Inducible gene expression in tumors colonized by modified oncolytic vaccinia virus strains.
Exogenous gene induction of therapeutic, diagnostic, and safety mechanisms could be a considerable improvement in oncolytic virotherapy. Here, we introduced a doxycycline-inducible promoter system (comprised of a tetracycline repressor, several promoter constructs, and a tet operator sequence) into oncolytic recombinant vaccinia viruses (rVACV), which were further characterized in detail. Experiments in cell cultures as well as in tumor-bearing mice were analyzed to determine the role of the inducible-system components. To accomplish this, we took advantage of the optical reporter construct, which resulted in the production of click-beetle luciferase as well as a red fluorescent protein. The results indicated that each of the system components could be used to optimize the induction rates and had an influence on the background expression levels. Depending on the given gene to be induced in rVACV-colonized tumors of patients, we discuss the doxycycline-inducible promoter system adjustment and further optimization. Oncolytic virotherapy of cancer can greatly benefit from the expression of heterologous genes. It is reasonable that some of those heterologous gene products could have detrimental effects either on the cancer patient or on the oncolytic virus itself if they are expressed at the wrong time or if the expression levels are too high. Therefore, exogenous control of gene expression levels by administration of a nontoxic inducer will have positive effects on the safety as well as the therapeutic outcome of oncolytic virotherapy. In addition, it paves the way for the introduction of new therapeutic genes into the genome of oncolytic viruses that could not have been tested otherwise.


Prognostic nutritional index and serum lactate dehydrogenase predict the prognosis of nasopharyngeal carcinoma patients who received intensity-modulated radiation therapy.
This research aimed to evaluate the prognostic significance of baseline prognostic nutritional index (PNI) and lactate dehydrogenase (LDH) for the outcome of individuals diagnosed with non-metastatic nasopharyngeal carcinoma (NPC). A retrospective analysis was conducted on data from 810 patients with non-metastatic NPC who underwent intensity-modulated radiation therapy (IMRT) with or without chemotherapy. The best cut-offs for PNI and LDH were identified by X-tile software to be 48.5 and 150, respectively. To find the independent prognostic factors for survival outcomes, univariate and multivariate regression analyses were conducted, and AUCs were used to compare their prognostic values. Multivariate analysis revealed that patients with PNI > 48.5 had better overall survival (OS) (HR: 0.502, P < 0.001), progression-free survival (PFS) (HR: 0.618, P < 0.001), and distant metastasis-free survival (DMFS) (HR: 0.637, P = 0.005). Higher LDH was associated with poorer OS (HR: 1.798, P < 0.001), PFS (HR: 1.671, P < 0.001), and DMFS (HR: 1.756, P < 0.001). The combination of low PNI and high LDH in non-metastatic NPC patients was correlated with poor OS (P < 0.001), PFS (P < 0.001), and DMFS (P < 0.001). The combination of PNI and LDH had the highest AUCs for predicting OS, PFS, and DMFS. PNI and LDH might become valuable predictors of the prognosis of non-metastatic NPC patients undergoing IMRT with or without chemotherapy. Prognostic accuracy can be enhanced by combining PNI and LDH.


Growth of hepatitis A virus in a mouse liver cell line.
Hepatitis A virus (HAV) has been adapted to grow efficiently in primate and some nonprimate cell lines but not in cells of murine origin. To understand the inability of the virus to grow in mouse cells, we studied the replication of HAV in immortalized and nontransformed MMH-D3 mouse liver cells, which require growth factors and collagen to maintain their phenotype. HAV grew in MMH-D3 cells transfected with virion RNA but not in those infected with viral particles, indicating a cell entry block for HAV. However, MMH-D3 cells cultured under suboptimal conditions in the absence of growth factors acquired susceptibility to HAV infection. Serial passages of the virus in MMH-D3 cells under suboptimal growth conditions resulted in the selection of HAV variants that grew efficiently in MMH-D3 cells cultured under both optimal and suboptimal conditions. Nucleotide sequence analysis of the MMH-D3 cell-adapted HAV revealed that N1237D and D2132G substitutions were present in the capsid regions of six viral clones. These two mutations are most likely located on the surface of the virion and may play a role in the entry of HAV into the mouse liver cells. Our results demonstrate that mouse hepatocyte-like cells code for all factors required for the efficient growth of HAV in cell culture.


Combined Intimate Partner Violence and HIV/AIDS Prevention in Rural Uganda: Design of the SHARE Intervention Strategy.
Intimate partner violence (IPV) has a bidirectional relationship with HIV infection. Researchers from the Rakai Health Sciences Program (RHSP), an HIV research and services organization in rural Uganda, conducted a combination IPV and HIV prevention intervention called the Safe Homes and Respect for Everyone (SHARE) Project between 2005 and 2009. SHARE was associated with significant declines in physical and sexual IPV and overall HIV incidence, and its model could be adopted as a promising practice in other settings. In this article we describe how SHARE's IPV-prevention strategies were integrated into RHSP's existing HIV programming and provide recommendations for replication of the approach.


Genomic analysis and characterization of new loci associated with seed protein and oil content in soybeans.
Breeding for increased protein without a reduction in oil content in soybeans [Glycine max (L.) Merr.] is a challenge for soybean breeders but an expected goal. Many efforts have been made to develop new soybean varieties with high yield in combination with desirable protein and/or oil traits. An elite line, R05-1415, was reported to be high yielding, high protein, and low oil. Several significant quantitative trait loci (QTL) for protein and oil were reported in this line, but many of them were unstable across environments or genetic backgrounds. Thus, a new study under multiple field environments using the Infinium BARCSoySNP6K BeadChips was conducted to detect and confirm stable genomic loci for these traits. Genetic analyses consistently detected a single major genomic locus conveying these two traits with remarkably high phenotypic variation explained (R2 ), varying between 24.2% and 43.5%. This new genomic locus is located between 25.0 and 26.7 Mb, distant from the previously reported QTL and did not overlap with other commonly reported QTL and the recently cloned gene Glyma.20G085100. Homolog analysis indicated that this QTL did not result from the paracentric chromosome inversion with an adjacent genomic fragment that harbors the reported QTL. The pleiotropic effect of this QTL could be a challenge for improving protein and oil simultaneously; however, a further study of four candidate genes with significant expressions in the seed developmental stages coupled with haplotype analysis may be able to pinpoint causative genes. The functionality and roles of these genes can be determined and characterized, which lay a solid foundation for the improvement of protein and oil content in soybeans.


Liner exchange and bone grafting: rare option to treat wear & lysis of stable TKAs.
Liner exchange and bone grafting are commonly performed for wear and osteolysis around well-fixed modular acetabular components that otherwise would require structural allografting and revision THA. However, liner exchange in the face of substantial lysis around TKA has been performed rarely with reports of failure rates of up to 25% at 3 year followup. We therefore evaluated the technique of liner exchange and bone grafting for cases of wear and extensive osteolysis around TKAs in which the components were well-fixed and well-aligned to determine (1) rerevision rates; (2) fate of the bone graft; (3) radiographic loosening rates; and (4) functional scores. We retrospectively reviewed 22 patients (25 knees) who underwent revision TKA with exchange of the modular polyethylene insert and bone grafting in cases with well-fixed components and large areas of osteolysis (up to 54 cm(2) on a single projection) at the time of revision. The average area of osteolysis was 21 cm(2) and 10 cm(2) on the AP projection of the femur and tibia, respectively. On the lateral projection, the average area of osteolysis for the femur and tibia was 22 cm(2) and 9.3 cm(2). Minimum clinical and radiographic followup was 22 and 22 months (average, 61 and 59; range, 22-142 and 22-130, respectively). One of the 25 knees was revised for aseptic loosening or recurrence of osteolysis. On radiographs, 84.6% and 70% of femoral and tibial osteolytic lesions, respectively, showed evidence of complete or near complete graft incorporation. The remaining lesions showed evidence of partial graft incorporation with the exception of one tibial lesion, which was in the revised case. All other components were well fixed with no evidence of radiographic loosening. In this selected series of cases with extensive osteolysis around well-fixed and well-aligned TKAs, liner exchange and bone grafting provided durable midterm results with extensive graft incorporation. Level IV, therapeutic study. See Guidelines for Authors for a complete description of levels of evidence.


Virtual Laparoscopy Simulation: a Promising Pedagogic Tool in Gynecology.
Virtual simulators have played a vital role in preparing surgeons for laparoscopic and robotic procedures in gynecologic surgery. The efficacy of the simulator was evaluated to improve basic (trainee) laparoscopic skills and assess training levels. This prospective, comparative study was conducted in volunteer residents in the obstetrics and gynecology training program of Université Laval. Study participants performed 9 laparoscopic simulator tasks on 2 different occasions. Skills improvement between sessions and differences between junior and senior residents were examined. Thirteen junior and 11 senior residents participated in the study. Junior trainees significantly improved their speed of execution, accuracy, and maintenance of horizontal view. Senior trainees mainly accelerated their rapidity in completing different tasks. They performed better than junior trainees, with economy of movements, and tended toward greater precision, speed of execution, and safe retraction in various tasks. Virtual simulators are useful pedagogic tools that could benefit both junior and senior residents. Integration into the residency curricula should be considered.


Human research ethics committees in technical universities.
Human research ethics has developed in both theory and practice mostly from experiences in medical research. Human participants, however, are used in a much broader range of research than ethics committees oversee, including both basic and applied research at technical universities. Although mandated in the United States, the United Kingdom, Canada, and Australia, non-medical research involving humans need not receive ethics review in much of Europe, Asia, Latin America, and Africa. Our survey of the top 50 technical universities in the world shows that, where not specifically mandated by law, most technical universities do not employ ethics committees to review human studies. As the domains of basic and applied sciences expand, ethics committees are increasingly needed to guide and oversee all such research regardless of legal requirements. We offer as examples, from our experience as an ethics committee in a major European technical university, ways in which such a committee provides needed services and can help ensure more ethical studies involving humans outside the standard medical context. We provide some arguments for creating such committees, and in our supplemental article, we provide specific examples of cases and concerns that may confront technical, engineering, and design research, as well as outline the general framework we have used in creating our committee.


