Cell-free nucleic acids as a non-invasive route for investigating atherosclerosis.
Metabolic syndrome is directly linked with atherosclerotic burden and cell-free nucleic acids (cf-NA) analysis has recently emerged as a novel research tool in atherosclerosis practice and research. cf-NA are nucleic acids (DNA, mRNA, miRNA, mitochondrial DNA) found in plasma and cell-free fractions of various other biological fluids. They have all the characteristics of the nucleic acids in the cells of their origin, thus constituting an emerging field for non-invasive assessment. Initially, quantitative and qualitative analysis of cf-NA has been accepted as clinically useful in non-invasive prenatal diagnosis, and in the diagnosis and monitoring of numerous cancers. As to atherosclerosis, cf-NA analysis poses an important challenge in diagnosis and prognostic evaluation of acute coronary syndrome, in prediction of cardiovascular disease, in non-invasive early detection of atherosclerosis and understanding its pathological mechanism in vivo, in assessing various issues of treatment for atherosclerosis in vivo, and in the unique simultaneous measurement of mRNA levels and protein concentrations in a single sample of plasma. Examples of its use are presented in this review. Besides the advances in technologies, the precise evaluation and optimization of pre-analytical and analytical aspects of cf-NA analysis have impacted importantly on the reliability of test results. We have, therefore, reviewed the most important analytical considerations. Further clinical studies and analytical improvements will answer the question as to whether cf-NA, as novel biomarkers, can be reliably applied clinically in non-invasive, early diagnosis and monitoring of the vulnerable atherosclerotic plaques of patients who could suffer from acute coronary syndrome.


Feasibility of mirabegron in the treatment of overactive bladder in patients affected by Parkinson's disease: A pilot study.
We investigated the effectiveness and safety of mirabegron oral treatment in a group of patients with Parkinson's disease (PD) and overactive bladder (OAB), refractory to antimuscarinics. Thirty patients with PD and refractory OAB were prospectively included in the study. At baseline, motor symptoms, severity of disease and cognitive status were assessed with the Hoehn-Yahr Scale, the Unified Parkinson's disease Rating Scale, the Mini Mental State examination and the Montreal Cognitive Assessment. At baseline, urinary symptoms, satisfaction with treatment and the impact of urinary incontinence on quality of life (QoL) were assessed with the 3-day voiding diary, the Visual Analogue Scale (VAS), the Incontinence-QoL questionnaire and urodynamics. Patients started assuming mirabegron 50 mg tablets once daily. Evaluation of urinary symptoms and related questionnaires, motor symptoms, severity of PD and uroflowmetry with postvoid residual volume measurement were then repeated at the 3- and 6-month follow up. Side effects were also noted. At baseline, the most frequently reported urinary symptoms were: urinary urgency (present in all the patients), urge urinary incontinence in 28/30 (93.3%) and increased daytime urinary frequency in 25 (83.3%) patients. At the 3-month follow up, 7 out of the 30 patients achieved a complete urinary continence. Significant improvements in VAS and Incontinence-QoL scores were observed in 24 patients. These benefits were maintained for the whole observation period. Four patients discontinued treatment due to poor efficacy, and two due to the cost of the drug. Mirabegron is a safe and effective treatment in patients with PD and OAB refractory to anticholinergics in the short-term follow up.


Holding back, intimacy, and psychological and relationship outcomes among couples coping with prostate cancer.
The present study evaluated intimacy as a mechanism for the effects of holding back sharing concerns about cancer on couples' psychological distress, well-being, and marital satisfaction using the actor-partner interdependence model (APIM), and evaluated 2 possible moderators of these associations: the number of patient and spouse cancer concerns. We had 139 men treated for localized prostate cancer in the past year and their spouses complete surveys about holding back sharing cancer concerns, intimacy, distress, and relationship satisfaction, as well as patient and spouse cancer concerns. APIM-indicated that the association between holding back sharing concerns, and patient and spouse distress, well-being, and relationship satisfaction could be partially accounted for by their influence on patient and spouse perceptions of relationship intimacy. The number of cancer concerns did not moderate the mediational model. Holding back has strong associations with both partners' well-being and distress. Holding back sharing concerns was particularly detrimental for couples' intimacy and relationship satisfaction.


