Online fluorescent dye detection method for the characterization of immunoglobulin G aggregation by size exclusion chromatography and asymmetrical flow field flow fractionation.
The aim of this study was to develop an online fluorescent dye detection method suitable for high-pressure size exclusion chromatography (HP-SEC) and asymmetrical flow field flow fractionation (AF4). The noncovalent extrinsic fluorescent dye 4,4'-dianilino-1,1'-binaphthyl-5,5'-disulfonic acid (Bis-ANS) was added to the mobile phase or the sample, and the fluorescence emission at 488nm was recorded on excitation at 385nm. By combining HP-SEC and AF4 with online dye detection, it was possible to simultaneously detect heat-induced aggregation and structural changes of monomeric and aggregated immunoglobulin G (IgG); an increase in Bis-ANS fluorescence was observed in both the aggregate and monomer fractions. These structural changes of individual fractions, which were not detectable by online UV and multiangle laser light scattering (MALLS) or by stand-alone dynamic light scattering (DLS), intrinsic IgG fluorescence, and far-UV circular dichroism (CD), resulted in progressive aggregation on storage. The developed online fluorescent dye detection for HP-SEC or AF4 with Bis-ANS is a powerful method to detect both aggregation and structural changes of both monomeric and aggregated IgG in heat-stressed formulations.


Efficient goal attainment and engagement in a care manager system using unstructured notes.
To improve efficient goal attainment of patients by analyzing the unstructured text in care manager (CM) notes (CMNs). Our task is to determine whether the goal assigned by the CM can be achieved in a timely manner. Our data consists of CM structured and unstructured records from a private firm in Orlando, FL. The CM data is based on phone interactions between the CM and the patient. A portion of the data has been manually annotated to indicate engagement. We present 2 machine learning classifiers: an engagement model and a goal attainment model. We can successfully distinguish automatically between engagement and lack of engagement. Subsequently, incorporating engagement and features from textual information from the unstructured notes significantly improves goal attainment classification. Two key challenges in this task were the time-consuming annotation effort for engagement classification and the limited amount of data for the more difficult goal attainment class (specifically, for people who take a long time to achieve their goals). We successfully explore domain adaptation and transfer learning techniques to improve performance on the under-represented classes. We also explore the value of using features from unstructured notes to improve the model and interpretability. Unstructured CMNs can be used to improve accuracy of our classification models for predicting patient self-management goal attainment. This work can be used to help identify patients who may require special attention from CMs to improve engagement in self-management.


New pseudophase structure for alpha-Pu.
We propose a new pseudophase crystal structure, based on an orthorhombic distortion of the diamond structure, for the ground-state alpha phase of plutonium. Electronic-structure calculations in the generalized-gradient approximation give approximately the same total energy for the two structures. Interestingly, our new pseudophase structure is the same as the gamma-Pu structure except with very different b/a and c/a ratios. We show how the contraction relative to the gamma phase, principally in the z direction, leads to an alpha-like structure in the [0,1,1] plane and reproduces the short range bonds of the alpha phase. This is an important link between two complex structures of Pu and opens new possibilities for exploring the very rich phase diagram of Pu through theoretical calculations.


Nephrogenic diabetes insipidus.
Nephrogenic diabetes insipidus (NDI), which can be inherited or acquired, is characterized by an inability to concentrate urine despite normal or elevated plasma concentrations of the antidiuretic hormone arginine vasopressin (AVP). Polyuria, with hyposthenuria, and polydipsia are the cardinal clinical manifestations of the disease. About 90% of patients with congenital NDI are males with X-linked recessive NDI (OMIM 304800) who have mutations in the arginine-vasopressin receptor 2 (AVPR2) gene that codes for the vasopressin V2 receptor. In about 10% of the families studied, congenital NDI has an autosomal recessive or autosomal dominant mode of inheritance (OMIM 222000 and 125800). In these families, mutations have been identified in the aquaporin-2 gene (AQP2) (OMIM 107777), which codes for the vasopressin-sensitive water channel. Most missense AVPR2 mutations lead to receptors that are trapped intracellularly; a few mutant receptors reach the cell surface but are unable to bind AVP or to properly trigger an intracellular cyclic adenosine monophosphate signal. Similarly, most AQP2 mutant proteins are also misrouted. Prior knowledge of AVPR2 or AQP2 mutations in NDI families and perinatal mutation testing is of direct clinical value because early diagnosis and treatment can avert the physical and mental retardation associated with repeated episodes of dehydration.


Idiopathic hereditary pancreatitis: experience with surgical treatment.
Idiopathic hereditary pancreatitis is a rate form of primary chronic pancreatitis transmitted as an autosomal dominant trait with a variable clinical penetrance. We studied a Hispanic kindred of 23, seven of whom had the disease. In each of seven cases, abdominal pain had started when the patient was a teenager. The disease was confirmed surgically in three patients, biochemically in three, and roentgenographically in one. No causes were determined. The three patients who had surgery each had a ten- to 30-year history of recurrent severe abdominal pain requiring multiple hospital admissions. In each the pancreatic duct had a "chain-of-lakes" appearance on endoscopic retrograde cholangiopancreatographic examination and was drained by longitudinal pancreatojejunostomy. There was no recurrence of symptoms after surgery. Retrograde drainage of the pancreatic duct reliably relieves the symptoms of idiopathic hereditary pancreatitis.


Caribbean-wide decline in carbonate production threatens coral reef growth.
Global-scale deteriorations in coral reef health have caused major shifts in species composition. One projected consequence is a lowering of reef carbonate production rates, potentially impairing reef growth, compromising ecosystem functionality and ultimately leading to net reef erosion. Here, using measures of gross and net carbonate production and erosion from 19 Caribbean reefs, we show that contemporary carbonate production rates are now substantially below historical (mid- to late-Holocene) values. On average, current production rates are reduced by at least 50%, and 37% of surveyed sites were net erosional. Calculated accretion rates (mm year(-1)) for shallow fore-reef habitats are also close to an order of magnitude lower than Holocene averages. A live coral cover threshold of ~10% appears critical to maintaining positive production states. Below this ecological threshold carbonate budgets typically become net negative and threaten reef accretion. Collectively, these data suggest that recent ecological declines are now suppressing Caribbean reef growth potential.


Role of candidate modifier genes on the phenotypic expression of hypertrophy in patients with hypertrophic cardiomyopathy.
The phenotypic expression of left ventricular hypertrophy (LVH) in patients with hypertrophic cardiomyopathy (HCM) is variable. This phenotypic variability is not completely explained by the responsible mutations or other known factors. Recent data denote a role for the modifier genes and environmental factors. We studied the role of 3 potential modifier genes, i.e., angiotensinogen (AGT), angiotensin II receptor 1a (AT1a), and endothelin-1 (END1) on the phenotypic expression of LVH in patients with hypertrophic cardiomyopathy (HCM). The study population was comprised of 108 genetically independent patients with HCM. Left ventricular mass index (LVMI) and LVH score were determined per published protocols. The genotypes of AGT (M235T, T174M, and G-6A), AT1a, and END1 were determined by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) or mutation-specific PCR (MS-PCR). Male patients had higher mean LVMI and LVH score than female patients (146.0 +/- 33.5 vs 129.4 +/- 33.6, p = 0.01 and 6.0 vs 5.0, p = 0.010, respectively). Gender accounted for 4.8% and 5.4% of the variability of LVMI and LVH score, respectively. The END1 genotypes also had a significant influence on LVH scores accounting for 2.9% of their variability (p = 0.042). The median LVH score was greater in patients with the AA and AG genotypes, as compared to patients with the GG genotype (7.0 vs 5.0, p = 0.034). Neither the AGT nor the AT1 genotypes had a significant influence on the expression of LVH. In multivariate regression analysis, END1 and gender accounted for 7.3% of the variability of the LVH score (p = 0.007). Our results show that gender and the END1 gene modify the phenotypic expression of hypertrophy in patients with HCM.


HDAC8 Inhibition Blocks SMC3 Deacetylation and Delays Cell Cycle Progression without Affecting Cohesin-dependent Transcription in MCF7 Cancer Cells.
Cohesin, a multi-subunit protein complex involved in chromosome organization, is frequently mutated or aberrantly expressed in cancer. Multiple functions of cohesin, including cell division and gene expression, highlight its potential as a novel therapeutic target. The SMC3 subunit of cohesin is acetylated (ac) during S phase to establish cohesion between replicated chromosomes. Following anaphase, ac-SMC3 is deacetylated by HDAC8. Reversal of SMC3 acetylation is imperative for recycling cohesin so that it can be reloaded in interphase for both non-mitotic and mitotic functions. We blocked deacetylation of ac-SMC3 using an HDAC8-specific inhibitor PCI-34051 in MCF7 breast cancer cells, and examined the effects on transcription of cohesin-dependent genes that respond to estrogen. HDAC8 inhibition led to accumulation of ac-SMC3 as expected, but surprisingly, had no influence on the transcription of estrogen-responsive genes that are altered by siRNA targeting of RAD21 or SMC3. Knockdown of RAD21 altered estrogen receptor α (ER) recruitment at SOX4 and IL20, and affected transcription of these genes, while HDAC8 inhibition did not. Rather, inhibition of HDAC8 delayed cell cycle progression, suppressed proliferation and induced apoptosis in a concentration-dependent manner. We conclude that HDAC8 inhibition does not change the estrogen-specific transcriptional role of cohesin in MCF7 cells, but instead, compromises cell cycle progression and cell survival. Our results argue that candidate inhibitors of cohesin function may differ in their effects depending on the cellular genotype and should be thoroughly tested for predicted effects on cohesin's mechanistic roles.


