to the continuous-quality equation for soil organic matter turnover.
All living systems depend on transformations of elements between different states. In particular, the transformation of dead organic matter in the soil (SOM) by decomposers (microbes) releases elements incorporated in SOM and makes the elements available anew to plants. A major problem in analysing and describing this process is that SOM, as the result of the decomposer activity, is a mixture of a very large number of molecules with widely differing chemical and physical properties. The continuous-quality equation (CQE) is a general equation describing this complexity by assigning a continuous-quality variable to each carbon atom in SOM. The use of CQE has been impeded by its complicated mathematics. Here, we show by deriving exact solutions that, at least for some specific cases, there exist solutions to CQE. These exact solutions show that previous approximations have overestimated the rate by which litter decomposes and as a consequence underestimated steady state SOM amounts. The exact and approximate solutions also differ with respect to the parameter space in which they yield finite steady-state SOM amounts. The latter point is important because temperature is one of the parameters and climatic change may move the solution from a region of the parameter space with infinite steady-state SOM to a region of finite steady-state SOM, with potentially large changes in soil carbon stores. We also show that the solution satisfies the Chapman-Kolmogorov theorem. The importance of this is that it provides efficient algorithms for numerical solutions.


Women's Participation in a Savings Group and Depression: a Community-Based Financial Capability Intervention in Mozambique.
As one of the fastest growing community-based financial capability approaches, village savings and loan group (VSLG) is an organized group and formal entity that creates opportunities for participants to save and access financial assets. VSLG has potentially positive impacts on increasing women's financial resources and social support and further improves their mental health. Participation in a VSLG not only increases women's opportunities for asset-building and income generation, but also facilitates trust and promotes social capital development. However, few studies have examined the association between the VSLG participation and women's depression status. To fill the knowledge gap, we examined the association between the VSLG participation and depressive symptoms among low-income women in Mozambique. The data was collected as part of the VSLG program evaluation. The study applied a posttest-only comparison group quasi-experimental design and sampled female VSLG participants and non-participants from three sub-villages in the Sofala province, Mozambique. A multi-stage sampling technique was employed, and a total of 205 women were randomly selected, including 105 VSLG participants and 100 non-participants. Depressive symptoms were measured using the short version of the Edinburgh Depression Scale (EDS) with a summative score ranging from 7 to 28. Using the cutoff value at a score of 14, we created a dichotomous depression indicator. Linear and logit models were used to examine the associations of the VSLG participation and the participation duration with the depression score and the presence of depression controlling for demographic variables. The VSLG participants had a statistically lower mean depression score of 12.2 (SD = 4.4) compared to non-participants (15.0, SD = 4.0, p < .001). The regression analysis suggested that the VSLG participants had a mean depression score of 2.7 lower than the non-participants (p < .001). Nearly 60% of the non-participants reported the presence of depression; however, this percentage is 31% for participants (p < .001). The multivariate logit model indicated the odds of the presence of depression for participants are .34 of that for non-participants. Similar results were obtained when the VSLG program duration was used as an independent variable. Study findings showed a positive association between the VSLG participation and women's mental health. Future research should further explore the intervention mechanisms and assess how the VSLG participation affects women's mental health. Findings also provided important insights into developing community-based financial capability interventions to improve low-income women's mental health.


miR-196a downregulation increases the expression of type I and III collagens in keloid fibroblasts.
Keloids are a fibroproliferative disease due to abnormal wound healing process after skin injury. They are characterized by overproduction of extracellular matrix (ECM) such as collagens. MicroRNAs (miRNAs) are noncoding small RNAs and negatively regulate protein expression. Several miRNAs that have critical roles in tissue fibrosis and ECM metabolism have been reported. However, regulation and function of miRNAs in keloid remain to be explored. The purpose of this study was to identify miRNAs involved in keloid pathogenesis. We performed miRNA microarray analysis to compare miRNA expression profiles between keloid-derived fibroblasts (KFs) and normal fibroblasts (NFs). In all, 7 upregulated and 20 downregulated miRNAs were identified. Among these, we focused on miR-196a, which showed the highest fold change. Overexpression or knockdown of miR-196a led to a decreased or increased level of secreted type I/III collagens, respectively. Reporter analysis showed direct binding of miR-196a to the 3' untranslated region (UTR) of COL1A1 and COL3A1. In conclusion, we demonstrate for the first time that miRNA expression profile is altered in KFs compared with NFs. Downregulation of miR-196a may be one of the mechanisms by which collagens are highly deposited in keloid tissues. Our findings suggest that miR-196a could be a new therapeutic target for keloid lesions.


Cellular uptake and photosensitizing properties of anticancer porphyrins in cell membranes and low and high density lipoproteins.
The mechanisms of the phototoxic effect of anticancer porphyrins used in the photodynamic therapy (PDT) of tumours are not yet completely understood. Irradiation of porphyrins gives rise to singlet oxygen which reacts with key residues of proteins, polyunsaturated fatty acids and cholesterol in membranes, leading to inactivation of various enzymes and transporters. Lipoproteins, mainly low density lipoproteins (LDL), are efficient carriers of anticancer porphyrins in blood and can deliver these photosensitizers to tissues through the apolipoprotein (apo) B/E specific LDL receptor pathway. In this review, we discuss some aspects of anticancer porphyrin transport, cellular uptake and photosensitizing properties in cell membranes and lipoproteins.


Developmentally regulated processing and replication of the Tetrahymena rDNA minichromosome.
The ribosomal DNA locus of Tetrahymena thermophila undergoes a dramatic series of developmentally regulated processing events to generate the amplified rDNA minichromosome during formation of the somatic macronucleus. DNA transformation and classical genetic approaches have identified cis-acting elements that regulate rDNA processing in the developing macronucleus and subsequent vegetative rDNA maintenance.


Necator americanus: optimization of the golden hamster model for testing anthelmintic drugs.
Laboratory golden hamsters (Mesocricetus auratus) were infected with Necator americanus under several different parasite and host conditions to optimize the model for testing anthelminthic drugs. The results confirmed that male hamsters were more susceptible to infection than females. Host age in the range of 5-15 weeks was not a factor that impacted on adult worm burden, and similar worm burdens were achieved using doses of 150, 250 or 500 N. americanus L3 (NaL3). The largest numbers of adult hookworms were recovered on days 21-28 post-infection, with a significant decrease at days 40-50 post-infection. Therefore adult worm recovery is maximal approximately 11-18 days prior to patency and host blood loss. From these studies a drug evaluation protocol was developed using 150 NaL3 as the infectious dose and then evaluating the anthelminthic effects of the drugs albendazole, tribendimidine, and pyrantel pamoate on days 21-28 post-infection. The model confirms the anthelminthic activity of albendazole, tribendimidine, and pyrantel pamoate and has the potential as a laboratory animal model to detect emerging drug resistance.


Seizure-induced rhabdomyolysis accompanied by acute renal failure in a dog.
Secondary complications from extreme and prolonged convulsive seizures occurred in a young dog. Physical exertion resulted in necrosis of a substantial amount of skeletal muscle. The release of large quantities of myoglobin into the circulation caused severe impairment of renal function. Anoxia, apparently associated with the inability to breathe during seizures, resulted in brain damage. It was concluded that, although primary therapy must be directed toward alleviation of intense seizure activity, subsequent screening for renal disease and corollary supportive therapy would be appropriate.


Effectiveness of dinoprostone gel, misoprostol vaginal insert and dinoprostone vaginal insert for induction of labour in twin pregnancies.
To compare the effectiveness and safety of Dinoprostone Gel (DG), Misoprostol Vaginal Insert (MVI) and Dinoprostone Vaginal Insert (DVI) for induction of labour (IOL) in twin pregnancies. Retrospective cohort study of twin pregnancies > 34 + 0 weeks gestation that underwent induction of labour (IOL) with DG, MVI or DVI between December 2016 and November 2019 in a Tertiary NHS hospital, North West England, UK. Delivery characteristics, maternal complications and neonatal outcomes were compared between the three groups. A total of 87 twin pregnancies were included for analysis. 27 women received DG, 34 received MVI and 26 DVI. The MVI cohort had a higher proportion of nulliparous women (55.9%) compared to the DG and DVI cohorts, 29.6% and 38.5% respectively. No other differences amongst demographic characteristics were considered clinically significant. DG demonstrated a significantly quicker time to delivery (minutes) compared to DVI (1021 ± 556 versus 1649 ± 852; P = 0.0026). Significantly fewer women required terbutaline for hyperstimulation/tachysystole in the DG group compared to MVI (0% vs 32%; RR 0.05; 95% CI 0.003-0.88). Both DG and MVI groups required significantly less oxytocin following artificial rupture of membranes compared to DVI (33% vs 65%; RR 0.51; 95% CI 0.28-0.93) and (29% vs 65%; RR 0.45; 95% CI 0.25-0.81). There were no significant differences in mode of delivery, maternal complications and neonatal outcomes. Our data suggests that for women with a twin pregnancy considering a planned labour that induction with DG, MVI and DVI appear to be equally safe and effective IOL methods. These results should be interpreted with caution due to the study being underpowered to detect significant adverse outcomes. In order to determine the optimal method of IOL in twins, direct randomised comparison is needed.


The effect of environment on the recognition and binding of vancomycin to native and resistant forms of lipid II.
Molecular dynamics simulations and free energy calculations have been used to examine in detail the mechanism by which a receptor molecule (the glycopeptide antibiotic vancomycin) recognizes and binds to a target molecule (lipid II) embedded within a membrane environment. The simulations show that the direct interaction of vancomycin with lipid II, as opposed to initial binding to the membrane, leads most readily to the formation of a stable complex. The recognition of lipid II by vancomycin occurred via the N-terminal amine group of vancomycin and the C-terminal carboxyl group of lipid II. Despite lying at the membrane-water interface, the interaction of vancomycin with lipid II was found to be essentially identical to that of soluble tripeptide analogs of lipid II (Ac-d-Ala-d-Ala; root mean-square deviation 0.11 nm). Free energy calculations also suggest that the relative binding affinity of vancomycin for native, resistant, and synthetic forms of membrane-bound lipid II was unaffected by the membrane environment. The effect of the dimerization of vancomycin on the binding of lipid II, the position of lipid II within a biological membrane, and the effect of the isoamylene tail of lipid II on membrane fluidity have also been examined.