Activation of human neutrophil procollagenase by nitrogen dioxide and peroxynitrite: a novel mechanism for procollagenase activation involving nitric oxide.
The involvement of nitric oxide (NO) and its reactive intermediates such as nitrogen dioxide (NO2) and peroxynitrite (ONOO-) in the activation of matrix metallo-proteinase was investigated. The human neutrophil procollagenase (matrix metalloproteinase-8) (M(r), 85 kDa) was purified to homogeneity from human neutrophils by using column chromatography. After incubation of human neutrophil procollagenase with various nitrogen oxide-generating systems, collagenolytic activity in each reaction system was measured. In addition, neutrophil collagenase activity was determined by assessment of proteolysis of human alpha 1-protease inhibitor. NO was formed by the propylamine NONOate, and NO2 was generated by oxidation of NO with 2-(4-carboxyphenyl)-4,4,5,5-tetramethylimidazoline-1-oxyl 3-oxide (carboxy-PTIO). NO2, formed by NONOate and carboxy-PTIO, and the synthetic ONOO- exhibited strong activation of the procollagenase at 1-20 microM. Significant activation of the procollagenase was observed with use of authentic NO2 gas as well. Constant flux infusion of ONOO- into the procollagenase solution resulted in stronger procollagenase activation than did a bolus addition of ONOO- to the reaction mixture. However, NO showed only weak activating potential under the aerobic (ambient) condition; an NO concentration of more than 10 mM was needed for appreciable activation of the procollagenase. Of considerable importance was the fact that NO participates in activation of the neutrophil collagenase through its conversion to NO2 or ONOO- in human neutrophils. These results suggest that NO2 and ONOO- may be potent activators of human neutrophil procollagenase.


Solving the multiple competitive facilities location and design problem on the plane.
A continuous location problem in which a firm wants to set up two or more new facilities in a competitive environment is considered. Other facilities offering the same product or service already exist in the area. Both the locations and the qualities of the new facilities are to be found so as to maximize the profit obtained by the firm. This is a global optimization problem, with many local optima. In this paper we analyze several approaches to solve it, namely, three multistart local search heuristics, a multistart simulated annealing algorithm, and two variants of an evolutionary algorithm. Through a comprehensive computational study it is shown that the evolutionary algorithms are the heuristics that provide the best solutions. Furthermore, using a set of problems for which the optimal solutions are known, only the evolutionary algorithms were able to find the optimal solutions for all the instances. The evolutionary strategies presented in this paper can be easily adapted to handle other continuous location problems.


Occurrence of spinal extradural arachnoid cysts in a child with concomitant intracranial midline abnormalities: case report.
Spinal extradural arachnoid cysts (SEACs) are uncommon spinal lesions that may cause myelopathy, most frequently in the 2nd decade of life. There are multiple theories of their pathogenesis, and associated entities include spinal trauma, spina bifida, and the lymphedema-distichiasis syndrome. The authors report the case of an otherwise healthy, developmentally normal 13-year-old boy who presented with multiple SEACs. Upon further neuroimaging workup, he was found to have an asymptomatic retrocerebellar arachnoid cyst, cavum septi pellucidi, and cavum vergae. Three contiguous but separate spinal cysts were identified intraoperatively, and they were completely excised with closure of the dural defects. The patient recovered motor and sensory function of the lower extremities. This collection of uncommon neuroimaging findings provides important clues to the pathogenesis of the disease and guides the optimal management of patients with SEACs. The unusual presentation of SEACs, together with uncommon midline abnormalities, provides further evidence of their congenital, midline origin. Therefore, it is reasonable to pursue imaging of the brain in atypical cases of SEACs.


Children's perceptions of injuries: a qualitative study in Sweden.
To understand one of the major public health problems for children, it is important to consider the children's perspective. The purpose of this qualitative study was to explore, describe, and categorize children's perceptions of injury severity and children's explanations of the injuries they experience. A total of 29 students from six randomly selected schools were interviewed in age groups of 9, 13, and 17 years. Manifest content analysis according to Graneheim and Lundman (2004) was used to categorize children's own statements. Need of medical attention, long-term consequences, and familiarity with the injury risk situation were identified as important determinants of children's perception of injury severity. Three categories emerged from children's explanations of their injuries: "Because of Me" (beliefs, lack of concentration, health conditions, and lack of awareness of risk), "Because of the Situation" (rain, ice, wind, animals, inanimate objects, constructions, and the children's games), and "Just Inexplicable" to the children. Findings suggest that children have a wide perception of injury severity and that children's beliefs of injury causation, as well as children's familiarity with injury risk situations, need to be considered in future studies focusing on the development of childhood injury prevention strategies. Additionally, results suggest that sometimes children cannot or do not want to explain their injuries.