Functional Characterization of an Interferon Gamma Receptor-Like Protein on Entamoeba histolytica.
Entamoeba histolytica is an anaerobic parasitic protozoan and the causative agent of amoebiasis. E. histolytica expresses proteins that are structurally homologous to human proteins and uses them as virulence factors. We have previously shown that E. histolytica binds exogenous interferon gamma (IFN-γ) on its surface, and in this study, we explored whether exogenous IFN-γ could modulate parasite virulence. We identified an IFN-γ receptor-like protein on the surface of E. histolytica trophozoites by using anti-IFN-γ receptor 1 (IFN-γR1) antibody and performing immunofluorescence, Western blot, protein sequencing, and in silico analyses. Coupling of human IFN-γ to the IFN-γ receptor-like protein on live E. histolytica trophozoites significantly upregulated the expression of E. histolytica cysteine protease A1 (EhCP-A1), EhCP-A2, EhCP-A4, EhCP-A5, amebapore A (APA), cyclooxygenase 1 (Cox-1), Gal-lectin (Hgl), and peroxiredoxin (Prx) in a time-dependent fashion. IFN-γ signaling via the IFN-γ receptor-like protein enhanced E. histolytica's erythrophagocytosis of human red blood cells, which was abrogated by the STAT1 inhibitor fludarabine. Exogenous IFN-γ enhanced chemotaxis of E. histolytica, its killing of Caco-2 colonic and Hep G2 liver cells, and amebic liver abscess formation in hamsters. These results demonstrate that E. histolytica expresses a surface IFN-γ receptor-like protein that is functional and may play a role in disease pathogenesis and/or immune evasion.


The enhancer of the immunoglobulin heavy chain locus is flanked by presumptive chromosomal loop anchorage elements.
We have located presumptive chromosomal loop anchorage elements within the mouse heavy chain immunoglobulin locus. Analysis of 31 kilobases spanning diversity, joining, enhancer, switch, and the mu and delta constant regions reveals that only a single 1-kilobase segment exhibits specific binding to nuclear matrices. It is of particular significance that the transcriptional enhancer element resides within this matrix association region (MAR). Fine structure mapping indicates that binding is mediated by A+T-rich approximately 350-base pair segments that reside on either side of the enhancer. The MAR sequences residing 5' of the enhancer contain topoisomerase II consensus sequences like the MAR located upstream of the kappa light chain gene enhancer. The heavy chain gene MARs, however, exhibit a lower affinity for matrix association compared to the kappa gene MAR. Significantly, the juxtaposition of enhancer elements with MARs appears to be evolutionarily conserved within the immunoglobulin genes, suggesting that MARs may act as positive and/or negative regulators of enhancer function.


Adsorption of mesitylene via mesoporous adsorbents.
Mesitylene (or 1,3,5-trimethylbenzene) is a volatile organic compound emitted from various industrial processes, e.g., spray coating. Its emissions have become a critical issue because mesitylene is toxic and cannot be removed using traditional adsorbents, e.g., zeolite (H-ZSM-5; the diameter of mesitylene molecules is greater than the pore size of H-ZSM-5). Hence, an adsorbent with a large pore size, MCM-41, is used in this study to investigate its adsorption capacity for mesitylene and compare with that of H-ZSM-5. Experimental results reveal that MCM-41 without Al2O3 exhibits a good adsorption capacity (184 mg/g) for the gas stream containing 100 ppm of mesitylene at a relative humidity of 10%. The adsorption kinetics is well described by the Freundlich isotherm. Furthermore, experimental results reveal that MCM-41 is effective for the adsorption of low concentrations (10 ppm) of mesitylene. In addition, adsorption-desorption tests revealed that the sample MCM-41-AS is stable to sustain the adsorption capacity after 10 adsorption-desorption cycles. After 10 adsorption-desorption cycles, MCM-41-AS retains 92.4% of its initial adsorption capacity (170 vs. 184 mg/g). Finally, MCM-41 and H-ZSM-5 in series are effective for the simultaneous removal of mesitylene and toluene in the gas stream. This study aims to improve the performance of adsorbent for mesitylene, which is typically applied in the spray-coating industry. The zeolite MCM-41-AS is selected as a candidate for the investigation. Experimental results reveal that MCM-41-AS exhibits a good adsorption capacity for mesitylene and that it can be integrated with H-ZSM-5-25 for the simultaneous adsorption of mesitylene and toluene.


Canadian Society of Transplantation Members' Views on Anonymity in Organ Donation and Transplantation.
Anonymity has been central to medical, psychosocial, and societal practices in organ donation and transplantation. The purpose of this investigation was to explore transplant professionals' views on anonymity in the context of organ transplantation. The study consisted of an electronic 18-item survey distributed to the Canadian Society of Transplantation membership, asking about anonymity vs open communication/contact between organ recipients and donor families. Of the 541 members surveyed, 106 replied. Among respondents, 71% felt that organ recipients and donor families should only communicate anonymously, yet 47% felt that identifying information could be included in correspondence between consenting recipients and donor families. When asked whether organ recipients and donor families should be allowed to meet, 53% of respondents agreed, 27% disagreed, and 20% neither agreed nor disagreed. With social media facilitating communication and eliminating the ability to maintain donor/recipient anonymity, 38% of respondents felt that a reexamination of current policies and practices pertaining to anonymity was necessary. In conclusion, there was no dominant position on the issue of anonymity/communication between donor families and transplant recipients. Further research and discussion concerning the views of healthcare professionals, organ recipients, and donor families on the mandate of anonymity is needed and may influence future policy.


Positive and Healthy Living Program Manual Development for Young People Living With HIV at the Comprehensive Care Center at the Kenyatta National Hospital: An Open Pilot Implementation Trial.
Background: Human Immunodeficiency Virus/Acquired Immune Deficiency Syndrome (HIV/AIDS) remains a great challenge among young people in Kenya. Young people living with HIV are faced with a lot of challenges that are often overlooked and may have an impact on their treatment adherence and overall well-being. This calls for interventions that are age-appropriate and which tap into the psychosocial problems they experience. This is a protocol of a proposed study aimed at developing a facilitator-led peer support manual called the "Positive and Healthy Living Program" that will be the basis for running support groups with young people at the Comprehensive Care Center (CCC) at the Kenyatta National Hospital (KNH). Methods: We will carry out our study in two phases. The first phase will focus on the development of the manual and training of peer-facilitators. The second phase will make use of a pilot trial research design using both qualitative and quantitative approaches. It will be carried out among 10-24 year-olds attending CCC at KNH, and will consist of three groups: Tumaini Group (10-14 years), Amani Group (15-19 years), and Hodari Group (20-24 years). The groups will participate in an eight-session support group, whose activities will focus on four domains: social-recreation, psychotherapy, peer-modeling, and psychoeducation. Quantitative data will be collected using laboratory measures of Viral Load and CD4 as well as socio-psychological assessment tools. Qualitative data will be collected through interviews with the young people and peer facilitators. We will conduct a descriptive analysis which will describe the key features of the dataset and bivariate analyses will examine the association between variables. The change will be measured at baseline and post-treatment. The interviews will be coded into themes and we will generate experiential categories from the data around the effectiveness of the program, the peer facilitators' experience of providing support, how the young people respond to the program, and its influence on their overall well-being. Discussion: We expect that the peer facilitators will find this manualized treatment acceptable and the eight-sessions group intervention will be feasible for the three age groups. We hypothesize that there will be improvements detected with regards to reported adherence and viral load, self-esteem, depression, and psychological functioning.


Intravenous injection of MVA virus targets CD8+ lymphocytes to tumors to control tumor growth upon combinatorial treatment with a TLR9 agonist.
Effector T-cell access to tumor tissue is a limiting step for clinical efficacy of antigen-specific T cell-based immunotherapies. Ectopic mouse tumor models, in which a subcutaneously (s.c.) implanted tumor is treated with s.c. or intramuscular therapeutic immunization, may not be optimal for targeting effector T cells to an organ-borne tumor. We used an orthotopic renal carcinoma model to evaluate the impact of injection routes on therapeutic efficacy of a Modified Vaccinia virus Ankara viral vector expressing the human mucin 1 tumor-associated xeno-antigen (MVA-MUC1). We show that intravenous (i.v.) administration of MVA-MUC1 displayed enhanced efficacy when compared with s.c. injection. Therapeutic efficacy of MVA-MUC1 was further enhanced by i.v. injection of a TLR9 agonist. In all cases, infiltration of tumor-bearing kidney by CD8(+) lymphocytes was associated with control of tumor growth. Biodistribution experiments indicate that, following i.v. injection, MVA-encoded antigens are quickly expressed in visceral organs and, in particular, in splenic antigen-presenting cells, compared with those following s.c. injection. This appears to result in a faster generation of MUC1-specific CD8(+) T cells. Lymphocytes infiltrating tumor-bearing kidneys are characterized by an effector memory phenotype and express PD-1 and Tim3 immune checkpoint molecules. Therapeutic efficacy was associated with a modification of the tumor microenvironment toward a Th1-type immune response and recruitment of activated lymphocytes. This study supports the clinical evaluation of MVA-based immunotherapies via the i.v. route.


[Level of evidence for therapeutic drug monitoring of carboplatin].
Carboplatin is an anticancer agent widely used in different types of cancer in adults and paediatrics patients. Carboplatin undergoes an high protein binding percentage in plasma and presents a major hematotoxicity. Some studies determined unbound and bound platin concentrations using flameless atomic absorption spectrophotometry or chromatography. These studies showed a relationship between carboplatin exposition expressed using AUC of ultrafiltrable concentrations with time and myelotoxicity. However, the relationship between carboplatin exposition and its efficacy is not yet established. The population approach using a nonlinear mixed effects model showed a low variability of drug exposition parameters. In some cases, Bayesian adaptative dosing were proposed. However, some particular populations will have to be taken into account such as intensive doses of carboplatin, children, patients with hemodialysis-dependent renal insufficiency or obese patients. Therapeutic carboplatin monitoring is today recommended measuring AUC ultrafiltrable concentrations in time and using described adjusted-modelling.


