Altered Actions of Memantine and NMDA-Induced Currents in a New Grid2-Deleted Mouse Line.
Memantine is a non-competitive antagonist of the N-methyl-D-aspartate (NMDA) receptor, and is an approved drug for the treatment of moderate-to-severe Alzheimer's disease. We identified a mouse strain with a naturally occurring mutation and an ataxic phenotype that presents with severe leg cramps. To investigate the phenotypes of these mutant mice, we screened several phenotype-modulating drugs and found that memantine (10 mg/kg) disrupted the sense of balance in the mutants. Moreover, the mutant mice showed an attenuated optokinetic response (OKR) and impaired OKR learning, which was also observed in wild-type mice treated with memantine. Microsatellite analyses indicated that the Grid2 gene-deletion is responsible for these phenotypes. Patch-clamp analysis showed a relatively small change in NMDA-dependent current in cultured granule cells from Grid2 gene-deleted mice, suggesting that GRID2 is important for correct NMDA receptor function. In general, NMDA receptors are activated after the activation of non-NMDA receptors, such as AMPA receptors, and AMPA receptor dysregulation also occurs in Grid2 mutant mice. Indeed, the AMPA treatment enhanced memantine susceptibility in wild-type mice, which was indicated by balance sense and OKR impairments. The present study explores a new role for GRID2 and highlights the adverse effects of memantine in different genetic backgrounds.


Characterization of an improved 1-3 piezoelectric composite by simulation and experiment.
To increase electromechanical coupling factor of 1-3 piezoelectric composite and reduce its bending deformation under external stress, an improved 1-3 piezoelectric composite is developed. In the improved structure, both epoxy resin and silicone rubber are used as polymer material. The simulation model of the improved 1-3 piezoelectric composite was established using the finite element software ANSYS. The relationship of the performance of the improved composite to the volume percentage of silicone rubber was determined by harmonic response analysis and the bending deformation under external stress was simulated by static analysis. The improved composite samples were prepared by cutting and filling methods, and the performance was tested. The feasibility of the improved structure was verified by finite element simulation and experiment. The electromechanical coupling factor of the improved composite can reach 0.67 and meanwhile the characteristic impedance can decline to 13 MRayl. The electromechanical coupling factor of the improved composite is higher than that of the composite with only epoxy resin as the polymer and the improved composite can reduce bending deformation. Comparison of simulation and experiment, the results of the experiment are in general agreement with those from the simulation. However, most experimental values were higher than the simulation results, and the abnormality of the test results was also more obvious than that of the simulation. These findings may be attributed to slight difference in the material parameters of simulation and experiment.


5-HT2A receptor promoter polymorphism, -1438G/A and negative symptom response to olanzapine in schizophrenia.
The 5-HT2A receptor promoter -1438G/A polymorphism, which is in complete linkage disequilibrium with the 5-HT2A 102T/C polymorphism, may be related to antipsychotic response. The aim of this paper is to determine relationships between the -1438G/A polymorphism and olanzapine negative symptom response. DNA from 41 subjects with schizophrenia (as defined by the Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition) was analyzed for the -1438G/A polymorphism. Olanzapine (7.5-20 mg/day) was given for 6 weeks and response was assessed using the Scale for Assessment of Negative Symptoms (SANS). A linear regression used a dependent variable of percent change in SANS. Independent variables included 5-HT2A polymorphisms and interactions. The -1438G/A polymorphism and percent change in SANS showed a significant trend (P=.0542). The A/A genotype group had a 45% reduction in SANS compared with 19% in the other groups. We conclude that the A/A genotype may be associated with olanzapine negative symptom response, seen as a 2-fold greater percent reduction in SANS, and may be clinically relevant.


MprA and DosR coregulate a Mycobacterium tuberculosis virulence operon encoding Rv1813c and Rv1812c.
Mycobacterium tuberculosis remains a significant global pathogen, causing extensive morbidity and mortality worldwide. This bacterium persists within granulomatous lesions in a poorly characterized, nonreplicating state. The two-component signal transduction systems MprAB and DosRS-DosT (DevRS-Rv2027c) are responsive to conditions likely to be present within granulomatous lesions and mediate aspects of M. tuberculosis persistence in vitro and in vivo. Here, we describe a previously uncharacterized locus, Rv1813c-Rv1812c, that is coregulated by both MprA and DosR. We demonstrate that MprA and DosR bind to adjacent and overlapping sequences within the promoter region of Rv1813c and direct transcription from an initiation site located several hundred base pairs upstream of the Rv1813 translation start site. We further show that Rv1813c and Rv1812c are cotranscribed, and that the genomic organization of this operon is specific to M. tuberculosis and Mycobacterium bovis. Although Rv1813c is not required for survival of M. tuberculosis in vitro, including under conditions in which MprAB and DosRST signaling are activated, an M. tuberculosis ΔRv1813c mutant is attenuated in the low-dose aerosol model of murine tuberculosis, where it exhibits a lower bacterial burden, delayed time to death, and decreased ability to stimulate proinflammatory cytokines interleukin-1β (IL-1β) and IL-12. Interestingly, overcomplementation of these phenotypes is observed in the M. tuberculosis ΔRv1813c mutant expressing both Rv1813c and Rv1812c, but not Rv1813c alone, in trans. Therefore, Rv1813c and Rv1812c may represent general stress-responsive elements that are necessary for aspects of M. tuberculosis virulence and the host immune response to infection.


Improved self-inactivating retroviral vectors derived from spleen necrosis virus.
Self-inactivating (SIN) retroviral vectors contain a deletion spanning most of the right long terminal repeat's (LTR's) U3 region. Reverse transcription copies this deletion to both LTRs. As a result, there is no transcription from the 5' LTR, preventing further replication. Many previously developed SIN vectors, however, had reduced titers or were genetically unstable. Earlier, we reported that certain SIN vectors derived from spleen necrosis virus (SNV) experienced reconstitution of the U3-deleted LTR at high frequencies. This reconstitution occurred on the DNA level and appeared to be dependent on defined vector sequences. To study this phenomenon in more detail, we developed an almost completely U3-free retroviral vector. The promoter and enhancer of the left LTR were replaced with those of the cytomegalovirus immediate-early genes. This promoter swap did not impair the level of transcription or alter its start site. Our data indicate that SNV contains a strong initiator which resembles that of human immunodeficiency virus. We show that the vectors replicate with efficiencies similar to those of vectors possessing two wild-type LTRs. U3-deleted vectors carrying the hygromycin B phosphotransferase gene did not observably undergo LTR reconstitution, even when replicated in helper cells containing SNV-LTR sequences. However, vectors carrying the neomycin resistance gene did undergo LTR reconstitution with the use of homologous helper cell LTR sequences as template. This supports our earlier finding that sequences within the neomycin resistance gene can trigger recombination.


Thoracoscopic lobectomy versus open lobectomy in stage I non-small cell lung cancer: a meta-analysis.
The objective of the present meta-analysis was to evaluate the survival, recurrence rate, and complications in patients with stage I non-small cell lung cancer (NSCLC) who received video-assisted thoracoscopic surgery (VATS) or open lobectomy. A literature search was conducted on June 31, 2012 using combinations of the search terms video-assisted thoracic surgery, open thoracotomy, lobectomy, and non-small-cell lung cancer (NSCLC). Inclusion criteria were: 1) Compared video-assisted thoracic surgery (VATS) lobectomy with open lobectomy. 2) Stage I NSCLC. 2) No previous treatment for lung cancer. 4) Outcome data included 5-year survival rate, complication, and recurrence rate. Tests of heterogeneity, sensitivity, and publication bias were performed. A total of 23 studies (21 retrospective and 2 prospective) met the inclusion criteria. VATS was associated with a longer 5-year survival (odds ratio [OR] = 1.622, 95% confidence interval [CI] 1.272 to 2.069; P<0.001), higher local recurrence rate (OR = 2.152, 95% CI 1.349 to 3.434; P = 0.001), similar distant recurrence rate (OR = 0.91, 95% CI 0.33 to 2.48; P = 0.8560), and lower total complication rate (OR = 0.45, 95% CI 0.24 to 0.84; P = 0.013) compared to open lobectomy. VATS was also associated with lower rates arrhythmias, prolonged air leakage, and pneumonia but it did not show any statistical significance. Patients with stage I NSCLC undergoing VATS lobectomy had longer survival and fewer complications than those who received open lobectomy.


Mycoplasma hominis periaortic abscess following heart-lung transplantation.
We report the first case of Mycoplasma hominis periaortic abscess after heart-lung transplantation. The absence of sternal wound infection delayed the diagnosis, but the patient successfully recovered with debridement surgeries and long-term antibiotic therapy. Owing to the difficulty in detection and the intrinsic resistance to beta-lactams, M. hominis infections are prone to being misdiagnosed and undertreated. M. hominis should be suspected in cases where conventional microbiological identification and treatment approaches fail.


Sorafenib, Lenvatinib, or Lenvatinib Combining PD-1 Inhibitors Plus TACE in Unresectable Hepatocellular Carcinoma: A Retrospective Analysis.
Introduction: This retrospective study aimed to compare the efficacy and safety of transarterial chemoembolization plus lenvatinib and programmed death 1 (PD-1) inhibitors versus transarterial chemoembolization plus lenvatinib or sorafenib in patients with unresectable hepatocellular carcinoma. Methods: Consecutive patients with unresectable hepatocellular carcinoma who received transarterial chemoembolization plus lenvatinib and PD-1 inhibitors, lenvatinib, or sorafenib were retrospectively identified in our institution between January 2018 and August 2020. The primary endpoint was overall survival. Results: A total of 84 patients were included in this analysis. The median overall survival was significantly improved in the transarterial chemoembolization plus lenvatinib and PD-1 inhibitor group compared with the transarterial chemoembolization plus sorafenib group (26.7 months [95% confidence interval 25.2-31.6] vs 14.4 months [95% confidence interval 9.5-18.9]; hazard ratio 0.39 [95% confidence interval 0.17-0.72]; P = .007) or the transarterial chemoembolization plus lenvatinib group (26.7 months [95% confidence interval 25.2-31.6] vs 17.9 [95% confidence interval 13.4-22.2] months; hazard ratio 0.45 [95% confidence interval 0.17-0.87]; P = .031). Transarterial chemoembolization plus lenvatinib and PD-1 inhibitor also significantly prolonged median progression-free survival compared with transarterial chemoembolization plus sorafenib group (8.2 months [95% confidence interval 3.3-13.0] vs 6.0 months [95% confidence interval 4.2-7.8]; hazard ratio 0.47 [95% confidence interval 0.24-0.74]; P = .005) or the transarterial chemoembolization plus lenvatinib group (8.2 months [95% confidence interval 3.3-13.0] vs 6.6 [95% confidence interval 4.3-7.9] months; hazard ratio 0.58 [95% confidence interval 0.31-0.96]; P = .047). No significant difference was seen between groups in the incidence of an adverse event or grade 3 or higher adverse event. Conclusion: Transarterial chemoembolization plus lenvatinib, and PD-1 inhibitor was associated with better survival benefits and acceptable toxicities, which may provide an additional therapeutic option for unresectable hepatocellular carcinoma.