Antimicrobial stewardship, therapeutic drug monitoring and infection management in the ICU: results from the international A- TEAMICU survey.
Severe infections and multidrug-resistant pathogens are common in critically ill patients. Antimicrobial stewardship (AMS) and therapeutic drug monitoring (TDM) are contemporary tools to optimize the use of antimicrobials. The A-TEAMICU survey was initiated to gain contemporary insights into dissemination and structure of AMS programs and TDM practices in intensive care units. This study involved online survey of members of ESICM and six national professional intensive care societies. Data of 812 respondents from mostly European high- and middle-income countries were available for analysis. 63% had AMS rounds available in their ICU, where 78% performed rounds weekly or more often. While 82% had local guidelines for treatment of infections, only 70% had cumulative antimicrobial susceptibility reports and 56% monitored the quantity of antimicrobials administered. A restriction of antimicrobials was reported by 62%. TDM of antimicrobial agents was used in 61% of ICUs, mostly glycopeptides (89%), aminoglycosides (77%), carbapenems (32%), penicillins (30%), azole antifungals (27%), cephalosporins (17%), and linezolid (16%). 76% of respondents used prolonged/continuous infusion of antimicrobials. The availability of an AMS had a significant association with the use of TDM. Many respondents of the survey have AMS in their ICUs. TDM of antimicrobials and optimized administration of antibiotics are broadly used among respondents. The availability of antimicrobial susceptibility reports and a surveillance of antimicrobial use should be actively sought by intensivists where unavailable. Results of this survey may inform further research and educational activities.


Fragile X syndrome and model organisms: identifying potential routes of therapeutic intervention.
Fragile X syndrome (FXS) is a cognitive disorder caused by silencing of the fragile X mental retardation 1 gene (FMR1). Since the discovery of the gene almost two decades ago, most scientific contributions have focused on identifying the molecular function of the fragile X mental retardation protein (FMRP) and understanding how absence of FMR1 gene expression gives rise to the disease phenotypes. The use of model organisms has allowed rapid progression in the FXS field and has given insight into the molecular basis of the disease. The mouse and fly FXS models have enabled studies to identify potential targets and pathways for pharmacological treatment. Here, we briefly review the two primary FXS model systems and describe how studies in these organisms have led us closer to therapeutic treatments for patients afflicted with FXS.


Anosmia/Hyposmia is a Good Predictor of Coronavirus Disease 2019 (COVID-19) Infection: A Meta-Analysis.
Introduction The number of positive cases and deaths from the coronavirus disease 2019 (COVID-19) is still increasing. The early detection of the disease is very important. Olfactory dysfunction has been reported as the main symptom in part of the patients. Objective To analyze the potential usefulness of anosmia or hyposmia in the detection of the COVID-19 infection. Data Synthesis We systematically searched the PubMed Central database using specific keywords related to our aims until July 31st, 2020. All articles published on COVID-19 and anosmia or hyposmia were retrieved. A statistical analysis was performed using the Review Manager (RevMan, Cochrane, London, UK) software, version 5.4. A total of 10 studies involving 21,638 patients were included in the present analysis. The meta-analysis showed that anosmia or hyposmia is significantly associated with positive COVID-19 infections (risk ratio [RR]: 4.56; 95% confidence interval [95%CI]: 3.32-6.24; p < 0.00001; I 2 = 78%, random-effects modeling). Conclusion The presence of anosmia or hyposmia is a good predictor of positive COVID-19 infections. Patients with onset of anosmia or hyposmia should take the test or undergo screening for the possibility of COVID-19 infection.


Thapsigargin--from Thapsia L. to mipsagargin.
The sesquiterpene lactone thapsigargin is found in the plant Thapsia garganica L., and is one of the major constituents of the roots and fruits of this Mediterranean species. In 1978, the first pharmacological effects of thapsigargin were established and the full structure was elucidated in 1985. Shortly after, the overall mechanism of the Sarco-endoplasmic reticulum Ca2+-ATPase (SERCA) inhibition that leads to apoptosis was discovered. Thapsigargin has a potent antagonistic effect on the SERCA and is widely used to study Ca2+-signaling. The effect on SERCA has also been utilized in the treatment of solid tumors. A prodrug has been designed to target the blood vessels of cancer cells; the death of these blood vessels then leads to tumor necrosis. The first clinical trials of this drug were initiated in 2008, and the potent drug is expected to enter the market in the near future under the generic name Mipsagargin (G-202). This review will describe the discovery of the new drug, the on-going elucidation of the biosynthesis of thapsigargin in the plant and attempts to supply the global market with a novel potent anti-cancer drug.