Efficacy of static stretching and proprioceptive neuromuscular facilitation stretch on hamstrings length after a single session.
A number of studies have investigated the efficacy of several repetitions of proprioceptive neuromuscular facilitation stretching (PNF) and static stretching (SS). However, there is limited research comparing the effects of a single bout of these stretching maneuvers. The aim of this study was to compare the effectiveness of a single bout of a therapist-applied 30-second SS vs. a single bout of therapist-applied 6-second hamstring (agonist) contract PNF. Forty-five healthy subjects between the ages of 21 and 35 were randomly allocated to 1 of the 2 stretching groups or a control group, in which no stretching was received. The flexibility of the hamstring was determined by a range of passive knee extension, measured using a universal goniometer, with the subject in the supine position and the hip at 90° flexion, before and after intervention. A significant increase in knee extension was found for both intervention groups after a single stretch (SS group = 7.53°, p < 0.01 and PNF group = 11.80°, p < 0.01). Both interventions resulted in a significantly greater increase in knee extension when compared to the control group (p < 0.01). The PNF group demonstrated significantly greater gains in knee extension compared to the SS group (mean difference 4.27°, p < 0.01). It can be concluded that a therapist applied SS or PNF results in a significant increase in hamstring flexibility. A hamstring (agonist) contract PNF is more effective than an SS in a single stretching session. These findings are important to physiotherapists or trainers working in clinical and sporting environments. Where in the past therapists may have spent time conducting multiple repetitions of a PNF and an SS, a single bout of either technique may be considered just as effective. A key component of the study methodology was the exclusion of a warm-up period before stretching. Therefore, the findings of efficacy of a single PNF are of particular relevance in sporting environments and busy clinical settings where time may be limited.


The enteroviruses: recent advances.
New information accrues periodically in the ledger accounting for infections and diseases associated with the human enteroviruses. The discoveries of "new" serotypes and how they affect people are subjects of continuing attention. Some other relevant information on "old" serotypes relates to variations in age-specific attack rates and associated morbidity and mortality for neonates and older infants. Among the morbidity reports are recounts during outbreaks of virus-positive cerebrospinal fluids (CSFs) that initially may not have cytological or biochemical abnormalities. Prolonged enterovirus infections may develop in persons having agammaglobulinemia. Lastly, some provocative associations concern the pathologic expressions of enteroviruses in the development and persistence of injury to the heart (myocardiopathies) and the pancreas (insulin-dependent diabetes mellitus).


Spontaneous motility of passive emulsion droplets in polar active gels.
We study by computer simulations the dynamics of a droplet of passive, isotropic fluid, embedded in a polar active gel. The latter represents a fluid of active force dipoles, which exert either contractile or extensile stresses on their surroundings, modelling for instance a suspension of cytoskeletal filaments and molecular motors. When the polarisation of the active gel is anchored normal to the droplet at its surface, the nematic elasticity of the active gel drives the formation of a hedgehog defect; this defect then drives an active flow which propels the droplet forward. In an extensile gel, motility can occur even with tangential anchoring, which is compatible with a defect-free polarisation pattern. In this case, upon increasing activity the droplet first rotates uniformly, and then undergoes a discontinuous nonequilibrium transition into a translationally motile state, powered by bending deformations in the surrounding active medium.


Student Perceptions of Veterinary Anatomy Practical Classes: A Longitudinal Study.
Using cadaveric material to teach veterinary students poses many challenges. However, little research exists on the contribution of this traditional approach to student learning. This longitudinal study aimed to investigate student perceptions of cadaver-based anatomy classes in a vertically integrated veterinary curriculum at the University of Nottingham's School of Veterinary Medicine and Science. Likert-scale statements and free-text boxes were used in a questionnaire distributed to second-year veterinary students (response rate 59%, 61/103). The same questionnaire was subsequently distributed to the same cohort 2 years later, in the students' fourth year of study (response rate 68%, 67/98). Students agreed that cadaver-based activities aid their learning, and they particularly value opportunities to develop practical skills while learning anatomy. There are few changes in perception as undergraduates progress to clinical years of teaching. Students perceive anatomy to be important, and feel that their learning has prepared them for clinical placements. This study emphasizes the importance of using cadaveric materials effectively in anatomy teaching and, in particular, using clinical skills training to enhance the anatomy curriculum.


Abuse of isobutyl nitrite inhalation (Rush) by adolescents.
Isobutyl nitrite is a volatile liquid sold without a prescription as a "room odorizer" but is widely used as an inhalant to produce feelings of euphoria. Of the 173 13-22-year-olds (mean age, 16 years) clients of a long-term drug treatment facility who completed a questionnaire related to drug use, 74 (43%) admitted to having used isobutyl nitrite at least once, 22 (13%) had used the substance ten or more times, and eight (4%) used the substance 50 or more times. Inhalation of this malodorous substance leads almost universally to dizziness and lightheadedness and usually to severe symptoms of vasodilatation such as "pounding of the heart," blurred vision, and a "warm feeling." The feeling was unpleasant to 44 percent of the users in this study, and most also experienced pulsatile headache. In addition, contact dermatitis and irritation of the tracheobronchial tree and eyes occurred in some users. The primary use of this substance as an euphoric agent warrants consideration of banning its sale altogether.


A 24-week open-label extension study of olanzapine-fluoxetine combination and olanzapine monotherapy in the treatment of bipolar depression.
Olanzapine-fluoxetine combination has shown efficacy in the acute treatment of depressive episodes in patients with bipolar I disorder. The present analyses examined the efficacy and safety of longer term treatment with olanzapine-fluoxetine combination or olanzapine monotherapy in a 6-month open-label extension study. 376 patients with DSM-IV bipolar I disorder, depressed, who completed an acute trial entered the open-label study and received 1 week of olanzapine monotherapy (5-20 mg/day). At all subsequent visits, patients could choose between olanzapine monotherapy or olanzapine-fluoxetine combination (6/25, 6/50, or 12/50 mg/day). Three treatment groups were defined retrospectively according to the medication course taken from week 1: olanzapine, olanzapine-fluoxetine combination, or switched. The efficacy measures were the Montgomery-Asberg Depression Rating Scale (MADRS), Clinical Global Impressions-Bipolar Version, and Young Mania Rating Scale. The study was conducted from July 2000 to May 2002. Among patients who started in remission, MADRS total scores did not change significantly from baseline to endpoint in the olanzapine-fluoxetine combination (0.8) or olanzapine (0.3) groups, but increased slightly in the switched (2.3, p = .02) group. For patients who started in nonremission, MADRS total scores decreased significantly in all groups (olanzapine-fluoxetine combination: -5.7, p = .001; olanzapine: -11.6, p = .004; switched: -6.4, p = .015). The majority of patients who entered the study in nonremission achieved remission (MADRS total score < or = 12) during the trial (olanzapine-fluoxetine combination: 66.7%, olanzapine: 64.7%, switched: 62.5%). The overall rate of depressive relapse was 27.4%, and the overall incidence of mania emergence was 5.9%. The present findings suggest that long-term treatment with olanzapine-fluoxetine combination may be a useful option for the management of depressive symptoms and carries a low risk of mania emergence.


Analysis of arsenical metabolites in biological samples.
Quantitation of iAs and its methylated metabolites in biological samples provides dosimetric information needed to understand dose-response relations. Here, methods are described for separation of inorganic and mono-, di-, and trimethylated arsenicals by thin layer chromatography. This method has been extensively used to track the metabolism of the radionuclide [(73)As] in a variety of in vitro assay systems. In addition, a hydride generation-cryotrapping-gas chromatography-atomic absorption spectrometric method is described for the quantitation of arsenicals in biological samples. This method uses pH-selective hydride generation to differentiate among arsenicals containing trivalent or pentavalent arsenic.


The Science of Selection: Using Best Practices From Industry to Improve Success in Surgery Training.
The selection of high-quality applicants is critical to the future of surgery. However, it is unclear if current practices meet industry criteria of a successful selection system, as measured by administrative efficiency and performance and attrition of those selected. We performed a modified systematic review process to gain an understanding of current selection processes, remediation practices, and attrition rates in surgery residency training programs in the United States. We also conducted semistructured interviews with local residency program directors and coordinators to obtain a specific snapshot of the amount of time and resources dedicated to these activities in various sized programs. The associated financial costs of these activities are also presented. The administrative costs for current residency selection processes are substantial, ranging from $45,000 to $148,000 for each program per year. Approximately 30% of residents require at least 1 remediation intervention, costing programs $3400 to $5300 per episode, and typically involve concerns around nontechnical skills. Attrition rates range from 20% to 40%. This review suggests that additional methodologies may allow surgery residency programs to identify best-fit candidates more efficiently and effectively, while also decreasing remediation and attrition rates. Possible solutions include incorporation of structured interviews, personality inventories, and situational judgment tests. Resources dedicated to current interview practices, remediation efforts, and attrition management can be redirected to support these methodologies. By applying the science of selection and assessment to the recruitment process, programs may be able to make more data-driven decisions to identify candidates who will be successful at their institution.


Wet granulation in rotary processor and fluid bed: comparison of granule and tablet properties.
The aim of the present study was to investigate and compare granule and tablet properties of granules prepared by wet granulation in a rotary processor or a conventional fluid bed. For this purpose the working range of selected process variables was determined and a factorial study with 3 factors (equipment type, filler type, and liquid addition rate) and 1 covariate (fluidizing air flow rate) was performed. Two grades of calcium carbonate with different size and shape characteristics were applied, and the liquid addition and fluidizing air flow rates were investigated in the widest possible range. Dry mixtures of microcrystalline cellulose, polyvinyl povidone, calcium carbonate, and riboflavin, in a 10:5:84:1 ratio, were granulated in both types of equipment. The granulation end point was determined manually in the fluid bed and by torque measurements in the rotary processor. The filler type had a more pronounced effect on granular properties in the fluid bed, but the rotary processor showed a higher dependency on the investigated process variables. The rotary processor gave rise to more dense granules with better flow properties, but the fluid bed granules had slightly better compressional properties. Furthermore, the distribution of a low-dose drug was found to be more homogeneous in the rotary processor granules and tablets. Generally, wet granulation in a rotary processor was found to be a good alternative to conventional fluid bed granulation, especially when cohesive powders with poor flow properties or formulations with low drug content are to be granulated by a fluidizing air technique.