Occurrence and impact of intracranial pressure elevation during treatment of severe intraventricular hemorrhage.
Elevated intracranial pressure is one of the proposed mechanisms leading to poor outcomes in patients with intraventricular hemorrhage. We sought to characterize the occurrence and significance of intracranial hypertension in severe intraventricular hemorrhage requiring extraventricular drainage. Prospective analysis from two randomized, multicenter, clinical trials. Intensive care units of 23 academic hospitals. One hundred patients with obstructive intraventricular hemorrhage and intracerebral hemorrhage volume <30 mL requiring emergency extraventricular drainage from two randomized multicenter studies comparing intraventricular recombinant tissue plasminogen activator (n=78) to placebo (n=22). Intracranial pressure was recorded every 4 hrs in all patients and before and after a 1-hr extraventricular drainage closure period after injection. Intracranial pressure readings were analyzed at predefined thresholds and compared between treatment groups, before and after injection of study agent, and before and after opening of third and fourth ventricles on computed tomography. Impact on 30-day outcomes was assessed. Initial intracranial pressure ranged from -2 to 60 mm Hg (median; interquartile range, 11;10). Of 2576 intracranial pressure readings, 91.5% (2359) were ≤20 mm Hg, 1.6% were >30, 0.5% were >40, and 0.2% were >50 mm Hg. In a multivariate analysis, threshold events>20 mm Hg and >30 mm Hg were more frequent in placebo vs. recombinant tissue plasminogen activator-treated groups (p=.03 and p=.08, respectively). Intracranial pressure elevation>20 mm Hg occurred during a required 1-hr extraventricular drainage closure interval in 207 of 868 (23.8%) injections of study agent, although early reopening of the extraventricular drainage only occurred in 7.9%. After radiographic opening of the lower ventricular system, intracranial pressure events>20 mm Hg remained significantly associated with initial intraventricular hemorrhage volume (p=.002) and extraventricular drainage placement ipsilateral to the largest intraventricular hemorrhage volume (p=.001), but not with thrombolytic treatment (p=.05) or intracerebral hemorrhage volume (p=.14). Ventriculoperitoneal shunts were required in 13.6% of placebo and 6.4% of recombinant tissue plasminogen activator-treated patients (p=.37). Percentage of intracranial pressure readings per patient>30 mm Hg and initial intracerebral hemorrhage and intraventricular hemorrhage volumes were independent predictors of 30-day mortality after adjustment for other outcome predictors (p=.003, p=.03, and p<.001, respectively). Independent predictors of poor modified Rankin Scale score at 30 days were percent of intracranial pressure events>30 mm Hg per patient (p=.01; but not >20 mm Hg), both intracerebral hemorrhage and intraventricular hemorrhage volume, and pulse pressure. Intracranial pressure is not frequently elevated during monitoring and drainage with an extraventricular drainage in patients with severe intraventricular hemorrhage, although intracranial pressure >30 mm Hg predicts higher short-term mortality. Thrombolytic therapy may reduce the frequency of high intracranial pressure events. Intracranial pressure elevation appears to be significantly correlated with extraventricular drainage placement in the ventricle with greatest clot volume.


Health-Related Quality of Life and Use of Hospital Services by Patients with Heart Failure and Their Family Caregivers: A Multicenter Case-Control Study.
Heart failure (HF) causes high rates of hospital admissions. It is known that disease progression impacts the health-related quality of life (HRQoL) of both patients and caregivers, yet to date, this finding is based on cross-sectional studies with limited samples. The study aim is to analyze the relationship between HF patients' use of hospital services (a proxy for disease progression) and the HRQoL of their family caregivers. This work is a multicenter nested case-control study on a population of patients admitted to hospitals in southern Spain due to heart failure. The sample comprised 530 patient-caregiver dyads. Hospital admission data were retrospectively collected for the 5 years prior to inclusion in the study. Bivariate analyses and multivariate logistic regression were used to determine associations between patient deterioration and caregivers' quality of life. Patients' use of hospital services was associated with worsened quality of life for family caregivers, with an overall OR of 1.48 (95% CI: 1.23-1.79). A positive correlation was found between patients' perceptions of their physical health and the perceived mental health of caregivers (r = 0.127, p = 0.004) and between the perceived mental health of both (r = 0.291; p <0.0001). Greater use of hospital services by patients with HF is an independent predictor of deterioration of family caregivers' HRQoL. The physical and mental components of patients' and their family caregivers' HRQoL interact and influence each other. Additional factors, such as the nature and intensity of care provided, also determine the worsening of a family caregiver's HRQoL. These results can be used to identify family caregivers of people with heart failure at risk of suffering a deterioration in their health-related quality of life. Increased use of hospital services is an independent predictor of the deterioration of the family caregivers' health-related quality of life. Since clinical nurses are the main provider who gives support and education to family caregivers, they should be alert to this situation and individualize interventions to prevent this deterioration.


Design and rationale of the atrial fibrillation occurring transiently with stress (AFOTS) incidence study.
Atrial fibrillation (AF) is often detected for the first time in patients hospitalized for medical illness or non-cardiovascular surgery. AF occurring transiently with stress (AFOTS) describes this manifestation of AF, which may either be the result of a non-cardiac stressor, or existing paroxysmal AF that was not previously detected. Current estimates of AFOTS incidence are imprecise: ranging from 1 to 44%, owing to the marked heterogeneity in patient populations, identification and methods used to detect AFOTS. The prospective, two-centre epidemiological AFOTS Incidence study will enroll 250 consecutive participants without a history of AF but with at increased risk of AF (Age ≥ 65 or >50 with one risk factor for AF) admitted to intensive care units (ICUs) for medical illness or non-cardiac surgery. Upon admission, participants will wear an ECG patch monitor that will remain in place for 14 days, or until discharge from hospital. Patients' consent to participation is deferred for up to 72 h after admission. The primary endpoint is the incidence of AF lasting ≥30 s. The study is powered to detect an AF incidence of 17% ± 5%. We conducted a vanguard feasibility study, and 55 participants have completed participation. The median duration of monitoring was seven days. AF was detected by the clinical team in 8 participants (14%; 95% Confidence Interval 7-26%). The AFOTS Incidence study will employ a systematic and highly sensitive protocol for detecting AFOTS in medical illness and non-cardiac surgery ICU patients. This study is feasible and will provide a reliable estimate of the true incidence of AFOTS in this population.


Closing the gap in postfracture care at the population level: a randomized controlled trial.
Postfracture care is suboptimal, and strategies to address this major gap in care are necessary. We investigated whether notifications sent by mail to physicians and patients would lead to improved postfracture care. We conducted a randomized controlled trial (ClinicalTrials.gov identifier NCT00594789) in the province of Manitoba, Canada, from June 2008 to May 2010. Using medical claims data, we identified 4264 men and women age 50 years or older who recently reported major fractures, and who had not undergone recent bone mineral density testing or treatment for osteoporosis. Participants were randomized to three groups: group 1 received usual care (n = 1480), patients in group 2 had mailed notification of the fracture sent to their primary care physicians (n = 1363), and group 3 had notifications sent to both physicians and patients (n = 1421). Bone mineral density testing and the start of pharmacologic treatment for osteoporosis within the following 12 months were documented. Among participants in group 1 (usual care), 15.8% of women and 7.6% of men underwent testing for bone mineral density or started pharmacologic treatment for osteoporosis. Outcome measures improved among participants in group 2 (30.3% of women and 19.0% of men, both p < 0.001) and group 3 (34.0% of women and 19.8% of men, both p < 0.001). No additional benefit was seen with patient notification in addition to physician notification. Combining groups 2 and 3, the absolute increase for the combined end point of bone mineral density testing or pharmacologic treatment was 14.9% (16.4% among women, 11.8% among men). The number needed to notify to change patient care was 7 (6 for women, 6 for men). The adjusted odds ratio (OR) to change patient care in group 2 was 2.45 (95% confidence interval [CI] 2.01-2.98); for group 3 the OR was 2.82 (95% CI 2.33-3.43). This notification system provides a relatively simple way to enhance post-fracture care.


Active detection of chronic obstructive pulmonary disease and asthma in the general population. Results and economic consequences of the DIMCA program.
The aim of this prospective study was to detect subjects in the general population with objective signs of chronic obstructive pulmonary disease (COPD) or asthma at an early stage. This was done by means of a two-stage protocol involving screening and a subsequent 2-yr monitoring of all subjects with positive results of screening. The study was done in 10 general practices located in the eastern part of the Netherlands. A random sample was taken from the general population aged 25 to 70 yr. All known COPD and asthma patients were excluded. A total of 1,749 subjects met the inclusion criteria: 1,155 subjects (66%) agreed to participate in the screening stage of the study. A total of 604 subjects (52.3%) showed symptoms or objective signs of COPD or asthma during the screening and were considered "positive." Of those with positive screening results, 384 subjects (64%) agreed to participate in the second, 2-yr monitoring stage of the study. The costs involved in detection were calculated for three different scenarios, as follows: (1) The detection of subjects with persistently decreased lung function or an increased level of bronchial hyperresponsiveness (BHR) during 6 mo of monitoring; (2) Scenario 1 plus the detection of subjects with a rapid decline in lung function with signs of BHR during 12 mo of monitoring; (3) Scenario 2 plus the detection of subjects with a moderate increase in the decline in lung function or signs of BHR during 24 mo of monitoring. The costs of lung function assessments, organization, transportation, and patient time were included. The costs were converted to United States dollars on the basis of purchasing power (1 United States dollar = 2.08 Netherlands guilders). During the second stage, 252 subjects were detected with objective signs of COPD or asthma at an early stage. Smoking status as a screening criterion was neither sensitive nor specific. Because there was no evidence of biased recruitment or selection during the program, the proportions of subjects found to have objective signs of COPD or asthma at an early stage could be extrapolated to the general population. Of the general population, 7.7% showed persistently reduced lung function or increased BHR. Another 12.5 % of the general population showed a rapid decline in lung function (> 80 ml/yr) in combination with signs of BHR, and a further 19.4% of the general population showed mild objective signs of COPD or asthma. The average costs per detected case varied from US$953 (Scenario 1) to US$469 (Scenario 3). In conclusion, detection of COPD or asthma at an early stage by means of a two-stage protocol was feasible at relatively little expense in comparison with other mass screening programs. Persistently decreased lung function or a rapid decline in lung function (Scenario 2) was observed in approximately 20% of the general adult population.


Diagnosis and management of monocular diplopia.
Monocular diplopia is a symptom which may range from a visual curiosity to being visually dehabilitating. Its proper diagnosis and management presents a challenge to the clinician. A comprehensive review of the literature illustrates the multiple etiologies which may produce this complaint. Differential diagnosis is an important factor in the ultimate successful management of the problem, since the latter is to a degree etiology specific. Method of differential diagnosis and management options are discussed.