Usability, Acceptability, and Satisfaction of a Wearable Activity Tracker in Older Adults: Observational Study in a Real-Life Context in Northern Portugal.
The use of activity trackers has significantly increased over the last few years. This technology has the potential to improve the levels of physical activity and health-related behaviors in older adults. However, despite the potential benefits, the rate of adoption remains low among older adults. Therefore, understanding how technology is perceived may potentially offer insight to promote its use. This study aimed to (1) assess acceptability, usability, and user satisfaction with the Xiaomi Mi Band 2 in Portuguese community-dwelling older adults in a real-world context; (2) explore the mediating effect of the usability on the relationship between user characteristics and satisfaction; and (3) examine the moderating effect of user characteristics on the relationship between usability and user satisfaction. Older adults used the Xiaomi Mi Band 2 over 15 days. The user experience was evaluated through the Technology Acceptance Model 3, System Usability Scale, and User Satisfaction Evaluation Questionnaire. An integrated framework for usability and user satisfaction was used to explore user experience. Statistical data analysis included descriptive data analysis, reliability analysis, confirmatory factor analysis, and mediation and moderation analyses. A sample of 110 older adults with an average age of 68.41 years (SD 3.11) completed the user experience questionnaires. Mean user acceptance was very high-perceived ease of use: 6.45 (SD 0.78); perceptions of external control: 6.74 (SD 0.55); computer anxiety: 6.85 (SD 0.47); and behavioral intention: 6.60 (SD 0.97). The usability was excellent with an average score of 92.70 (SD 10.73), and user satisfaction was classified as a good experience 23.30 (SD 2.40). The mediation analysis confirmed the direct positive effect of usability on satisfaction (β=.530; P<.01) and the direct negative effect of depression on usability (β=-.369; P<.01). Lastly, the indirect effect of usability on user satisfaction was higher in individuals with lower Geriatric Depression Scale levels. Findings demonstrate that the Xiaomi Mi Band 2 is suitable for older adults. Furthermore, the results confirmed usability as a determinant of satisfaction with the technology and extended the existing knowledge about wearable activity trackers in older adults.


Complete remission for 4 years with crizotinib in advanced ALK-positive non-small cell lung cancer after thoracostomy for empyema.
Anaplastic lymphoma kinase (ALK) gene translocation occurs in 3%-5% of patients with non-small cell lung cancer (NSCLC), typically in younger patients. Crizotinib (tyrosine kinase inhibitor) has been considered as the standard of care for advanced ALK-positive lung cancer but it only gives a median progression-free survival of 7.7-11 months. A 41-year-old old man, former smoker, was diagnosed with NSCLC in the right lung with manifest pleural effusion. This case was complicated by a pleural empyema and because of a trapped lung, there was an indication for the construction of a thoracostomy. After confirmation of the ALK translocation, therapy with crizotinib was started. After 8 weeks, there was excellent response, and 6 months later, all lesions were undetectable on CT scan. There was also complete healing of the thoracostomy wound. This case describes a relatively young patient with a poor prognosis but with a remarkable and long-term response to crizotinib monotherapy.


Citrullinemia: phenotypic variations.
An 18-month-old female infant was found to have citrullinemia on routine plasma screening by the Scriver Method at 5 days of age. At 10 days of age, plasma citrulline concentration was 0.704mumol/ml (normal, 0.010 to 0.030mumol/ml) and has remained 60 to 80 times higher than normal. Urine citrulline concentration was markedly elevated. Hyperammonemia occurred at 1 month of age. The serum ammonia concentration was 473mug/100 ml (normal, 50 to 250 mug/100 ml) and rose to 770mug/100 ml at 4 months of age. Dietary protein was restricted to 1.6 gm/kg/day. Without further change in protein intake, the serum ammonia concentration decreased to 280mug/100 ml and, since then, it has returned to normal. The addition of three synthetic L-amino acids was required for a short time during dietary therapy. At 10 months of age, the infant was given a normal diet. At 18 months of age, her physical and mental development is normal. Activity of argininosuccinic acid synthetase measured in skin fibroblasts was 0.0037mumol of radioactive carbon dioxide per milligram of protein per hour. To demonstrate heterozygosity, fasting plasma citrulline concentrations were measured in five members of the family. Comparison of findings in this patient with those reported in the literature suggests phenotypical variation of the disease, probably due to genetic heterogeneity.


The direct, indirect and intangible costs of visual impairment caused by neovascular age-related macular degeneration.
Neovascular age-related macular degeneration (nvAMD) is a chronic, progressive disease of the central retina, and its prevalence is expected to rise with the ageing population. Using a bottom-up approach based on retrospective data, this cross-sectional study estimated average annual direct costs of nvAMD to be £4,047, and average annual indirect costs to be £449. An attempt to measure intangible costs through willingness-to-pay yielded a lower response rate and estimated intangible costs to be 11.5% of monthly income. Direct costs were significantly higher for male participants, for those who have mild or moderate visual impairment in both eyes, and for those who have been diagnosed for a shorter time. The findings of this study suggest that the availability of early diagnosis, effective treatment, support services, and sustained research into the management of nvAMD may reduce the burden of visual impairment caused by nvAMD to affected individuals and the state.