[The differential diagnosis in Menières disease: the basilar impression (author's transl)].
We examined the cranio-cervical region in 180 patients with nonspecific dizziness and in most cases unilateral sensory-neural hearing loss. In 32 cases we found malformations of the cranio-cervical region. The sympatomatology of the basilar impression is probably caused by the compression of the vessels of the lower cerebellar regions and the brain stem as well as disturbances of the cerebro-spinal fluid circulation. The basilar impression is diagnosed by means of lateral X-rays of the skull base, tomography of this region and in some cases computertomography of the posterior cranial fossa. The importance of basilar impression in the differential diagnosis of Menières disease and acoustic neuroma is discussed.


Essential role for ERK mitogen-activated protein kinase in matrix metalloproteinase-9 regulation in rat cortical astrocytes.
Matrix metalloproteinases (MMPs) contribute to the pathophysiology of brain injury and inflammation but little is known about their regulatory signaling pathways in brain cells. Here we examine the role of mitogen-activated protein (MAP) kinase pathways in MMP-9 regulation in cortical rat astrocytes. The protein kinase C (PKC) activator phorbol 12-myristate 13-acetate (PMA) induced MMP-9 but not MMP-2 secretion as measured by gelatin zymography. Northern blot and RT-PCR analysis showed that MMP-9 responses occurred at the mRNA level. Although PMA increased phosphorylation in all three major MAP kinase pathways (ERK, p38 MAP kinase, and JNK), only inhibition of the ERK pathway by the MEK/ERK inhibitor U0126 (0.1-10 microM) significantly reduced MMP-9 upregulation, even when treatment was delayed for 4 h after PMA exposure. Inhibitors of p38 MAP kinase (SB203580) and JNK (SP600125) had no effect. This PKC pathway was compared to a cytokine response by exposing astrocytes to TNFalpha, which also activated MAP kinase and induced MMP-9 upregulation. But in this case, all three MAP kinase inhibitors (U0126, SB203580, and SP600125) reduced TNFalpha-induced MMP-9 upregulation. Taken together, these results suggest that the ERK MAP kinase is essential for MMP-9 upregulation via PKC and cytokine pathways in astrocytes.


Evaluating the role of fluorodeoxyglucose positron emission tomography-computed tomography in multi-disciplinary team recommendations for oesophago-gastric cancer.
National guidelines recommend that fluorodeoxyglucose positron emission tomography-computed tomography (PET-CT) is performed in all patients being considered for radical treatment of oesophageal or oesophago-gastric cancer without computerised tomography scan (CTS) evidence of metastasis. Guidance also mandates that all patients with cancer have treatment decisions made within the context of a multi-disciplinary team (MDT) meeting. Little is known, however, about the influence of PET-CT on decision making within MDTs. The aim of this study was to assess the role of PET-CT in oesophago-gastric cancer on MDT decision making. A retrospective analysis of a prospectively held database of all patients with biopsy-proven oesophageal or oesophago-gastric cancer discussed by a specialist MDT was interrogated. Patients selected for radical treatment without CTS evidence of M1 disease were identified. The influence of PET-CT on MDT decision making was examined by establishing whether the PET-CT confirmed CTS findings of M0 disease (and did not change the patient staging pathway) or whether the PET-CT changed the pathway by showing unsuspected M1 disease, refuting CTS suspicious metastases, or identifying another lesion (needing further investigation). In 102 MDT meetings, 418 patients were discussed, of whom 240 were initially considered for radical treatment and 238 undergoing PET-CT. The PET-CT confirmed CTS findings for 147 (61.8%) and changed MDT recommendations in 91 patients (38.2%) by (i) identifying M1 disease (n=43), (ii) refuting CTS suspicions of M1 disease (n=25), and (iii) identifying new lesions required for investigations (n=23). The addition of PET-CT to standard staging for oesophageal cancer led to changes in MDT recommendations in 93 (38.2%) patients, improving patient selection for radical treatment. The validity of the proposed methods for evaluating PET-CT on MDT decision making requires more work in other centres and teams.


System-wide assembly of pathways and modules hierarchically reveal metabolic mechanism of cerebral ischemia.
The relationship between cerebral ischemia and metabolic disorders is poorly understood, which is partly due to the lack of comparative fusing data for larger complete systems and to the complexity of metabolic cascade reactions. Based on the fusing maps of comprehensive serum metabolome, fatty acid and amino acid profiling, we identified 35 potential metabolic biomarkers for ischemic stroke. Our analyses revealed 8 significantly altered pathways by MetPA (Metabolomics Pathway Analysis, impact score >0.10) and 15 significantly rewired modules in a complex ischemic network using the Markov clustering (MCL) method; all of these pathways became more homologous as the number of overlapping nodes was increased. We then detected 24 extensive pathways based on the total modular nodes from the network analysis, 12 of which were new discovery pathways. We provided a new perspective from the viewpoint of abnormal metabolites for the overall study of ischemic stroke as well as a new method to simplify the network analysis by selecting the more closely connected edges and nodes to build a module map of stroke.


Investigation of anti-D antibody dependent cell mediated cytotoxicity in patients with thyroid cancer.
Anti-D dependent cellular cytotoxicity (ADCC) of peripheral mononuclear blood cells was measured in 69 thyroid cancer patients belonging to one of the main subtypes, and in 11 age-matched controls with benign thyroid disease. Independently of cancer subtype, tumor status, previous operation, age and sex, no differences in ADCC were observed. Further studies are warranted to clarify the cause of the constant ADCC levels.


Laminar organization of frequency-defined local axons within and between the inferior colliculi of the guinea pig.
We present a comprehensive description of the local (intrinsic and commissural) connections in the central nucleus of the inferior colliculi (CNICs) in guinea pig. Focal injections of the anterograde tracer biocytin were made into physiologically identified loci of the CNIC and the spatial organisation of the labeled fibres was revealed with computer-assisted three-dimensional (3-D) reconstruction. The intrinsic fibres form a series of V-shaped laminar plexuses composed of fibres bearing both terminal and en passant boutons. Each laminar plexus has a central wing located in the CNIC that extends into the dorsal cortex and an external wing located in the external cortex. The edge where the two wings intersect delimits the lateral border of the central nucleus with the external cortex. The density of labeled terminals was consistently lower in the cortices than in the CNIC. The laminar plexus connects points of similar frequency within the CNIC. Seen in 3-D, the location, orientation, shape, and area of the laminar plexus vary as a function of best frequency. The commissural fibres ending in the contralateral IC to the injection also form a laminar plexus which is symmetrical to the ipsilateral plexus. Electrolytic lesions placed in the contralateral IC at sites with best frequencies corresponding to those of the injection coincided with the terminals of the commissural fibres in most instances. Possible patterns for the organisation of these connections (point-to-point and diverging) are discussed. Three systems of peripheral axons to the laminar plexus are described: parallel, oblique, and perpendicular to the central wing. The novel parallel system has terminals in both ICs that run parallel to the central wing. It might constitute the anatomical basis for across-frequency interactions. The oblique and perpendicular systems are fibres of passage projecting to the commissure and brachium of the IC, respectively.


Analysis of enquiries to the National Pharmacy Association following major changes to controlled drug legislation in the UK.
The National Pharmacy Association (NPA) provides an advice service to community pharmacists in the UK, and keeps a database of the enquiries it receives. The aim of this research was to analyse the database for the period of October 2007 to March 2008 to gain an insight into how well pharmacists coped with legislative changes directly affecting pharmacy by identifying which changes generated the most enquiries during these 6 months and ascertaining in which months these queries were at their highest levels. Anonymised telephone enquiries regarding controlled drugs (CDs) received by the NPA from pharmacists during a 6-month period were reviewed and categorised according to the legislative change or other CD issue to which they related. A Poisson model was applied to determine whether there was a significant difference in the total number of CD queries generated each month. Altogether 6082 queries regarding CDs were received, of which 57% related to legislative changes. The three legislative changes that took place during the 6-month period all generated a significant increase in numbers of queries around the time of the change. Queries regarding the new form of CD register comprised the largest single category. Community pharmacists seek information regarding legislative changes when such changes come into force to a greater degree than when the legislation is drafted, consulted upon or enacted. The high number of queries received by the NPA does reflect that there was a difficulty in applying the various legislative changes in practice.


[Homozygous deletion of p16 and p15 genes in laryngeal squamous cell carcinoma].
To assess the relationship of homozygous deletion status of p16 (MTS1/INK4a/CDKN2A), p15(MTS2/INK4b/CDKN2B) genes and laryngeal squamous cell carcinoma(LSCC) progression. DNA was extracted from fresh tumors. Homozygous deletion of p16 exon 2(p16E2) in 80 cases of LSCC and p15 exon 2(p15E2) in 67 cases of LSCC were detected by the polymerase chain reaction technique. The p16E2 deletion rate in 80 cases was 12.5%(10/80); the p15E2 deletion rate in 67 cases was 11.94%(8/67); the p16E2 and p15E2 codeletion rate in 67 cases was 5.97%(4/67). Homozygous deletion of p16E2 and p15E2 is related with LSCC oncogenesis, and it may play a role to some extent in LSCC malignant progression.