Differential organization of gamma-aminobutyric acid type A and glycine receptors in the somatic and dendritic compartments of rat abducens motoneurons.
Premotor inhibitory neurons responsible for the decrease in the firing discharge during fast or slow eye movements selectively target the cell bodies and the dendrites of abducens motoneurons. Gamma-aminobutyric acid (GABA) and glycine, the main inhibitory synaptic neurotransmitters in the central nervous system, act via glycine and GABAA receptors, assembled from various types of subunits, which determine the kinetics of the currents mediated. Therefore, our hypothesis was that the expression of the inhibitory receptors on the somatic and the dendritic compartments, involved in different functions, may differ. In this study, we compared the subcellular patterns of expression of the main GABAA receptor subunits (GABAARalpha1, alpha2, alpha3, alpha5), glycine receptors (GlyRalpha1), and gephyrin in the somatic and dendritic compartments of rat abducens motoneurons, using double or triple immunocytochemical experiments with confocal microscopy. Significant differences exist in the patterns of organization and the synaptic expression of the GlyR and GABAAR subunits in the cell bodies and dendrites of abducens motoneurons. In the somata, only the GABAARalpha1 subunit was expressed, whereas both GABAARalpha1 and GABAARalpha3 were present in the dendrites. The GlyRalpha1 to GABAARalpha1 density ratio was reversed in the somatic and dendritic compartments (0.9 vs. 2.3). A quantitative electron microscopy study showed that the modes whereby gephyrin reaches its postsynaptic inhibitory synaptic target differ between the somata and the dendrites. Therefore, our results support the idea that a structure-function adaptation occurs at the single-neuron level.


Clinical use of high-frequency oscillatory ventilation in adult patients with acute respiratory distress syndrome.
High-frequency oscillatory ventilation (HFOV) is an emerging ventilatory strategy for adults that has been used successfully in the neonatal and pediatric population. This modality utilizes high mean airway pressures to maintain an open lung and low tidal volumes at a high frequency that allow for adequate ventilation while at the same time preventing alveolar overdistension. With the current understanding that excessive lung stretch and inadequate end-expiratory ventilatory volume may be injurious to the lungs, HFOV seems to be the ideal lung-protective ventilatory mode. During the past 8 yrs, there have been increasing numbers of studies describing its use in adult patients with acute respiratory distress syndrome. This article aims to review the published studies of HFOV in adults with acute respiratory distress syndrome with regard to its safety and efficacy. To assist us with our review, we did a search of MEDLINE (from 1966 to present) and EMBASE (1980 to present) databases to identify adult, clinical, English-language, research articles related to HFOV use. In addition, we reviewed relevant animal and mechanical ventilation studies. We did not perform a formal systematic review. The application of HFOV was mainly reported as a rescue ventilatory mode in adult patients with acute respiratory distress syndrome who were thought to have failed conventional ventilation. In these patients, HFOV has consistently been shown to improve oxygenation without obvious increases in complications measured. There was only one randomized, controlled trial comparing HFOV with conventional ventilation. This study showed that there was a nonsignificant trend toward a lower mortality rate in the HFOV group. In addition, HFOV was as effective and safe as conventional ventilation. Although there are limitations, multiple studies have shown that earlier initiation of HFOV in patients with severe acute respiratory distress syndrome may also be associated with a lower mortality. HFOV seems to be safe and effective for adults with severe acute respiratory distress syndrome who have failed conventional ventilation. Further research is needed to determine the ideal patients, timing, and optimal technique with which to provide HFOV. When considering HFOV as an early, lung-protective mode of ventilation, there is still a need to perform an adequately powered, randomized, controlled trial comparing it with the best available form of conventional ventilation. However, we believe that such a trial should wait until we have a better understanding of HFOV in adults.


Left atrial enlargement and reduced physical function during aging.
Diastolic dysfunction, often seen with increasing age, is associated with reduced exercise capacity and increased mortality. Mortality rates in older individuals are linked to the development of disability, which may be preceded by functional limitations. The goal of this study was to identify which echocardiographic measures of diastolic function correlate with physical function in older subjects. A total of 36 men and women from the Louisiana Healthy Aging Study, age 62-101 yr, received a complete echocardiographic exam and performed the 10-item continuous-scale physical-functional performance test (CS-PFP-10). After adjustment for age and gender, left atrial volume index (ρ = -0.59; p = .0005) correlated with the total CS-PFP-10 score. Increased left atrial volume index may be a marker of impaired performance of activities of daily living in older individuals.


Luteinizing hormone-releasing hormone agonist effects on skeletal muscle: how hormonal therapy in prostate cancer affects muscular strength.
Since the discovery of Huggins in 1941 demonstrating the androgen dependence of prostate cancer cells, the use of pharmacological therapies to decrease systemic androgen concentrations has been one of the main treatment options for prostate cancer. Despite their efficacy luteinizing hormone releasing hormone agonists (LHRHas) have a number of side effects, of which many have not been fully investigated in humans. This review focuses on the effects of LHRHas on skeletal muscle in 3 main areas, namely effects at the androgen receptor, at the neuromuscular junction and on skeletal muscle myofibers. Since prostate cancer is predominantly a disease of elderly individuals, the aging effects of LHRHa therapy on skeletal muscle are magnified and of clinical importance. A comprehensive MEDLINE search was performed of pertinent studies in the literature relating to the use of LHRHa and skeletal muscle. LHRHas affect 3 primary sites within the skeletal muscle system, namely androgen receptor, the neuromuscular junction and second messenger systems, including insulin-like growth factor-1. All sites have been demonstrated to lead to a decrease in isokinetic exercise strength in large muscle groups. The musculoskeletal effects of LHRHas for the treatment of prostate cancer should be counteracted via a program of exercise strength training to decrease the morbidity associated with skeletal muscle weakness.


Quality of life measures in children with Down syndrome with disorders of gut-brain interaction.
Down syndrome (DS) is associated with multiple medical comorbidities. Perhaps related to such, caregivers of individuals with DS report lower quality of life (QoL) compared to individuals without DS. It has been shown that disorders of gut-brain interaction (DGBI) such as functional constipation (FC) and irritable bowel syndrome (IBS) are common in individuals with DS. We measured caregiver-reported QoL in individuals with DS with a DGBI and compared them to individuals with DS without a DGBI via a cross-sectional national survey. All measures of QoL were lower in those with DS who meet criteria for a DGBI compared to those with DS without a DGBI. Males and females with DS and at least one DGBI had similar QoL scores. While FC was the most common DGBI seen in individuals with DS, there was no difference in any aspect of QoL in subjects with FC when compared to individuals with other DGBIs. However, all measures of QoL were lower in those with IBS compared to individuals with other DGBIs. These findings suggest that management of gastrointestinal symptoms from DGBIs, particularly IBS, may serve as a target for increasing QoL in a notable subset of individuals with DS.


Detecting drawdowns masked by environmental stresses with water-level models.
Detecting and quantifying small drawdown at observation wells distant from the pumping well greatly expands the characterized aquifer volume. However, this detection is often obscured by water level fluctuations such as barometric and tidal effects. A reliable analytical approach for distinguishing drawdown from nonpumping water-level fluctuations is presented and tested here. Drawdown is distinguished by analytically simulating all pumping and nonpumping water-level stresses simultaneously during the period of record. Pumping signals are generated with Theis models, where the pumping schedule is translated into water-level change with the Theis solution. This approach closely matched drawdowns simulated with a complex three-dimensional, hypothetical model and reasonably estimated drawdowns from an aquifer test conducted in a complex hydrogeologic system. Pumping-induced changes generated with a numerical model and analytical Theis model agreed (RMS as low as 0.007 m) in cases where pumping signals traveled more than 1 km across confining units and fault structures. Maximum drawdowns of about 0.05 m were analytically estimated from field investigations where environmental fluctuations approached 0.2 m during the analysis period.


Pronounced cytoplasmic pH gradients are not required for tip growth in plant and fungal cells.
The existence of pronounced cytoplasmic pH gradients within the apices of tip-growing cells, and the role of cytoplasmic pH in regulating tip growth, were investigated in three different cell types: vegetative hyphae of Neurospora crassa; pollen tubes of Agapanthus umbellatus; and rhizoids of Dryopteris affinis gametophytes. Examination of cytoplasmic pH in growing cells was performed by simultaneous, dual emission confocal ratio imaging of the pH-sensitive probe carboxy SNARF-1. Considerable attention was paid to the fine tuning of dye loading and imaging parameters to minimise cellular perturbation and assess the extent of dye partitioning into organelles. With optimal conditions, cytoplasmic pH was measured routinely with a precision of between +/-0.03 and +/-0.06 of a pH unit and a spatial resolution of 2.3 microm2. Based on in vitro calibration, estimated values of mean cytoplasmic pH for cells loaded with dye-ester were between 7.15 and 7.25 for the three cell types. After pressure injecting Neurospora hyphae with dextran-conjugated dye, however, the mean cytoplasmic pH was estimated to be 7.57. Dextran dyes are believed to give a better estimate of cytoplasmic pH because of their superior localisation and retention within the cytosol. No significant cytoplasmic pH gradient (delta pH of >0.1 unit) was observed within the apical 50 microm in growing cells of any of the three cell types. Acidification or alkalinisation of the cytoplasm in Neurospora hyphae, using a cell permeant weak acid (propionic acid at pH 7.0) or weak base (trimethylamine at pH 8.0), slowed down but did not abolish growth. However, similar manipulation of the cytoplasmic pH of Agapanthus pollen tubes and Dryopteris rhizoids completely inhibited growth. Modification of external pH affected the growth pattern of all cell types. In hyphae and pollen tubes, changes in external pH were found to have a small transient effect on cytoplasmic pH but the cells rapidly readjusted towards their original pH. Our results suggest that pronounced longitudinal gradients in cytoplasmic pH are not essential for the regulation of tip growth.


Revised equations for estimating glomerular filtration rate based on the Lund-Malmö Study cohort.
To increase the accuracy of estimated GFR (eGFR) from creatinine overall and at measured GFR ≥90 mL/min per 1.73 m(2) by revising the Lund-Malmö (LM) equations, to elaborate on more complex forms to improve the LM and CKD-EPI equations further, and to assess benefits of adding lean body mass (LBM). Swedish Caucasians (n = 850, 376 women; median 60, range 18-95 years) referred for GFR measurement (plasma iohexol-clearance: median 55, range 5-173 mL/min/1.73 m(2)) constituted the Lund-Malmö Study cohort. Bias, precision, accuracy, expressed as median absolute percentage difference and percentage of estimates ±10% (P(10)) and ±30% (P(30)) of measured GFR, and classification ability with respect to five GFR stages were compared with the original LM, CKD-EPI and MDRD equations. LM Revised overall performed better than LM Original without LBM due to increased accuracy at measured GFR ≥90 mL/min/1.73 m(2). Further extensions of the CKD-EPI or LM equations did not substantially improve overall performance. In particular, the performance of LM Revised at measured GFR ≥90 mL/min/1.73 m(2) could not be improved further without decreasing accuracy and classification ability at lower GFR-levels. Adding LBM to the equations had no strong effect on accuracy. Comparisons with the CKD-EPI and MDRD equations suggest that the LM equations are superior for the present Swedish population, due to markedly higher accuracy of the LM equations at measured GFR <30 mL/min/1.73 m(2). However, the LM equations cannot be recommended for use in general clinical practice until validated in other populations.