Further limiting bisphenol a in food uses could provide health and economic benefits.
There is mounting evidence that bisphenol A (BPA), a chemical used in the production of polycarbonate plastics and the linings of aluminum cans, may have adverse health consequences. The Food and Drug Administration has banned BPA from baby bottles and sippy cups but has deferred further action on other food uses-that is, uses in metal-based food and beverage containers. This article quantifies the potential social costs of childhood obesity and adult coronary heart disease attributable to BPA exposure in the United States in 2008 and models the potential health and economic benefits associated with replacing BPA in all food uses. BPA exposure was estimated to be associated with 12,404 cases of childhood obesity and 33,863 cases of newly incident coronary heart disease, with estimated social costs of $2.98 billion in 2008. Removing BPA from food uses might prevent 6,236 cases of childhood obesity and 22,350 cases of newly incident coronary heart disease per year, with potential annual economic benefits of $1.74 billion (sensitivity analysis: $889 million-$13.8 billion per year). Although more data are needed, these potentially large health and economic benefits could outweigh the costs of using a safer substitute for BPA.


Effect of hypoxia/hypercapnia on metabolism of 6-[(18)F]fluoro-L-DOPA in newborn piglets.
There is evidence that the dopaminergic system is sensitive to altered p(O(2)) in the immature brain. However, the respective enzyme activities have not been measured in the living neonatal brain together with brain oxidative metabolism. Therefore 18F-labelled 6-fluoro-L-3,4-dihydroxyphenylalanine (FDOPA) together with positron emission tomography (PET) was used to estimate the activity of the aromatic amino acid decarboxylase (AADC) in the brain of fifteen newborn piglets (2-5 days old). Two PET scans were performed in each piglet. Eleven animals underwent a period of normoxia and moderate hypoxia/hypercapnia (H/H). The remaining four animals were used as an untreated control group. Simultaneously, the brain tissue p(O(2)) was recorded, the regional cerebral blood flow (CBF) was measured with colored microspheres and the cerebral metabolic rate of oxygen (CMRO(2)) was determined. In addition, in four untreated and six H/H treated piglets the relative amounts of fluorodopamine and the respective metabolites were determined in brain tissue samples using HPLC analysis. H/H conditions were induced by lowering the inspired fraction of oxygen from 0.35 to 0.10 and adding CO(2) to the inspired gas resulting in an arterial p(CO(2)) between 74 and 79 mmHg. H/H elicited a more than 3-fold increase of the CBF (P<0.05) so that the CMRO(2) remained unchanged throughout the H/H period. Despite this, the brain tissue p(O(2)) was reduced from 19+/-4 to 6+/-3 mmHg (P<0.05). The permeability-surface area product of FDOPA (PS(FDOPA)) was unchanged. However, the transfer rate of FDOPA (k(3)(FDOPA)) of the nigrostriatal dopaminergic system and the relative amounts of fluorodopamine and the respective metabolites were significantly increased (P<0.05). It is suggested that H/H induces an increase of AADC activity. However, an H/H-induced CBF increase maintains bulk O(2) delivery and preserves CMRO(2).


Advancements in wearable technology for monitoring lactate levels using lactate oxidase enzyme and free enzyme as analytical approaches: A review.
Lactate is a metabolite that holds significant importance in human healthcare, biotechnology, and the food industry. The need for lactate monitoring has led to the development of various devices for measuring lactate concentration. Traditional laboratory methods, which involve extracting blood samples through invasive techniques such as needles, are costly, time-consuming, and require in-person sampling. To overcome these limitations, new technologies for lactate monitoring have emerged. Wearable biosensors are a promising approach that offers non-invasiveness, low cost, and short response times. They can be easily attached to the skin and provide continuous monitoring. In this review, we evaluate different types of wearable biosensors for lactate monitoring using lactate oxidase enzyme as biological recognition element and free enzyme systems.


Shaping Health Behavior across Generations: Evidence from Time Use data in the Panel Study of Income Dynamics and its Supplements.
This paper examines ways that families use time to shape their children's health behaviors. Specifically, it explores ways that parents can prepare children to make health-enhancing choices as adults. It also analyzes ways that offspring manage their time during young adulthood, when they are old enough to make independent decisions about whether they will spend their time producing health, as the Grossman model argues that individuals can do (Grossman 2000). The empirical research uses time diary data from the Child Development Supplement CDS), data from the Panel Study of Income Dynamics (PSID) core and the PSID's Transition to Adulthood (TA) module. By examining the intergenerational transmission of healthy behaviors, it is possible to better understand decisions individuals make in adulthood.


Amending the Law for Licensing Medical Practitioners of China in 2021: A Commentary.
The Law for Licensing Medical Practitioners of the People's Republic of China, enacted in 1999, was amended in 2021. This commentary reviews the key points of the amendment and raises doubts as to one of its points. Specifically, we argue that the minimum education level required to take the physicians' licensing examination should be set to completion of a bachelor degree, instead of a vocational diploma or junior college graduation as in the 2021 amendment. China adopted a system of multi-tiered medical education more than 70 years ago. This policy has resulted in a threshold of entry-level medical education far below the global standards. The highly heterogeneous education background of physicians in China has led to low standards of practicing physicians, which in turn have significantly negative impacts on the health care market. We illustrate changes over time in the educational distribution and regional distribution of practicing physicians in China, and present reasons to improve entry-level educational standards, by setting the physician licensing threshold at an appropriate level. This will not only improve the overall quality of physicians but will also help address equity and efficiency issues in the health care market.