Regulation of autophagy by extracellular signal-regulated protein kinases during 1-methyl-4-phenylpyridinium-induced cell death.
Increased autophagic vacuoles (AVs) occur in injured or degenerating neurons, under both developmental and pathological situations. Although regulation of starvation-induced autophagy has been extensively studied, less is known about autophagic responses to pathological damage. The neurotoxin 1-methyl-4-phenylpyridinium (MPP(+)) produces mitochondria-targeted injury, which contributes to parkinsonism induced by 1-methyl-4-phenyl-1,2,3,6-tetrahydro-pyridine in mammals. Here, we demonstrate that MPP(+) elicited increased autophagy in SH-SY5Y cells, as assessed by electron microscopy, immunofluorescence for the autophagy protein LC3/Atg8, LC3 electrophoretic mobility shift, mitochondrial degradation, and monodansylcadaverine staining for late AVs/autolysosomes. During nutrient deprivation, class III phosphatidylinositol-3 kinase (PI3K) stimulates autophagy in concert with the autophagy-regulatory protein beclin 1/Atg6. Although PI3K inhibitors and RNA interference knockdown of beclin 1 effectively inhibited autophagy elicited by amino acid deprivation, neither reduced MPP+-induced autophagic stress. In contrast, inhibition of mitogen-activated protein kinase/extracellular signal-regulated protein kinase kinase reduced AV content, mitochondrial degradation, and cell death in MPP+-treated cells. RNA interference studies targeting core Atg proteins also reduced AV content and cell death. Likewise, in primary midbrain dopaminergic neurons, MPP+ elicited increased AV content, which was reversed by inhibition of mitogen-activated protein kinase/extracellular signal-regulated protein kinase kinase but not PI3K. These results implicate a role for extracellular signal-regulated protein kinase (ERK) signaling upstream of MPP+-elicited autophagic stress. Moreover, pathological stimulation of beclin 1-independent autophagy is associated with neuronal cell death.


Seroprevalence of three paramyxoviruses; Hendra virus, Tioman virus, Cedar virus and a rhabdovirus, Australian bat lyssavirus, in a range expanding fruit bat, the Grey-headed flying fox (Pteropus poliocephalus).
Habitat-mediated global change is driving shifts in species' distributions which can alter the spatial risks associated with emerging zoonotic pathogens. Many emerging infectious pathogens are transmitted by highly mobile species, including bats, which can act as spill-over hosts for pathogenic viruses. Over three years, we investigated the seroepidemiology of paramyxoviruses and Australian bat lyssavirus in a range-expanding fruit bat, the Grey-headed flying fox (Pteropus poliocephalus), in a new camp in Adelaide, South Australia. Over six, biannual, sampling sessions, we quantified median florescent intensity (MFI) antibody levels for four viruses for a total of 297 individual bats using a multiplex Luminex binding assay. Where appropriate, florescence thresholds were determined using finite mixture modelling to classify bats' serological status. Overall, apparent seroprevalence of antibodies directed at Hendra, Cedar and Tioman virus antigens was 43.2%, 26.6% and 95.7%, respectively. We used hurdle models to explore correlates of seropositivity and antibody levels when seropositive. Increased body condition was significantly associated with Hendra seropositivity (Odds ratio = 3.67; p = 0.002) and Hendra virus levels were significantly higher in pregnant females (p = 0.002). While most bats were seropositive for Tioman virus, antibody levels for this virus were significantly higher in adults (p < 0.001). Unexpectedly, all sera were negative for Australian bat lyssavirus. Temporal variation in antibody levels suggests that antibodies to Hendra virus and Tioman virus may wax and wane on a seasonal basis. These findings suggest a common exposure to Hendra virus and other paramyxoviruses in this flying fox camp in South Australia.


The level of HER2 expression is a predictor of antibody-HER2 trafficking behavior in cancer cells.
The receptor tyrosine kinase HER2 is known to play a central role in mitogenic signaling, motivating the development of targeted, HER2-specific therapies. However, despite the longstanding use of antibodies to target HER2, controversies remain concerning antibody/HER2 trafficking behavior in cancer cells. Understanding this behavior has direct relevance to the mechanism of action and effective design of such antibodies. In the current study, we analyzed the intracellular dynamics of trastuzumab, a marketed HER2-targeting antibody, in a panel of breast and prostate cancer cell lines that have a wide range of HER2 expression levels. Our results reveal distinct post-endocytic trafficking behavior of antibody-HER2 complexes in cells with different HER2 expression levels. In particular, HER2-overexpressing cells exhibit efficient HER2 recycling and limited reductions in HER2 levels upon antibody treatment, and consequently display a high level of antibody persistence on their plasma membrane. By contrast, in cells with low HER2 expression, trastuzumab treatment results in rapid antibody clearance from the plasma membrane combined with substantial decreases in HER2 levels and undetectable levels of recycling. A cell line with intermediate levels of HER2 expression exhibits both antibody recycling and clearance from the cell surface. Significantly, these analyses demonstrate that HER2 expression levels, rather than cell origin (breast or prostate), is a determinant of subcellular trafficking properties. Such studies have relevance to optimizing the design of antibodies to target HER2.


Immune system and cholangiocytes: A puzzling affair in primary biliary cholangitis.
Primary biliary cholangitis (PBC) is a cholestatic liver disease characterized by the destruction of the small and medium bile ducts. Its pathogenesis is still unknown. Despite the genome wide association study findings, the therapies targeting the cytokines pathway, tested so far, have failed. The concept of the biliary epithelium as a key player of the PBC pathogenesis has emerged over the last few years. It is now well accepted that the biliary epithelial cells (BECs) actively participate to the genesis of the damage. The chronic stimulation of BECs via microbes and bile changes the cell phenotype toward an active state, which, across the production of proinflammatory mediators, can recruit, retain, and activate immune cells. The consequent immune system activation can in turn damage BECs. Thus, the crosstalk between both innate and adaptive immune cells and the biliary epithelium creates a paracrine loop responsible for the disease progression. In this review, we summarize the evidence provided in literature about the role of BECs and the immune system in the pathogenesis of PBC. We also dissect the relationship between the immune system and the BECs, focusing on the unanswered questions and the future potential directions of the translational research and the cellular therapy in this area.


Peripheral-type benzodiazepine receptors: a second site of action for benzodiazepines.
Benzodiazepines are among the most widely used therapeutic drugs because of their sedative and anxiolytic effects mediated through modulation of GABAA receptors. Another recognition site for these drugs, termed the peripheral-type (or mitochondrial) benzodiazepine receptor, is much more prevalent throughout the body for which a physiologic and pharmacologic role has just been found. This drug receptor plays a central role in the regulation of steroidogenesis by mediating the rate-limiting step in this biosynthetic pathway, which is transport of cholesterol to inner mitochondrial membranes. Although once considered by many to be an insignificant drug-binding site because a specific function remained elusive for many years, peripheral-type benzodiazepine receptors are now viewed with renewed interest because certain benzodiazepines such as diazepam may exert secondary effects on steroid production under appropriate physiologic conditions. Elucidation of this receptorial role should initiate new studies to examine in more detail the pharmacologic profile of drugs that bind to these sites and provides a novel target for the treatment of certain types of endocrine disorders.


Active epilepsy as an index of burden of neurocysticercosis in Vellore district, India.
To determine the contribution of neurocysticercosis (NCC) to the causation of active epilepsy (AE) in a south Indian community. We conducted a door-to-door survey of 50,617 people between the ages of 2 and 60 years in a rural (38,105 people) and urban setting (12,512 people) in the Vellore district of the south Indian state of Tamil Nadu to identify patients with AE. Patients with AE were investigated with a contrast-enhanced CT scan and serologic study using enzyme-linked immunotransfer blot (EITB) for cysticercal antibodies. We identified 194 patients with AE. The prevalence of AE was 3.83 per 1,000 people, with the prevalence in the urban clusters more than twice that in the rural clusters (6.23 vs 3.04 per 1,000) (p < 0.0001). A diagnosis of NCC was made in 46 (28.4%) of the 162 patients undergoing a CT scan, and EITB was positive in 21 (13%) patients. Overall, 55 (34%) patients were diagnosed with NCC (11 definitive NCC and 44 probable NCC). There was no significant difference in the prevalence of NCC causing AE in the urban (1.28 per 1,000) and rural (1.02 per 1,000) communities. NCC is the cause of nearly one-third of all cases of AE in both the urban and rural regions. Extrapolating our results to the country as a whole leads to an estimated disease burden of 1 million patients in India with AE attributable to NCC.


Dynamics of lead (Pb) in different soil conditions.
Four soil samples were collected from various locations having wider range of pH and EC for incubation experiment. These soil samples were added with Pb (NO3)2 salts, and samples were taken on 1st, 35th, 45thand 60th day. Soils samples were fractioned by sequential extraction to estimate the concentration of lead in different fractions viz, water soluble, exchangeable +adsorbed, organic, carbonate and residual fraction. The result reveals that concentration of organic and carbonate bound-Pb was high in waterlogged condition and soil pH has been comprehensively identified as the single most important soil factor controlling the availability of lead (Pb) in soil. Low content of Pb in exchangeable + adsorbed (KNO3) and water soluble (H2O) fraction in all soils (except in S1) could signify low availability of Pb to plants. Bioavailable fractions, viz. water soluble and exchangeable + adsorbed, were low in all soils (except S1) well below critical limits, which may not pose any toxicity in the food chain.


Analysis of five tests commonly used in determining the ability to control single motor units.
A correlation regression analysis was used to analyze five individual test and two types of scores in terms of their predictive value in determining overall performance in single motor unit (SMU) control. Fine wire electrodes were inserted into the biceps brachii muscle of twenty-five women subjects. Each subject was tested for electrical silence, motor unit isolation, on-off control, frequency control and rhythm control. A detailed protocol and scoring criteria are described for each test. Ten predictors were analyzed--a time score and an error score for each of the five tests. A correlation regression analysis showed that the On-Off Control, the Rhythm Control and the Motor Unit Isolation tests have greater predictive value than the remaining tests. Furthermore, the error scores obtained from these tests are more reliable than the time scores in predicting overall performance in SMU control. The implications that the present findings may have on future studies are discussed.