A thermophilic glucoamylase fromCephalosporium eichhorniae.
A novel exocellular glucoamylase produced by a thermophilic fungus,Cephalosporium eichhorniae, was purified by a combination of membrane filtration and Sephadex chromatography. The enzyme was a glycoprotein, 28% carbohydrate by weight. It was composed of a single polypeptide chain with a molecular weight of 26,850. The enzyme was thermostable with optimum activity between 45 and 62°C. It had a substrate preference of amylose>amylopectin. Analysis by thin-layer and gas-liquid chromatography showed the major hydrolytic product of starch was glucose, classifying this enzyme as a thermophilic glucoamylase.


Biochemical and functional characterization of an exonuclease from Chaetomium thermophilum.
Exonucleases are often found associated with polymerase or helicase domains in the same enzyme or can function as autonomous entities to maintain genome stability. Here, we uncovered Chaetomium thermophilum RecQ family proteins that also have exonuclease activity in addition to their main helicase function. The novel exonuclease activity is separate from the helical core domain and coexists with the latter two enzymatic activities on the same polypeptide. The CtRecQ121-366 exonuclease region performs independently as an exonuclease. We describe its catalytic mechanism and biological characteristics. We demonstrate unequivocally that CtRecQ121-366 exclusively displays exonuclease activity and that this activity has a 3'-5' polarity that can both hydrolyze ssDNA and cleave dsDNA substrates. The hydrolytic activity of majority exonuclease is driven by bimetal ions, and this appears to be the case for the CtRecQ121-366 exonuclease as well. Additionally, the maximum activity of CtRecQ121-366 was observed at pH 8.0-9.0, low salt with Mg2+. The two helices in the structure, a6 and a7, play significant roles in the execution by anticipating their shape and changing essential amino acids.


Citrate anticoagulation for CRRT: don't always trust the postfilter iCa results!
Citrate has been recommended as the first-line anticoagulant for continuous renal replacement therapy (CRRT) in critically ill patients. Compared with heparin, citrate anticoagulation is safer and more efficacious. Citrate inhibits the coagulation cascade by lowering the ionized calcium (iCa) concentration in the filter. Monitoring of systemic iCa concentrations is inherent to the protocol, and monitoring of postfilter iCa is recommended to adjust citrate flow and optimize anticoagulation. While systemic iCa targets are in the physiological range, postfilter iCa concentrations are targeted between 0.20 and 0.35 mmol/l. In a previous issue of Critical Care, Schwarzer et al. compared systemic and postfilter iCa measurements of patients receiving citrate-based CRRT between six devices. They highlight the unreliability of iCa concentrations in the postfilter range, because the instruments cannot be validated in the low iCa range. The maximum mean difference between two instruments was as high as 0.33 mmol/l (range 0.21-0.50 mmol/l). The authors call for dialysis companies to revise their protocols. However, the first implication of their study is that the accuracy of blood gas analyzers to measure iCa in the low range needs to improve; and, secondly, clinicians using citrate anticoagulation need to be aware that the postfilter iCa result may be falsely high or low. This is particularly relevant when frequent premature filter clotting is observed despite postfilter iCa results in the seemingly target range. In these situations, citrate flow can be safely increased up to 4 mmol/l blood flow under monitoring of signs of citrate accumulation.


Giant congenital aneurysm of the right atrium: echocardiographic diagnosis and surgical management.
A rare case of giant congenital aneurysm of the right atrium in a 30-year-old woman is described. The patient had no history of cardiac diseases or trauma. Transthoracic 2-dimensional echocardiography with injection of isotonic saline enabled complete definition of the abnormal anatomy and allowed subsequent successful surgical correction.


On the capacity of attention: its estimation and its role in working memory and cognitive aptitudes.
Working memory (WM) is the set of mental processes holding limited information in a temporarily accessible state in service of cognition. We provide a theoretical framework to understand the relation between WM and aptitude measures. The WM measures that have yielded high correlations with aptitudes include separate storage-and-processing task components, on the assumption that WM involves both storage and processing. We argue that the critical aspect of successful WM measures is that rehearsal and grouping processes are prevented, allowing a clearer estimate of how many separate chunks of information the focus of attention circumscribes at once. Storage-and-processing tasks correlate with aptitudes, according to this view, largely because the processing task prevents rehearsal and grouping of items to be recalled. In a developmental study, we document that several scope-of-attention measures that do not include a separate processing component, but nevertheless prevent efficient rehearsal or grouping, also correlate well with aptitudes and with storage-and-processing measures. So does digit span in children too young to rehearse.


Sestrin2 is induced by glucose starvation via the unfolded protein response and protects cells from non-canonical necroptotic cell death.
Sestrin2 is a member of a family of stress responsive proteins, which controls cell viability via antioxidant activity and regulation of the mammalian target of rapamycin protein kinase (mTOR). Sestrin2 is induced by different stress insults, which diminish ATP production and induce energetic stress in the cells. Glucose is a critical substrate for ATP production utilized via glycolysis and mitochondrial respiration as well as for glycosylation of newly synthesized proteins in the endoplasmic reticulum (ER) and Golgi. Thus, glucose starvation causes both energy deficiency and activation of ER stress followed by the unfolding protein response (UPR). Here, we show that UPR induces Sestrin2 via ATF4 and NRF2 transcription factors and demonstrate that Sestrin2 protects cells from glucose starvation-induced cell death. Sestrin2 inactivation sensitizes cells to necroptotic cell death that is associated with a decline in ATP levels and can be suppressed by Necrostatin 7. We propose that Sestrin2 protects cells from glucose starvation-induced cell death via regulation of mitochondrial homeostasis.


Gratitude journals can improve nurses' mental well-being.
With nursing burnout on the rise, effective self-care interventions such as gratitude journals are needed to help nurses cope with symptoms of burnout. Gratitude journals can support nurses' emotional health by improving stress management and offering an opportunity for overall self-reflection.


Risk factors for urethral stricture following external beam radiotherapy and HDR brachytherapy for prostate cancer.
A potential late side effect of high-dose-rate (HDR) prostate brachytherapy combined with external beam radiotherapy (EBRT) is urethral stricture. The purpose of this study was to evaluate dosimetric parameters possibly contributing to stricture development, including dose to bladder base and D2cc(Gy) within 10 mm of prostatic urethra (D2cc 10 mm), which has, so far, not been reported in the literature. Patients developing urethral stricture, and matched controls, were identified from a prospective database of those receiving 46 Gy in 23 fractions of EBRT, followed by a single 15 Gy HDR brachytherapy dose. Patients had locally advanced, high- and intermediate-risk prostate cancer. Brachytherapy treatment planning parameters (planning target volume (PTV) size (cm 3), V110(%) bladder base, D2cc 10mm, number of HDR catheters, number of source dwell positions, and total source dwell time within 10 mm of the prostatic urethra) were analyzed to determine potential risk factors for urethral stricture. Seventy-two patients were treated, 22 of whom developed a urethral stricture. Univariate logistic regression performed on the planning parameters identified increased PTV size and D2cc 10 mm, with decreased V110(%) bladder base as risk factors for stricture formation. It is suggested that PTV size, V110(%) bladder base, and D2cc 10mm are predictive of urethral stricture formation following EBRT and brachytherapy to the prostate. This study demonstrates the importance of minimizing high dwell times and hot spots close to organs at risk and also the correction of any craniocaudal movement of catheters to avoid potential hot spots in the bulbomembranous urethra where there may be increased dose sensitivity.


JAX2, an ethanol extract of Hyssopus cuspidatus Boriss, can prevent bronchial asthma by inhibiting MAPK/NF-κB inflammatory signaling.
Hyssopus cuspidatus Boriss has been used to treat bronchial asthma for many years in Uighur medicine. JAX2, an ethanol extract from this plant, has effectiveness against bronchial asthma. However, the molecular basis for the anti-inflammatory effects of JAX2 remains unclear. This study aimed to evaluate the mechanism of JAX2 against bronchial asthma. We established an asthma model in rats using ovalbumin (OVA), and an inflammatory model in RAW264.7 cells using lipopolysaccharide (LPS) stimulation. To evaluate the anti-inflammatory effects of JAX2, the concentrations of tumor necrosis factor (TNF)-α, interleukin (IL)-4, IL-6, IL-17, eotaxin and immunoglobulin (Ig)E were measured by enzyme-linked immunosorbent assay (ELISA). Cell viability was investigated by the 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)2H-tetrazolium, inner salt (MTS) assay. Further, nitric oxide (NO) and reactive oxygen species (ROS) were determined using Griess reagent and 2,7-dichlorofluorescein diacetate. The phosphorylation of p-extracellular signal-regulated kinase (ERK), p-Jun-NH2-terminal kinase (JNK), p38 kinases (p38) and p-inhibitor of nuclear factor kappa-B kinase (IKK), and nuclear translocation of p-p65 kinases (p-p65) were determined by immunofluorescence to uncover the effects of JAX2 on the Mitogen-activated protein kinase (MAPK) and nuclear factor-kappa B (NF-κB) signaling pathways. After JAX2 administration to rats, Interferon (IFN)-γ concentrations in BALF increased significantly. Further, the concentrations of TNF-α, IL-4, IL-6, IL-17 and eotaxin in BALF, and IgE in serum decreased. JAX2 decreased TNF-α, IL-6 and NO in cell supernatant, and reduced ROS intracellularly. Concurrently, IFN-γ concentrations increased in cell supernatant significantly. In LPS-induced RAW264.7 cells, JAX2 inhibited phosphorylation of p-ERK, p-JNK and p-38 MAPK. The subsequent phosphorylation of p-IKK and nuclear translocation of the p-p65 subunit of NF-κB were also suppressed. Based on these findings, we believe that JAX2 has both preventive and treatment effects in bronchial asthma. Furthermore, in the RAW264.7 cell inflammatory model, JAX2 also inhibited NF-κB and MAPK signaling pathways.