Detection of anti-preS1 antibodies for recovery of hepatitis B patients by immunoassay.
To establish a convenient immunoassay method based on recombinant antigen preS1(21-119aa) to detect anti-preS1 antibodies and evaluate the clinical significance of antibodies in hepatitis B. The expression plasmid pET-28a-preS1 was constructed, and a large quantity of preS1(21-119aa) fragment of the large HBsAg protein was obtained. The preS1 fragment purified by Ni(2+)-IDA affinity chromatography was used as coated antigen to establish the indirect ELISA based on streptavidin-biotin system for detection of the anti-preS1 antibodies in sera from HBV-infected patients. For follow-up study, serial sera were collected during the clinical course of 21 HBV-infected patients and anti-preS1 antibodies, preS1 antigen, HBV-DNA and other serological HBV markers were analyzed. preS1(21-119aa) fragment was highly expressed from the plasmid pET-28a-preS1 in a soluble form in E.Coli (30mg.L(-1)), and easily purified to high purity over 90% by one step of Ni(2+)-IDA-sepharose 6B affinity chromatography. The purity and antigenicity of the purified preS1(21-119aa) protein was determined by 150g.L(-1) SDS-PAGE, Western blot and a direct ELISA. Recombinant preS1(21-119aa) protein was successfully applied in the immunoassay which could sensitively detect the anti-preS1 antibodies in serum specimens of acute or chronic hepatitis B patients. Results showed that more than half of 19 acute hepatitis B patients produced anti-preS1 antibodies during recovery of the disease, however, the response was only found in a few of chronic patients. In the clinical follow-up study of 11 patients with anti-preS1 positive serological profile, HBsAg and HBV-DNA clearance occurred in 6 of 10 acute hepatitis B patients in 5-6 months, and seroconversion of HBeAg and disappearance of HBV-DNA occurred in 1 chronic patients treated with lavumidine, a antiviral agent. The high-purity preS1(21-119aa) coated antigen was successfully prepared by gene expression and affinity chromatography. Using this antigen, a conveniently detective system of anti-preS1 antibodies in sera was established. Preliminarily clinical trial the occurrence of anti-preS1 antibodies in acute hepatitis B patients suggests the clearance of HBV from serum in a short-term time, and anti-preS1 positive in chronic patients means health improvement or recovery from the disease.


Primary prophylaxis of variceal bleeding in cirrhotic patients: a cohort study.
Current guidelines recommend beta-blockers for primary prevention of variceal haemorrhage in cirrhotic patients, and band ligation for patients with contraindications or intolerance to beta-blockers. However, it has been suggested that these patients may respond poorly to band ligation. We evaluated the usefulness of a strategy in which band ligation was used to treat patients with contraindications or intolerance and patients not responding to beta-blockers identified by hepatic vein pressure gradient measurement. Haemodynamic responders and patients refusing hepatic vein pressure gradient measurement were given long-term beta-blockers. One hundred and thirty-five consecutive patients with high-risk oesophageal varices and no prior bleeding were enrolled. Twenty-five patients with contraindications (group A), 26 with intolerance to beta-blockers (group B) and 25 showing a poor haemodynamic response (Group C) underwent band ligation. Twenty-two haemodynamic responders (Group D) and 37 refusing hepatic vein pressure gradient measurement (Group E) were treated with beta-blockers. Median follow-up was 32 months. 12/135 patients (8.9%) bled: 3/25 (12%) in group A, 1/26 (3.8%) in group B, 0/25 (0%) in group C, 0/22 (0%) in group D and 8/37 (22.2%) in group E. Mortality was 8/135 (5.9%). Patients with contraindications, intolerance or not responding to beta-blockers treated with band ligation achieve protection from variceal bleeding comparable to that of good responders to beta-blockers.


Quantifying temporal isolation: a modelling approach assessing the effect of flowering time differences on crop-to-weed pollen flow in sunflower.
Flowering time divergence can be a crucial component of reproductive isolation between sympatric populations, but few studies have quantified its actual contribution to the reduction of gene flow. In this study, we aimed at estimating pollen-mediated gene flow between cultivated sunflower and a weedy conspecific sunflower population growing in the same field and at quantifying, how it is affected by the weeds' flowering time. For that purpose, we extended an existing mating model by including a temporal distance (i.e. flowering time difference between potential parents) effect on mating probabilities. Using phenological and genotypic data gathered on the crop and on a sample of the weedy population and its offspring, we estimated an average hybridization rate of approximately 10%. This rate varied strongly from 30% on average for weeds flowering at the crop flowering peak to 0% when the crop finished flowering and was affected by the local density of weeds. Our result also suggested the occurrence of other factors limiting crop-to-weed gene flow. This level of gene flow and its dependence on flowering time might influence the evolutionary fate of weedy sunflower populations sympatric to their crop relative.


Psychophysiological arousal and biased perception of bodily anxiety symptoms in socially anxious children and adolescents: a systematic review.
Cognitive models of social anxiety [Clark and Wells, Social phobia: Diagnosis, assessment, and treatment, Guilford Press, New York, 1995], diagnostic criteria and studies on adult samples suggest that both an overestimation of bodily anxiety symptoms and psychophysiological abnormalities play an important role in social anxiety. To date, less is known about such a perception bias and physiological characteristics in children and adolescents with social anxiety. We performed a systematic review of the literature in the electronic databases Medline, PsycINFO, and PSYNDEX. Additional studies were identified by hand search using the ancestry approach. We identified 1,461 studies, screened their titles and abstracts, viewed 94 papers, and included 28 of these. Study samples were heterogeneous and consisted of socially phobic, high socially anxious, shy and test anxious children and adolescents. Regarding a biased perception, most studies in the review suggest that bodily symptoms of anxiety were overestimated by children and adolescents across the social anxiety spectrum when compared with control groups. An elevated psychophysiological reactivity to social stress was present in samples of high social anxiety, shyness, and test anxiety. In clinical samples with social phobia, by contrast, no differences or an even lower physiological responding compared with healthy control groups were reported. In addition, some evidence for a chronic psychophysiological hyperarousal was found across all sample types. The results are discussed with regard to current models of social anxiety, psychophysiological theories, and treatment implications.


Photoinduced Electron Transfer and Hole Migration in Nanosized Helical Aromatic Oligoamide Foldamers.
A series of photoactive triads have been synthesized and investigated in order to elucidate photoinduced electron transfer and hole migration mechanism across nanosized, rigid helical foldamers. The triads are comprised of a central helical oligoamide foldamer bridge with 9, 14, 18, 19, or 34 8-amino-2-quinolinecarboxylic acid repeat units, and of two chromophores, an N-terminal oligo(para-phenylenevinylene) electron donor and a C-terminal perylene bis-imide electron acceptor. Time-resolved fluorescence and transient absorption spectroscopic studies showed that, following photoexcitation of the electron acceptor, fast electron transfer occurs initially from the oligoquinoline bridge to the acceptor chromophore on the picosecond time scale. The oligo(para-phenylenevinylene) electron donor is oxidized after a time delay during which the hole migrates across the foldamer from the acceptor to the donor. The charge separated state that is finally generated was found to be remarkably long-lived (>80 μs). While the initial charge injection rate is largely invariant for all foldamer lengths (ca. 60 ps), the subsequent hole transfer to the donor varies from 1 × 109 s-1 for the longest sequence to 17 × 109 s-1 for the shortest. In all cases, charge transfer is very fast considering the foldamer length. Detailed analysis of the process in different media and at varying temperatures is consistent with a hopping mechanism of hole transport through the foldamer helix, with individual hops occurring on the subpicosecond time scale (kET = 2.5 × 1012 s-1 in CH2Cl2). This work demonstrates the possibility of fast long-range hole transfer over 300 Å (through bonds) across a synthetic modular bridge, an achievement that had been previously observed principally with DNA structures.


K+ channels and the microglial respiratory burst.
Microglial activation following central nervous system damage or disease often culminates in a respiratory burst that is necessary for antimicrobial function, but, paradoxically, can damage bystander cells. We show that several K+ channels are expressed and play a role in the respiratory burst of cultured rat microglia. Three pharmacologically separable K+ currents had properties of Kv1.3 and the Ca2+/calmodulin-gated channels, SK2, SK3, and SK4. mRNA was detected for Kv1.3, Kv1.5, SK2, and/or SK3, and SK4. Protein was detected for Kv1.3, Kv1.5, and SK3 (selective SK2 and SK4 antibodies not available). No Kv1.5-like current was detected, and confocal immunofluorescence showed the protein to be subcellular, in contrast to the robust membrane localization of Kv1.3. To determine whether any of these channels play a role in microglial activation, a respiratory burst was stimulated with phorbol 12-myristate 13-acetate and measured using a single cell, fluorescence-based dihydrorhodamine 123 assay. The respiratory burst was markedly inhibited by blockers of SK2 (apamin) and SK4 channels (clotrimazole and charybdotoxin), and to a lesser extent, by the potent Kv1.3 blocker agitoxin-2.


Evidence for community structuring associated with the actinorhizal shrub Ceanothus americanus in tallgrass prairies in Illinois, USA.
The role of the actinorhizal shrub Ceanothus americanus L. in community organisation of native grasslands was examined in three Illinois tallgrass prairie remnants. We asked can C. americanus structure tallgrass prairie at the patch scale through nitrogen fixation leading to differences in species assemblages and diversity? Expectations were that warm season (C4) graminoid species would be more abundant and species diversity would be greater outside Ceanothus patches than in patches associated with C. americanus where, based on results from numerous nitrogen augmentation and deposition studies, we expected greater abundance of cool season (C3) graminoid species and lower species diversity. Plots with and without association to C. americanus were compared for differences in floristic similarity, diversity, and C3 versus C4 graminoid species abundance. C3 graminoid species, including the adventive Poa pratensis L., were significantly more abundant in prairie associated with C. americanus than were C4 species. C4 species, such as Andropogon gerardi Vitman and Schizachyrium scopariumii (Michx.) Nash., were more abundant than C3 graminoid species in prairie patches without association to C. americanus. Means for diversity among plots associated with C. americanus were significantly lower than for other plots, nearly so for evenness and species density, but not different for sum total cover or total species richness. There were no statistically-significant differences in measures of mean total soil nitrogen at points along linear transects radiating from the base of C. americanus shrubs nor between mean soil amino-sugar-nitrogen concentrations within and outside of C. americanus patches. Nonetheless, comparative evidence supports the hypothesis that C. americanus structures composition and diversity at the local patch scale in tallgrass prairie.