Predicting remission of diabetes after RYGB surgery following intensive management to optimize preoperative glucose control.
The purpose of the study is to investigate the association of preoperative glucose optimization prior to a Roux-en-Y gastric bypass (RYGB) and diabetes remission. The study is a retrospective review of 245 patients with a history of diabetes type II and a RYGB from 2008 to 2012 at UMass Memorial Hospital. Patients that benefited from glucose optimization prior to RYGB were more likely to achieve diabetes remission 1 year after surgery. The preoperative glucose optimization intervention demonstrated that when patients decreased their HbA1c prior to surgery by 1 %, these individuals were 68 % more likely to remit (p = 0.015). Duration of diabetes (p = 0.005) and insulin use (p < 0.001) were also significant predictors of remission, whereas age, race, and gender were not. Our study results indicate that a greater degree of glycemic improvement in response to presurgical medical intervention is associated with higher rates of diabetes remission post-operatively among obese adults with diabetes type II. Conversely, the lack of favorable glycemic response to intensification of medical management predicts a poor glycemic response to bariatric surgery. Further research is needed to determine if this difference is due to physiological factors or is simply an indicator of patient behavior.


The Role of Palliative Care in Heart Failure, Part 3: Facilitators and Barriers to Cardiac Palliative Care Clinic Development.
Background: Patients with heart failure frequently have significant disease burden and complex psychosocial needs. The integration of palliative care into the management of these patients can decrease symptom burden throughout their course of illness. Therefore, in 2009, we established a cardiac palliative care clinic colocated with heart failure providers in a large academic heart hospital. Objective: To better understand the facilitators and barriers to integrating palliative care into our heart failure management service. Design: Qualitative study using a semistructured interview guide. Setting, Subjects: Between October 2020 and January 2021, we invited all 25 primary cardiac providers at our academic medical center in the midwestern United States to participate in semistructured qualitative interviews to discuss their experiences with the cardiac palliative care clinic. Measurements: Interview transcripts were analyzed using a deductive-dominant thematic analysis approach to reveal emerging themes. Results: Providers noted that the integration of palliative care into the treatment of patients with heart failure was helped and hindered primarily by issues related to operations and communications. Operational themes about clinic proximity and the use of telehealth as well as communication themes around provider-provider communication and the understanding of palliative care were particularly salient. Conclusions: The facilitators and barriers identified have broad applicability that are independent of the etiological nature (e.g., cancer, pulmonary, neurological) of any specialty or palliative care clinic. Moreover, the strategies we used to implement improvements in our clinic may be of benefit to other practice models such as independent and embedded clinics.


Expression of the transcription factors snail, slug, and twist and their clinical significance in human breast cancer.
Slug, Snail, and Twist are transcription factors that regulate the expression of tumor suppressors such as E-cadherin. We examined the distribution and expression of these three molecules together with the methylation of the Twist gene promoter in human breast cancer to elucidate their clinical significance. Frozen sections from breast cancer primary tumors (tumor, n = 114; background, n = 30) were immunostained with Slug, Snail, and Twist antibodies. RNA was reverse-transcribed, quantified, and analyzed by quantitative polymerase chain reaction (Q-PCR). Results were expressed as copy number of transcript per 50 ng of RNA (standardized against beta-actin). Immunohistochemistry revealed that all three molecules were stained in mammary tissues, with an increase in Twist within tumor tissues; this was supported by Q-PCR analysis. Q-PCR analysis showed that Slug was elevated with increasing tumor grade and prognostic indices. Twist was elevated with increasing nodal involvement (tumor-node-metastasis status). Conversely, Snail was reduced in expression corresponding with prognostic indices and tumor grade. Increased levels of Slug were associated with tumors from patients with metastatic disease or disease recurrence, and increased expression of Twist was associated with tumors from patients who had died from breast cancer. It is interesting to note that Snail expression was significantly reduced in patients with a poor outcome and those who had node-positive tumors. In addition, tumors exhibited methylation of the Twist promoter. These data demonstrate that all three transcription factors have inappropriate expression in breast cancer and that this may play a part in the progression of human breast tumors.