Target discovery screens using pooled shRNA libraries and next-generation sequencing: A model workflow and analytical algorithm.
In the search for novel therapeutic targets, RNA interference screening has become a valuable tool. High-throughput technologies are now broadly accessible but their assay development from baseline remains resource-intensive and challenging. Focusing on this assay development process, we here describe a target discovery screen using pooled shRNA libraries and next-generation sequencing (NGS) deconvolution in a cell line model of Ewing sarcoma. In a strategy designed for comparative and synthetic lethal studies, we screened for targets specific to the A673 Ewing sarcoma cell line. Methods, results and pitfalls are described for the entire multi-step screening procedure, from lentiviral shRNA delivery to bioinformatics analysis, illustrating a complete model workflow. We demonstrate that successful studies are feasible from the first assay performance and independent of specialized screening units. Furthermore, we show that a resource-saving screen depth of 100-fold average shRNA representation can suffice to generate reproducible target hits despite heterogeneity in the derived datasets. Because statistical analysis methods are debatable for such datasets, we created ProFED, an analysis package designed to facilitate descriptive data analysis and hit calling using an aim-oriented profile filtering approach. In its versatile design, this open-source online tool provides fast and easy analysis of shRNA and other count-based datasets to complement other analytical algorithms.


The construction and psychometric properties of an instrument measuring support for sexual rights.
The purpose of this paper is to report the findings of an attempt to construct and examine the psychometric properties of an instrument that measures support for the 11 sexual rights formulated by the World Association of Sexology. Since international conferences held in the mid-1990s, a new approach to sexual issues has been promoted by health professionals, sex educators, and human rights advocates. This approach emphasizes the empowerment of individuals and examines sexual health concerns within a human rights framework. Goals of a sexual rights approach have been to promote awareness and advocacy of sexual issues and especially to facilitate the work toward better sex education and sexual health services. Analysis of reliability and validity data from two university samples (n = 388 and n = 175) indicated that the majority of the sexual rights scales have acceptable psychometric properties. This paper concludes with a discussion of ways to improve the scales for future use and the need to validate the scales using samples that allow generalizability of findings.


T Cell Repertoire Evolution after Allogeneic Bone Marrow Transplantation: An Organizational Perspective.
High-throughput sequencing (HTS) of human T cell receptors has revealed a high level of complexity in the T cell repertoire, which makes it difficult to correlate T cell reconstitution with clinical outcomes. The associations identified thus far are of a broadly statistical nature, precluding precise modeling of outcomes based on T cell repertoire development following bone marrow transplantation (BMT). Previous work has demonstrated an inherent, mathematically definable order observed in the T cells from a diverse group of donors, which is perturbed in recipients following BMT. In this study, T cell receptor (TCR)-β sequences from HLA-matched related donor and recipient pairs are analyzed to further develop this methodology. TCR-β sequencing from unsorted and sorted T cell subsets isolated from the peripheral blood samples of BMT donors and recipients show conservation and symmetry of VJ segment usage in the clonal frequencies, linked to the organization of the gene segments along the TCR locus. This TCR-β VJ segment translational symmetry is preserved post-transplantation and even in cases of acute graft-versus-host disease (aGVHD), suggesting that GVHD occurrence represents a polyclonal donor T cell response to recipient antigens. The complexity of the repertoire is significantly diminished after BMT, and the T cell clonal hierarchy is altered post-transplantation. Low-frequency donor clones tended to take on a higher rank in the recipients following BMT, especially in patients with aGVHD. Over time, the repertoire evolves to a more donor-like state in the recipients who did not develop GVHD as opposed to those who did. The results presented here support new methods of quantifying and characterizing post-transplantation T cell repertoire reconstitution.


From Genome to Phenotype: An Integrative Approach to Evaluate the Biodiversity of Lactococcus lactis.
Lactococcus lactis is one of the most extensively used lactic acid bacteria for the manufacture of dairy products. Exploring the biodiversity of L. lactis is extremely promising both to acquire new knowledge and for food and health-driven applications. L. lactis is divided into four subspecies: lactis, cremoris, hordniae and tructae, but only subsp. lactis and subsp. cremoris are of industrial interest. Due to its various biotopes, Lactococcus subsp. lactis is considered the most diverse. The diversity of L. lactis subsp. lactis has been assessed at genetic, genomic and phenotypic levels. Multi-Locus Sequence Type (MLST) analysis of strains from different origins revealed that the subsp. lactis can be classified in two groups: "domesticated" strains with low genetic diversity, and "environmental" strains that are the main contributors of the genetic diversity of the subsp. lactis. As expected, the phenotype investigation of L. lactis strains reported here revealed highly diverse carbohydrate metabolism, especially in plant- and gut-derived carbohydrates, diacetyl production and stress survival. The integration of genotypic and phenotypic studies could improve the relevance of screening culture collections for the selection of strains dedicated to specific functions and applications.


The Influence of Sonic and Ultrasonic Vibration on the Shear Bond Strength of a Selected Resin Luting Cement.
This study determined the effect of sonic and ultrasonic instrumentation on the shear bond strengths of Panavia 21, a popular cement for the luting of resin-bonded restorations. 84 Ni-Cr cylinders were cemented to randomly selected resin composite substrates using Panavia 21 following the manufacturer's instructions. The Ni-Cr-composite specimens were divided into 7 groups of 12 specimens each based upon the procedure used for removing the excess cement. For Group 1 (Co) specimens the excess cement was removed with microbrushes immediately after cementation. Groups 2 through 7 were based on the use of vibrating instrument and the time period after which the excess material was removed. These included the cement, Panavia 21, three vibrating instruments, Sonic with a universal tip (So), Piezoelectric ultrasonic with a USPIS tip (Pu), Magnetorestrive ultrasonic with a FS1-100 tip (Mu) and two different time periods, soon after cementation (9m) and one hour after cementation (1h). Once excess cement REMOVAL WAS COMPLETED, THE SPECIMENS WERE SUBJECTED TO SHEAR TESTING. Mean Shear Bond strengths ranged from 16.03 MPa (Co) to 19.91 MPa (So 1h). Statistical analysis demonstrated that interaction of the main effects were significant (F = 4.27, p = 0.042). Post-hoc analysis demonstrated that the effect of timing was significant in all the instrumented groups. The majority of the tested specimens failed cohesively compared to mainly adhesive failures for the control group. The effect of type of instrumentation immediately following polymerization setting had no effect on the shear bond strengths however a delay of 1 hour for all types of instrumentation had a beneficial effect of improving observed shear bond strengths.


Nitrendipine-induced gingival overgrowth in dogs.
This study assessed the development of gingival overgrowth in dogs given nitrendipine, a new antihypertensive dihydropyridine. Nine male Beagle dogs with established plaque and gingivitis were used. Following a baseline examination, which involved assessment of plaque, gingivitis, and gingival enlargement, six dogs (test) received nitrendipine twice daily in a high dose, while 3 dogs (control) received placebo. Clinical scorings were repeated after 10 and 20 weeks. At termination of the study gingival biopsies were excised and examined morphometrically. Already at the 10-week examination definite changes in gingival size had occurred, and following 20 weeks of nitrendipine treatment the gingival enlargement had markedly increased. In none of the control dogs were there any signs of gingival size changes. The histopathological examination showed that the only principal changes in histopathological morphology were that areas of non-infiltrated connective tissue in test specimens showed an increase in vascularity and appeared more loose compared with the dense tissue in control specimens. The morphometric analysis demonstrated minor differences between test and control specimens in regard to all tissue fractions observed; however, such differences were numerically very small, and were not considered biologically significant. Thus, the results demonstrated that nitrendipine administered to Beagle dogs during a 20-week period causes marked overgrowth of gingival tissue of apparently normal composition. The fact that cases of gingival overgrowth have been reported among patients receiving other dihydropyridines suggest that the observed gingival overgrowth is an effect of this class of drugs.


Two new linear acetogenins from the fruits of Goniothalamus gracilipes.
Two new linear acetogenins, gracilipin A (1) and methylsaccopetrin A (2) along with seven known compounds, saccopetrin A (3), 7,3',4'-trimethylquercetin (4), rhamnazin (5), casticin (6), isokanugin (7), melisimplexin (8) and 5-hydroxy-3,7-dimethoxy-3',4'-methylenedioxyflavone (9) were isolated from the fruits of Goniothalamus gracilipes Bân. Their structures were established by spectral analysis, such as mass spectrometry, 1D-NMR, 2D-NMR and circular dichroism (CD). Compounds 1 and 3 showed cytotoxic activity against KB cell line with IC50 values of 14.6 and 15.3 μM, respectively.


Cardiovascular biomarker discovery by SELDI-TOF mass spectrometry.
Surface-enhanced laser desorption/ionization time-of-flight mass spectrometry (TOFMS) allows for rapid differential proteomic analysis of various experimental conditions. In the first step of retention chromatography, the proteome to be analyzed is fractionated based on the biochemical properties of the proteins or peptides of which it is composed. TOF-MS separates the proteins according to their molecular mass and charge (m/z in which conditions are optimized to obtain single charged proteins: z = 1). The most discriminating conditions are used to validate differential peaks on a larger number of samples. Once the biomarker is statistically validated, its identification is performed by reproducing the chromatographic conditions that are appropriate for its retention on spin-columns, followed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, trypsin digestion of the band of interest, and subsequent peptide mapping after MS. It may be necessary to determine part of the amino acid sequence of the protein of interest to confirm its identity.


Suppression of experimental allergic encephalomyelitis with thoracic duct lymphocytes.
Thoracic duct lymphocytes (TDL) from Lewis rats immunized 9-10 days previously with basic protein in complete Freund's adjuvant (BP-CFA) failed to induce experimental allergic encephalomyelitis (EAE) in syngeneic recipients. This contrasts with the successful transfer of EAE by lymph node cell suspensions from donors immunized 9 days previously with BP-CFA. Only minor EAE was induced passively by TDL from rats immunized 11-12 days before with BP-CFA. TDL collected 9-20 days after BP-CFA immunization, however, were successful in transferring specific suppression of EAE tested by the lack of disease in the recipients immunized actively with BP-CFA 1 week after the TDL transfer. The data indicate that the thoracic duct contains specific suppressor cells shortly before, during and after the development of clinical EAE.