Perivascular Adipose Tissue and Coronary Atherosclerosis: from Biology to Imaging Phenotyping.
Perivascular adipose tissue (PVAT) has a complex, bidirectional relationship with the vascular wall. In disease states, PVAT secretes pro-inflammatory adipocytokines which may contribute to atherosclerosis. Recent evidence demonstrates that pericoronary adipose tissue (PCAT) may also function as a sensor of coronary inflammation. This review details PVAT biology and its clinical translation to current imaging phenotyping. PCAT attenuation derived from routine coronary computed tomography (CT) angiography is a novel noninvasive imaging biomarker of coronary inflammation. Pro-inflammatory cytokines released from the arterial wall diffuse directly into the surrounding PCAT and inhibit adipocyte lipid accumulation in a paracrine manner. This can be detected as an increased PCAT CT attenuation, a metric which associates with high-risk plaque features and independently predicts cardiac mortality. There is also evidence that PCAT attenuation relates to coronary plaque progression and is modified by systemic anti-inflammatory therapies. Due to its proximity to the coronary arteries, PCAT has emerged as an important fat depot in cardiovascular research. PCAT CT attenuation has the potential to improve cardiovascular risk stratification, and future clinical studies should examine its role in guiding targeted medical therapy.


Lactobacillus fermentum, a probiotic capable to release glutathione, prevents colonic inflammation in the TNBS model of rat colitis.
Inflammatory bowel disease is associated with intestinal oxidative stress. In the present study we test the preventative effect of Lactobacillus fermentum, a probiotic that produces per se glutathione, in the trinitrobenzenesulphonic acid (TNBS) model of rat colitis. Colitis was induced in rats by intracolonic administration of 10 mg of TNBS dissolved in 0.25 ml of 50% ethanol. L. fermentum was administered orally (5x10(8) CFU suspended in 0.5 ml of skim milk) to a group of rats for 3 weeks, starting 2 weeks before colitis induction. Colonic damage was evaluated both histologically and biochemically, and the colonic luminal contents were used for bacterial studies as well as for short chain fatty acid (SCFA) production. L. fermentum treatment resulted in an amelioration of the inflammatory response in colitic rats as evidenced histologically and by a significant reduction of colonic MPO activity (P<0.05). The probiotic partially counteracted the colonic glutathione depletion induced by the inflammatory process. In addition, probiotic-treated colitic rats showed significant lower colonic tumour necrosis factor (TNF)alpha levels (P<0.01) and inducible nitric oxide synthase (iNOS) expression when compared to non-treated rats. Finally, the probiotic induced growth of Lactobacilli species and production of SCFA in colonic contents in comparison with control colitic rats. Administration of the probiotic L. fermentum facilitates the recovery of the inflamed tissue in the TNBS model of rat colitis, an effect associated with increased levels of glutathione as well as with amelioration of the production of some of the mediators involved in the inflammatory response of the intestine, such as TNFalpha and NO.


A validated HPTLC method for determination of terbutaline sulfate in biological samples: Application to pharmacokinetic study.
Terbutaline sulfate (TBS) was assayed in biological samples by validated HPTLC method. Densitometric analysis of TBS was carried out at 366 nm on precoated TLC aluminum plates with silica gel 60F254 as a stationary phase and chloroform-methanol (9.0:1.0, v/v) as a mobile phase. TBS was well resolved at RF 0.34 ± 0.02. In all matrices, the calibration curve appeared linear (r (2) ⩾ 0.9943) in the tested range of 100-1000 ng spot(-1) with a limit of quantification of 18.35 ng spot(-1). Drug recovery from biological fluids averaged ⩾95.92%. In both matrices, rapid degradation of drug favored and the T 0.5 of drug ranged from 9.92 to 12.41 h at 4 °C and from 6.31 to 9.13 h at 20 °C. Frozen at -20 °C, this drug was stable for at least 2 months (without losses >10%). The maximum plasma concentration (Cpmax) was found to be 5875.03 ± 114 ng mL(-1), which is significantly higher than the maximum saliva concentration (Csmax, 1501.69 ± 96 ng mL(-1)). Therefore, the validated method could be used to carry out pharmacokinetic studies of the TBS from novel drug delivery systems.


METTL3 stabilizes HDAC5 mRNA in an m6A-dependent manner to facilitate malignant proliferation of osteosarcoma cells.
Osteosarcoma (OS) is a prevalent primary bone sarcoma. Methyltransferase-like 3 (METTL3) is dysregulated in human malignancies. This study explored the mechanism of METTL3 in OS cell proliferation. Our results demonstrated that METTL3 was highly expressed in OS, and correlated with the tumor size, clinical stage, and distant metastasis of OS patients. Higher METTL3 expression indicated poorer prognosis. METTL3 silencing inhibited the malignant proliferation of OS cells, while METTL3 overexpression led to an opposite trend. METTL3 upregulated histone deacetylase 5 (HDAC5) expression in OS cells by increasing the m6A level. HDAC5 reduced the enrichment of H3K9/K14ac on miR-142 promoter, thus suppressing miR-142-5p expression and upregulating armadillo-repeat-containing 8 (ARMC8) level. HDAC5 overexpression or miR-142-5p silencing attenuated the inhibitory effect of METTL3 silencing on OS cell proliferation. Xenograft tumor experiment in nude mice confirmed that METTL3 silencing repressed OS cell proliferation in vivo via the HDAC5/miR-142-5p/ARMC8 axis. Collectively, METTL3-mediated m6A modification facilitated OS cell proliferation via the HDAC5/miR-142-5p/ARMC8 axis.


Influence of Er:YAG laser irradiation distance on the bond strength of a restorative system to enamel.
The aim of the present study was to investigate in vitro the effect of Er:YAG laser on bonding to enamel, varying the irradiation distance. Tensile bond strength of an adhesive restorative system to non-irradiated and irradiated enamel surfaces was evaluated. Thirty caries-free human third molars were sectioned in mesio-distal direction and embedded in acrylic resin. Enamel was flattened, and a 3-mm-diameter bonding area was demarcated. Specimens were randomly assigned into six groups: groups I-V were treated with the Er:YAG laser (80 mJ/2 Hz), varying the irradiation distance (11, 12 mm-focused, 14, 16 and 17 mm, respectively), followed by 35% phosphoric acid etching. Control group (VI) received treatment with phosphoric acid alone. Single Bond adhesive system was applied on the conditioned enamel, and composite resin cones, bonded to enamel, were fabricated with Z250. After storage, samples were tested in tensile to failure (50 kgf and 0.5 mm/min). Means in MPa were: I-9.67 (+/-3.44); II-13.29 (+/-2.65); III-13.33 (+/-2.22); IV-14.87 (+/-3.58); V-16.43 (+/-4.52); VI-22.90 (+/-3.03). ANOVA and Tukey test revealed statistically significant decrease of bond strength in group I (P < 0.05). Groups II-IV presented similar results, as did groups IV and V. Control group (VI) yielded the best overall performance (P < 0.05). Er:YAG laser irradiation adversely affected adhesion to enamel. However, bond strength was influenced by the irradiation distance, thus being stronger with the increase of distance to the target tissue.


Mid-mantle deformation inferred from seismic anisotropy.
With time, convective processes in the Earth's mantle will tend to align crystals, grains and inclusions. This mantle fabric is detectable seismologically, as it produces an anisotropy in material properties--in particular, a directional dependence in seismic-wave velocity. This alignment is enhanced at the boundaries of the mantle where there are rapid changes in the direction and magnitude of mantle flow, and therefore most observations of anisotropy are confined to the uppermost mantle or lithosphere and the lowermost-mantle analogue of the lithosphere, the D" region. Here we present evidence from shear-wave splitting measurements for mid-mantle anisotropy in the vicinity of the 660-km discontinuity, the boundary between the upper and lower mantle. Deep-focus earthquakes in the Tonga-Kermadec and New Hebrides subduction zones recorded at Australian seismograph stations record some of the largest values of shear-wave splitting hitherto reported. The results suggest that, at least locally, there may exist a mid-mantle boundary layer, which could indicate the impediment of flow between the upper and lower mantle in this region.


Effect of the incorporation of β-galactosidase in the GOS production during manufacture of soft cheese.
Galactooligosaccharides (GOS) are non-digestible oligosaccharides with recognized prebiotic role. The present study aims to evaluate a β-galactosidase from K. lactis during soft cheese making and to analyse the impact on carbohydrates metabolism, proteolysis, and volatile compounds production, physicochemical and microbiological characteristics of the product. The enzyme was added to cheese milk (fluid milk plus whey powder) before (40 min.) or simultaneously of the starter addition (Ep and E treatments, respectively); cheese without enzyme addition was also made (C treatment). Also, we characterized fresh and soft commercial cheeses from the point of view of carbohydrate fraction, highlighting GOS, and organic acid profiles. The inclusion of the enzyme in soft cheese making produced a delay in reaching the target pH (~5.2). Carbohydrate fermentation profiles differed among treatments during cheese making and ripening. GOS were only detected in Ep and E cheeses (0.88 and 0.51 g/100 g, respectively). Lactose content was lower, and glucose and galactose levels were higher in E and Ep than C. No differences in physicochemical and microbial composition and organic acids profiles among samples were observed. Bioformation of volatile compounds belonging to the chemical families of aldehydes, ketones, alcohols, esters and acids, was not substantially affected by the modification in the carbohydrate profile. GOS were not detected in any of the commercial cheeses; great variations in the carbohydrate contents and organic acids were found. The results obtained demonstrate the feasibility of obtaining cheeses with GOS. Although the GOS values achieved are not adequate enough for the desired effect, the proposed technological approach turned out to be satisfying and original. Cheeses with prebiotic fiber are not still widespread in the market.