G16-mediated activation of nuclear factor kappaB by the adenosine A1 receptor involves c-Src, protein kinase C, and ERK signaling.
The G(i)-linked adenosine A1 receptor has been shown to mediate anti-inflammatory actions, possibly via modulation of the transcription factor nuclear factor-kappaB (NFkappaB). Here we demonstrate that an adenosine A1 agonist, N(6)-cyclohexyladenosine (CHA), activated IKKalpha/beta phosphorylation through PTX-insensitive G proteins in human lymphoblastoma Reh cells. To delineate the mechanism of action, different PTX-insensitive G proteins were expressed in human embryonic kidney 293 cells. Only Galpha(16) supported the CHA-induced IKK phosphorylation and NFkappaB-driven luciferase activity in time-dependent, dose-dependent, and PTX-insensitive manners. Gbetagamma subunits also modulated IKK/NFkappaB, as indicated by the stimulatory actions of Gbeta(1)gamma(2) and the abrogation of CHA-induced response by transducin. The participation of phospholipase Cbeta, protein kinase C, and calmodulin-dependent kinase II in CHA-induced IKK/NFkappaB activation were demonstrated by employing specific inhibitors and dominant-negative mutants. Inhibition of c-Src and numerous intermediates along the extracellular signal-regulated (ERK) kinase cascade including Ras, Raf-1 kinase, and MEK1/2 abolished the CHA-induced IKK/NFkappaB activation. Although c-Jun N-terminal kinase and p38 MAPK were also activated by CHA, they were not required for the IKK/NFkappaB regulation. Similar results were obtained using Reh cells. These data suggest that the G(16)-mediated activation of IKK/NFkappaB by CHA required a complex signaling network composed of multiple intermediates.


Size limit of molecules permeating the junctional membrane channels.
The permeability of the cell-to-cell membrane channels in salivary gland cell junction (Chironomus thummi) was probed with fluorescent-labeled amino acids and synthetic or natural peptides. Molecules up to 1200 daltons pass through the channels with velocities depending on molecular size. Molecules of 1900 daltons or greater do not pass. This passage failure seems to reflect the normal size limit for junctional channel permeation; the channels continue to be permeated by the molecules up to 1200 daltons when these are mixed with the nonpermeant molecules. From this size limit a channel diameter of 10 to 14 angstroms is estimated.


Fast motif recognition via application of statistical thresholds.
Improving the accuracy and efficiency of motif recognition is an important computational challenge that has application to detecting transcription factor binding sites in genomic data. Closely related to motif recognition is the CONSENSUS STRING decision problem that asks, given a parameter d and a set of l-length strings S = {s1, ..., sn}, whether there exists a consensus string that has Hamming distance at most d from any string in S. A set of strings S is pairwise bounded if the Hamming distance between any pair of strings in S is at most 2d. It is trivial to determine whether a set is pairwise bounded, and a set cannot have a consensus string unless it is pairwise bounded. We use CONSENSUS STRING to determine whether or not a pairwise bounded set has a consensus. Unfortunately, CONSENSUS STRING is NP-complete. The lack of an efficient method to solve the CONSENSUS STRING problem has caused it to become a computational bottleneck in MCL-WMR, a motif recognition program capable of solving difficult motif recognition problem instances. We focus on the development of a method for solving CONSENSUS STRING quickly with a small probability of error. We apply this heuristic to develop a new motif recognition program, sMCL-WMR, which has impressive accuracy and efficiency. We demonstrate the performance of sMCL-WMR in detecting weak motifs in large data sets and in real genomic data sets, and compare the performance to other leading motif recognition programs. In our preliminary discussion of our CONSENSUS STRING algorithm we give insight into the issue of sampling pairwise bounded sets, and discuss its relevance to motif recognition. Our novel heuristic gives birth to a state of the art program, sMCL-WMR, that is capable of detecting weak motifs in data sets with a large number of strings. sMCL-WMR is orders of magnitude faster than its predecessor MCL-WMR and is capable of solving previously unsolved synthetic motif recognition problems. Lastly, sMCL-WMR shows impressive accuracy in detecting transcription factor binding sites in the genomic data and used in the assessment of Tompa et al.


Neural mechanisms of reading facial emotions in young and older adults.
The ability to read and appropriately respond to emotions in others is central for successful social interaction. Young and older adults are better at identifying positive than negative facial expressions and also expressions of young than older faces. Little, however, is known about the neural processes associated with reading different emotions, particularly in faces of different ages, in samples of young and older adults. During fMRI, young and older participants identified expressions in happy, neutral, and angry young and older faces. The results suggest a functional dissociation of ventromedial prefrontal cortex (vmPFC) and dorsomedial prefrontal cortex (dmPFC) in reading facial emotions that is largely comparable in young and older adults: Both age groups showed greater vmPFC activity to happy compared to angry or neutral faces, which was positively correlated with expression identification for happy compared to angry faces. In contrast, both age groups showed greater activity in dmPFC to neutral or angry than happy faces which was negatively correlated with expression identification for neutral compared to happy faces. A similar region of dmPFC showed greater activity for older than young faces, but no brain-behavior correlations. Greater vmPFC activity in the present study may reflect greater affective processing involved in reading happy compared to neutral or angry faces. Greater dmPFC activity may reflect more cognitive control involved in decoding and/or regulating negative emotions associated with neutral or angry than happy, and older than young, faces.