COVID-19 diagnosis and management: a comprehensive review.
Severe acute respiratory syndrome coronavirus (SARS-CoV)-2, a novel coronavirus from the same family as SARS-CoV and Middle East respiratory syndrome coronavirus, has spread worldwide leading the World Health Organization to declare a pandemic. The disease caused by SARS-CoV-2, coronavirus disease 2019 (COVID-19), presents flu-like symptoms which can become serious in high-risk individuals. Here, we provide an overview of the known clinical features and treatment options for COVID-19. We carried out a systematic literature search using the main online databases (PubMed, Google Scholar, MEDLINE, UpToDate, Embase and Web of Science) with the following keywords: 'COVID-19', '2019-nCoV', 'coronavirus' and 'SARS-CoV-2'. We included publications from 1 January 2019 to 3 April 2020 which focused on clinical features and treatments. We found that infection is transmitted from human to human and through contact with contaminated environmental surfaces. Hand hygiene is fundamental to prevent contamination. Wearing personal protective equipment is recommended in specific environments. The main symptoms of COVID-19 are fever, cough, fatigue, slight dyspnoea, sore throat, headache, conjunctivitis and gastrointestinal issues. Real-time PCR is used as a diagnostic tool using nasal swab, tracheal aspirate or bronchoalveolar lavage samples. Computed tomography findings are important for both diagnosis and follow-up. To date, there is no evidence of any effective treatment for COVID-19. The main therapies being used to treat the disease are antiviral drugs, chloroquine/hydroxychloroquine and respiratory therapy. In conclusion, although many therapies have been proposed, quarantine is the only intervention that appears to be effective in decreasing the contagion rate. Specifically designed randomized clinical trials are needed to determine the most appropriate evidence-based treatment modality.


A trans-theoretical approach to alcohol abuse profile in the general population of an islamic country - Mashhad, Iran.
Background:From a public health perspective, alcohol-related problems have enormous social and individual consequences. Objectives:The aim of the present study was to apply the TTM on the general population of Mashhad city to evaluate the change levels and possible relative factors regarding alcohol abuse. Methods:: In a cross-sectional design, a total number of 564 people from the General Population of Mashhad, Iran participated. Stages of change questionnaire based on trans-theoretical model (TTM) and the checklist including socio-demographic characteristics as well as possible related factors were used. SPSS 11.5 software was used for all statistical analyses. Results: Among 564 people who took part in this survey, 245 (43.43%) had the history of alcohol consumption or they were current alcohol users. The analysis showed that 19.2% of the participants were in pre-contemplation stage, 3.3% in contemplation, 1.2% in preparation, 2.9% in action, 2% in maintenance and 71.4% were in termination phase. Age, job, smoking, and hookah smoking were identified as predictors of pre-contemplation stage. Marital status, job, and smoking were predictors of termination phase. Conclusion: This picture is from an Islamic holy city where assumed religious beliefs have cramped drinking patterns. According to harm of alcohol abuse, it is necessary to tailor the intervention for target populations. Factors identified as predictors of alcohol abuse such as age, marital status and occupation, as well as being cigarette and hookah smokers should be taken into account in the design of future interventions.


Implicit measures of "wanting" and "liking" in humans.
Incentive Sensitization Theory (IST; e.g., Robinson and Berridge, 1993. Brain Res. Rev., 18, 291; Robinson and Berridge, 2003 Trends Neurosci., 26, 507) suggests that a common dopamine system that deals with incentive salience attribution is affected by different types of drugs. Repeated drug use will sensitize this neural system, which means that drugs increasingly trigger the experience of incentive salience or "wanting". Importantly, Robinson and Berridge stress that there is a dissociation between drug "wanting" (the unconscious attribution of incentive salience) and drug "liking" (the unconscious hedonic experience when one consumes drugs). Whereas the former plays an essential role in the development and maintenance of drug addiction, the latter does not. Although this model was based mainly on research with non-human animals, more recently the dissociation between "wanting" and "liking" has been examined in humans as well. A widely used and promising means of studying these processes are behavioral implicit measures such as the Implicit Association Test (IAT), the Approach-Avoidance Task (AAT), different types of Stimulus-Response Compatibility (SRC) tasks, and Affective Simon Tasks (AST). IST makes the clear prediction that (1) there should be a positive correlation between indices of "wanting" (e.g., drug consumption) and implicit "wanting" scores. Similarly, there should be a positive correlation between indices of "liking" (e.g., various expressions of subjective pleasure) and implicit "liking" scores; (2) there should be higher "wanting" scores in substance abusers or frequent substance users compared to non-users or infrequent users, and there should be no differences in "liking" between these groups (or even less "liking" in frequent substance users); (3) manipulations of "wanting" should affect implicit "wanting" scores whereas manipulations of "liking" should affect implicit "liking" scores. However, studies that tested these hypotheses did not produce equivocal results. To shed light on these discrepancies, we first discuss the different definitions of "wanting" and "liking" and the different tests that have been used to assess these processes. Then, we discuss whether it is reasonable to assume that these tests are valid measures of "wanting" and "liking" and we review correlational, quasi-experimental, and experimental studies that inform us about this issue. Finally, we discuss the future potential of implicit measures in research on IST and make several recommendations to improve both theory and methodology.


RNF157 attenuates CD4+ T cell-mediated autoimmune response by promoting HDAC1 ubiquitination and degradation.
Background: CD4+ T cells play an important role in body development and homeostasis. Quantitative and functional changes in CD4+ T cells result in abnormal immune responses, which lead to inflammation, cancer, or autoimmune diseases, such as multiple sclerosis (MS). Ubiquitination plays an essential role in the differentiation and functioning of CD4+ T cells. However, the function of several E3 ubiquitin ligases in CD4+ T cell differentiation and T cell-mediated pathological diseases remains unclear. Methods: RNA sequencing data were analyzed to identify the E3 ubiquitin ligases that participate in the pathogenesis of MS. Furthermore, conditional knockout mice were generated. Specifically, flow cytometry, qPCR, western blot, CO-IP and cell transfer adoptive experiments were performed. Results: In this study, we identified The RING finger 157 (RNF157) as a vital regulator of CD4+ T cell differentiation; it promoted Th1 differentiation but attenuated Th17 differentiation and CCR4 and CXCR3 expressions in CD4+ T cells, thereby limiting experimental autoimmune encephalomyelitis development. Mechanistically, RNF157 in CD4+ T cells targeted HDAC1 for K48-linked ubiquitination and degradation. Notably, RNF157 expression was significantly decreased and showed a significant negative correlation with RORγt expression in patients with MS. Conclusions: Our study highlights the critical role of RNF157 in regulating CD4+ T cell functions in autoimmune diseases and suggests RNF157 as a potential target in adaptive immune responses against MS and other autoimmune disorders.


Auditory adaptation: notes on power and parabolic curve fitting across intensity and time. I.
The classic procedure for the study of auditory adaptation has been the method of Simultaneous Dichotic Loudness Balance ( SDLB ). Data from four publications ( Jerger , 1957; Palva and Karja , 1969; Weiler , Loeb and Alluisi , 1972 and Weiler and Hood, 1977) were combined to yield values extending from 20 dB to 80 dB average sensation level. A growth curve was fitted to the data by means of the Gompertz curve-fitting technique. A close fit was obtained, and a somewhat closer fit was found using a parabolic curve. In addition to the average data for SDLB adaptation across test intensities, the Gompertz method was used to fit the course of adaptation over time for one additional study ( Thwing , 1955).


The use of functionally graded dental crowns to improve biocompatibility: a finite element analysis.
In post-core crown restorations, the significant mismatch between stiffness of artificial crowns and dental tissues leads to stress concentration at the interfaces. The aim of the present study was to reduce the destructive stresses by using a class of inhomogeneous materials called functionally graded materials (FGMs). For the purpose of the study, a 3-dimentional computer model of a premolar tooth and its surrounding tissues were generated. A post-core crown restoration with various crown materials, homogenous and FGM materials, were simulated and analyzed by finite element method. Finite element and statistical analysis showed that, in case of oblique loading, a significant difference (p < 0.05) was found at the maximum von Mises stresses of the crown margin between FGM and homogeneous crowns. The maximum von Mises stresses of the crown margin generated by FGM crowns were lower than those generated by homogenous crowns (70.8 vs. 46.3 MPa) and alumina crown resulted in the highest von Mises stress at the crown margin (77.7 MPa). Crown materials of high modulus of elasticity produced high stresses at the cervical region. FGM crowns may reduce the stress concentration at the cervical margins and consequently reduce the possibility of fracture.


Predictive Value of School-Aged Children's Schistosomiasis Prevalence and Egg Intensity for Other Age Groups in Western Kenya.
World Health Organization recommendations for the timing and target population for mass drug administration (MDA) for schistosomiasis are based on the prevalence of infection in school children within a given community. In a large study comparing MDA approaches for Schistosoma mansoni control, we evaluated whether prevalence of infection and egg burdens in 9- to 12-year-old students reflected infection levels in young children and adults in the same community. Cross-sectional surveys of preadolescents (9-12 years old) were compared with those of first year students (5-8 years old) in 225 villages and adults (20-55 years old) in 150 villages along the Kenyan shores of Lake Victoria. Village schistosomiasis prevalence and intensity levels in preadolescents strongly correlated (P < 0.0001) with prevalence and infection intensity for other age groups in the community. Our findings suggest that S. mansoni prevalence and intensity among 9- to 12-year-olds are valid for community sampling purposes in mapping for MDAs.


Production of interleukin 1 in glomerular cell cultures from rats with nephrotoxic serum nephritis.
To investigate the role of cytokines in the pathogenesis of experimental glomerulonephritis (GN), we examined interleukin 1 (IL-1) activity in isolated glomeruli of rats with an accelerated autologous form of nephrotoxic serum nephritis (NTSN). Glomeruli were isolated and explanted in tissue culture. The prominent feature of the glomerular outgrowth of the glomeruli in the NTSN was the presence of large numbers of type III cells (macrophages). In addition, there were significantly greater numbers of type II (mesangial) cells in culture from the NTSN rats as compared with glomeruli from normal rats, though the numbers of type I (epithelial) cells were the same. IL-1 bioactivity was significantly higher in culture supernatants generated by glomeruli isolated from NTSN animals versus normal animals early in the course of the disease. When glomerular culture supernatants and purified IL-1 were pre-incubated with an anti-IL-1 beta antibody, a parallel decrease of the biological activity was found, suggesting that the biologically active binding sites of the culture supernatants and monocyte-produced IL-1 share some common structure. The administration of a rabbit anti-rat macrophage serum (AMS) prevented the outgrowths of type III cells (macrophages) and type II (mesangial) cells and reduced the production of IL-1 activity in the NTSN rats. In this experimental model IL-1 synthesis, probably by multiple cell types, is present early in the disease process and perhaps may be an important mediator of glomerular immune injury.