Epidemiological aspects of ovarian malignancies in North Sardinia in the period 1992-2001.
Malignant ovarian tumors have been continuously increasing in Western countries and represent the leading cause of death for gynecological cancer. In fact, the mortality for malignant ovarian tumors remains very high with a low percentage of 5-year survival in the advanced stage of disease. The aim of this study was to evaluate the incidence trend and epidemiological characteristics of malignant ovarian tumors in the Province of Sassari, Sardinia (Italy) in the period 1992-2001 and to report the variations in comparison to the 1974-1985 period. The analysis of our data regarding the period 1992-2001, if compared with those of the period 1974-85, showed an increase of malignant ovarian tumors which triplicated achieving an incidence of 11.99/100,000 vs 4.27/100,000. The analysis of our epidemiologic data showed an increase of the age of first diagnosis (mean 60.9 years for epithelial ovarian tumors), the occurrence of the cancer in women at low socio-economic levels and a family history of cancer among the patients with malignant ovarian tumors. These data suggest that both local environmental factors combined with genetic characteristics play a role in the pathogenesis of ovarian tumors. The genetic characteristics could be of particular interest because Sardinia has been through the centuries a geographical area with little population migration. The marked increase in the incidence of ovarian tumors in the last several years points out the need to organize systematic screening by ultrasonography in our population.


MicroRNA-203 inhibits malignant melanoma cell migration by targeting versican.
MicroRNA (miR)-203 has been demonstrated to function as a suppressor in tumorigenesis. Recently, miR-203 was reported to play a role in malignant melanoma (MM); however, the detailed function of miR-203 in MM remains unclear. In the present study, the expression of miR-203 was shown to be significantly downregulated in MM tissues when compared with normal adjacent tissues. Based on a bioinformatic prediction, versican was further identified as a novel target of miR-203, and the expression of versican was markedly increased in MM tissues. Inhibition of miR-203 increased the protein expression of versican, while upregulation of miR-203 inhibited the protein expression of versican in MM A375 cells. In addition, the upregulation of versican significantly promoted A375 cell migration; however, upregulation of miR-203 suppressed A375 cell migration. The present study further investigated whether miR-203 was involved in versican-mediated A375 cell migration, and the results indicated that upregulation of miR-203 significantly inhibited A375 cell migration, which was impaired by overexpression of versican. These observations indicated that versican functions as a downstream effector in miR-203-mediated MM cell migration. Therefore, the results demonstrated that miR-203 exhibited an inhibitory effect on MM cell migration via directly targeting versican, thus, may become an effective inhibitor for MM metastasis.


Variant estrogen receptor alpha mRNAs in human breast cancer specimens.
A panel of human breast cancer specimens was examined for single base change mutations by DNA sequencing and for larger deletions using a PCR-based assay. In the cancer specimens examined, no sequencing variants were detected other than a previously characterized polymorphism. Although most of the specimens contained estrogen receptor (ER) variants at a low level, 2 of 118 specimens exhibited variants which, after amplification, constituted most of the amplified ER cDNA. One specimen contained a single variant form, and there was little evidence of the wild-type ER mRNA by PCR, Northern blotting or immunocytochemistry. The second specimen, despite the presence of a normal-sized mRNA by Northern blotting and normal immunocytochemical staining for ER, contained at least 5 different variant forms as well as the wild-type ER. All but 1 of the variant forms were processing variants, and 3 of these processing variants have not been described before. One variant, although lacking exons 2-4, has break points in exons 1 and 5 that do not correspond to intron-exon boundaries. This variant might reflect more widespread damage to the genome in this breast cancer specimen. The low level of occurrence of variants suggests that ER variant forms, at least in the coding region, do not contribute generally to the progression of breast cancer.


Resident perceptions of radiosurgical training and the effect of a focused resident training seminar.
Neurosurgical training is critical in providing residents with the skill set, knowledge, and confidence to perform challenging neurosurgical procedures. Radiosurgery, which neurosurgeons helped define and refine, differs from more traditional, open neurosurgical approaches. This study evaluates the opinions of residents on current radiosurgical training and the effect of a focused educational course on those residents. The American Association of Neurological Surgeons sponsored a 3-day course focused on intracranial and spinal radiosurgery. Senior-level residents were nominated by US program directors to participate in the course. Twenty-eight residents from distinct training programs were surveyed before and after the course to discern current training practices in radiosurgery and the effect of the focused educational program. The median training level of the participants was postgraduate Year 5 (mean 5.3 years, range 3-7 years). Two-thirds of residents reported that their training institutions had no formal radiosurgery rotation. Twenty-five percent planned to obtain postresidency fellowship training that would include radiosurgery. Before the course, 79% of the residents expected to include radiosurgery in their practice. However, prior to the course, those describing themselves as "very uncomfortable" with performing intracranial or spinal radiosurgery were 33.3 and 45.8%, respectively. After the course, mean self-assessment scores for understanding the indications and performing intracranial radiosurgery increased by 43 and 89%, respectively. The mean scores for understanding the indications and comfort with performing spinal radiosurgery increased by 79 and 200%, respectively. Following the course, there was a 12.3% increase in the number of residents planning to perform radiosurgery following residency. Current neurosurgical residents appear uneasy about their grasp of radiosurgical indications and their ability to perform the procedure. Focused training courses sponsored by professional societies may improve resident education and training in this area of neurosurgery, which has a skill set and basis of knowledge different from traditional open neurosurgical procedures. Further evaluation of the radiosurgical training process for residents must be performed so as to ensure competency and sufficient workforce to meet expanding demands for neurosurgeons performing radiosurgery in a multidisciplinary climate.


Somatic alterations in juvenile polyps from BMPR1A and SMAD4 mutation carriers.
Juvenile polyposis syndrome (JPS) is a rare autosomal dominant disorder predisposing to gastrointestinal hamartomatous polyps and cancer with a pathogenic SMAD4 or BMPR1A germline mutation (1st-hit) being identified in about 40-50% of patients. Little is known, however, about the occurrence and nature of somatic alterations (2nd-hit) in SMAD4-/BMPR1A-related juvenile polyps. In this study, we screened 25 polyps from three patients carrying either a pathogenic SMAD4 (c.1244-1247delACAG) or BMPR1A (c.583C>T; p.Gln195*) germline mutation for somatic alterations. The SMAD4-related polyps were also analyzed for SMAD4 protein expression by immunohistochemistry. Despite comprehensive screening for loss of heterozygosity (LOH), mutations in the coding sequence, chromosomal rearrangements, and promoter methylation, no somatic alterations could be identified in 14 SMAD4-related polyps. SMAD4 protein expression, however, was lost in 8 (57%) of 14 juvenile polyps with 6 showing concomitant loss in both, the epithelial and stromal, compartments. In the BMPR1A-related polyps, five out of nine (56%) displayed LOH. Further analysis of selected polyps revealed that LOH was gene copy number neutral and had occurred in the epithelial compartment. The heterogeneity of genetic mutations and protein expression levels indicates that different modes of gene inactivation can be operational in SMAD4- and BMPR1A-related polyp formation. Our observation, that about half of BMPR1A-related polyps displayed LOH, predominantly in the epithelial compartment, is compatible with BMPR1A acting as a tumour suppressor gene. Still, it remains to be determined whether juvenile polyp development generally requires loss of BMPR1A expression or, as observed in some SMAD4-related polyps, can occur despite normal protein expression.


para-Quinodimethane-bridged perylene dimers and pericondensed quaterrylenes: the effect of the fusion mode on the ground states and physical properties.
Polycyclic hydrocarbon compounds with a singlet biradical ground state show unique physical properties and promising material applications; therefore, it is important to understand the fundamental structure/biradical character/physical properties relationships. In this study, para-quinodimethane (p-QDM)-bridged quinoidal perylene dimers 4 and 5 with different fusion modes and their corresponding aromatic counterparts, the pericondensed quaterrylenes 6 and 7, were synthesized. Their ground-state electronic structures and physical properties were studied by using various experiments assisted with DFT calculations. The proaromatic p-QDM-bridged perylene monoimide dimer 4 has a singlet biradical ground state with a small singlet/triplet energy gap (-2.97 kcal mol(-1)), whereas the antiaromatic s-indacene-bridged N-annulated perylene dimer 5 exists as a closed-shell quinoid with an obvious intramolecular charge-transfer character. Both of these dimers showed shorter singlet excited-state lifetimes, larger two-photon-absorption cross sections, and smaller energy gaps than the corresponding aromatic quaterrylene derivatives 6 and 7, respectively. Our studies revealed how the fusion mode and aromaticity affect the ground state and, consequently, the photophysical properties and electronic properties of a series of extended polycyclic hydrocarbon compounds.


Microtopographical assembly of cardiomyocytes.
One of the central challenges in cardiac tissue engineering is the control of the assembly and organization of functional cardiac tissue. Maintenance of a three-dimensional tissue architecture is key to myocardial function in vivo, and a variety of studies hint that provision of topological cues within scaffolds can facilitate the engineering of functional myocardial tissue by promoting this architecture. To explore this possibility in an isolated and well-defined fashion, we have designed scaffolds of polydimethylsiloxane (PDMS) with microtopographic pillars ("micropegs") to provide cells with defined structures with which to interact in three dimensions. We show that these surfaces permit HL-1 cardiomyocytes to grow, form myofibrillar structures and cell-cell adhesions, and beat spontaneously. Additionally, the cells and their nuclei interact with the full length of the micropegs, indicating that the micropegs promote a three-dimensional cytoarchitecture in the context of a two-dimensional scaffold. We also show that the number of cells interacting with a micropeg can be controlled by manipulating incubation time, micropeg spatial arrangement, or micropeg diameter. Western blots reveal that the expression of the junctional markers N-cadherin and connexin 43 is upregulated in the presence of specific arrangements of micropegs, suggesting that micropegs can enhance cardiomyocyte function. Together, these data show that microtopography can be used to provide three-dimensional adhesion and control the assembly of functional cardiac tissue on a two-dimensional surface.


Fiber composite alloys: preparation by controlled dissociation of metallic solid solutions.
Fiber-reinforced alloys have been prepared by the controlled dissociation of gold-nickel and aluminum-zinc solid solutions. To cause the dissociation, the alloys were taken from a one-phase field to a two-phase field by lowering the temperature. The new two-phase structure was in each case formed by diffusion controlled processes, and under certain conditions one of the new equilibrium phases appeared in the form of fibers, a few tenths of a micron in diameter, embedded in a matrix of the other. The precipitated phase first appeared in the form of spherical particles, but these aligned and then coalesced to form the fibers.