Biselectivity of isoDGR peptides for fibronectin binding integrin subtypes α5β1 and αvβ6: conformational control through flanking amino acids.
Integrins are the major class of cell adhesion proteins. Their interaction with different ligands of the extracellular matrix is diverse. To get more insight into these interactions, artificial ligands endowed with a well-defined activity/selectivity profile are necessary. Herein, we present a library of cyclic pentapeptides, based on our previously reported peptide motif c(-phg-isoDGR-X-), in which high activity toward fibronectin binding integrins α5β1 and αvβ6 and not on vitronectin binding integrins αvβ3 and αvβ5 has been achieved by changing the flanking amino acids. The structure of the most promising candidates has been determined using a combined approach of NMR, distance geometry, and molecular dynamics simulations, and docking studies have been further used to elucidate the peptide-integrin interactions at the molecular level. The peptides' binding affinity has been characterized by enzyme linked immunosorbent assay experiments, and the results have been verified by cell adhesion experiments on specifically functionalized surfaces.


Automated alignment and pattern recognition of single-molecule force spectroscopy data.
Recently, direct measurements of forces stabilizing single proteins or individual receptor-ligand bonds became possible with ultra-sensitive force probe methods like the atomic force microscope (AFM). In force spectroscopy experiments using AFM, a single molecule or receptor-ligand pair is tethered between the tip of a micromachined cantilever and a supporting surface. While the molecule is stretched, forces are measured by the deflection of the cantilever and plotted against extension, yielding a force spectrum characteristic for each biomolecular system. In order to obtain statistically relevant results, several hundred to thousand single-molecule experiments have to be performed, each resulting in a unique force spectrum. We developed software and algorithms to analyse large numbers of force spectra. Our algorithms include the fitting polymer extension models to force peaks as well as the automatic alignment of spectra. The aligned spectra allowed recognition of patterns of peaks across different spectra. We demonstrate the capabilities of our software by analysing force spectra that were recorded by unfolding single transmembrane proteins such as bacteriorhodopsin and NhaA. Different unfolding pathways were detected by classifying peak patterns. Deviant spectra, e.g. those with no attachment or erratic peaks, can be easily identified. The software is based on the programming language C++, the GNU Scientific Library (GSL), the software WaveMetrics IGOR Pro and available open-source at http://bioinformatics.org/fskit/.


The role of locus coeruleus in the antiepileptic activity induced by vagus nerve stimulation.
Stimulation of the vagus nerve produces antiepileptic effects. This is used clinically to treat drug-refractory epilepsies. The mechanisms responsible for these effects depend on the activation of vagal afferents reaching the nucleus of the solitary tract. This review focuses on the neuroanatomy of the nucleus of the solitary tract and its relation with the nucleus locus coeruleus as a preferential anatomical substrate in producing antiepileptic effects. In fact, following the transient or permanent inactivation of locus coeruleus neurons, some antiepileptic effects of vagus nerve stimulation are lost. The activation of locus coeruleus per se is known to limit the spread of a seizure and the duration of a variety of seizure types. This is due to the fine chemical neuroanatomy of norepinephrine pathways that arise from the locus coeruleus, which produce widespread changes in cortical areas. These changes may be sustained by norepinephrine alone, or in combination with its co-transmitters. In addition, vagus nerve stimulation may prevent seizures by activating the serotonin-containing dorsal raphe neurons.


[Assessment of natriuretic peptide indices and oxidative stress in patients with chronic heart failure].
The authors have studied indices of natriuretic peptide and oxidative stress in patients with chronic heart failure (CHF). 52 male patients with postinfarction cardiosclerosis (PICS) who have developed CHF have been observed. The age of the patients varied from 38 till 60. It was established that CHF patients with progression of the disease had worsening of their clinical condition together with an increase of oxidative stress which was characterized through decrease of NO metabolites, NADPH--diaphorase (eNOS), increase of nitrite reductase (iNOS) and peroxinitrite (ONOO), correlative increase the level of brain natriuretic peptide in blood plazma. Reliable connection between considerable increase of oxidative stress and the level of NT-pro BNP was noted in CHF patients, which demands necessity of correction of observed disorders.


Precipitation isoscapes for New Zealand: enhanced temporal detail using precipitation-weighted daily climatology.
Predictive understanding of precipitation δ(2)H and δ(18)O in New Zealand faces unique challenges, including high spatial variability in precipitation amounts, alternation between subtropical and sub-Antarctic precipitation sources, and a compressed latitudinal range of 34 to 47 °S. To map the precipitation isotope ratios across New Zealand, three years of integrated monthly precipitation samples were acquired from >50 stations. Conventional mean-annual precipitation δ(2)H and δ(18)O maps were produced by regressions using geographic and annual climate variables. Incomplete data and short-term variation in climate and precipitation sources limited the utility of this approach. We overcome these difficulties by calculating precipitation-weighted monthly climate parameters using national 5-km-gridded daily climate data. This data plus geographic variables were regressed to predict δ(2)H, δ(18)O, and d-excess at all sites. The procedure yields statistically-valid predictions of the isotope composition of precipitation (long-term average root mean square error (RMSE) for δ(18)O = 0.6 ‰; δ(2)H = 5.5 ‰); and monthly RMSE δ(18)O = 1.9 ‰, δ(2)H = 16 ‰. This approach has substantial benefits for studies that require the isotope composition of precipitation during specific time intervals, and may be further improved by comparison to daily and event-based precipitation samples as well as the use of back-trajectory calculations.


TCF4 induces enzalutamide resistance via neuroendocrine differentiation in prostate cancer.
In treating patients with castration resistant prostate cancer (CRPC), enzalutamide, the second-generation androgen receptor (AR) antagonist, is an accepted standard of care. However, clinical benefits are limited to a median time of 4.8 months because resistance inevitably emerges. To determine the mechanism of treatment resistance, we carried out a RNA sequence analysis and found increased expression levels of neuroendocrine markers in the enzalutamide-resistant LNCaP human prostate cancer (CaP) cell line when compared to the parental cell line. Subsequent studies demonstrated that Transcription Factor-4 (TCF4), a transcription factor implicated in WNT signaling, mediated neuroendocrine differentiation (NED) in response to enzalutamide treatment and was elevated in the enzalutamide-resistant LNCaP. In addition, we observed that PTHrP mediated enzalutamide resistance in tissue culture and inducible TCF4 overexpression resulted in enzalutamide-resistance in a mouse xenograft model. Finally, small molecule inhibitors of TCF4 or PTHrP partially reversed enzalutamide resistance in CaP cells. When tissues obtained from men who died of metastatic CaP were examined, a positive correlation was found between the expression levels of TCF4 and PTHrP. Taken together, the current results indicate that TCF4 induces enzalutamide resistance via NED in CaP.


Clinical utility of urine specific gravity, electrical conductivity, and color as on-farm methods for evaluating urine concentration in dairy cattle.
Urine concentration (UC) provides clinically useful information concerning hydration status and renal function of animals. To characterize the clinical performance of urine specific gravity measured by optical refractometry (USG-R ) or Multistix-SG urine reagent dipstick (USG-D ), urine electrical conductivity using an OAKTON Con 6 conductivity handheld meter (UEC ), urine color (UColor ) using a custom-designed 8-point color chart, and urine creatinine concentration (UCreat ) for assessing UC in dairy cattle. 20 periparturient Holstein-Friesian cows. Urine was obtained by perineal stimulation or urethral catheterization and urine osmolality (UOsm , reference method), USG-R , USG-D , UEC , UColor , and UCreat determined. Diagnostic test performance was evaluated using Spearman's rho and logistic regression to determine the area under the receiver operating curve (AUC) and optimal cut point for diagnosing hypohydration (UOsm ≥800 mOsm/kg). P < .05 was considered significant. The best performing test for diagnosing hypohydration was USG-R (AUC = 0.90) at an optimal cut point ≥1.030. The second-best performing test was UEC (AUC = 0.82) at a cut point of ≥23.7 mS/cm, followed by UCreat (AUC = 0.76) at a cut point of ≥95.3 mg/dL, and UColor (AUC = 0.74) at a cut point of ≥4 on an 8-point scale. Urine specific gravity measured by dipstick performed poorly (AUC = 0.63). USG-R and UEC provide practical and sufficiently accurate methods for measuring UC in dairy cattle. Urine color had moderate clinical utility as a no-cost cow-side method for assessing UC, whereas dipstick refractometry is not recommended for assessing UC.


Energetic preference of 8-oxoG eversion pathways in a DNA glycosylase.
Base eversion is a fundamental process in the biochemistry of nucleic acids, allowing proteins engaged in DNA repair and epigenetic modifications to access target bases in DNA. Crystal structures reveal end points of these processes, but not the pathways involved in the dynamic process of base recognition. To elucidate the pathway taken by 8-oxoguanine during base excision repair by Fpg, we calculated free energy surfaces during eversion of the damaged base through the major and minor grooves. The minor groove pathway and free energy barrier (6-7 kcal/mol) are consistent with previously reported results (Qi, Y.; Spong, M. C.; Nam, K.; Banerjee, A.; Jiralerspong, S.; Karplus, M.; Verdine, G. L. Nature 2009, 462, 762.) However, eversion of 8-oxoG through the major groove encounters a significantly lower barrier (3-4 kcal/mol) more consistent with experimentally determined rates of enzymatic sliding during lesion search (Blainey, P. C.; van Oijent, A. M.; Banerjee, A.; Verdine, G. L.; Xie, X. S. Proc. Natl. Acad. Sci. U.S.A. 2006, 103, 5752.). Major groove eversion has been suggested for other glycosylases, suggesting that in addition to function, dynamics of base eversion may also be conserved.


[Lysosomal proteinase activity in the small intestine in postnatal ontogeny].
A lysosomal fraction was isolated from various parts of small intestinal mucose of rats at postnatal periods of growth as well as from adult animals. Activity of marked lysosomal enzymes (acid phosphatase and cathepsins) was 3-fold higher in ileum than the jejunum of young rats at the lactation periods of growth. In adult animals the enzymatic activity was similar both in proximal and distal parts of small intestines. At the postnatal period of growth the lysosomal enzymes appear to carry out a compensatory function, participating in final steps of milk protein digestion.