Automated analysis of intraoperative phase in laparoscopic cholecystectomy: A comparison of one attending surgeon and their residents.
This study compares the intraoperative phase times in laparoscopic cholecystectomy performed by an attending surgeon and supervised residents over 10-years to assess operative times as a marker of performance and any impact of case severity on times. Laparoscopic cholecystectomy videos were uploaded to Touch Surgery™ Enterprise, a combined software and hardware solution for securely recording, storing, and analysing surgical videos, which provide analytics of intraoperative phase times. Case severity and visualisation of the critical view of safety (CVS) were manually assessed using modified 10-point intraoperative gallbladder scoring system (mG10) and CVS scores, respectively. Attending and residents' times were compared unmatched and matched by mG10. Secondary analysis of anonymized laparoscopic cholecystectomy video, recorded as standard of care. Adult patients who underwent elective laparoscopic cholecystectomy a single UK hospital. Cases were performed by one attending and their residents. 159 (attending=96, resident=63) laparoscopic cholecystectomy videos and intraoperative phase times were reviewed on Touch Surgery™ Enterprise and analyzed. Attending cases were more challenging (p=0.037). Residents achieved higher CVS scores (p=0.034) and showed longer dissection of hepatocystic triangle (HCT) times (p=0.012) in more challenging cases. Residents' total operative time (p=0.001) and dissection of HCT (p=0.002) times exceeded the attending's in low-severity matched cases (mG10=1). Residents' total operative times (p<0.001), port insertion/gallbladder exposure (p=0.032), and dissection of HCT (p<0.001) exceeded the attending's in matched cases (mG10=2). Residents' total operative (p<0.001), dissection of HCT (p<0.001), and gallbladder dissection (p=0.010) times exceeded the attendings in unmatched cases. Residents' total operative and dissection of HCT times significantly exceeded the attending's unmatched cases and low-severity matched cases which could suggest training need, however, also reflects an expected assessment of competence, and validates time as a marker of performance.


Convulsive episodes in patients with group A xeroderma pigmentosum.
To clarify the incidence of convulsive episodes in patients with group A xeroderma pigmentosum (XPA). By investigating the history of convulsive episodes of our 33 XPA patients through either their medical charts or direct interviews with their caretakers. Five patients had several episodes of afebrile convulsion at ages older than 12. With the exception of one patient who began to show convulsive episodes at 13, no other XPA patients exhibited febrile seizures. As far as our 33 XPA patients were concerned, 15% exhibited epilepsy, and 3% experienced febrile seizures. Japanese XPA patients showed a lower incidence of febrile seizures, while exhibiting a higher incidence of epilepsy. It is assumed that the brain of young patients with XPA is difficult to develop convulsions.


Nicotine dependence and quitting stages of smokers in Nepal: A community based cross-sectional study.
Nicotine dependence is an addiction to tobacco products caused by the chemical nicotine present in tobacco. 80% of premature deaths due to nicotine dependence come from low-and middle-income countries. Since most of the public health studies have focused solely on psychological and behavioral factors associated with tobacco smoking, this study aims to assess the nicotine dependence and stages of change of quitting smoking. A community based quantitative cross-sectional study was conducted among 280 smokers aged 15-69 years in Bharatpur metropolitan, Nepal. A semi-structured and validated questionnaire was used during the face-to-face interviews. Nicotine dependence among participants was assessed using the six-item Fagerstrom Test for Nicotine Dependence (FTND). Chi-square test and multivariate logistic regression analysis were performed to assess the associations between variables at the significance level α = 0.05. In the study population, the mean score of FTND was 5.15 ± 2.34. 33.9% participants had a high level of nicotine dependence and nearly half of the participants felt difficulty to refrain smoking even in No-smoking areas. Almost three out of ten respondents were prepared for smoking cessation. It was found that age group 20-39 years were more likely to have nicotine dependence (AOR 3.308, 95% CI = 1.039-10.534), those who initiated smoking before age of 15 were associated with a greater risk of nicotine dependence (AOR 3.68, 95% CI = 1.826-7.446), participants spending more on tobacco products (more than Rs 2400 monthly) were associated with an increased risk of nicotine dependence (AOR 4.47, 95% CI = 2.225-8.991), those who initiated smoking due to mental stress were more likely to have nicotine addiction (AOR 2.522, 95% CI = 1.004-6.028), and those who had no thought of quitting smoking were more associated with nicotine dependence (AOR 4.935, 95% CI = 1.458-16.699). Our study showed that high level of nicotine dependence is a major public health problem in low-and middle-income countries like Nepal. It also highlights that effective smoking cessation programs should be developed considering the level of nicotine dependence with more focus on early interventions of its associated factors.


[The effect of intracerebral transplantation of mesenchymal stem cells on orientation and exploratory behavior of aged animals].
Male Wistar-Kyoto rats (12 and 22-24 months old) were intracerebrally transplantated with syngeneic mesenchymal stem cells (MSC). Three weeks or one year later, orientation and exploratory behavior of the same animals (not 2 years old) was assessed using an «open field» test. Older rats (22-24 months old) were inhibited in the basic behavioral acts in comparison to younger animals (2-3 months old). Transplantation of MSC did not cause improvement of an orientation and exploratory behavior of aged rats. Morphological and immunohistochemical analysis suggests intracerebral MSC transplantation led to the injury of the cortex of the ipsilateral hemisphere.


Inhibition of Long Noncoding RNA Linc-Pint by Hepatitis C Virus in Infected Hepatocytes Enhances Lipogenesis.
HCV often causes chronic infection in liver, cirrhosis, and, in some instances, HCC. HCV encodes several factors' those impair host genes for establishment of chronic infection. The long noncoding RNAs (lncRNAs) display diverse effects on biological regulations. However, their role in virus replication and underlying diseases is poorly understood. In this study, we have shown that HCV exploits lncRNA long intergenic nonprotein-coding RNA, p53 induced transcript (Linc-Pint) in hepatocytes for enhancement of lipogenesis. We identified a lncRNA, Linc-Pint, which is significantly down-regulated in HCV-replicating hepatocytes and liver specimens from HCV infected patients. Using RNA pull-down proteomics, we identified serine/arginine protein specific kinase 2 (SRPK2) as an interacting partner of Linc-Pint. A subsequent study demonstrated that overexpression of Linc-Pint inhibits the expression of lipogenesis-related genes, such as fatty acid synthase and ATP-citrate lyase. We also observed that Linc-Pint significantly inhibits HCV replication. Furthermore, HCV-mediated enhanced lipogenesis can be controlled by exogenous Linc-Pint expression. Together, our results suggested that HCV-mediated down-regulation of Linc-Pint enhances lipogenesis favoring virus replication and liver disease progression. We have shown that SRPK2 is a direct target of Linc-Pint and that depletion of SRPK2 inhibits lipogenesis. Our study contributes to the mechanistic understanding of the role of Linc-Pint in HCV-associated liver pathogenesis.


Synthesis, oligomerization, and biological activity of the human immunodeficiency virus type 2 envelope glycoprotein expressed by a recombinant vaccinia virus.
The full-length envelope gene from an infectious human immunodeficiency virus type 2 (HIV-2) molecular clone was expressed in CD4+ and CD4- cells by a recombinant vaccinia virus vector. Pulse-chase experiments indicated that gp160 was processed into gp120 and gp41 subunits. Although large amounts of gp120 were shed into the medium, the recombinant vaccinia virus-infected cells fused with uninfected CD4+ cells. The receptor binding of HIV-2 gp120 was further analyzed using a panel composed of nine soluble CD4 mutants containing insertions of 2 amino acids within the first and second immunoglobulin-like domains. Of three mutations previously shown to interfere with HIV-1 gp120 binding, two also interfered with binding of the HIV-2 glycoprotein indicating use of the same binding site. Chemical crosslinking, sucrose gradient sedimentation, and sodium dodecyl sulfate-polyacrylamide gel electrophoresis were employed to study the oligomerization of the envelope protein. The data indicated that gp160 assembles posttranslationally into dimers and higher oligomers that are probably tetramers.


Evaluation of the detoxifying effect of yeast glucomannan on aflatoxicosis in broilers as assessed by gross examination and histopathology.
We evaluated the efficacy of yeast glucomannan (Mycosorb), incorporated into the diet at 0.5 and 1 g/kg, in reducing the detrimental effects of 2 mg aflatoxin/kg diet on growing broiler chicks from 1 to 21 d of age. A total of 240 male broiler chicks (Ross-308) was divided into 6 treatment groups [Control, Aflatoxin (AF), Yeast glucomannan (YG; 0.5 g/kg), AF plus YG (0.5 g/kg), YG (1 g/kg), and AF plus YG (1 g/kg)]. Ten chicks from each of the 6 groups were slaughtered and pathological examinations were performed on the liver, bursa of Fabricius, thymus, spleen and kidney. The aflatoxin treatment caused moderate to severe hydropic/fatty degeneration in the hepatocytes of the liver and the tubular epithelium of the kidneys, and follicular depletion in the bursa of Fabricius, thymus and spleen. Yeast glucomannan added to the aflatoxin-containing diet at 0.5 and 1 g/kg diminished the severity of pathological changes, slightly and moderately, respectively. The number of affected organs was also reduced in the group given 1 g/kg yeast glucomannan, compared to the aflatoxin group. These results show that yeast glucomannan effectively diminished the adverse effects of aflatoxin on the pathological changes and that the higher concentration of yeast glucomannan (1 g/kg) was more effective than the lower concentration (0.5 g/kg) and itself had no adverse effect.


Transient Th1/Th2 disbalance indicates postoperative effusions and edema after cardiopulmonary bypass in children.
Cardiac surgery with cardiopulmonary bypass (CPB) induces substantial release of IL-10, indicating increased Th2 cell response. Therefore, in this study, we wanted to verify if this response is due to CPB or surgical trauma, and to study its relation to postoperative effusions and edema (POEE) in children. Th1/Th2 reaction was monitored in children undergoing cardiovascular surgery with (n = 75) and without CPB (n = 29). Surgery with CPB compared to surgery without CPB induced a transient shift towards Th2. Elevated Th2 response was related to increased vascular permeability and POEE. The immune suppression/Th2 response is typical for CPB, and at intermediate level is tolerable but at high level could be adverse for the patients.