Accelerated cellular senescence phenotype of GAPDH-depleted human lung carcinoma cells.
Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) is a pivotal glycolytic enzyme, and a signaling molecule which acts at the interface between stress factors and the cellular apoptotic machinery. Earlier, we found that knockdown of GAPDH in human carcinoma cell lines resulted in cell proliferation arrest and chemoresistance to S phase-specific cytotoxic agents. To elucidate the mechanism by which GAPDH depletion arrests cell proliferation, we examined the effect of GAPDH knockdown on human carcinoma cells A549. Our results show that GAPDH-depleted cells establish senescence phenotype, as revealed by proliferation arrest, changes in morphology, SA-β-galactosidase staining, and more than 2-fold up-regulation of senescence-associated genes DEC1 and GLB1. Accelerated senescence following GAPDH depletion results from compromised glycolysis and energy crisis leading to the sustained AMPK activation via phosphorylation of α subunit at Thr172. Our findings demonstrate that GAPDH depletion switches human tumor cells to senescent phenotype via AMPK network, in the absence of DNA damage. Rescue experiments using metabolic and genetic models confirmed that GAPDH has important regulatory functions linking the energy metabolism and the cell cycle networks. Induction of senescence in LKB1-deficient non-small cell lung cancer cells via GAPDH depletion suggests a novel strategy to control tumor cell proliferation.


T Cell Proliferative Responses and IgG Antibodies to β2GPI in Patients with Diabetes and Atherosclerosis.
Diabetes increases the risk of myocardial infarction (MI) by 2 to 3 folds. Tlymphocytes play a role in atherosclerosis, which is the main pathology behind MI. Cellular immune responses to beta-2 glycoprotein I (β2GPI) are shown in carotid atherosclerosis. To investigate the self-reactive, β2GPI-specific T-lymphocytes in patients with and without diabetes and atherosclerosis. Collectively, 164 subjects with and without diabetes that underwent coronary angiography were divided into four groups based on their diabetes status and coronary stenosis. Group I=Diabetic with ≥50% stenosis: A+D+ (n=66); Group II=Non-diabetic with ≥50% stenosis, A+D- (n=39); Group III=Diabetic with <50% stenosis: A-D+ (n=28); and Group IV=Non-diabetic with <50% stenosis: AD- (n=31). All groups were evaluated for anti-β2GPI IgG antibody by ELISA method. Then, PBMCs were isolated from 18 subjects and were stimulated with β2GPI-derived peptides to assess their proliferation in accordance with their HLA-DRB1 alleles. Mean β2GPI IgG levels were higher in groups with ≥50% stenosis (A+) compared to those with <50% stenosis (A-), (P=0.02). The co-presence of diabetes in A+ individuals increased mean β2GPI-specific IgG. Auto-reactive β2GPI-specific T cells were detected in the repertoire of T-lymphocytes in all groups. β2GPI-peptides showed promiscuous restriction by various HLADRB1. β2GPI is the target of cellular and humoral immune responses in patients with atherosclerosis. Since the T cell responses but not antibodies were detectable in A-D+ and A-D- groups, it is reasonable to assume that cellular responses preceded the humoral responses. Post-translation modifications of β2GPI under oxidative and glycemic stresses may have increased the IgG levels in patients with diabetes. Finally, identification of antigens that trigger immuno-pathogenesis in atherosclerosis and diabetes may help the development of immunomodulation methods to prevent or treat these debilitating diseases.


Temperature distributions during thermoradiotherapy: a sensitivity study with a transient numerical model of the rabbit eye.
An approach to the treatment of medium-sized choroidal melanomas combines radiation with ferromagnetic hyperthermia. The study herein discusses results with a numerical thermal model of a choroidal melanoma in the rabbit eye as treated with episcleral, thermoradiotherapy plaques. The sensitivity of a temperature-dependent blood perfusion model is investigated.


Generation of an E. coli platform strain for improved sucrose utilization using adaptive laboratory evolution.
Sucrose is an attractive industrial carbon source due to its abundance and the fact that it can be cheaply generated from sources such as sugarcane. However, only a few characterized Escherichia coli strains are able to metabolize sucrose, and those that can are typically slow growing or pathogenic strains. To generate a platform strain capable of efficiently utilizing sucrose with a high growth rate, adaptive laboratory evolution (ALE) was utilized to evolve engineered E. coli K-12 MG1655 strains containing the sucrose utilizing csc genes (cscB, cscK, cscA) alongside the native sucrose consuming E. coli W. Evolved K-12 clones displayed an increase in growth and sucrose uptake rates of 1.72- and 1.40-fold on sugarcane juice as compared to the original engineered strains, respectively, while E. coli W clones showed a 1.4-fold increase in sucrose uptake rate without a significant increase in growth rate. Whole genome sequencing of evolved clones and populations revealed that two genetic regions were frequently mutated in the K-12 strains; the global transcription regulatory genes rpoB and rpoC, and the metabolic region related to a pyrimidine biosynthetic deficiency in K-12 attributed to pyrE expression. These two mutated regions have been characterized to confer a similar benefit when glucose is the main carbon source, and reverse engineering revealed the same causal advantages on M9 sucrose. Additionally, the most prevalent mutation found in the evolved E. coli W lineages was the inactivation of the cscR gene, the transcriptional repression of sucrose uptake genes. The generated K-12 and W platform strains, and the specific sets of mutations that enable their phenotypes, are available as valuable tools for sucrose-based industrial bioproduction in the facile E. coli chassis.


Frequency and spectrum of mutations at codons 12 and 13 of the c-K-ras gene in human tumors.
The frequency of point mutations at codons 12 and 13 of the c-K-ras gene has been determined in a panel of more than 400 human tumors. Mutant c-K-ras genes were detected in about 75% of adenocarcinomas of the pancreas (n = 84); 40% of adenomas (n = 72) and carcinomas (n = 244) of the colon end rectum; 30% of carcinomas of the bile duct (n = 19); 25% of carcinomas of the lung (n = 92), and in lower frequency in other carcinomas, including liver, stomach, and kidney. No mutations were found in carcinomas of the breast, prostate, esophagus, and gall bladder, among others. Comparative analysis of the spectrum of mutations show that while G to A transitions were the most frequent mutations in pancreatic and colo-rectal tumors, G to T transversions were more prevalent in lung carcinomas. The aspartic acid mutation at codon 13 (GGC----GAC) was relatively frequent in colo-rectal tumors but rare in pancreatic and lung carcinomas. The differences in the mutation spectrum of the c-K-ras gene in cancers of the gastrointestinal and respiratory tracts are suggestive of differential exposure to genotoxic agents.


Hyper-reactive malarial splenomegaly in the absence of raised IgM antibodies.
Hyper-reactive malarial splenomegaly (HMS) is diagnosed by the presence of massive splenomegaly, raised IgM and antimalarial antibodies and a response to antimalarial therapy. Although malaria is endemic to Pakistan, HMS is uncommon. We report on HMS in a patient with massive splenomegaly, positive Plasmodium falciparum polymerase chain reaction but normal immunoglobulin M antibody levels. The investigations were not consistent with any other diagnosis. HMS is also briefly reviewed.


Implementation of Evidence-Based Presurgical Testing Guidelines in Patients Undergoing Ambulatory Surgery for Endometrial Cancer.
The aim of this quality improvement intervention was to evaluate the safety and cost savings of presurgical testing (PST) guidelines for patients undergoing hysterectomy for endometrial pathology in the ambulatory setting. Evidence-based presurgical testing (PST) guidelines were developed by a multidisciplinary team. These guidelines were implemented on the gynecologic surgery service of a comprehensive cancer center in January 2016. All patients with a diagnosis of endometrial pathology who underwent ambulatory surgery during the specified time periods were included in this analysis. A pre-post analysis was performed (preperiod, July 2014-December 2015; postperiod, July 2016-December 2017). Rates of completed presurgical tests and perioperative adverse events were compared between time periods. Cost savings related to the reduction in PST were calculated using the direct cost of testing and reported in percentage cost reduction. A total of 749 hysterectomies were completed in the preperiod and 775 in the postperiod. After implementation of PST guidelines, complete blood counts, coagulation testing, comprehensive metabolic panels, chest x-rays, and electrocardiograms were reduced by 13.4%, 78.1%, 36.8%, 39.0%, and 15.5%, respectively (all P < .001). Rates of perioperative cardiopulmonary adverse events (0% v 0%) and hematologic adverse events (3.3% v 2.0%; P = .10) were stable between time periods. There were no deaths within 90 days of surgery. There was a 41.4% reduction in direct costs related to PST in the postperiod. The use of evidence-based PST guidelines for patients with endometrial pathology undergoing hysterectomy in the ambulatory setting is safe and cost-effective. A multidisciplinary approach is essential for successful development and implementation.


X-ray parametric scattering by a diamond crystal.
Spontaneous X-ray parametric scattering from a diamond single crystal has been observed at the Photon Factory. The high perfection of the diamond single crystal and the small angular divergence of synchrotron radiation has enabled down-converted X-ray photon pairs to scatter into two small solid angles which satisfy the phase-matching condition. High-efficiency noise reduction was performed using avalanche photodiode detectors with good time resolution.


Variation in morphology of the valve of the great cardiac vein in the human hearts.
The research was carried on 70 hearts of adult humans. The hearts were subjected to direct analyses on the autopsy material. The presence or absence of the valve of the great cardiac vein was analysed with reference to sex. The terminal valve of the great cardiac vein was found in 92% of the examined hearts. To define the location of the valve of the great cardiac vein the osmium of the left atrium oblique vein was treated as the reference point. In 3% of cases the presence of the valve was observed independently of the opening of the left atrium oblique vein. The valve of the great cardiac vein was placed proximally from the opening of the left atrium oblique vein. The valve of the great cardiac vein placed distally in relation to the beginning of the coronary sinus was never observed. The three types of the great cardiac vein valves were found: 1. single, semilunar cusp; 2. endothelial fold; 3. double pouches.