Conversion of androstenedione to 3 beta-hydroxy-5-androsten-17-one and 3 beta-hydroxy-4-androsten-17-one by the testicular microsomal fraction of Sprague-Dawley rats.
When androstenedione was incubated with testicular microsomes of Sprague-Dawley rats in the presence of reduced nicotinamide-adenine dinucleotide (NADH), unknown metabolites were produced, in addition to testosterone and 7 alpha-hydroxyandrostenedione. The metabolites were identified as 3 beta-hydroxy-4-androsten-17-one and 3 beta-hydroxy-5-androsten-17-one (3:1) by biochemical and radiochemical methods. These results confirmed the occurrence of the reverse reactions from androstenedione to 3 beta-hydroxy-4-androsten-17-one and 3 beta-hydroxy-5-androsten-17-one catalyzed by the 3 beta-hydroxysteroid dehydrogenase and 5-ene-4-ene isomerase in the microsomal fraction of Sprague-Dawley rat testes.


Histopathologic features of the tumor budding in adenocarcinoma of the lung: tumor budding as an index to predict the potential aggressiveness.
The term tumor budding has been applied to single cells or small clusters of up to four cells within the stromal tissue at the invasive margin of colorectal cancers. This morphologic feature is increasingly being recognized as an adverse prognostic factor. The purpose of this study was to evaluate the clinicopathologic significance of tumor budding in adenocarcinomas of the lung. We investigated the relationship between tumor budding and clinicopathologic parameters of adenocarcinomas of the lung and the prognostic significance of tumor budding by reviewing the cases of 201 consecutive patients who had undergone complete resection of adenocarcinoma of the lung measuring 30 mm or less in diameter. We examined immunohistochemical profile of budding cells (BCs) by immunohistochemical staining with 14 antibodies. Tumor budding was observed in 78 (43.1%) of the 181 cases with invasive adenocarcinoma. The presence of tumor budding was significantly associated with lymph node metastasis (p = 0.005), pathologic stage (p < 0.001), vascular invasion (p = 0.003), lymphatic invasion (p = 0.009), and pleural invasion (p = 0.029). Examination of the relation between the presence of tumor budding and the predominant histologic subtype revealed that the predominant papillary subtype was significantly associated with the presence of tumor budding (p = 0.0023), whereas the predominant bronchioloalveolar carcinoma subtype was significantly associated with the absence of tumor budding (p < 0.001). The overall 5-year survival rates of the group with budding and the group without budding was 67.5% and 88.3%, respectively, and difference was significant (p = 0.0057). Compared with cancer cells forming nests, BCs displayed reduced expression of cellular adhesion molecule, E-cadherin, and beta-catenin (p < 0.05 and p < 0.05, respectively) and increased expression of laminin5-gamma2 (p < 0.05). However, BCs displayed reduced expression of differentiation marker, surfactant protein A (p < 0.05). Multivariate analysis revealed that tumor budding was significant independent prognostic factor of the small-sized adenocarcinoma of the lung. Our data showed that tumor budding in adenocarcinoma of the lung is a distinct morphologic feature that has biologic and prognostic significance.


Clinical implications of matrix metalloproteinases.
Matrix metalloproteinases (MMPs) are a family of neutral proteinases that are important for normal development, wound healing, and a wide variety of pathological processes, including the spread of metastatic cancer cells, arthritic destruction of joints, atherosclerosis, pulmonary fibrosis, emphysema and neuroinflammation. In the central nervous system (CNS), MMPs have been shown to degrade components of the basal lamina, leading to disruption of the blood brain barrier and to contribute to the neuroinflammatory responses in many neurological diseases. Inhibition of MMPs have been shown to prevent progression of these diseases. Currently, certain MMP inhibitors have entered into clinical trials. A goal to the future should be to design selective synthetic inhibitors of MMPs that have minimum side effects. MMP inhibitors are designed in such a way that these can not only bind at the active site of the proteinases but also to have the characteristics to bind to other sites of MMPs which might be a promising route for therapy. To name a few: catechins, a component isolated from green tea; and Novastal, derived from extracts of shark cartilage are currently in clinical trials for the treatment of MMP-mediated diseases.


Closed-Loop Behavioral Control Increases Coherence in the Fly Brain.
A crucial function of the brain is to be able to distinguish whether or not changes in the environment are caused by one's own actions. Even the smallest brains appear to be capable of making this distinction, as has been shown by closed-loop behavioral experiments in flies controlling visual stimuli in virtual reality paradigms. We questioned whether activity in the fruit fly brain is different during such closed-loop behavior, compared with passive viewing of a stimulus. To address this question, we used a procedure to record local field potential (LFP) activity across the fly brain while flies were controlling a virtual object through their movement on an air-supported ball. The virtual object was flickered at a precise frequency (7 Hz), creating a frequency tag that allowed us to track brain responses to the object while animals were behaving. Following experiments under closed-loop control, we replayed the same stimulus to the fly in open loop, such that it could no longer control the stimulus. We found identical receptive fields and similar strength of frequency tags across the brain for the virtual object under closed loop and replay. However, when comparing central versus peripheral brain regions, we found that brain responses were differentially modulated depending on whether flies were in control or not. Additionally, coherence of LFP activity in the brain increased when flies were in control, compared with replay, even if motor behavior was similar. This suggests that processes associated with closed-loop control promote temporal coordination in the insect brain. We show that closed-loop control of a visual stimulus promotes temporal coordination across the Drosophila brain, compared with open-loop replay of the same visual sequences. This is significant because it suggests that, to understand goal-directed behavior or visual attention in flies, it may be most informative to sample neural activity from multiple regions across the brain simultaneously, and to examine temporal relationships (e.g., coherence) between these regions.


Estimation of the fraction of cancer cells in a tumor DNA sample using DNA methylation.
Contamination of normal cells is almost always present in tumor samples and affects their molecular analyses. DNA methylation, a stable epigenetic modification, is cell type-dependent, and different between cancer and normal cells. Here, we aimed to demonstrate that DNA methylation can be used to estimate the fraction of cancer cells in a tumor DNA sample, using esophageal squamous cell carcinoma (ESCC) as an example. First, by an Infinium HumanMethylation450 BeadChip array, we isolated three genomic regions (TFAP2B, ARHGEF4, and RAPGEFL1) i) highly methylated in four ESCC cell lines, ii) hardly methylated in a pooled sample of non-cancerous mucosae, a pooled sample of normal esophageal mucosae, and peripheral leukocytes, and iii) frequently methylated in 28 ESCCs (TFAP2B, 24/28; ARHGEF4, 20/28; and RAPGEFL1, 19/28). Second, using eight pairs of cancer and non-cancer cell samples prepared by laser capture microdissection, we confirmed that at least one of the three regions was almost completely methylated in ESCC cells, and all the three regions were almost completely unmethylated in non-cancer cells. We also confirmed that DNA copy number alterations of the three regions in 15 ESCC samples were rare, and did not affect the estimation of the fraction of cancer cells. Then, the fraction of cancer cells in a tumor DNA sample was defined as the highest methylation level of the three regions, and we confirmed a high correlation between the fraction assessed by the DNA methylation fraction marker and the fraction assessed by a pathologist (r=0.85; p<0.001). Finally, we observed that, by correction of the cancer cell content, CpG islands in promoter regions of tumor-suppressor genes were almost completely methylated. These results demonstrate that DNA methylation can be used to estimate the fraction of cancer cells in a tumor DNA sample.


Extracellular Vesicles in Health and Disease.
The journal, Aging and Disease, has released a special issue on "Extracellular Vesicles (EVs) in Health and Disease." The special issue comprises review and original research articles discussing the role of EVs in aging and senescence, the utility of evaluating EVs in body fluids for understanding the pathophysiology or progression of various diseases such as Parkinson's Disease, Multiple Sclerosis, Chronic Traumatic Encephalopathy, and Morphine induced amyloidopathy. Also, a series of articles discussed the promise of stem cell-derived EVs for treating Parkinson's Disease, Sjogren's Syndrome, and Inflammatory Bowel Disease, and advancements in loading EVs to deliver nucleic acid therapies. This editorial discusses the highlights from these articles.


A shuttling-based two-qubit logic gate for linking distant silicon quantum processors.
Control of entanglement between qubits at distant quantum processors using a two-qubit gate is an essential function of a scalable, modular implementation of quantum computation. Among the many qubit platforms, spin qubits in silicon quantum dots are promising for large-scale integration along with their nanofabrication capability. However, linking distant silicon quantum processors is challenging as two-qubit gates in spin qubits typically utilize short-range exchange coupling, which is only effective between nearest-neighbor quantum dots. Here we demonstrate a two-qubit gate between spin qubits via coherent spin shuttling, a key technology for linking distant silicon quantum processors. Coherent shuttling of a spin qubit enables efficient switching of the exchange coupling with an on/off ratio exceeding 1000, while preserving the spin coherence by 99.6% for the single shuttling between neighboring dots. With this shuttling-mode exchange control, we demonstrate a two-qubit controlled-phase gate with a fidelity of 93%, assessed via randomized benchmarking. Combination of our technique and a phase coherent shuttling of a qubit across a large quantum dot array will provide feasible path toward a quantum link between distant silicon quantum processors, a key requirement for large-scale quantum computation.


The Role of Cognitive Factors in Predicting Balance and Fall Risk in a Neuro-Rehabilitation Setting.
There is a consistent body of evidence supporting the role of cognitive functions, particularly executive function, in the elderly and in neurological conditions which become more frequent with ageing. The aim of our study was to assess the role of different domains of cognitive functions to predict balance and fall risk in a sample of adults with various neurological conditions in a rehabilitation setting. This was a prospective, cohort study conducted in a single centre in the UK. 114 participants consecutively admitted to a Neuro-Rehabilitation Unit were prospectively assessed for fall accidents. Baseline assessment included a measure of balance (Berg Balance Scale) and a battery of standard cognitive tests measuring executive function, speed of information processing, verbal and visual memory, visual perception and intellectual function. The outcomes of interest were the risk of becoming a faller, balance and fall rate. Two tests of executive function were significantly associated with fall risk, the Stroop Colour Word Test (IRR 1.01, 95% CI 1.00-1.03) and the number of errors on part B of the Trail Making Test (IRR 1.23, 95% CI 1.03-1.49). Composite scores of executive function, speed of information processing and visual memory domains resulted in 2 to 3 times increased likelihood of having better balance (OR 2.74 95% CI 1.08 to 6.94, OR 2.72 95% CI 1.16 to 6.36 and OR 2.44 95% CI 1.11 to 5.35 respectively). Our results show that specific subcomponents of executive functions are able to predict fall risk, while a more global cognitive dysfunction is associated with poorer balance.