Recent developments in drug abuse and doping control in sport.
Since September 1988 when Ben Johnson was tested positive for a banned substance at the Seoul Olympic Games, many national and international initiatives have been taken to improve the control of doping in sport. This paper summarizes the revised provisions of the International Olympic Committee list of doping classes and methods. Recent events are also reviewed, including the investigation into the use of performance-enhancing drugs by Australian sportsmen and women (the Black Enquiry), by the Canadians (the Dubin Enquiry) and by the British (the Coni Enquiry and the Jacobs Enquiry). Recent political initiatives are considered, in particular the major international agreements such as the Soviet-American Joint Commission, the International Olympic Antidoping Charter and the European Anti-Doping Convention. Sportsmen and women who, in the past, have been tested positive for banned substances have been the focus of much attention from the sporting community, the general public and the media, but others including coaches, physicians, scientists and others, must share the blame for the spread of drug abuse in sport. In the UK the new policies and programmes of the Sports Council (and the Sports Councils in Wales, Scotland and Northern Ireland) should provide a better programme for doping control than that previously used, and the new international agreements should provide a better framework for international cooperation on education, detection and research.


Polyaromatic Bis(indolyl)methane Derivatives with Antiproliferative and Antiparasitic Activity.
Bis(indolyl)methanes (BIMs) are a class of compounds that have been recognized as an important core in the design of drugs with important pharmacological properties, such as promising anticancer and antiparasitic activities. Here, we explored the biological activity of the BIM core functionalized with different (hetero)aromatic moieties. We synthesized substituted BIM derivatives with triphenylamine, N,N-dimethyl-1-naphthylamine and 8-hydroxylquinolyl groups, studied their photophysical properties and evaluated their in vitro antiproliferative and antiparasitic activities. The triphenylamine BIM derivative 2a displayed an IC50 of 3.21, 3.30 and 3.93 μM against Trypanosoma brucei, Leishmania major and HT-29 cancer cell line, respectively. The selectivity index demonstrated that compound 2a was up to eight-fold more active against the parasites and HT-29 than against the healthy cell line MRC-5. Fluorescence microscopy studies with MRC-5 cells and T. brucei parasites incubated with derivative 2a indicate that the compound seems to accumulate in the cell's mitochondria and in the parasite's nucleus. In conclusion, the BIM scaffold functionalized with the triphenylamine moiety proved to be the most promising antiparasitic and anticancer agent of this series.


Dynamics of a specialized and complex health care system: Exploring general practitioners' management of multimorbidity.
To explore general practitioners' (GPs') experiences of cooperation with hospital-based physicians regarding multimorbid patients and to identify challenges as well as strategies in managing such challenges.Study setting: Three medical practices in a provincial town in Denmark. A qualitative methodological design was used with explorative data collection among GPs.Data collection/Extraction methods: Participant observation, qualitative interviews and a focus group interview were conducted. Interpretive description was used as the analytical framework.Principal findings: The GPs appreciated cooperating with physicians in optimizing treatment of multimorbid patients. However, three main challenges were experienced: insufficient communication and coordination; unclear divisions of roles and responsibilities; and differences in the way of approaching patients. The GPs navigated these challenges and complexities by taking advantage of their personal relationships and by developing creative and patient-centred ad hoc solutions to difficulties in cross-sectorial cooperation. A hospital initiative to support care for multimorbid patients has not been adopted by the GPs as a preferred strategy. The structures of the health care system severely challenged cooperation regarding multimorbid patients; nevertheless, these GPs were aware of the advantages of cooperation, and their mainstay strategy in this involved personalized solutions and flexibility.


How does methylation suppress the electron-induced decomposition of 1-methyl-nitroimidazoles?
The efficient decomposition of nitroimidazoles (NIs) by low energy electrons is believed to underlie their radiosensitizing properties. Recent dissociative electron attachment (DEA) measurements showed that methylation at the N1 site unexpectedly suppresses the electron-induced reactions in 4(5)-NI. We report theoretical results that provide a clear interpretation of that astounding finding. Around 1.5 eV, DEA reactions into several fragments are initiated by a π* resonance, not considered in previous studies. The autoionization lifetime of this anion state, which limits the predissociation dynamics, is considerably shorter in the methylated species, thereby suppressing the DEA signals. On the other hand, the lifetime of the π* resonance located around 3 eV is less affected by methylation, which explains why DEA is still observed at these energies. Our results demonstrate how even a simple methylation can significantly modify the probabilities for DEA reactions, which may be significant for NI-based cancer therapy.


Spatial distribution and characteristics of plastic pollution in the salt marshes of Bahía Blanca Estuary, Argentina.
This study delves into the magnitude and attributes of plastic pollution in the salt marshes of the Bahía Blanca Estuary, Argentina, with a specific focus on its spatial distribution. The investigation included the evaluation of microplastics (1-5 mm), mesoplastics (5-25 mm) and macroplastics (25-100 mm), discovering elevated levels along the high salt marsh strandline compared to low salt marsh and mudflat areas. Notably, the abundance of plastic reached staggering levels, reaching up to 20,060 items/m2 in the vicinity of an illegal dumpsite. Microplastics, particularly in the 2-4 mm range, were dominant, and the main plastic components were high-density polyethylene (HDPE), low-density polyethylene (LDPE), polypropylene (PP), and polystyrene (PS). Plastic films emerged as the predominant plastic type, while the presence of pellets hinted at potential sources such as illegal dumping and port-related activities. This contamination could be largely attributed to inappropriate waste management practices and urban runoff, which pose a substantial ecological threat to these ecosystems. Urgent remedial action is essential to protect these marshes, underscoring the critical need for comprehensive wetland management and educational initiatives to ensure their long-term sustainability.


Efficacy of flumethrin 1% pour-on against ticks on cattle under field conditions in Ethiopia.
The efficacy of a flumethrin 1% pour-on (Bayticol, Bayer AH) was evaluated against natural infestations of ticks on cattle on a dairy farm in Ethiopia during 1997/98. The cattle, (n = 92), which were Friesian/ Zebu crosses, were heavily infested with Boophilus decoloratus. Dry cows (n = 8) were randomly selected and allocated either into a treatment or a control group. Flumethrin 1 % pour-on was applied to the treatment group according to the manufacturer's recommendation, i.e. along the dorsal mid-line from the head to the base of the tail. Ticks were counted and identified in situ on treated and control animals. When the total tick counts of the treated group were similar to those on the control, the trial was discontinued. Excellent results were achieved with flumethrin against heavy tick infestations. There was a rapid kill after 24 h post-treatment, and from day 4 onwards 100% control was achieved and maintained for a further 29 days. The results obtained indicate that there was a significant difference between the mean tick counts of the control group compared with those of the treatment group (P < 0,05). The evidence presented here suggests that the flumethrin 1 % pour-on may be applied at 45 day intervals and should provide effective protection against the economically important ticks if it is used at the recommended dosage rate and applied correctly.


Balanced networks under spike-time dependent plasticity.
The dynamics of local cortical networks are irregular, but correlated. Dynamic excitatory-inhibitory balance is a plausible mechanism that generates such irregular activity, but it remains unclear how balance is achieved and maintained in plastic neural networks. In particular, it is not fully understood how plasticity induced changes in the network affect balance, and in turn, how correlated, balanced activity impacts learning. How do the dynamics of balanced networks change under different plasticity rules? How does correlated spiking activity in recurrent networks change the evolution of weights, their eventual magnitude, and structure across the network? To address these questions, we develop a theory of spike-timing dependent plasticity in balanced networks. We show that balance can be attained and maintained under plasticity-induced weight changes. We find that correlations in the input mildly affect the evolution of synaptic weights. Under certain plasticity rules, we find an emergence of correlations between firing rates and synaptic weights. Under these rules, synaptic weights converge to a stable manifold in weight space with their final configuration dependent on the initial state of the network. Lastly, we show that our framework can also describe the dynamics of plastic balanced networks when subsets of neurons receive targeted optogenetic input.


Gene-environment interaction models to unmask susceptibility mechanisms in Parkinson's disease.
Lipoxygenase (LOX) activity has been implicated in neurodegenerative disorders such as Alzheimer's disease, but its effects in Parkinson's disease (PD) pathogenesis are less understood. Gene-environment interaction models have utility in unmasking the impact of specific cellular pathways in toxicity that may not be observed using a solely genetic or toxicant disease model alone. To evaluate if distinct LOX isozymes selectively contribute to PD-related neurodegeneration, transgenic (i.e. 5-LOX and 12/15-LOX deficient) mice can be challenged with a toxin that mimics cell injury and death in the disorder. Here we describe the use of a neurotoxin, 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP), which produces a nigrostriatal lesion to elucidate the distinct contributions of LOX isozymes to neurodegeneration related to PD. The use of MPTP in mouse, and nonhuman primate, is well-established to recapitulate the nigrostriatal damage in PD. The extent of MPTP-induced lesioning is measured by HPLC analysis of dopamine and its metabolites and semi-quantitative Western blot analysis of striatum for tyrosine hydroxylase (TH), the rate-limiting enzyme for the synthesis of dopamine. To assess inflammatory markers, which may demonstrate LOX isozyme-selective sensitivity, glial fibrillary acidic protein (GFAP) and Iba-1 immunohistochemistry are performed on brain sections containing substantia nigra, and GFAP Western blot analysis is performed on striatal homogenates. This experimental approach can provide novel insights into gene-environment interactions underlying nigrostriatal degeneration and PD.


Formate metabolism in health and disease.
Formate is a one-carbon molecule at the crossroad between cellular and whole body metabolism, between host and microbiome metabolism, and between nutrition and toxicology. This centrality confers formate with a key role in human physiology and disease that is currently unappreciated. Here we review the scientific literature on formate metabolism, highlighting cellular pathways, whole body metabolism, and interactions with the diet and the gut microbiome. We will discuss the relevance of formate metabolism in the context of embryonic development, cancer, obesity, immunometabolism, and neurodegeneration. We will conclude with an outlook of some open questions bringing formate metabolism into the spotlight.