Electric quadrupole transitions in carbon dioxide.
Recent advances in high sensitivity spectroscopy have made it possible, in combination with accurate theoretical predictions, to observe, for the first time, very weak electric quadrupole transitions in a polar polyatomic molecule of water. Here, we present accurate theoretical predictions of the complete quadrupole rovibrational spectrum of a non-polar molecule CO2, important in atmospheric and astrophysical applications. Our predictions are validated by recent cavity enhanced absorption spectroscopy measurements and are used to assign few weak features in the recent ExoMars Atmospheric Chemistry Suite mid-infrared spectroscopic observations of the Martian atmosphere. Predicted quadrupole transitions appear in some of the mid-infrared CO2 and water vapor transparency regions, making them important for detection and characterization of the minor absorbers in water- and CO2-rich environments, such as those present in the atmospheres of Earth, Venus, and Mars.


Stigma resistance and its association with internalised stigma and psychosocial outcomes among psychiatric outpatients.
Studies have suggested that stigma resistance plays an important role in the recovery from mental illness. However, there has been limited research in Asian countries that has examined the benefits of stigma resistance among the mentally ill in Asian populations. Hence, this study aimed to assess the prevalence of stigma resistance and establish the socio-demographic correlates of stigma resistance, as well as its association with internalised stigma and psychosocial outcomes among a multi-ethnic population of 280 outpatients with obsessive compulsive disorder (OCD), schizophrenia, depressive disorders and anxiety disorders in Singapore. Prevalence of stigma resistance measured using the Stigma Resistance subscale of the Internalised Stigma of Mental Illness Scale was 82.9%. ANOVA and logistic regressions were conducted and results revealed that: (i) Stigma resistance was positively associated with being separated/divorced/widowed but negatively associated with depression diagnosis; (ii) Psychosocial outcomes such as self-esteem and psychological health were positively associated with stigma resistance; and (iii) Internalised stigma was negatively associated with stigma resistance. Moving forward, treatments could emphasize on improving the self-esteem and psychological health of patients to increase their stigma resistance for counteracting effects of public and internalised stigma.


[Acute Postoperative Negative Pressure Pulmonary Edema Caused by the Compression of Brachiocephalic Artery].
We report a case of negative-pressure pulmonary edema occurring by tracheal obstruction caused by the brachiocephalic artery. The patient had deformed thorax with cerebral palsy, which deformed thorax placing the brachiocephalic artery high over the trachea, resulting in close and tight contact between the artery and trachea. Additional deformity of the thorax associated with myotonic attacks after general anesthesia might shorten the distance between the sternal notch and the vertebral body, resulting in the tracheal obstruction by the artery.


Sperm-bound amidase activity as a marker for acrosomal integrity in bull spermatozoa.
An assay for sperm-bound amidase activity was validated using bovine spermatozoa and N-benzoyl-DL-arginine p-nitroanilide as substrate. The assay had intra- and interassay coefficients of variations of 5 and 12%, respectively. It is an inexpensive, simple and rapid assay since 100 samples can be evaluated in 2 hours and it requires only 4 X 10(6) spermatozoa per sample. Sperm-bound amidase activity was proportional (r = 0.95) to the percentage of spermatozoa with an intact acrosome, as determined by differential interference-contrast microscopy. A change of five percentage units in the incidence of damaged spermatozoa was detectable. Using this procedure, assessment of sperm-bound amidase activity is therefore a sensitive and efficient means of evaluating acrosomal integrity.


Combination of citicoline and physiotherapy in children with cerebral palsy.
The most common cause of physical disability in children is cerebral palsy. This study was aimed to evaluate the effect of citicoline in combination to physiotherapy versus physiotherapy alone, to improve the functional outcome in pediatric cerebral palsy. The clinical trial was performed on 50 pediatric patients aged 18-75 months with spastic diplegia or quadriplegic cerebral palsy. Patients were assessed in two groups: case group, under treatment, using injection of citicoline (10 mg/kg) every other day for 3 months and physiotherapy. Gross motor function classification system (GMFCS) levels were assessed in all patients before and after treatment. Patient's mean age was 38.7 ± 17.2 months, and 52% were girls. Differences in the frequency of GMFCS levels between groups were not statistically significant, before (P = 0.09) and after (P = 0.47) treatment. In case group improving in GMFCS, level was occurred in 9/11 with level 2 to level 1, 3/5 with level 3 to other levels and 3/7 with level 4 to other levels. In the control group improving in GMFCS, level was occurred in 3/9 with level 2 to level 1, 3/10 with level 3 to other levels, and 2/4 with level 4 other levels. GMFCS level in 64% of cases was improved, whereas in control group, 32% was improved (P = 0.02). Results demonstrated that citicoline in combination to physiotherapy appears to be a promising agent to improve gross motor function in patients with cerebral palsy versus physiotherapy alone. Although, further studies are need to be done.


Efficacy and safety of angiotensin II receptor blockade in elderly patients with diabetes.
While national guidelines recommend ACE inhibitor (ACEI) or angiotensin receptor blocker (ARB) therapy in patients with diabetes and nephropathy, guidelines concerning elderly patients with diabetes have not endorsed these drugs. We sought to assess the nephroprotective efficacy and safety of ARB therapy in elderly patients by conducting age-specific subgroup analyses using data from the Reduction of Endpoints in NIDDM with the Angiotensin II Antagonist Losartan (RENAAL) study. We studied 1,513 patients with type 2 diabetes and nephropathy who randomly received either losartan or placebo. We tested for effect modification by age of the effect of losartan on the incidence of the predefined end points (doubling of serum creatinine, end-stage renal disease [ESRD], or death) and the risk of adverse events. Of 1,513 participants, 421 (27.8%) were aged >65 years (maximum age 74 years). Age did not modify the efficacy of losartan in reducing the risk of the primary outcome, a composite of doubling of serum creatinine, ESRD, or death (P(interaction) = 0.66) or its individual components (all P(interaction) > 0.44). In patients aged >65 years, losartan reduced the risk of ESRD by 50% (95% CI 30-81, P = 0.005). We found no evidence that older patients were more likely to experience adverse events from losartan such as a rise in serum creatinine or hyperkalemia than younger patients. Elderly patients had the same level of benefits and risks as younger patients from treatment with losartan. Underuse of ACEI and ARB therapy in elderly patients because of the perceived lack of efficacy or a greater risk of adverse events appears unjustified.


Dexmedetomidine maintains blood-brain barrier integrity by inhibiting Drp1-related endothelial mitochondrial dysfunction in ischemic stroke.
Stroke is the second leading cause of death and long-term disability worldwide, which lacks effective treatment. Perioperative stroke is associated with much higher rates of mortality and disability. The neuroprotective role of dexmedetomidine (Dex), a highly selective agonist of alpha2-adrenergic receptor, has been reported in a stroke rat model, and it was found that pretreatment of Dex before stroke could alleviate blood-brain barrier (BBB) breakdown. However, the underlying mechanisms are still unknown. As the brain endothelial cells are the main constituents of BBB and in high demand of energy, mitochondrial function of endothelial cells plays an important role in the maintenance of BBB. Given that dynamin-related protein 1 (Drp1) is a protein mediating mitochondrial fission, with mitochondrial fusion that balances mitochondrial morphology and ensures mitochondria function, the present study was designed to investigate the possible role of Drp1 in endothelial cells involved in the neuroprotective effects of Dex in ischemic stroke. Our results showed that preconditioning with Dex reduced infarction volume, alleviated brain water content and BBB damage, and improved neurological scores in middle cerebral artery occlusion rats. Meanwhile, Dex enhanced cell activity and decreased cell apoptosis in oxygen-glucose deprivation human brain microvascular endothelial cells in vitro. These protective effects of Dex were correlated with the mitochondrial morphology integrality of endothelial cells, mediated by increased phosphorylation of serine 637 in Drp1, and could be reversed by α2-adrenergic receptor antagonist Yohimbine and AMP-activated protein kinase inhibitor Compound C. These findings suggest new molecular pathways involved in the neuroprotective effects of Dex in ischemic stroke. As Dex is routinely used as a sedative drug clinically, our findings provide molecular evidence that it has perioperative neuroprotection from ischemic stroke.


My journey to DNA repair.
I completed my medical studies at the Karolinska Institute in Stockholm but have always been devoted to basic research. My longstanding interest is to understand fundamental DNA repair mechanisms in the fields of cancer therapy, inherited human genetic disorders and ancient DNA. I initially measured DNA decay, including rates of base loss and cytosine deamination. I have discovered several important DNA repair proteins and determined their mechanisms of action. The discovery of uracil-DNA glycosylase defined a new category of repair enzymes with each specialized for different types of DNA damage. The base excision repair pathway was first reconstituted with human proteins in my group. Cell-free analysis for mammalian nucleotide excision repair of DNA was also developed in my laboratory. I found multiple distinct DNA ligases in mammalian cells, and led the first genetic and biochemical work on DNA ligases I, III and IV. I discovered the mammalian exonucleases DNase III (TREX1) and IV (FEN1). Interestingly, expression of TREX1 was altered in some human autoimmune diseases. I also showed that the mutagenic DNA adduct O(6)-methylguanine (O(6)mG) is repaired without removing the guanine from DNA, identifying a surprising mechanism by which the methyl group is transferred to a residue in the repair protein itself. A further novel process of DNA repair discovered by my research group is the action of AlkB as an iron-dependent enzyme carrying out oxidative demethylation.


Dissociative Experiences are Associated with Obsessive-Compulsive Symptoms in a Non-clinical Sample: A Latent Profile Analysis.
There has been a burgeoning literature considering the significant associations between obsessive-compulsive symptoms and dissociative experiences. In this study, the relationsips between dissociative symtomotology and dimensions of obsessive-compulsive symptoms were examined in homogeneous sub-groups obtained with latent class algorithm in an undergraduate Turkish sample. Latent profile analysis, a recently developed classification method based on latent class analysis, was applied to the Dissociative Experiences Scale (DES) item-response data from 2976 undergraduates. Differences in severity of obsessive-compulsive symptoms, anxiety and depression across groups were evaluated by running multinomial logistic regression analyses. Associations between latent class probabilities and psychological variables in terms of obsessive-compulsive sub-types, anxiety, and depression were assessed by computing Pearson's product-moment correlation coefficients. The findings of the latent profile analysis supported further evidence for discontinuity model of dissociative experiences. The analysis empirically justified the distinction among three sub-groups based on the DES items. A marked proportion of the sample (42%) was assigned to the high dissociative class. In the further analyses, all sub-types of obsessive-compulsive symptoms significantly differed across latent classes. Regarding the relationships between obsessive-compulsive symptoms and dissociative symptomatology, low dissociation appeared to be a buffering factor dealing with obsessive-compulsive symptoms; whereas high dissociation appeared to be significantly associated with high levels of obsessive-compulsive symptoms. It is concluded that the concept of dissociation can be best understood in a typological approach that dissociative symptomatology not only exacerbates obsessive-compulsive symptoms but also serves as an adaptive coping mechanism.