Cell walls of the dimorphic fungal pathogens Sporothrix schenckii and Sporothrix brasiliensis exhibit bilaminate structures and sloughing of extensive and intact layers.
Sporotrichosis is a subcutaneous mycosis caused by pathogenic species of the Sporothrix genus. A new emerging species, Sporothrix brasiliensis, is related to cat-transmitted sporotrichosis and has severe clinical manifestations. The cell wall of pathogenic fungi is a unique structure and impacts directly on the host immune response. We reveal and compare the cell wall structures of Sporothrix schenckii and S. brasiliensis using high-pressure freezing electron microscopy to study the cell wall organization of both species. To analyze the components of the cell wall, we also used infrared and 13C and 1H NMR spectroscopy and the sugar composition was determined by quantitative high-performance anion-exchange chromatography. Our ultrastructural data revealed a bi-layered cell wall structure for both species, including an external microfibrillar layer and an inner electron-dense layer. The inner and outer layers of the S. brasiliensis cell wall were thicker than those of S. schenckii, correlating with an increase in the chitin and rhamnose contents. Moreover, the outer microfibrillar layer of the S. brasiliensis cell wall had longer microfibrils interconnecting yeast cells. Distinct from those of other dimorphic fungi, the cell wall of Sporothrix spp. lacked α-glucan component. Interestingly, glycogen α-particles were identified in the cytoplasm close to the cell wall and the plasma membrane. The cell wall structure as well as the presence of glycogen α-particles varied over time during cell culture. The structural differences observed in the cell wall of these Sporothrix species seemed to impact its uptake by monocyte-derived human macrophages. The data presented here show a unique cell wall structure of S. brasiliensis and S. schenckii during the yeast parasitic phase. A new cell wall model for Sporothrix spp. is therefore proposed that suggests that these fungi molt sheets of intact cell wall layers. This observation may have significant effects on localized and disseminated immunopathology.


Celecoxib inhibits serum amyloid a-induced matrix metalloproteinase-10 expression in human endothelial cells.
Although serum amyloid A (SAA) is an established biomarker of coronary artery disease (CAD), its direct role in matrix degradation is obscure. This study investigated the effect of SAA on the expression of matrix metalloproteinase-10 (MMP-10) in endothelial cells. The effect of celecoxib on SAA-dependent MMP-10 expression and its possible mediator were also investigated. From our time course microarray screening, SAA (20 microg/ml) was found to increase MMP-10 mRNA expression over time (4-48 h) in human endothelial cells. Quantitative real-time PCR confirmed this transcriptional induction. Correspondingly, secreted MMP-10 protein was also markedly induced by SAA treatment for 24 h in a dose-dependent manner. We further examined cyclooxygenase-2 (COX-2) and its major product, prostaglandin E(2) (PGE(2)), as possible mediators of MMP-10 induction. Direct PGE(2) treatment could result in MMP-10 induction. Celecoxib, a selective COX-2 inhibitor, suppressed MMP-10 secretion induced by SAA. SAA induced MMP-10 expression and celecoxib prevented its induction. MMP-10 induction was at least partly mediated by PGE(2).


Protease-activated receptor stimulation activates a Ca2+-independent phospholipase A2 in bladder microvascular endothelial cells.
Increased mast cell numbers and mast cell activation represent one of the prevalent etiologic theories for interstitial cystitis, an inflammatory condition in the bladder. This study was designed primarily to determine whether increased mast cell tryptase in the bladder wall may play a role in activating bladder endothelial cell phospholipase A(2) (PLA(2)), leading to increased inflammatory phospholipid metabolite accumulation, which may propagate the inflammatory process. We stimulated human bladder microvascular endothelial cells with thrombin or tryptase and measured the activation of PLA(2) and the production of multiple membrane phospholipid-derived inflammatory mediators. Thrombin and tryptase stimulation resulted in activation of a Ca(2+)-independent PLA(2), leading to increased release of arachidonic acid and prostacyclin and increased production of platelet-activating factor. These responses were blocked completely by pretreatment of human bladder microvascular endothelial cells with the Ca(2+)-independent PLA(2)-selective inhibitor bromoenol lactone. The combination of increased prostacyclin and platelet-activating factor in the bladder circulation may result in vasodilation and increased polymorphonuclear leukocyte adherence to the endothelium and may facilitate recruitment of polymorphonuclear leukocytes to the bladder wall of patients with interstitial cystitis.


Recent and rapid anthropogenic habitat fragmentation increases extinction risk for freshwater biodiversity.
Anthropogenic habitat fragmentation is often implicated as driving the current global extinction crisis, particularly in freshwater ecosystems. The genetic signal of recent population isolation can be confounded by the complex spatial arrangement of dendritic river systems. Consequently, many populations may presently be managed separately based on an incorrect assumption that they have evolved in isolation. Integrating landscape genomics data with models of connectivity that account for landscape structure, we show that the cumulative effects of multiple in-stream barriers have contributed to the recent decline of a freshwater fish from the Murray-Darling Basin, Australia. In addition, individual-based eco-evolutionary simulations further demonstrate that contemporary inferences about population isolation are consistent with the 160-year time frame since construction of in-stream barriers began in the region. Our findings suggest that the impact of very recent fragmentation may be often underestimated for freshwater biodiversity. We argue that proactive conservation measures to reconnect many riverine populations are urgently needed.


The p47 GTPases Iigp2 and Irgb10 regulate innate immunity and inflammation to murine Chlamydia psittaci infection.
C57BL/6J mice were 10(5)-fold more resistant to Chlamydia psittaci infection than DBA/2J mice by LD(100) determinations. Linkage analysis using BXD recombinant inbred strains revealed a single effector locus at a 1.5-Mbp region on chromosome 11 encoding a cluster of three p47 GTPases (Irgb10, Igtp, and Iigp2). Western blots of infected tissue showed that Irgb10 was elevated in resistant mice and one of the two possible Iigp2 protein isoforms was preferentially expressed in susceptible mice. The BXD39 strain, susceptible at Irgb10 and resistant at Iigp2, had an intermediate phenotype implicating the nonredundant role of these p47 GTPases. C57BL/6J and DBA/2J exhibited a difference in IFN-gamma-dependent chlamydial control, which was reversible by Iigp2 small interfering RNA knockdown. Microarrays of infected peritoneal lavage revealed >10-fold up-regulation of neutrophil-recruiting chemokines in susceptible mice and >100-fold increase in macrophage differentiation genes in resistant mice, indicating that the susceptibility pattern involves the stimulation of different inflammatory cell-recruiting pathways. Massive neutrophil recruitment was seen in susceptible mice by histology and flow cytometry, and neutrophil chemokine receptor (CXCR2) knockout mice on a susceptible background survived a lethal challenge, confirming that neutrophil recruitment was required for susceptibility. Congenic Igtp knockout mice also susceptible at Irgb10 and Iigp2 on a resistant background recruited neutrophils and succumbed to infection. We conclude that Irgb10 and Iigp2 act together to confer differential susceptibility against murine chlamydial infection. Data indicate that these p47 GTPases have cell-autonomous effects that result in vastly different inflammatory stimulations, leading to either recovery or death.


Comparison of olanzapine long-acting injection and oral olanzapine: a 2-year, randomized, open-label study in outpatients with schizophrenia.
We compared long-term treatment effectiveness of monthly olanzapine long-acting injection (LAI) with that of oral olanzapine. Outpatients with 2 or more episodes of psychotic worsening in the past 24 months with Positive and Negative Syndrome Scale total score of lower than 70 were randomized to 405 mg/4 weeks of olanzapine LAI (n = 264) or 10 mg/d of oral olanzapine (n = 260) for 2 years of open-label treatment. Dosing thereafter was flexible (150-405 mg/4 weeks of LAI vs 5-20 mg/d of oral). Primary outcome was time to all-cause discontinuation. At baseline, patients were clinically stable (mean Positive and Negative Syndrome Scale total score of 57). Seventeen percent of patients had been psychiatrically hospitalized in the previous 6 months, and 4.6% were rated nonadherent in the month before study entry. The groups did not differ significantly in median time to all-cause discontinuation (645 days for LAI, 678 days for oral; P = 0.61), discontinuation rate (53.8% for LAI, 51.2% for oral; P = 0.60), or relapse rate (20.1% for LAI, 18.5% for oral; P = 0.66). Postbaseline psychiatric hospitalization rate was low for both groups (7.6% for LAI, 9.2% for oral), but mean hospitalization duration was significantly longer for oral patients (1.80 days [20 for those hospitalized] vs 0.43 days [6 for those hospitalized], P = 0.02). There were no clinically significant group differences in adverse events or safety measures. No post-injection delirium/sedation syndrome events occurred. In conclusion, olanzapine LAI and oral olanzapine were similarly effective and well tolerated for up to 2 years of treatment in patients with schizophrenia. Treatment discontinuation for olanzapine LAI was similar to that of oral olanzapine, despite the 3-hour post-injection observation period and other precautionary procedures related to risk of post-injection delirium/sedation syndrome.


Effect of amplitude modulation on profile detection.
The effect of amplitude modulation on profile detection was examined in three experiments. The observer's task was to determine in which observation interval an increment was added to the 1000-Hz target component of a multitone complex in which the components were equally spaced on a logarithmic frequency scale from 200 to 5000 Hz. The target was unmodulated throughout the study. In some conditions, all nontarget components of the standard were modulated in phase; in other conditions, they were modulated with random phase. In experiment 1, the threshold was measured as a function of the modulation rate. The results show that, at low modulation rates, 5 Hz for example, modulation elevates threshold by about 13 dB. The threshold decreases as the modulation rate increases, with the threshold elevation being only 3 dB at 80 Hz. In experiment 2, threshold was measured as a function of modulation depth for both 21- and 5-component complexes. The results show that for a 5-Hz modulation rate the threshold decreases as the modulation depth decreases, and that the rate of decrease is greater for the 21-component complex than for the 5-component complex. In experiment 3, the effects of random-phase modulation were explored; the phase of the modulation waveform was randomly chosen for each component. The results show that there is no difference between in-phase and random-phase modulation when each component occupies a different critical band. If, however, two or more components occupy the same critical band, then randomizing the phase of modulation reduces the effective depth of modulation within that critical band, and the effect of modulation is thereby lessened.