Calf-level factors associated with bovine neonatal pancytopenia--a multi-country case-control study.
Bovine neonatal pancytopenia (BNP), a high fatality condition causing haemorrhages in calves aged less than 4 weeks, was first reported in 2007 in Germany and subsequently observed at low incidence in other European countries and New Zealand. A multi-country matched case-control study was conducted in 2011 to identify calf-level risk factors for BNP. 405 BNP cases were recruited from 330 farms in Belgium, France, Germany and the Netherlands by laboratory confirmation of farmer-reported cases. Up to four calves of similar age from the same farm were selected as controls (1154 calves). Risk factor data were collected by questionnaire. Multivariable modelling using conditional logistic regression indicated that PregSure®BVD (PregSure, Pfizer Animal Health) vaccination of the dam was strongly associated with BNP cases (adjusted matched Odds Ratio - amOR 17.8 first lactation dams; 95% confidence interval - ci 2.4, 134.4; p = 0.005), and second or more lactation PregSure-vaccinated dams were more likely to have a case than first lactation vaccinated dams (amOR 2.2 second lactation; ci 1.1, 4.3; p = 0.024; amOR 5.3 third or more lactation; ci 2.9, 9.8; p = <0.001). Feeding colostrum from other cows was strongly associated with BNP if the dam was not PregSure-vaccinated (amOR 30.5; ci 2.1, 440.5; p = 0.012), but the effect was less if the dam was PregSure-vaccinated (amOR 2.1; ci 1.1, 4.0; p = 0.024). Feeding exclusively dam's milk was a higher risk than other types of milk (amOR 3.4; ci 1.6, 7.5; p = 0.002). The population attributable fractions were 0.84 (ci 0.68, 0.92) for PregSure vaccination, 0.13 (ci 0.06, 0.19) for feeding other cows' colostrum, and 0.15 (ci 0.08, 0.22) for feeding dam's milk. No other calf-level factors were identified, suggesting that there are other important factors that are outside the scope of this study, such as genetics, which explain why BNP develops in some PregSure-colostrum-exposed calves but not in others.


Strong Polyamide-6 Nanocomposites with Cellulose Nanofibers Mediated by Green Solvent Mixtures.
Cellulose nanofiber (CNF) as a bio-based reinforcement has attracted tremendous interests in engineering polymer composites. This study developed a sustainable approach to reinforce polyamide-6 or nylon-6 (PA6) with CNFs through solvent casting in formic acid/water mixtures. The methodology provides an energy-efficient pathway towards well-dispersed high-CNF content PA6 biocomposites. Nanocomposite formulations up to 50 wt.% of CNFs were prepared, and excellent improvements in the tensile properties were observed, with an increase in the elastic modulus from 1.5 to 4.2 GPa, and in the tensile strength from 46.3 to 124 MPa. The experimental tensile values were compared with the analytical values obtained by micromechanical models. Fractured surfaces were observed using scanning electron microscopy to examine the interface morphology. FTIR revealed strong hydrogen bonding at the interface, and the thermal parameters were determined using TGA and DSC, where the nanocomposites' crystallinity tended to reduce with the increase in the CNF content. In addition, nanocomposites showed good thermomechanical stability for all formulations. Overall, this work provides a facile fabrication pathway for high-CNF content nanocomposites of PA6 for high-performance and advanced material applications.


Hip sonography: 20 years experience and results.
Hip sonography in the "Graf technique" is an important tool to detect early DDH cases and dysplastic hips in the German speaking countries, although hip sonography is still controversially discussed in the English speaking countries. The reason may be different techniques with different classification and outcome. Exact anatomical identification, measurement technique, typing, difference between instability and elasticity with a reproducible "standard plane" is absolutely necessary to make hip ultrasound (US) comparable and reliable. US screening had reduced open reductions in Germany to 1:4000, in Austria to 0.23 per thousand. Because of the early screening and earlier less aggressive treatment, the overall costs could be reduced by 1/3 in comparison to the pre-ultrasound area. US screening seems to prevent many, but not all operations and can be recommended as an important tool for prevention.


Xanthohumol induces paraptosis of leukemia cells through p38 mitogen activated protein kinase signaling pathway.
Xanthohumol as a natural polyphenol demonstrates an anticancer activity, but its underlying mechanism remains unclear. In this study, we showed that xanthohumol (XN) induces paraptosis of leukemia cells. The paraptosis is one cell death which is characterized by dilation of the endoplasmic reticulum and/or mitochondria. The results demonstrated that XN treatment significantly inhibited cell proliferation and triggered extensive cytoplasmic vacuolation of HL-60 leukemia cells, but it did not cause the cleavage of caspase-3 protein or apoptosis. In contrast, XN treatment resulted in LC3-II accumulation through blocking of autophagosome maturation. Interestingly, the induction of cytoplasmic vacuolization by XN is not associated with autophagy modulated by XN, therefore, XN-induced cell death of HL-60 leukemia cells is not the classical apoptotic cell death. Intriguingly, XN treatment triggered the dilatation of endoplasma reticulum (ER) and induced ER stress by upregulating C/EBP homologous protein and unfolded protein response regulator Grp78/Bip. Furthermore, XN treatment triggered p38 mitogen activated protein kinase and its specific inhibitor inhibited the paraptosis of HL-60 leukemia cells by XN. In conclusion, we for the first time demonstrated that XN treatment can induce paraptosis of leukemia cells through activation of p38 MAPK signaling.


The interaction among donor characteristics, severity of liver disease, and the cost of liver transplantation.
Accurate assessment of the impact of donor quality on liver transplant (LT) costs has been limited by the lack of a large, multicenter study of detailed clinical and economic data. A novel, retrospective database linking information from the University HealthSystem Consortium and the Organ Procurement and Transplantation Network registry was analyzed using multivariate regression to determine the relationship between donor quality (assessed through the Donor Risk Index [DRI]), recipient illness severity, and total inpatient costs (transplant and all readmissions) for 1 year following LT. Cost data were available for 9059 LT recipients. Increasing MELD score, higher DRI, simultaneous liver-kidney transplant, female sex, and prior liver transplant were associated with increasing cost of LT (P < 0.05). MELD and DRI interact to synergistically increase the cost of LT (P < 0.05). Donors in the highest DRI quartile added close to $12,000 to the cost of transplantation and nearly $22,000 to posttransplant costs in comparison to the lowest risk donors. Among the individual components of the DRI, donation after cardiac death (increased costs by $20,769 versus brain dead donors) had the greatest impact on transplant costs. Overall, 1-year costs were increased in older donors, minority donors, nationally shared organs, and those with cold ischemic times of 7-13 hours (P < 0.05 for all). In conclusion, donor quality, as measured by the DRI, is an independent predictor of LT costs in the perioperative and postoperative periods. Centers in highly competitive regions that perform transplantation on higher MELD patients with high DRI livers may be particularly affected by the synergistic impact of these factors.


[Concordances between autism spectrum disorders and attention deficit hyperactivity disorder].
The current literature acknowledges an overlap of genetic, clinical and neuropsychological aspects between autism spectrum disorder (ASD) and attention deficit hyperactivity disorder (ADHD), suggesting that there may be a common pattern that covers features ranging from the common genetic and structural aetiology to shared patterns of symptoms. To review the current advances in these common aspects. Several studies have pointed out preschool attentional difficulties as the basis of both disorders. From the genetic perspective, it is estimated that 50-72% of the genetic factors overlap between the two disorders. They also share a decrease in the volume of the corpus callosum and left frontal grey matter, as well as functional alterations such as dorsolateral prefrontal, striato-thalamic and superior parietal hypoactivation. Results are also found regarding executive functioning, with differential profiles for the two conditions, and also concerning the relationship between the repetitive and impulsive behaviours in the early stages of ASD and ensuing problems of hyperactivity. This new conception of the ASD-ADHD continuum, with a common neurodevelopmental basis and associated clinical features, could be of great use in clinical practice. It is suggested that this association should be taken into account when it comes to deciding on the treatment.


Analysis of calpain-3 protein in muscle biopsies of different muscular dystrophies from India.
Calpain-3, a Ca [2]+ -dependent protease has been implicated in the pathology of neuromuscular disorders (NMDs). The current study aimed to analyze calpain-3 expression in cases diagnosed as muscular dystrophy from the Indian population. Calpain-3 Western blot analysis in muscle biopsies of immunohistochemically confirmed cases of Duchenne muscular dystrophy (DMD) (n=10), dysferlinopathy (n=30) and sarcoglycanopathy (n=8) was carried out. Calpain-3 Western blotting was also used in a blinded study to identify cases of calpain-3 deficiency in 28 NMD patients with potential muscular dystrophy. Calpain-3 appeared as a full length 94 kDa band with an autolytic product (~60 kDa) on Western blots with antibody NCL-CALP-12A2 (Ab-2). Eight of the 10 DMD samples showed absence of 94 kDa band but presence of 60 kDa band while one case of sarcoglycanopathy showed absence of both. Twenty one of the 30 dysferlinopathy samples showed both bands while six showed only the 60 kDa band and three showed absence of both. In the blinded study, five NMD cases with potential muscular dystrophy that showed complete absence of both bands in retrospect exhibited clinical features of limb girdle muscular dystrophy 2A (LGMD2A). While the study revealed a consistent pattern of calpain-3 in DMD, one sarcoglycanopathy and three dysferlinopathy samples exhibited secondary reduction in calpain-3. It was recognized that both calpain-3 bands should be considered to confirm calpain deficiency. Further, western blot offers an economical and fast preliminary screening method for LGMD2A especially in cases of complete absence of calpain-3 prior to conclusive diagnosis by genetic testing.


The interactions between accessibility and crash risk from a social equity perspective: A case study at the Rotterdam-The Hague metropolitan region.
Transport policies generally prioritize improving safety and accessibility levels, as they are regarded as the most important indicators of the quality of the transport system serving the public. However, inequalities associated with safety and accessibility issues are generally overlooked in these policies. Despite the importance and necessity of transport policies to address equity issues, there is still scarce knowledge on the interactions between equity, safety, and accessibility. This research aims to address this gap in the literature by creating a better understanding of the relationships between accessibility levels and traffic safety with a focus on social equity perspectives. A crash risk evaluation method and a Gravity model are utilized to analyze cycling safety and accessibility to jobs by bicycle. Two linear regression models (LM) were conducted to investigate the statistical correlations between cycling crash risk and accessibility. Moreover, the Bivariate local Moran's I method was employed to assess the spatial inequalities of distribution of crash risk and job accessibility over different income-level populations. The analyses showed that low-income people are not only disadvantaged in terms of job accessibility by bicycle but are also exposed to higher cycling crash risks, compared to high-income groups. Furthermore, most disadvantaged zones that have the highest need for road safety and accessibility improvements are identified as areas where low-income populations are exposed to higher crash risk and/or have lower access to jobs by bicycle. This study contributes to the transport literature by investigating the interactions between safety and accessibility and the impacts on transport equity. The findings of the statistical and spatial analysis are beneficial for the decision-makers, considering the probable mutual implications of land-use and transport developments and projects aiming to improve safety, accessibility, or both for different population groups.