The development of a real-time PCR to detect pathogenic Leptospira species in kidney tissue.
A LightCycler real-time PCR hybridization probe-based assay that detects a conserved region of the16S rRNA gene of pathogenic but not saprophytic Leptospira species was developed for the rapid detection of pathogenic leptospires directly from processed tissue samples. In addition, a differential PCR specific for saprophytic leptospires and a control PCR targeting the porcine beta-actin gene were developed. To assess the suitability of these PCR methods for diagnosis, a trial was performed on kidneys taken from adult pigs with evidence of leptospiral infection, primarily a history of reproductive disease and serological evidence of exposure to pathogenic leptospires (n=180) and aborted pig foetuses (n=24). Leptospire DNA was detected by the 'pathogenic' specific PCR in 25 tissues (14%) and the control beta-actin PCR was positive in all 204 samples confirming DNA was extracted from all samples. No leptospires were isolated from these samples by culture and no positives were detected with the 'saprophytic' PCR. In a subsidiary experiment, the 'pathogenic' PCR was used to analyse kidney samples from rodents (n=7) collected as part of vermin control in a zoo, with show animals with high microagglutination titres to Leptospira species, and five were positive. Fifteen PCR amplicons from 1 mouse, 2 rat and 14 pig kidney samples, were selected at random from positive PCRs (n=30) and sequenced. Sequence data indicated L. interrogans DNA in the pig and rat samples and L. inadai DNA, which is considered of intermediate pathogenicity, in the mouse sample. The only successful culture was from this mouse kidney and the isolate was confirmed to be L. inadai by classical serology. These data suggest this suite of PCRs is suitable for testing for the presence of pathogenic leptospires in pig herds where abortions and infertility occur and potentially in other animals such as rodents.


Metastatic melanoma within and to the conjunctiva.
Two patients with epibulbar juxtalimbal primary conjunctival melanomas experienced local intralymphatic metastases to the inferior cul-de-sac, and a hematogenous metastasis to the conjunctiva developed in five other patients with cutaneous melanomas. Whether reflective of a local or distant metastasis, all of the lesions histopathologically were located in the substantia propria, and were separated from the overlying epithelium by a thin mantle of collagen. There was no evidence of atypical intraepithelial melanocytic proliferation, as would be expected in association with a primary conjunctival melanoma. Two of the cutaneous metastases exhibited a binodular or multinodular appearance that correlated histopathologically with variably confluent micronodules suggestive of the origin of the clinical lesion from a shower of tumor cell emboli. Patients with local intralymphatic spread from a primary conjunctival melanoma may experience additional lesions in the conjunctival sac or eyelid skin and are at risk for regional or distant metastases. They should be examined closely several times a year. The patients with the distant metastases all had their previously diagnosed primary cutaneous tumors on the truncal skin (a similar tendency emerges from a review of previous ocular cases), typically had myriad other cutaneous lesions, and two of them had a neoplastic iridocyclitis and vitreitis. These patients tended to die of the disseminated tumors within 1 year after conjunctival metastases developed.


Simultaneous separations of cations and anions by capillary electrophoresis with contactless conductivity detection employing a sequential injection analysis manifold for flexible manipulation of sample plugs.
Different schemes for simultaneous separations of both cations and anions were implemented in capillary electrophoresis employing a sequential injection analysis manifold for flexible manipulation of the background electrolyte and samples. The utilization of contactless conductivity allowed sensitive detection in narrow capillaries of 10 μm internal diameter, which in turn enabled the use of hydrodynamic pumping without significant penalty in dispersion otherwise caused by the laminar nature of the flow. This allowed to position sample plugs at a desired place along the entire length of the capillary prior to electrophoretic separation or to carry out concurrent pumping during the separation itself for optimization of resolution. Also implemented was dual single-end injection, the injection of two sample plugs from one end of the capillary, and the small size of the contactless conductivity detector allowed the concurrent use of two cells at different points along the capillary for added flexibility. Equally considered was the effect of the pH-value of the background electrolyte as this strongly influences the electroosmotic flow, which also has bearing on the protocol to be used. For dual single-end injection reproducibilities in separation time of about 1-3% were obtained and the peak areas could be reproduced to within about 3-5% (both standard deviations). The detection limits for inorganic ions were all about 1 μM.


Hypothesis: biological role for J-C intronic matrix attachment regions in the molecular mechanism of antigen-driven somatic hypermutation.
A major function of J-C intronic matrix attachment regions (MAR) during immune diversification via somatic hypermutation (SHM) at immunoglobulin loci may be to manipulate the topology of DNA within the upstream target domain. The suggestion that SHM induction requires MAR-induced torsional strain, in conjunction with DNA remodelling at the J-C intron, completes the definition of a cogent paradigm within which all extant molecular data on the issue may be interpreted. Moreover, the suggestion that a mutagenic mechanism relieves MAR-generated superhelicity could provide an indication as to the evolutionary basis of SHM.


Detection of dopamine overflow and diffusion with voltammetry in slices of rat brain.
Voltammetric electrodes have been used to monitor extracellular dopamine in rat brain slices. The electrode tips are small enough to be immersed inside the slice. Specificity for dopamine is increased through the use of voltammetry and a cation exchange membrane at the electrode tip. Dopamine overflow is observed in the caudate nucleus following electrical stimulation (60 Hz, 1 s, 3 V) with an adjacent bipolar electrode. The amount of overflow observed is increased when the tissue is perfused with 10 microM cocaine or nomifensine, both recognized inhibitors of dopamine uptake. The ability of dopamine in the perfusion buffer to permeate the slice was monitored with two voltammetric electrodes, one in the cerebral cortex and the other in the caudate nucleus. At a high concentration (100 microM), dopamine rapidly appeared (2.7 +/- 0.4 min) in the interior of the cortex, but dopamine was not observed in the caudate until a significantly later time (8.9 +/- 1.0 min). To examine whether this difference is a reflection of the presence of different uptake systems in the two regions, pressure ejection was employed. In this experiment a double-barrelled pipette was used to eject dopamine or DOPAC at a fixed distance (approximately 70 micron) from the voltammetric electrode. Ejection of small amounts of both substances could be detected in the cortex. When the ejector-detector assembly was moved to the caudate, dopamine could only be observed following pressure ejection after perfusion of the slice with 10 microM nomifensine. Detection of DOPAC was unaffected. All of these experiments indicate that uptake systems in the caudate keep dopamine concentrations very low in the extracellular fluid of the slice.


Accelerated study on lysozyme deposition on poly(HEMA) contact lenses.
A technique was developed to accelerate lysozyme deposition on poly(HEMA) contact lenses and measure the amounts of the deposited lysozyme. This technique was for evaluation of bendazac lysine solution, a contact lens cleaning and wetting solution. Effect of temperature on lysozyme deposition on poly(HEMA) contact lenses was examined. Five temperatures ranging from 25 degrees C to 90 degrees C were chosen to examine the temperature effect. The amounts of lysozyme deposited on poly(HEMA) contact lenses at 25 C and 60 C were 0.27 microg/lens and 0.61 microg/lens, respectively. The amount increased sharply to 23 microg/lens at 70 degrees C with the maximum of 31 microg/lens at 90 degrees C. Kinetics of lysozyme deposition on poly(HEMA) contact lenses was examined at 80 degrees C. Lysozyme deposition increased sharply during the first 2 h and reached a plateau after 2 h. Effectiveness of various cleaning procedures was examined using bendazac lysine solution. When the contact lenses were washed without rubbing with fingers, the bendazac lysine reduced the amount of deposited lysozyme by more than 40% from 18.3 microg/lens to 10.6 microg/lens. The effect of bendazac lysine was most prominent when the contact lenses were shaken during storage in the presence of lysozyme in solution. If the contact lenses were cleaned by rubbing with fingers, the effect of bendazac lysine solution on the prevention of lysozyme deposition was negligible.


The molecular evolution of signal peptides.
Signal peptides direct mature peptides to their appropriate cellular location, after which they are cleaved off. Very many random alternatives can serve the same function. Of all coding sequences, therefore, signal peptides might come closest to being neutrally evolving. Here we consider this issue by examining the molecular evolution of 76 mouse-rat orthologues, each with defined signal peptides. Although they do evolve rapidly, they evolve about half as fast as neutral sequences. This indicates that a substantial proportion of mutations must be under stabilizing selection. A few putative signal sequences lack a hydrophobic core and these tend to be more slowly evolving than others, indicating even stronger stabilizing selection. However, closer scrutiny suggests that some of these represent mis-annotations in GenBank. It is also likely that some of the substitutions are not neutral. We find, for example, that the rate of protein evolution correlates with that of the mature peptide. This may be a result of compensatory evolution. We also find that signal peptides of immune genes tend to be faster evolving than the average, which suggests an association with antagonistic co-evolution. Previous reports also indicated that the signal peptide of the imprinted gene, Igf2r, is also unusually fast evolving. This, it was hypothesized, might also be indicative of antagonistic co-evolution. Comparison of Igf2r's signal peptide evolution shows that, although it is not an outlier, its rate of evolution is comparable to that of many of the faster evolving immune system signal sequences and 5/6 of the amino acid changes do not conserve hydrophobicity. This is at least suggestive that there is something unusual about Igf2r's signal sequence.