Long non-coding RNA ZEB1-AS1 indicates poor prognosis and promotes melanoma progression through targeting miR-1224-5p.
Long non-coding RNAs (lncRNAs) have critical roles in various types of cancer, but their roles in the development of melanoma and the underlying molecular mechanisms remain to be fully elucidated. In the present study, the role of zinc finger E-box binding homeobox 1 antisense RNA 1 (ZEB1-AS1) in melanoma was assessed. The expression levels of ZEB1-AS1 were increased in melanoma cell lines and tumor tissues as indicated by reverse transcription-quantitative polymerase chain reaction analysis. Kaplan-Meier analysis suggested that higher expression of ZEB1-AS1 predicts poor prognosis of melanoma patients. Furthermore, knockdown of ZEB1-AS1 inhibited the proliferation, migration and invasion of melanoma, suggesting a role of ZEB1-AS1 in the development and progression of melanoma. In addition, a luciferase reporter assay confirmed that the expression of miR-1224-5p was directly regulated by ZEB1-AS1. Transfection with miR-1224-5p mimics reduced the levels of ZEB1-AS1 and the proliferation, migration and invasion of melanoma. In conclusion, ZEB1-AS1 enhances the proliferation, migration and invasion of melanoma, at least in part by inhibiting the expression of miR-1224-5p, and its overexpression is associated with poor survival of melanoma patients. In addition, the ZEB1-AS1/miR-1224-5p interaction may be a promising therapeutic target for melanoma treatment.


DEDB: a database of Drosophila melanogaster exons in splicing graph form.
A wealth of quality genomic and mRNA/EST sequences in recent years has provided the data required for large-scale genome-wide analysis of alternative splicing. We have capitalized on this by constructing a database that contains alternative splicing information organized as splicing graphs, where all transcripts arising from a single gene are collected, organized and classified. The splicing graph then serves as the basis for the classification of the various types of alternative splicing events. DEDB http://proline.bic.nus.edu.sg/dedb/index.html is a database of Drosophila melanogaster exons obtained from FlyBase arranged in a splicing graph form that permits the creation of simple rules allowing for the classification of alternative splicing events. Pfam domains were also mapped onto the protein sequences allowing users to access the impact of alternative splicing events on domain organization. DEDB's catalogue of splicing graphs facilitates genome-wide classification of alternative splicing events for genome analysis. The splicing graph viewer brings together genome, transcript, protein and domain information to facilitate biologists in understanding the implications of alternative splicing.


Fascial organisation of motor synergies: a hypothesis.
In the field of biomechanics and motor control understanding movement coordination is paramount. Motor synergies represent the coordination of neural and physical elements embedded in our bodies in order to optimize the solutions to motor problems. Although we are able to measure and quantify the movement made manifested, we do not have confidence in explaining the anatomical bases of its organisation at different levels. It is our contention that the flexible hierarchical organization of movement relies on the fascial structurers to create functional linkages at different levels, and this concept attunes with the neural control of synergies. At the base of movement organization there is a (somatic) equilibrium point that exists on the fascia where the neurologically- and mechanically-generated tensions dynamically balance out. This somatic equilibrium point is at the base of postural control, afferent flow of information to the nervous system about the state of the muscles, and of the coordinative pre-activation of muscular contraction sequences specific for a synergy. Implications are discussed and suggestions for research and clinical applications are made.


Simultaneous infection of cattle with different Anaplasma phagocytophilum variants.
Anaplasma phagocytophilum is a tick-transmitted Gram-negative obligate intracellular bacterium that replicates in neutrophil granulocytes. It causes tick-borne fever in cattle and sheep. We report here the case of a 5-year-old cow from Germany with clinically overt granulocytic anaplasmosis presenting with fever, lower limb oedema and drop in milk-yield. The herd encompassed 10 animals, 8 other animals showed subclinical infection. The strains from the 9 A. phagocytophilum positive cows were molecularly characterized using ankA gene-based and multilocus sequence typing (MLST). Seven of 9 (78%) animals were infected simultaneously with different ankA variants belonging to ankA clusters I and IV. MLST analysis also revealed the presence of multiple strain types. This could be due to co-transmission or superinfection. Hosts harboring diverse A. phagocytophilum strains might enable the emergence of new ankA variants and/or MLST sequence types via bacterial recombination.


Cell Cycle Analysis by Mass Cytometry.
The regulated progression of cells through the cell cycle during proliferation is a critical factor in tumor progression, anti-neoplastic therapy response, immune system regulation, and developmental biology. While flow cytometric measurement of cell cycle progression is well established, mass cytometry assays allow the cell cycle to be measured along with up to 39 other antigens enabling characterization of the complex interactions between the cell cycle and a wide variety of cellular processes. This method describes the use of mass cytometry for the analysis of cell cycle state for cells from three different sources: in vitro cultured cell lines, ex vivo human blood or bone marrow, and in vivo labeling and ex vivo analysis of murine tissues. The method utilizes incorporation of 5-Iodo-2'-deoxyuridine (IdU), combined with measurement of phosphorylated retinoblastoma protein (pRb), cyclin B1, and phosphorylated histone H3 (p-HH3). These measurements can be integrated into a gating strategy that allows for clear separation of all five phases of the cell cycle.


Skin infection caused by Scedosporium apiospermum.
A woman with a skin infection because of Scedosporium apiospermum, in the interdigital spaces of her feet is presented. The minimum inhibition concentration values (MIC, microg ml(-1)) of this isolated mould for itraconazole, amphotericin B and terbinafine after 48 h were determined as 1, 8 and 16, respectively. The patient was treated successfully with oral terbinafine and topical clotrimazole.


Fusobacterium hwasookii sp. nov., Isolated from a Human Periodontitis Lesion.
In this study, we classified the five strains (ChDC F128(T), ChDC F145, ChDC F174, ChDC F206, and ChDC F300) as a novel species of genus Fusobacterium by DNA-DNA hybridization and multi-locus phylogenetic analysis (MLPA), based on a single sequence (24,715 bp) of 22 concatenated housekeeping genes, with morphological and chemotaxonomic characteristics. DNA-DNA hybridization data showed that the values of genomic relatedness between ChDC F128(T) and each of the other novel strains were ranged from 79.0 to 82.6 %, while those of genomic relatedness between ChDC F128(T) and type strain of each of subspecies of F. nucleatum or Fusobacterium periodonticum were ranged from 40.9 to 54.4 %. MLPA revealed that the 5 strains were clustered as one group and clearly discriminated with F. nucleatum and F. periodonticum with 100 % bootstrap value. The DNA G+C content of the five novel strains were ranged from 26.9 to 27.0 mol%. The cellular fatty acid analysis of clinical isolates and type strains revealed C14:0, C16:0, and cis-9 C16:1 as the major fatty acids. The cell wall peptidoglycan of the 5 strains was comprised of meso-lanthionine. These results show that the 5 strains are novel species and belong to the genus Fusobacterium. Strain ChDC F128(T) (=KCOM 1249(T) = KCTC 5108(T) = JCM 30218(T)) is suggested to be the type strain of a novel species of genus Fusobacterium, for which the name Fusobacterium hwasookii sp. nov. is proposed.


Human leukemic intermediate DNA components.
The DNA extracted from human leukemic leucocytes has been fractionated on a methylated albumin kieselguhr (MAK) column. The different fractions obtained have been reannealed to a Cot value of 20 mol times sec/1 to study the distribution of the intermediate DNA on the MAK column. Intermediate DNA contains two components, one (CsCl density after reannealing, 1.703 g/ml) obtained by reannealing high molecular weight DNA, the other (CsCl density after reannealing, 1.707 g/ml) obtained only by reannealing sonicated low molecular weight DNA. High molecular weight intermediate DNA (1.703 component) is eluted early from the MAK column in the fractions corresponding to the main DNA peak, while low molecular weight intermediate DNA (1.707 component) is more widespread on the MAK column, but appears to be enriched in the fractions eluted later. The possibility is discussed that the latter component is interspersed in that part of the genome which is apparently more homogeneous in density in an analytical CsCl gradient, and is absent on the skewed, more heterogeneous, heavy side of the main DNA in CsCl.


MALDI-TOF MS-based identification of black yeasts of the genus Exophiala.
In this study, we investigated the applicability of matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) for the identification of Exophiala species. The analysis included a total of 110 Exophiala isolates, including 15 CBS strains representing 4 species, Exophiala dermatitidis (61), E. phaeomuriformis (36), E. crusticola (9), and E. heteromorpha (4), that had been previously identified based on internal transcribed spacer (ITS) regions. We also compared the relative efficacies of Sabouraud glucose agar (SGA) and Columbia agar (CA) for use in MALDI-TOF MS. Remarkably, we obtained a log-score value ≥2.0 by using either SGA or CA for all 15 CBS strains, indicating species-level identification. The remaining 95 Exophiala strains were identified to the genus or species levels, with identification rates of 96.8% and 90.5%, using SGA or CA, respectively. Most of the E. dermatitidis (100% and 92.9%), E. phaeomuriformis (80.6% and 83.9%), E. crusticola (50% and 100%), and E. heteromorpha (100% and 100%) isolates were correctly identified using SGA or CA, respectively. Furthermore, 58.9% and 26.3% of the strains had log-score values of ≥2.0 by using SGA and CA, respectively. Our results indicate that MALDI-TOF MS is a rapid and reliable technique with high rates of correct taxonomic identification.


Effect of third ventricular injection of beta-endorphin on the electrophysiological responses of some regions of endocrine hypothalamus.
Multiunit activity (MUA) of arcuate nucleus and cortical EEG were recorded in the regularly cycling female rats on the day of proestrous under urethane anaesthesia. The MUA was compared before and after injection of beta-endorphin in third ventricle. In some animals MUA increased after 30-40 min and persisted for 3-4 hr, in others MUA got inhibited within 5-10 min of injection of beta-Endorphin and effect lasted for 5-6 hr. There was no change in frontoparietal EEG activity. In another group of animals medial pre-optic responses (MPO) to stimulation of medial amygdala were tested before and after ventricular infusion of beta-endorphin. Most of the facilitatory MPO responses got blocked. These observations suggest the involvement of opioid receptors in the mediation of neuroendocrine control of preovulatory events through the amygdalo-preoptico-medial basal hypothalamic axis. There seems to be heterogeneity of beta-endorphin receptors in the arcuate nucleus.