Patterns of nucleotide sequence variation in ICAM1 and TNF genes in twelve ethnic groups of India: roles of demographic history and natural selection.
We have studied DNA sequence variation in and around the genes ICAM1 and TNF, which play functional and correlated roles in inflammatory processes and immune cell responses, in 12 diverse ethnic groups of India, with a view to investigating the relative roles of demographic history and natural selection in shaping the observed patterns of variation. The total numbers of single nucleotide polymorphisms (SNPs) detected at the ICAM1 and TNF loci were 29 and 12, respectively. Haplotype and allele frequencies differed significantly across populations. The site frequency spectra at these loci were significantly different from those expected under neutrality, and showed an excess of intermediate-frequency variants consistent with balancing selection. However, as expected under balancing selection, there was no significant reduction of F(ST) values compared to neutral autosomal loci. Mismatch distributions were consistent with population expansion for both loci. On the other hand, the phylogenetic network among haplotypes for the TNF locus was similar to expectations under population expansion, while that for the ICAM1 was as expected under balancing selection. Nucleotide diversity at the ICAM1 locus was an order of magnitude lower in the promoter region, compared to the introns or exons, but no such difference was noted for the TNF gene. Thus, we conclude that the pattern of nucleotide variation in these genes has been modulated by both demographic history and selection. This is not surprising in view of the known allelic associations of several polymorphisms in these genes with various diseases, both infectious and noninfectious.


Effectiveness and clinical predictors of drug survival in psoriasis patients receiving apremilast: A registry analysis.
Little is known about the effectiveness and drug survival associated with apremilast under real-world conditions. To investigate the influence of patient and disease characteristics on drug survival associated with apremilast and to elucidate clinical effectiveness with regard to the psoriasis area and severity index (PASI) reduction. This was an observational, retrospective, multicenter analysis from the Austrian Psoriasis Registry. Data from 367 patients were eligible for analysis. The 12-month drug survival rate associated with apremilast (ie, the proportion of patients on the drug) was 57.3% and decreased significantly in patients younger than 40 years (relative hazard ratio = 1.49, P = .007918). Sex; concomitant arthritis; previous biologic therapy; obesity; and palmoplantar, scalp, nail, and intertriginous involvement did not significantly affect drug survival. At 12 months, the response rates in patients receiving apremilast per protocol with a PASI of 50, 75, 90, and 100 were 80.0%, 56.4%, 38.2%, and 22.7%, respectively. Inclusion of a substantial number of patients with no record of absolute PASI at study entry and lack of PASI reduction follow-up data of 103 patients (28.1%) after starting apremilast treatment. Apremilast is a robust antipsoriatic drug for which the drug survival is not strongly influenced by most patient- or disease-related factors except age. Drug survival is significantly shorter in patients younger than 40 years.


Multiple functions of Saccharomyces cerevisiae splicing protein Prp24 in U6 RNA structural rearrangements.
U6 spliceosomal RNA has a complex secondary structure that includes a highly conserved stemloop near the 3' end. The 3' stem is unwound when U6 RNA base-pairs with U4 RNA during spliceosome assembly, but likely reforms when U4 RNA leaves the spliceosome prior to the catalysis of splicing. A mutation in yeast U6 RNA that hyperstabilizes the 3' stem confers cold sensitivity and inhibits U4/U6 assembly as well as a later step in splicing. Here we show that extragenic suppressors of the 3' stem mutation map to the gene coding for splicing factor Prp24. The suppressor mutations are located in the second and third of three RNA-recognition motifs (RRMs) in Prp24 and are predicted to disrupt RNA binding. Mutations in U6 RNA predicted to destabilize a novel helix adjacent to the 3' stem also suppress the 3' stem mutation and enhance the growth defect of a suppressor mutation in RRM2 of Prp24. Both phenotypes are reverted by a compensatory mutation that restores pairing in the novel helix. These results are best explained by a model in which RRMs 2 and 3 of Prp24 stabilize an extended intramolecular structure in U6 RNA that competes with the U4/U6 RNA interaction, and thus influence both association and dissociation of U4 and U6 RNAs during the splicing cycle.


[Ophthalmologic peculiarities of sarcoidosis].
The present paper deals with the frequency of different ophthalmic sarcoid changes and their morphological and photographic characteristics. In addition, the diagnostic value of conjunctival biopsy is evaluated. The findings were based on repeated ophthalmological examinations of an unselected group of 281 patients with histologically confirmed sarcoidosis. Conjunctival (37/218) and lacrimal gland (33/254) changes were encountered more often than sarcoid uveitis (22/281), and a conjunctival granuloma was the most frequent single sarcoid ophthalmic finding. Sarcoid changes in the eyes and the adnexae were often characterized by a scarcity of symptoms and a typical outward appearance. In some cases of sarcoid uveitis fluorescein angiography revealed in the iris and the retina nodules which showed features suggesting the proliferative nature of the disease. The iris infrared transillumination technique illustrated the affinity of sarcoid nodules in the pupillary area. Conjunctival biopsy showed epithelioid cell granulomas compatible with sarcoidosis in nearly half of the patients in whom it was suspected in a slit-lamp examination of the conjunctivae. In the diagnosis of sarcoidosis, conjunctival biopsy is a procedure to be recommended before more demanding methods are tried.


[Circulatory effects of vecuronium as well as pancuronium under different conditions of anesthesia].
The cardiovascular effects of equipotent doses (1,25 X ED95) of vecuronium (70 micrograms/kg iv) and pancuronium (80 micrograms/kg iv) were studied in 16 patients scheduled for elective coronary artery bypass surgery during steady-state conditions of isoflurane (0,4-0,5 vol% end-tidal)-nitrous oxide anaesthesia. All patients were chronically treated with oral beta receptor-blocking agents. Vecuronium did not cause any significant cardiovascular changes whereas pancuronium produced increases in heart rate (13%), cardiac index (15%) and mean arterial pressure (4%) while systemic vascular resistance decreased (8%). In a second part of this study we analysed whether the magnitude of the vagolytic effects of pancuronium is influenced by the anaesthetic procedure and/or by preoperative beta-blocker therapy. A group of 8 patients who were pretreated with beta-receptor blockers and received fentanyl (7 micrograms/kg) during the anaesthetic procedure showed low control values of heart rate (HR), cardiac index (CI), mean arterial pressure (MAP) and the rate-pressure product (RPP) which were due to both, the antisympathetic effects of beta-blocker therapy and the central vagomimetic properties of fentanyl. The administration of pancuronium (80 micrograms/kg) caused the greatest percentage increases in HR (20%), CI (22%), MAP (8%) and RPP (31%) in this group of patients. In contrast, patients (n = 8) anaesthetized with isoflurane-nitrous oxide who were not on preoperative beta-receptor blocker medication, demonstrated higher haemodynamic control values and less increases in HR (10%), CI (10%) and RPP (15%), MAP did not change. The clinical significance of these findings is discussed.


Atypical Mycobacterial Infection after Abdominoplasty Overseas: A Case Report and Literature Review.
Increasing number of medical tourists travel internationally for cosmetic procedures. Lipotourism is a form of medical tourism becoming popular among patients of developed countries due to the cost efficiency of cosmetic procedures when performed in developing nations. There is a paucity of data on quality, safety, and risks involved with these surgeries. Many cases of infections have been documented in patients following cosmetic surgeries in developing countries. We present a case of a 34-year-old female who underwent abdominoplasty in Dominican Republic that was complicated with development of multiple abdominal wall abscesses due to infection from rapidly growing mycobacteria (RGM). In the absence of clear treatment guidelines, she was treated with a combination of intermittent surgical drainage and prolonged antibiotic course. This case is of interest as more than one species of RGM was isolated from the same patient. Our case highlights the fact that identification of these organisms can be difficult requiring referral of samples to specialized laboratories and treatment duration can last several months, which is determined by clinical and microbiological response.


Reassessment of small lymphocytic lymphoma in the era of monoclonal B-cell lymphocytosis.
In the 2008 World Health Organization classification, small lymphocytic lymphoma is defined as a neoplasm with the tissue morphology and immunophenotype of chronic lymphocytic leukemia, but with absence of leukemia. Minimal criteria of tissue involvement to separate small lymphocytic lymphoma from monoclonal B-cell lymphocytosis have not been defined. We reviewed the clinicopathological features of 36 patients with extramedullary tissue biopsies containing chronic lymphocytic leukemia-type cells and less than 5×10(9)/L peripheral blood monoclonal B cells. Pathological features (extent and patterns of involvement, architectural preservation, presence of proliferation centers) as well as cytogenetic and radiological findings were examined in relation to clinical outcome. The biopsies were performed to evaluate lymphadenopathy in 20 patients and for other reasons (most frequently staging of a non-hematologic neoplasm) in 16 patients. At latest follow-up (median 23 months), 21 untreated patients had no or stable lymphadenopathy, 3 had regressed lymphadenopathy, and 12 had developed progressive lymphadenopathy and/or received therapy for chronic lymphocytic leukemia/small lymphocytic lymphoma. Features associated with progression/treatment included lymph nodes 1.5 cm or greater on imaging studies (P=0.01) and presence of proliferation centers in the biopsied tissue (P=0.004). Neither the size nor extent of involvement of the excised lymph node correlated with progression/treatment. Our findings suggest that biopsies containing chronic lymphocytic leukemia-type cells, but lacking proliferation centers and with non-enlarged or only slightly enlarged lymph nodes on imaging, represent a very indolent disease that may best be considered a tissue equivalent of monoclonal B-cell lymphocytosis rather than overt small lymphocytic lymphoma. We propose that such cases be designated as tissue involvement by chronic lymphocytic leukemia/small lymphocytic lymphoma-like cells of uncertain significance.