Comprehensibility and neural substrate of communicative gestures in severe aphasia.
Communicative gestures can compensate incomprehensibility of oral speech in severe aphasia, but the brain damage that causes aphasia may also have an impact on the production of gestures. We compared the comprehensibility of gestural communication of persons with severe aphasia and non-aphasic persons and used voxel based lesion symptom mapping (VLSM) to determine lesion sites that are responsible for poor gestural expression in aphasia. On group level, persons with aphasia conveyed more information via gestures than controls indicating a compensatory use of gestures in persons with severe aphasia. However, individual analysis showed a broad range of gestural comprehensibility. VLSM suggested that poor gestural expression was associated with lesions in anterior temporal and inferior frontal regions. We hypothesize that likely functional correlates of these localizations are selection of and flexible changes between communication channels as well as between different types of gestures and between features of actions and objects that are expressed by gestures.


Electrophysiological and inotropic effects of H 234/09 (almokalant) in vitro: a comparison with two other novel IK blocking drugs, UK-68,798 (dofetilide) and E-4031.
The aim was to compare the electrophysiological and inotropic effects of the novel class III agents H 234/09, UK-68,798, and E-4031 in vitro. The electrophysiological effects were investigated by recording transmembrane action potentials in the isolated ventricular muscle and Purkinje fibres of the rabbit; effects on force (adjusted to the maximum isoprenaline response) and refractoriness were investigated in the isolated cat papillary muscle. It was shown that all the drugs induced a concentration dependent prolongation of the action potential duration, which was much more pronounced in the Purkinje fibres than in the ventricular muscle. However, when compared at concentrations giving a 15% increase of the action potential duration in ventricular muscle, H 234/09 was significantly less effective in the Purkinje fibres than the other two drugs. In the cat papillary muscle all drugs induced an increase in force development. This increase tended to parallel the increase in effective refractory period. However, at prolongations of effective refractory period of more than approximately 50% the increase in developed force levelled off. All the class III agents investigated showed a positive inotropic effect, which may be of advantage when compared to conventional class I antiarrhythmic agents, which have cardiodepressant actions. Compared to UK-68,798 and E-4031, H 234/09 showed a less unfavourable profile in terms of dispersion of repolarisation, which theoretically may reduce the risk of arrhythmias associated with delayed repolarisation. However, this less unfavourable profile must, like the positive inotropic effect, ultimately be investigated in clinical trials.


[Response surface methodology for optimizing the technology of induction and isolation of dendritic cells by multiple cytokines].
Objective To promote the induction and separation efficiency of bone marrow-derived dendritic cells (BMDCs) in vitro through optimizing the inducing and isolating process by multiple cytokines. Methods The factors to be optimized in single factor tests included recombinant mouse granulocyte macrophage-colony stimulating factor (rmGM-CSF), recombinant mouse interleukine 4 (rmIL-4), lipopolysaccharide (LPS), recombinant mouse tumor necrosis factor α (rmTNF-α) and inducing time. The numbers of immature dendritic cells (imDCs) and mature dendritic cells (mDCs) were investigated as the indicators. Box-Behnken experimental design-response surface methodology was used to analyze and verify the data. Morphological changes were observed using the inverted microscopy and the transmission electron microscopy. Surface molecules including CD11c and CD86 were detected using the flow cytometry. Results The optimum inducing conditions for imDCs were obtained as follows: rmGM-CSF was 46 ng/mL, rmIL-4 was 24 ng/mL, inducing time was 6 days, and the number of imDCs was (4.58±0.28)×106 cells, and the relative deviation was 4.00%. The optimum inducing conditions for mDCs were as follows: LPS was 1.4 μg/mL, rmTNF-α was 30 ng/mL, inducing time was 1 day, and the number of mDCs was (4.21±0.15)×106 cells, and the relative deviation was 3.80%. Sufficient typical imDCs and mDCs were obtained within 5-7 days of induction in vitro. Also, flow cytometry showed that the amplified imDCs had a high expression of CD11c (68.62%±2.3%) and a low expression of CD86 (37.95%±1.8%), and the mDCs had high expressions of both CD86 (90.34%±1.4%) and CD11c (82.05%±1.6%). Conclusion The combination of single factor tests and Box-Behnken design -response surface methodology could optimize the inducing and isolating method for DCs in vitro by multiple cytokines rapidly and efficiently, which provided basic experiment materials for further studies.


[Clinical application of the modified wisconsin card sorting test to children with attention deficit/hyperactivity disorder].
The cases with attention deficit/hyperactivity disorder (AD/HD) are known to have difficulties in performing various neuropsychological tests related to the executive function. Among them, the Wisconsin Card Sorting Test (WCST) is already applied to many children with AD/HD. There are, however, differences in the measurement of WCST, and also in the background conditions of the patients, such as the status of medication and the level of Intelligence Quotient (IQ), and presumably as a result, the outcome of WCST shows a diversity. The Keio version WCST (KWCST) is a modified WCST by reducing the number of cards and presenting subjects in two steps separated by a short pause, during which a brief instruction is given. This study was undertaken to compare the performances of children with AD/HD to normal controls using KWCST according to full-scale IQ (FIQ) and also the subtypes of AD/HD. Subjects in this study were 21 unmedicated children with AD/HD, ranging from 5 to 15 years of age, and 21 normal controls who were matched on sex and age. Children with AD/HD whose FIQ was above 80 showed significant low scores in such indices as categories achieved (CA), total errors (TE) , and nonperseverative errors of Nelson (NPEN) in the second step. As to the analysis according to the subtypes, the predominantly inattentive type showed significant low scores in CA, TE and NPEN of the second step, while the combined and predominantly hyperactive-impulsive types showed no significant differences compared with the normal controls. These results suggest that patients with AD/HD have difficulties in effectively utilizing their experiences in the first step as well as instructions which are given before the second step, and also indicate the importance of the second step from a view of the clinical applications.


[Influence of changing inhalation device from separate diskus of salmeterol and fluticasone propionate to combination diskus in asthma patients].
Salmeterol xinafoate (salmeterol) and fluticasone propionate (FP) combination (SFC) was reported to enhance improvement of lung function compared with concurrent administration of the two drugs from separate devices. However, there was no report of asthma control improvement comparing SFC and concurrent administration with separate inhalers in real-life general practice in Japan We evaluated asthma control improvement and patients' perception of SFC in asthmatic patients changing from concurrent therapy with salmeterol and FP with separate devices, Diskus. Treatment were changed to SFC in one hundred and three patients with mild to severe asthma receiving concurrent administration of salmeterol and FP with separate Diskus. Asthma control test (ACT), easy asthma program (EAP) and peak expiratory flow (PEF) were performed before and after 2 or 4 weeks of SFC treatment. SFC treatment increased ACT score by 1.6 point and improved asthma control status measured by EAP. Forty percent of the patients felt their asthmatic symptoms improved, of which 60% recognized their improvement started within a few days. The use of short-acting inhaled beta2-agonist was decreased in 67% of the patients, and exercise-induced dyspnea was improved in 30% of the patients. In addition, SFC was considered as convenient by 96% of the patients and made a good impression on 86% of the patients. This study demonstrates that SFC improves asthma control and better satisfaction than concurrent therapy with separate devices in asthmatic patients in real-life general practice.


Observation of a transient decrease in terahertz conductivity of single-layer graphene induced by ultrafast optical excitation.
We have measured the terahertz frequency-dependent sheet conductivity and its transient response following femtosecond optical excitation for single-layer graphene samples grown by chemical vapor deposition. The conductivity of the unexcited graphene sheet, which was spontaneously doped, showed a strong free-carrier response. The THz conductivity matched a Drude model over the available THz spectral range and yielded an average carrier scattering time of 70 fs. Upon photoexcitation, we observed a transient decrease in graphene conductivity. The THz frequency-dependence of the graphene photoresponse differs from that of the unexcited material but remains compatible with a Drude form. We show that the negative photoconductive response arises from an increase in the carrier scattering rate, with a minor offsetting increase in the Drude weight. This behavior, which differs in sign from that reported previously for epitaxial graphene, is expected for samples with relatively high mobilities and doping levels. The photoinduced conductivity transient has a picosecond lifetime and is associated with nonequilibrium excitation conditions in the graphene.


Host cell nuclear function and murine hepatitis virus replication.
Murine hepatitis virus strains A59 and JHM replicated with equal efficiency in both nucleated and enucleated L2 cells. In addition, treatment of the host cell with either actinomycin D or alpha-amanitin, both inhibitors of host cell RNA synthesis, had no effect on virus replication. Therefore, the replication of murine hepatitis virus did not appear to depend upon either the presence of the host cell nucleus or continued host cell RNA synthesis.


Histomorphometry and stability analysis of early loaded implants with two different surface conditions in beagle dogs.
Despite an improved bone reactions of Mg-incorporated implants in the animals, little yet has been carried out by the experimental investigations in functional loading conditions. This study investigated the clinical and histologic parameters of osseointegrated Mg-incorporated implants in early loading conditions. A total of 36 solid screw implants (diameter 3.75 mm, length 10 mm) were placed in the mandibles of 6 beagle dogs. Test groups included 18 Mg-incorporated implants. Turned titanium implants served as control. Gold crowns were inserted 4 weeks after implant placement and the dogs were immediately put on a food diet. Implants were observed for 10 weeks after loading. Radiographic assessments and stability tests were performed at the time of fixture installation, 2(nd) stage surgery, 4 weeks after loading, and 10 weeks after loading. Histological observations and morphometrical measurements were also performed. Of 36 implants, 33 displayed no discernible mobility, corresponding to successful clinical function. There was no statistically significant difference between test implants and controls in marginal bone levels (P = .46) and RFA values. The mean BIC% in the Mg-implants was 54.5 ± 8.4%. The mean BIC% in the turned implant was 45.3 ± 12.2%. These differences between the Mg-implant and control implant were statistically significant (P = .005). The anodized, Mg-incorporated implant demonstrated significantly more bone-to-implant contact (BIC) in early loading conditions. The results of this study in beagle dogs suggest the possibility of achieving predictable stability of early loaded free-standing dental implants with Mg-incorporated surface.