Usefulness of a 3D-Printed Thyroid Cancer Phantom for Clinician to Patient Communication.
Thyroid glands and surrounding structures are very complex, and this complexity can pose a challenge for clinicians when explaining and communicating to the patient the details of a proposed surgery for thyroid cancer. A three-dimensional (3D) thyroid cancer model could help and improve this communication. A 3D-printed phantom of a thyroid gland and its presenting cancer was produced from segmented head and neck contrast-enhanced computed tomography (CT) data from a patient with thyroid cancer. The phantom reflects the complex anatomy of the arteries, veins, nerves, and other surrounding organs, and the printing materials and techniques were adjusted to represent the texture and color of the actual structures. Using this phantom, patients and clinicians completed surveys on the usefulness of this 3D-printed thyroid cancer phantom. patients (n = 33) and clinicians (n = 10). In the patient survey, the patients communicated that the quality of understanding of their thyroid disease status was enhanced when clinicians explained using the phantom. The clinicians communicated that the 3D phantom was advantageous for explaining complex thyroid surgery procedures to patients, and that the 3D phantom was helpful in educating patients with relatively poor anatomical knowledge. Using 3D printing technology, we produced a CT-based 3D thyroid cancer phantom, and patient and clinician surveys on its utility indicated that it successfully helped educate patients, providing them with an improved understanding of the disease.


A critical review of bile acids and their receptors in hepatic encephalopathy.
Hepatic encephalopathy describes an array of neurological complications that arise due to liver insufficiency. The pathogenesis of hepatic encephalopathy shares a longstanding association with hyperammonemia and inflammation, and recently, aberrant bile acid signaling has been implicated in the development of key features of hepatic encephalopathy. These key features include neuronal dysfunction, neuroinflammation and blood-brain barrier permeability. This review summarizes the findings of recent studies demonstrating a role for bile acids in the pathogenesis of hepatic encephalopathy via one of three main bile acid receptors and speculates on the possible downstream consequences of aberrant bile acid signaling.


Community violence and posttraumatic stress disorder symptoms in urban youth: The moderating influence of friend and parent support.
Separate lines of research have demonstrated that community violence predicts posttraumatic stress disorder (PTSD) symptoms in youth and that social support is one protective factor against the development of PTSD symptoms. The current study sought to examine the associations between primary and secondary exposure to community violence and the moderating role of parent and friend support on these relations. Participants were 96 urban youths (aged 6-17 years; 58.4% racial/ethnic minority; 51% female) and a caregiver recruited from a university mental health clinic. Results indicated that both primary and secondary exposure to community violence predicted PTSD symptoms. Friend support, but not parent support, moderated the association between primary, but not secondary, exposure to community violence and PTSD symptoms. The findings suggest that friend support is a salient protective factor for urban youth who may be at risk of PTSD symptoms due to exposure to community violence.


Load monitoring, strength training, and recovery in futsal: Practitioners' perspectives.
This study aimed to describe the current practices in futsal regarding a variety of topics related to performance and injury risk mitigation. Thirty-seven coaches from Spain and Portugal completed a questionnaire consisting of 28 closed questions organized in four categories: a) background information; b) training load (TL) monitoring and assessment of players' physical qualities; c) strength training (ST) practices; and d) recovery (REC) methods. The results showed that coaches varied in experience (1-8 years) and age (from 20 years to >50 years). Overall, 97.3% of the participants declared monitoring TL, with rating of perceived exertion, heart rate monitors, and wearable technology being used by 86.5%, 40.5%, and 37.8%, respectively. Neuromuscular and strength testing are the most common practices to evaluate performance and fatigue during the season. ST is a significant component of futsal, being performed 3 times/week during the pre- and in-season. ST is prescribed via %1RM - XRM (59.5%), velocity-based training (21.7%), repetitions in reserve (18.9%), until failure (10.8%), and circuit training (2.7%). 'Better Monitoring', 'More Individualized', 'Better Facilities', 'More Staff', and 'More Time' were the main aspects to improve ST. Multiple post-match REC strategies are used, with durations ranging from 0-15 to 16-30 min independently of game location.


Physiological effects of microcurrent and its application for maximising acute responses and chronic adaptations to exercise.
Microcurrent is a non-invasive and safe electrotherapy applied through a series of sub-sensory electrical currents (less than 1 mA), which are of a similar magnitude to the currents generated endogenously by the human body. This review focuses on examining the physiological mechanisms mediating the effects of microcurrent when combined with different exercise modalities (e.g. endurance and strength) in healthy physically active individuals. The reviewed literature suggests the following candidate mechanisms could be involved in enhancing the effects of exercise when combined with microcurrent: (i) increased adenosine triphosphate resynthesis, (ii) maintenance of intercellular calcium homeostasis that in turn optimises exercise-induced structural and morphological adaptations, (iii) eliciting a hormone-like effect, which increases catecholamine secretion that in turn enhances exercise-induced lipolysis and (iv) enhanced muscle protein synthesis. In healthy individuals, despite a lack of standardisation on how microcurrent is combined with exercise (e.g. whether the microcurrent is pulsed or continuous), there is evidence concerning its effects in promoting body fat reduction, skeletal muscle remodelling and growth as well as attenuating delayed-onset muscle soreness. The greatest hindrance to understanding the combined effects of microcurrent and exercise is the variability of the implemented protocols, which adds further challenges to identifying the mechanisms, optimal patterns of current(s) and methodology of application. Future studies should standardise microcurrent protocols by accurately describing the used current [e.g. intensity (μA), frequency (Hz), application time (minutes) and treatment duration (e.g. weeks)] for specific exercise outcomes, e.g. strength and power, endurance, and gaining muscle mass or reducing body fat.


Psychometric analysis of a measure of acceptance of new technologies (UTAUT), applied to the use of haptic virtual simulators in dental students.
The aims of this study are (a) to assess the psychometric quality of an instrument of acceptance of new technologies adapted from the UTAUT model, (b) to validate the UTAUT model as a valid measure to be applied in dental education, and (c) to determine which factors of the UTAUT model predict the behavioural intention of using a haptic virtual simulator (HVS). A cross-sectional design study with a sample of 265 dentistry students was carried out. Using structural equation modelling, confirmatory factor analysis verified the adequacy of four-factor model, although the only factor that directly predicts behavioural intention is performance expectancy. Internal consistency coefficients of each factor ranged from 0.800 to 0.969. These findings, as well as the predictive power of performance expectancy on behaviour intention, are in line with previous evidence found in the literature. These findings suggest that the UTAUT scale has adequate reliability and construct factorial validity; therefore, UTAUT model could be a valuable approach to assess haptic virtual simulator acceptance in dental education.


Pten coordinates retinal neurogenesis by regulating Notch signalling.
Development of nervous tissue is a coordinated process of neural progenitor cell (NPC) proliferation and neuronal differentiation. Intracellular signalling events that regulate the balance between NPC proliferation and neuronal differentiation, therefore, determine the size and composition of nervous tissues. Here, we demonstrate that negative regulation of phosphoinosite 3-kinase (PI3K)-Akt signalling by phosphatase tensin homologue (Pten) is essential for maintaining NPC population in mouse retina. We found that mouse retinal progenitor cells (RPCs) lacking the Pten gene complete neurogenesis earlier than their normal developmental schedule, resulting in their premature depletion in the mature retina. We further discover that Notch intracellular domain (NICD) fails to form transcription activator complex in Pten-deficient RPCs, and thereby unable to support RPC maintenance. Taken together, our results suggest that Pten plays a pivotal role in retinal neurogenesis by supporting Notch-driven RPC maintenance against neurogenic PI3K-Akt signalling.


Total saponins from Panax japonicus attenuate acute alcoholic liver oxidative stress and hepatosteatosis by p62-related Nrf2 pathway and AMPK-ACC/PPARα axis in vivo and in vitro.
Panax japonicus (T. Nees) C.A. Mey. (PJ) has been used as a tonic traditional Chinese medicine (TCM) for years. Based on its meridian tropism in liver, spleen, and lung, PJ was popularly used to enhance the function of these organs. It is originally recorded with detoxicant effect on binge drink in Ben Cao Gang Mu Shi Yi, a persuasive Chinese materia medica. And binge dink has a close relationship with alcoholic liver disease (ALD). Hence, it's meaningful to investigate whether PJ exerts liver protection against binge drink toxicity. This investigation was carried out not only to emphasize the right recognition of total saponins from PJ (SPJ), but also to study on its sober-up effectiveness and defensive mechanism against acute alcoholic liver injury in vivo and in vitro. SPJ constituents were verified by HPLC-UV analysis. In vivo, acute alcoholic liver oxidative stress and hepatosteatosis were established by continuous ethanol gavage to C57BL/6 mice for 3 days. SPJ was pre-administered for 7 days to investigate its protective efficacy. Loss of righting reflex (LORR) assay was employed to assess anti-inebriation effect of SPJ. Transaminases levels and hematoxylin and eosin (H&E) staining were measured to indicate the alcoholic liver injury. Antioxidant enzymes were measured to evaluate the oxidative stress degree in liver. Measurement of hepatic lipid accumulation was based on Oil Red O staining. Levels of inflammatory cytokines were evaluated by enzyme-linked immunosorbent assay (ELISA). In vitro, HepG2 cells were treated with ethanol for 24 h, and SPJ was pre-administered for 2 h. 2,7-dichlorofluorescein diacetate (DCFH-DA) was used as a probe to indicate reactive oxygen species (ROS) generation. Nrf2 activation was verified by the favor of specific inhibitor, ML385. The nuclear translocation of Nrf2 was indicated with immunofluorescence analysis. Proteins expressions of related pathways were determined by Western blotting. Oleanane-type saponins are the most abundant constituents of SPJ. In this acute model, SPJ released inebriation of mice in a dose dependent manner. It decreased levels of serum ALT and AST, and hepatic TG. Besides, SPJ inhibited CYP2E1 expression and reduced MDA level in liver, with upregulations of antioxidant enzymes GSH, SOD and CAT. p62-related Nrf2 pathway was activated by SPJ with downstream upregulations of GCLC and NQO1 in liver. AMPK-ACC/PPARα axis was upregulated by SPJ to alleviate hepatic lipidosis. Hepatic IL-6 and TNF-α levels were downregulated by SPJ, which indicated a regressive lipid peroxidation in liver. In HepG2 cells, SPJ reduced ethanol-exposed ROS generation. Activated p62-related Nrf2 pathway was verified to contribute to the alleviation of alcohol-induced oxidative stress in hepatic cells. This attenuation of hepatic oxidative stress and steatosis suggested the therapeutic value of SPJ for ALD.