RNA editing of nuclear transcripts in Arabidopsis thaliana.
RNA editing is a transcript-based layer of gene regulation. To date, no systemic study on RNA editing of plant nuclear genes has been reported. Here, a transcriptome-wide search for editing sites in nuclear transcripts of Arabidopsis (Arabidopsis thaliana) was performed. MPSS (massively parallel signature sequencing) and PARE (parallel analysis of RNA ends) data retrieved from public databases were utilized, focusing on one-base-conversion editing. Besides cytidine (C)-to-uridine (U) editing in mitochondrial transcripts, many nuclear transcripts were found to be diversely edited. Interestingly, a sizable portion of these nuclear genes are involved in chloroplast- or mitochondrion-related functions, and many editing events are tissue-specific. Some editing sites, such as adenosine (A)-to-U editing loci, were found to be surrounded by peculiar elements. The editing events of some nuclear transcripts are highly enriched surrounding the borders between coding sequences (CDSs) and 3' untranslated regions (UTRs), suggesting site-specific editing. Furthermore, RNA editing is potentially implicated in new start or stop codon generation, and may affect alternative splicing of certain protein-coding transcripts. RNA editing in the precursor microRNAs (pre-miRNAs) of ath-miR854 family, resulting in secondary structure transformation, implies its potential role in microRNA (miRNA) maturation. To our knowledge, the results provide the first global view of RNA editing in plant nuclear transcripts.


Mesosiderite clasts with the most extreme positive europium anomalies among solar system rocks.
Pigeonite-plagioclase gabbros that occur as clasts in mesosiderites (brecciated stony-iron meteorites) show extreme fractionations of the rare-earth elements (REEs) with larger positive europium anomalies than any previously known for igneous rocks from the Earth, moon, or meteorite parent bodies and greater depletions of light REEs relative to heavy REEs than known for comparable cumulate gabbros. The REE pattern for merrillite in one of these clasts is depleted in light REEs and has a large positive europium anomaly as a result of metamorphic equilibration with the silicates. The extreme REE ratios exhibited by the mesosiderite clasts demonstrate that multistage igneous processes must have occurred on some asteroids in the early solar system. Melting of the crust by large-scale impacts or electrical induction from an early T-Tauri-phase sun may be responsible for these processes.


A remarkable effect of the combination of probabilistic peer-punishment and coevolutionary mechanism on the evolution of cooperation.
In the previous studies, the author has proposed the probabilistic peer-punishment based on the difference of payoff, and presented that the proposed peer-punishment utilizes its mechanism for preventing antisocial punishment like retaliation of a defector on a cooperator, effectively enhances the evolution of cooperation, and greatly increases the average payoff of all players in various parameters regarding static three types of topology of connections. Here, this study introduces both activities of breaking and creating connections of every player based on his/her preference to the model of the proposed peer-punishment. Every player will keep connections with his/her preferable players, whereas he/she will frequently break connections with his/her dissatisfied other players. Therefore, the new model of this study is the combination of probabilistic peer-punishment and coevolutionary mechanism that not only strategy of players but also connections between players evolve. This study discovers new knowledge that such combination induces high-level evolution of cooperation and great increase of the average payoff of all players in the condition where cooperation is hard to evolve.


B-cell leukemia in an adult sheep.
B-cell leukemia is a rare form of hematologic neoplasia in sheep, especially in adult animals. We present a case report of a 5-year-old WhiteFace Sheep wether with suspected acute lymphoblastic leukemia. The patient, a second-generation relative of ewes experimentally inoculated with atypical scrapie, exhibited acute lethargy and loss of appetite. Laboratory investigation revealed marked leukocytosis, lymphocytosis, and abnormal serum chemistry panel results. Microscopic examination of blood and bone marrow smears exhibited a high percentage of large neoplastic cells with lymphoid characteristics. Histopathologic analysis of the spleen, liver, lungs, and other organs confirmed the presence of widespread tissue infiltration by neoplastic cells. Immunohistochemical labeling demonstrated strong intracytoplasmic labeling for CD20, consistent with B-cell neoplasia. Flow cytometric analysis confirmed the B-cell lineage of the neoplastic cells. Screening for bovine leukemia virus, which can experimentally cause leukemia in sheep, yielded a negative result. In this case, the diagnosis of B-cell leukemia was supported by a comprehensive panel of diagnostic evaluations, including cytology, histopathology, immunohistochemistry, and immunophenotyping. This case report highlights the significance of accurate diagnosis and classification of hematologic neoplasia in sheep, emphasizing the need for immunophenotyping to aid in the diagnosis of B-cell leukemia. It also emphasizes the importance of considering spontaneous leukemia as a differential diagnosis in sheep with lymphoid neoplasia, especially in the absence of circulating infectious diseases.


Small fishes crossed a large mountain range: Quaternary stream capture events and freshwater fishes on both sides of the Taebaek Mountains.
The Taebaek Mountains in Korea serve as the most apparent biogeographic barrier for Korean freshwater fishes, resulting in 2 distinct ichthyofaunal assemblages on the eastern (East/Japan Sea slope) and western (Yellow Sea and Korea Strait slopes) sides of the mountain range. Of nearly 100 species of native primary freshwater fishes in Korea, only 18 species occur naturally on both sides of the mountain range. Interestingly, there are 5 rheophilic species (Phoxinus phoxinus, Coreoleuciscus splendidus, Ladislavia taczanowskii, Iksookimia koreensis and Koreocobitis rotundicaudata) found on both sides of the Taebaek Mountains that are geographically restricted to the Osip River (and several neighboring rivers, for L. taczanowskii and I. koreensis) on the eastern side of the mountain range. The Osip River and its neighboring rivers also shared a rheophilic freshwater fish, Liobagrus mediadiposalis, with the Nakdong River on the western side of the mountain range. We assessed historical biogeographic hypotheses on the presence of these rheophilic fishes, utilizing DNA sequence data from the mitochondrial cytochrome b gene. Results of our divergence time estimation indicate that ichthyofaunal transfers into the Osip River (and several neighboring rivers in East Sea slope) have occurred from the Han (Yellow Sea slope) and Nakdong (Korea Strait slope) Rivers since the Late Pleistocene. The inferred divergence times for the ichthyofaunal transfer across the Taebaek Mountains were consistent with the timing of hypothesized multiple reactivations of the Osip River Fault (Late Pleistocene), suggesting that the Osip River Fault reactivations may have caused stream capture events, followed by ichthyofaunal transfer, not only between the Osip and Nakdong Rivers, but also between the Osip and Han Rivers.


Anti-inflammatory agents and monoHER protect against DOX-induced cardiotoxicity and accumulation of CML in mice.
Cardiac damage is the major limiting factor for the clinical use of doxorubicin (DOX). Preclinical studies indicate that inflammatory effects may be involved in DOX-induced cardiotoxicity. Nepsilon-(carboxymethyl) lysine (CML) is suggested to be generated subsequent to oxidative stress, including inflammation. Therefore, the aim of this study was to investigate whether CML increased in the heart after DOX and whether anti-inflammatory agents reduced this effect in addition to their possible protection on DOX-induced cardiotoxicity. These effects were compared with those of the potential cardioprotector 7-monohydroxyethylrutoside (monoHER).BALB/c mice were treated with saline, DOX alone or DOX preceded by ketoprofen (KP), dexamethasone (DEX) or monoHER. Cardiac damage was evaluated according to Billingham. Nepsilon-(carboxymethyl) lysine was quantified immunohistochemically. Compared to saline, a 21.6-fold increase of damaged cardiomyocytes was observed in mice treated with DOX (P<0.001). Addition of KP, DEX or monoHER before DOX significantly reduced the mean ratio of abnormal cardiomyocytes in comparison to mice treated with DOX alone (P<or=0.02). In addition, DOX induced a significant increase in the number of CML-stained intramyocardial vessels per mm2 (P=0.001) and also in the intensity of CML staining (P=0.001) compared with the saline-treated group. Nepsilon-(carboxymethyl) lysine positivity was significantly reduced (P<or=0.01) by DOX-DEX, DOX-KP and DOX-monoHER. These results confirm that inflammation plays a role in DOX-induced cardiotoxicity, which is strengthened by the observed DOX-induced accumulation of CML, which can be reduced by anti-inflammatory agents and monoHER.


Preparation and performance of immobilized yeast cells in columns containing no inert carrier.
Schizosaccharomyces pombe was cultivated in a medium of glucose (10 g/L) malt extract (3 g/L), yeast extract (3 g/L), and bactopeptone (5 g/L) to form flocs. More than 95% of the cell population were flocculated. Variation in glucose concentration (from 10 to 100 g/L) did not affect flocculation. Yeast extract helped induce flocculation. Application of the immobilized yeast for the continuous production of ethanol was tested in a column reactor. Soft yeast flocs (50-200 mesh) underwent morphological changes to heavy particles (0.1-0.3 cm diameter) after continuously being fed with fresh substrates in the column. Productivity as high as 87 g EtOH L(-1) h(-1) was obtained when a 150 g/L glucose medium was fed. The performance of this yeast reactor was stable over a two-month period. The ethanol yield was 97% of the theoretical maximum based upon glucose consumed.


Nestin affects fusion pore dynamics in mouse astrocytes.
Astrocytes play a homeostatic role in the central nervous system and influence numerous aspects of neurophysiology via intracellular trafficking of vesicles. Intermediate filaments (IFs), also known as nanofilaments, regulate a number of cellular processes including organelle trafficking and adult hippocampal neurogenesis. We have recently demonstrated that the IF protein nestin, a marker of neural stem cells and immature and reactive astrocytes, is also expressed in some astrocytes in the unchallenged hippocampus and regulates neurogenesis through Notch signalling from astrocytes to neural stem cells, possibly via altered trafficking of vesicles containing the Notch ligand Jagged-1. We thus investigated whether nestin affects vesicle dynamics in astrocytes by examining single vesicle interactions with the plasmalemma and vesicle trafficking with high-resolution cell-attached membrane capacitance measurements and confocal microscopy. We used cell cultures of astrocytes from nestin-deficient (Nes-/- ) and wild-type (wt) mice, and fluorescent dextran and Fluo-2 to examine vesicle mobility and intracellular Ca2+ concentration respectively. Nes-/- astrocytes exhibited altered sizes of vesicles undergoing full fission and transient fusion, altered vesicle fusion pore geometry and kinetics, decreased spontaneous vesicle mobility and altered ATP-evoked mobility. Purinergic stimulation evoked Ca2+ signalling that was slightly attenuated in Nes-/- astrocytes, which exhibited more oscillatory Ca2+ responses than wt astrocytes. These results demonstrate at the single vesicle level that nestin regulates vesicle interactions with the plasmalemma and vesicle trafficking, indicating its potential role in astrocyte vesicle-based communication.