Associations between added sugars and micronutrient intakes and status: further analysis of data from the National Diet and Nutrition Survey of Young People aged 4 to 18 years.
Added sugars are often viewed as 'empty calories', negatively impacting micronutrient intakes, yet reviews consider the evidence inconclusive. This study aimed to quantify associations between dietary added sugars (as a percentage of energy) and micronutrient intake and biochemical status in the National Diet and Nutrition Survey. Using data from 1688 British children aged 4-18 years who completed 7 d weighed dietary records in 1997, micronutrient intakes were examined across quintiles of added sugars. After excluding low energy reporters, mean dietary intakes of most nutrients exceeded the reference nutrient intake, except for zinc. Compared with quintile 1 (9% added sugars), high consumers in quintile 5 (23% added sugars) had micronutrient intakes ranging from 24% lower to 6% higher (mean 14% lower). Zinc intakes in quintile 1 v. quintile 5 averaged 93% v. 78% of reference nutrient intake; magnesium 114% v. 94%; iron 115% v. 100%; and vitamin A 111% v. 92%, respectively. Plasma levels of magnesium, zinc and carotenoids did not vary across quintiles, but weak negative correlations were observed with serum ferritin and transferrin saturation. Plasma selenium was inversely correlated with added sugars (r -0.17; P < 0.0001) but there was no association with glutathione peroxidase. The impact of added sugars on micronutrient intakes appears modest overall but may have relevance for children consuming inadequate amounts of nutrient-rich foods coupled with a diet high in added sugars (approximately 23%). Further work is needed to explore the impact of different sources of added sugars and to refine assessments of inadequate intakes and status.


Fabrication and characterization of dual acting oleyl chitosan functionalised iron oxide/gold hybrid nanoparticles for MRI and CT imaging.
Bionanocomposites fabricated using metal nanoparticles serve a wide range of biomedical applications viz., site targeted drug delivery, imaging etc. Theranostics emerge as an important field of science, which focuses on the use of single entity for both disease diagnosis and treatment. The present work aimed at designing a multifunctional nanocomposite comprising of iron/gold hybrid nanoparticles, coated with oleyl chitosan and conjugated with methotrexate. The HR-TEM images revealed the spherical nature of the composite, while it's nontoxic and biocompatible property was proved by the MTT assay in NIH 3T3 cells and hemolysis assay. Though the VSM results exhibited the magnetic property, the MRI phantom images and X-ray contrast images demonstrated the potential of the composite to be used as contrast agent. Thus the prepared nanocomposite possess good cytocompatibility, magnetic property and also high X-ray attenuation, wherein it could serve as a novel platform for both MRI and CT diagnosis, as well as drug conjugation could aid in targeted drug delivery.


Emerging treatments for recurrent prostate cancer.
Despite radical treatment, many men with prostate cancer will develop recurrence of their disease. In an exciting era of new therapies for prostate cancer in general, we focus on how these will specifically benefit those men with recurrent disease. We consider salvage treatments aimed at those with local recurrence confined to the prostate gland, therapies for those presenting with metastatic recurrence and the approach to men presenting with a rising prostate-specific antigen but no demonstrable disease (M0). In general, men with recurrent disease are often under-represented in randomized clinical trials. Consequently, evidence to guide treatment for these men is often lacking and this needs to be addressed in order to improve and better define our approach to this problem in the future.


[A contrast with good intentions].
A seventy-five-year-old woman with moderate chronic renal failure was admitted to evaluate a complex renal cyst in the frame of acquired cystic kidney disease. Computed tomography (CT) was performed without contrast media due to the risk of radiocontrast-induced nephrotoxicity. Sonographic investigation at our ultrasound unit revealed a hypoechoic lesion measuring 20x20 mm in size by conventional B-mode sonography, confirmed by NTHI. The Hypoechoic lesion was consistent with complex renal cyst or renal tumour. This finding triggered investigation with CEUS.A sulphur hexafluoride-filled microbubble contrast medium was injected intravenously. The focal lesion CES pattern was characterized by intralesional enhancement in the arterial phase. Further diagnostic imaging including CT with contrast media confirmed a lesion consistent with renal tumour. The patient underwent right-sided nephrectomy; histopathological work-up revealed a renal cell carcinoma. Contrast-enhanced sonography could be clinically useful for the differential diagnosis of kidney lesions in patients with chronic renal failure.


Vascular and autonomic changes in adult cancer patients receiving anticancer chemotherapy.
Chemotherapy is associated with acute and long-term cardiotoxicity. To date, risk assessment has primarily focused on the heart; however, recent findings suggest that vascular and autonomic function may also be compromised. Whether this occurs during chemotherapy treatment remains unknown. Therefore, the present study evaluated carotid artery stiffness, cardiovagal baroreflex sensitivity (cBRS), and heart rate variability (HRV) in cancer patients currently being treated with adjuvant chemotherapy. Eleven current cancer patients receiving adjuvant chemotherapy and 11 matched (1:1) controls were studied. Carotid artery stiffness was assessed via two-dimensional ultrasonography. cBRS was assessed from the spontaneous changes in beat-to-beat time series of R-R interval and systolic blood pressure via the cross-correlation technique. HRV was assessed using the standard deviation of R-R intervals (SDNN) and low (LF) and high (HF) power frequencies. Carotid artery β-stiffness was significantly higher in the cancer patients compared with control participants (8.0 ± 0.8 vs. 6.3 ± 0.6 U, respectively; P = 0.02). cBRS was lower in the cancer patients compared with controls (4.3 ± 0.7 vs. 10.7 ± 1.9 ms/mmHg, respectively; P = 0.01), and all indices of HRV were lower in the cancer patients (SDNN, P = 0.02; LF, P = 0.01; HF, P = 0.02). There was no significant correlation between β-stiffness and cBRS ( P = 0.4). However, LF power was significantly correlated with cBRS (r = 0.66, P < 0.001). Compared with matched healthy controls, cancer patients undergoing chemotherapy demonstrated a significantly higher arterial stiffness and lower cBRS. The previously reported adverse effects of chemotherapy on the heart appear to also influence other aspects of cardiovascular health. NEW & NOTEWORTHY Patients treated with anticancer chemotherapy exhibit an impaired baroreflex control of arterial blood pressure and increased arterial stiffness. These findings hold significant value, in particular as part of a risk-stratification strategy in current cancer patients receiving chemotherapy. This is the first investigation, to our knowledge, to demonstrate an attenuated spontaneous baroreflex control of arterial blood pressure in cancer patients currently undergoing chemotherapy.


The influence of staining procedures on the assessment of cell proliferation as defined by the monoclonal antibody Ki-67.
We examined the influence of different staining techniques [(three-step immunoperoxidase technique (IP); alkaline phosphatase-anti-alkaline phosphatase technique (APAAP)] on the quantitative evaluation of Ki-67-labeled nuclei. We studied five melanocytic skin tumors. From each case, five parallel sections were prepared and stained using the peroxidase-antiperoxidase (PAP) technique (slide 1) and the APAAP technique once (slide 2). Slide 3 consisted of a single repetition of the APAAP technique, slide 4 was a double repetition, and slide 5 was a third repetition. We assessed the volume fraction (VV) of Ki-67-positive nuclei using computer-assisted image analysis. For each staining group, the mean value and standard deviation of VV were calculated. Comparing VV values obtained from the different staining groups we did not find a statistically significant difference between the IP and the various APAAP steps (Wilcoxon test, p = less than 0.05). However, the staining procedure influenced the quantitative results to some extent. The mean VV of the five staining groups ranged in our study from 0.10 to 0.17%, which is narrow compared with the overall variability among different cases (dermal melanocytic nevus, 0.01%; metastatic malignant melanoma, 0.43%). Therefore, we can state that for a rough evaluation of Ki-67-positive nuclei, the influence of different staining methods is negligible; for a subtle quantitative analysis, however, it would nevertheless be preferable to always apply the same staining technique.


Atomistic molecular-dynamics simulations enable prediction of the arginine permeation pathway through OccD1/OprD from Pseudomonas aeruginosa.
Pseudomonas aeruginosa is a Gram-negative bacterium that does not contain large, nonspecific porins in its outer membrane. Consequently, the outer membrane is highly impermeable to polar solutes and serves as a barrier against the penetration of antimicrobial agents. This is one of the reasons why such bacteria are intrinsically resistant to antibiotics. Polar molecules that permeate across the outer membrane do so through substrate-specific channels proteins. To design antibiotics that target substrate-channel proteins, it is essential to first identify the permeation pathways of their natural substrates. In P. aeruginosa, the largest family of substrate-specific proteins is the OccD (previously reported under the name OprD) family. Here, we employ equilibrium and steered molecular-dynamics simulations to study OccD1/OprD, the archetypical member of the OccD family. We study the permeation of arginine, one of the natural substrates of OccD1, through the protein. The combination of simulation methods allows us to predict the pathway taken by the amino acid, which is enabled by conformational rearrangements of the extracellular loops of the protein. Furthermore, we show that arginine adopts a specific orientation to form the molecular interactions that facilitate its passage through part of the protein. We predict a three-stage permeation process for arginine.


Surgical Treatment of Anomalous Origin of Right Coronary Artery in a Patient with Mitral Stenosis.
An anomalous origin of the right coronary artery is rarely observed, with a reported incidence between 0.026% and 0.25%. This condition is often completely asymptomatic and is found incidentally during angiographic evaluation for other cardiac diseases. However some patients present with exertion angina or sudden death. Surgical treatment in patients with anomalous RCA is still controversial. Treatment can be conservative, angioplasty or surgery. A 59-year-old man was admitted with severe mitral stenosis. He complained exertion and rest dyspnea, NYHA III class. He had sequels of embolic stroke, results of left atrial thrombus. Echocardiography showed calcified severe mitral stenosis with mitral orifice area of 1.1 square centimeters with PSPAP 60 mmHg and normal LV function. Routine coronary angiography before surgery showed aberrant origin of RCA from the left sinus of Valsalva with 90% stenosis at his origin. Multi-slice computed tomography proved the diagnosis of anomalous RCA arising from the left sinus of Valsalva and taking an inter-arterial course between the aorta and pulmonary artery. The patient underwent mitral valve replacement with mechanical St. Jude prosthesis No 29 and saphenous vein graft to RCA. We chose by-pass grafting techniques because after aortotomy, RCA was too close to LMCA, intramural course was too short and stenosis of RCA was outside of aortic wall. The patient's perioperative course was without complications and patient was discharged on the seventh postoperative day. Correction of anomalous of the origin of right coronary artery is mandatory in cases where patient has to be operated for other cardiac causes.