Protein kinase G-dependent heme oxygenase-1 induction by Agastache rugosa leaf extract protects RAW264.7 cells from hydrogen peroxide-induced injury.
It has been proposed that the inducible isoform of heme oxygenase (HO) protects cells against oxidant-mediated injury. Although components of Agastache rugosa showed antioxidant effect, it is unclear this effect is related with HO-1 activity. Thus, we investigated the effects of Agastache rugosa leaf extract (ALE) on HO-1 protein expression and enzyme activity, and its protective effect against H(2)O(2)-induced oxidative damage was also investigated using RAW264.7 macrophage cells. Results showed that ALE concentration dependently increased HO-1 protein and enzyme activity, and protected cells from H(2)O(2)-induced cytotoxicity, with an IC(50) of 0.526 mg/ml. Hemin, a HO-1 inducer, also showed similar effect to ALE. Furthermore, the protective effect of both ALE and hemin was inhibited by a HO inhibitor, zinc protoporphyrin IX. The expression of HO-1 protein by ALE was reduced by pretreatment with LY83583 and ODQ, specific inhibitors of guanylate cyclase, but not by PKA inhibitors, H89 and KT5720, indicating that PKG signaling pathway regulates HO-1 induction by ALE. Taken together, it is concluded that PKG-dependent HO-1 induction is one of the important antioxidant mechanisms by which ALE protects RAW264.7 cells from H(2)O(2). Thus, ALE along with other actions may be beneficial for the treatment of oxidant-induced cellular injuries.


Executive summary. Consensus statement of the National AIDS Plan Secretariat, Spanish Society of Emergency Medicine and AIDS Study Group of the Spanish Society of Infectious Diseases and Clinical Microbiology on Emergency and HIV Infection.
Emergency Services (ES) are the cornerstone of our health system and therefore it cannot remain indifferent to the HIV advances that have drastically changed the landscape of the disease, so, emergency specialist updating is not only necessary, it is also essential. The purpose of this paper is to support non-HIV specialist professionals in treating patients with urgent diseases resulting from HIV infection or related to it.


Ca2+-permeable AMPA receptors in mouse olfactory bulb astrocytes.
Ca2+ signaling in astrocytes is considered to be mainly mediated by metabotropic receptors linked to intracellular Ca2+ release. However, recent studies demonstrate a significant contribution of Ca2+ influx to spontaneous and evoked Ca2+ signaling in astrocytes, suggesting that Ca2+ influx might account for astrocytic Ca2+ signaling to a greater extent than previously thought. Here, we investigated AMPA-evoked Ca2+ influx into olfactory bulb astrocytes in mouse brain slices using Fluo-4 and GCaMP6s, respectively. Bath application of AMPA evoked Ca2+ transients in periglomerular astrocytes that persisted after neuronal transmitter release was inhibited by tetrodotoxin and bafilomycin A1. Withdrawal of external Ca2+ suppressed AMPA-evoked Ca2+ transients, whereas depletion of Ca2+ stores had no effect. Both Ca2+ transients and inward currents induced by AMPA receptor activation were partly reduced by Naspm, a blocker of Ca2+-permeable AMPA receptors lacking the GluA2 subunit. Antibody staining revealed a strong expression of GluA1 and GluA4 and a weak expression of GluA2 in periglomerular astrocytes. Our results indicate that Naspm-sensitive, Ca2+-permeable AMPA receptors contribute to Ca2+ signaling in periglomerular astrocytes in the olfactory bulb.


Formation of the astral mitotic spindle: ultrastructural basis for the centrosome-kinetochore interaction.
The formation of the astral mitotic spindle is initiated at the time of nuclear envelope breakdown from an interaction between the replicated spindle poles (i.e. centrosomes) and the chromosomes. As a result of this interaction bundles of microtubules are generated which firmly attach the kinetochores on each chromosome to opposite spindle poles. Since these kinetochore fibers are also involved in moving the chromosomes, the mechanism by which they are formed is of paramount importance to understanding the etiology of force production within the spindle. As a prelude to outlining such a mechanism, the dynamics of spindle formation and chromosome behavior are examined in the living cell. Next, the properties of centrosomes and kinetochores are reviewed with particular emphasis on the structural and functional changes that occur within these organelles as the cell transits from interphase to mitosis. Finally, a number of recent observations relevant to the mechanism by which these organelles interact are detailed and discussed. From these diverse data it can be concluded that kinetochore fiber microtubules are derived from dynamically unstable astral microtubules that grow into, or grow by and then interact laterally with, the kinetochore. Moreover, the data clearly demonstrate that the interaction of a single astral microtubule with one of the kinetochores on an unattached chromosome is sufficient to attach the chromosome to the spindle, orient it towards a pole, and initiate poleward motion. As the chromosomes move into the region of the forming spindle more astral microtubules become incorporated into the nascent kinetochore fibers and chromosome velocity decreases dramatically. During this time the distribution of spindle microtubules changes from two overlapping radial arrays to the fusiform array characteristic of metaphase cells.


Systolic and diastolic functions in elderly patients with and without heart failure.
A considerable number of patients with heart failure (HF) have a normal left ventricular ejection fraction (LVEF). In these subjects, HF has usually been related to diastolic heart failure (DHF), still a frequently overlooked clinical entity. This study reports the clinical, instrumental, and conventional echocardiographic evaluation of 159 consecutive, hospitalized elderly patients, 87 admitted with HF and 72 admitted for other reasons without overt HF. All of the 87 HF patients had signs of diastolic dysfunction (DDYS), yet 44.8% of them had a normal LVEF. Forty-four of the 72 patients admitted without overt HF (61.1%) had mild DDYS and 14 (19.5%) also had a reduced LVEF. There was a clear relationship between LVEF reduction and the severity of DDYS. HF is often a combination of diastolic and systolic function abnormalities. DHF may be difficult to detect in HF subjects with normal LVEF because their DDYS is often mild. However, there are signs of DDYS in all HF patients that increase in severity as LVEF decreases. DDYS could be considered a marker for all forms of HF, especially in elderly patients.


Metallothionein and apoptosis in primary human hepatocellular carcinoma (HCC) from northern China.
Metallothionein (MT), acting as an antioxidant and zinc binding protein, may play an important role in regulation of apoptosis. Its differential expression has been documented in various human tumours. MT expression by immunohistochemical staining with a polyclonal antibody and apoptotic cells (APC) by TUNEL technique were investigated in 20 cases of hepatocellular carcinoma (HCC) and 2 normal livers from Northern China. Adjacent normal liver was available from 9 of these cases and 6 had adjacent cirrhotic tissue. There was no difference for MT staining and incidence of APC between normal liver and adjacent normal liver, and thus both were used as control liver. Control liver had consistent MT staining with very low incidence of APC. Adjacent cirrhotic liver showed the same intensity of MT staining, with a similar incidence of APC to control liver. Twelve of 20 HCC cases (60%) showed no MT staining, and the rest also showed a low grade of MT staining as compared with control or adjacent cirrhotic livers. The incidence of APC in HCC was markedly higher than that in control liver or adjacent cirrhotic liver. The negative correlation of numbers of APC with MT expression was statistically significant (p < 0.005). The high incidence of APC in liver with a low MT expression was confirmed in double staining for MT and APC. The present investigation from Northern Chinese samples has shown that MT expression in HCC was different from that in other human tumours, such as breast carcinoma. This suggests a different pattern of expression of MT protein in these two kinds of cancer. This investigation is important in understanding the mechanisms of the drug resistance of tumour cells, and may help to design better treatment strategies.


A generalized model of a resource-population system : II. Stability analysis.
A system of nonlinear differential equations describing a resource-population system is analyzed in terms of the existence and characteristics of its equilibrium states.It is proved that, under the condition that k<αβ (necessary condition for the population being able to grow under optimal conditions), it is a necessary and sufficient condition for the system to have a steady state that the resource input rate to the system be constant.When the resource input rate is a constant different from zero, the system has only one equilibrium point, at M 0=β/0/k, A 0=-(β/0/kα)ln(1-k/αβ), and this equilibrium point is always stable. In other words, the system population-resource will always reach the steady state, either monotonically (node) or by damped oscillations (focus), from any arbitrary initial condition in the positive quadrant.When the resource input rate is equal to zero, the system has an infinite number of equilibrium points at M 0=0, A 0=constant. All these equilibrium points are unstable in the sense that any slight increase in M will move the system away from the equilibrium states, except for the point M 0=0, A 0=0, which is the only stable equilibrium point, to which the system will tend. This stable equilibrium point corresponds to the condition of complete annihilation of both resource and population.Finally, it is proved that the system does not have limit cycles in the positive quadrant and is therefore incapable of self-oscillations.


The neutral glyceroglucolipids of alveolar lavage from rabbit.
Three individual glycolipids have been isolated from the neutral lipid fraction of rabbit alveolar lavage. All three glycolipids contained glucose, glyceryl monoethers and fatty acids, and differed from each other primarily with respect to the number of glucose residues. The structures of these glycolipids were identified by mild alkaline methanolysis, oxidation with periodate and CrO3, and methylation studies, as: Glc(alpha 1 leads to 3)-1,(3)-O-alkyl-2-O-acylglycerol, Glc(alpha 1 leads to 6)Glc(alpha 1 leads to 6)Glc(alpha 1 leads to 6)Glc(alpha 1 lead to 6)Glc(alpha 1 leads to 3)-1,(3)-O-alkyl-2-O-acyglycerol, and Glc(alpha 1 leads to 6)Glc(alpha 1 leads to 6)Glc(alpha 1 leads to 6)Glc(alpha 1 leads to 6)Glc(alpha 1 leads to 6)Glc(alpha 1 leads to 3)-l,(3)-O-alkyl-2-O-acylglycerol.