Preparation of imprinted polymers at surface of magnetic nanoparticles for the selective extraction of tadalafil from medicines.
In this paper, highly selective core-shell molecularly imprinted polymers (MIPs) of tadalafil on the surface of magnetic nanoparticles (MNPs) were prepared. Three widely used functional monomers 2-(trifluoromethyl) acrylic acid (TFMAA), acrylic acid (AA), and methacrylic acid (MAA) were compared theoretically as the candidates for MIP preparation. MIP-coated magnetic nanoparticles (MIP-coated MNPs) showed large adsorption capacity, high recognition ability, and fast binding kinetics for tadalafil. Furthermore, because of the good magnetic properties, MIP-coated MNPs can achieve rapid and efficient separation with an external magnetic field simply. The resulting MIP-coated MNPs were used as dispersive solid-phase extraction (DSPE) materials coupled with HPLC-UV for the selective extraction and detection of tadalafil from medicines (herbal sexual health products). Encouraging results were obtained. The amounts of tadalafil that were detected from the herbal sexual health product was 43.46 nmol g(-1), and the recoveries were in the range of 87.36-90.93% with the RSD < 6.55%.


Mumps in the Czech Republic in 2013: Clinical Characteristics, Mumps Virus Genotyping, and Epidemiological Links.
The aim of the study was to map the incidence of mumps in the Czech Republic in terms of clinical symptoms, epidemiological links, and characteristics of circulating genotypes. Patients with suspected mumps examined in the Infectious Diseases Clinic of the Na Bulovce Hospital in 2013 were enrolled in the study. Buccal swab specimens were tested by means of nucleic acid detection (RT-qPCR) and when positive, they were cultured in tissue culture. Sequencing was carried out using the BigDye Terminator v3.1 Cycle Sequencing Kit and Genetic Analyzer 3500. The SeqScape software was used for the analysis of sequencing data and filtering out low quality reads. The phylogenetic analysis and genotyping were performed using the Mega 6 software. To generate the phylogenetic tree, all sequences were aligned by the MAFFT tool and the alignment obtained was edited using the BioEdit software. In all patients, selected biochemical markers (C-reactive protein, white blood cell count and serum amylase) were measured. The EPIDAT system used for reporting infectious diseases, record keeping, and data analysis in the Czech Republic was the source of statistical data. Eighty-nine patients with suspected mumps were examined in the Na Bulovce Hospital and 65 of them were laboratory confirmed with mumps: 40 males (61.5%) and 25 females (38.5%). The mean age of the study cohort was 25.9 years (median age of 23 years, age range from 10 to 73 years) and 14 patients were under 18 years of age. Thirty-four (52.3%) patients were vaccinated in childhood, 28 (43.1%) were unvaccinated, and for three persons, vaccination data were not available. A severe course of the disease was reported in 15 (23.1%) patients. Fourteen of them needed hospitalization because of orchitis (9 males) and meningitis (5 patients). One patient with orchitis was treated on an outpatient basis. The need for hospitalization tended to be lower in the unvaccinated patients (14.7% vs. 35.7%, p=0.076). In 2013, 1,553 cases of mumps were reported to the EPIDAT system. Of these, 640 were laboratory confirmed. The most often reported complications were orchitis (90 cases, i.e. 10.3%) and meningitis (21 cases, i.e. 1.4%). Orchitis was diagnosed in 30.3% of the unvaccinated and in 6.4% of the vaccinated males. Meningitis occurred in 3.1% of the unvaccinated and in 1.0% of the vaccinated patients. Despite the emergence of mumps among the vaccinated population, the present study has confirmed a positive effect of the vaccine, particularly on the incidence of complications and inflammatory markers. All 30 sequenced mumps virus strains were assigned to group G. A secondary vaccine failure due to waning immunity seems to be a plausible explanation for the rise in mumps cases.


Dynamic contrast enhanced magnetic resonance imaging of the terminal ileum: differentiation of activity of Crohn's disease.
To prospectively investigate a new high resolution MRI technique for dynamic evaluation of the enhancement kinetics of bowel parietal layers and to correlate it with CDAI, CRP, endoscopic activity and histologic features. About 16 consecutive patients with proven diagnosis of CD underwent ileocolonoscopy with biopsy and serial bowel dynamic contrasted-MRI (D-CE-MRI) evaluated in blind fashion. Quantitative analysis of bowel wall enhancement kinetics was performed basing on signal to noise ratio (SNR) of inner parietal layers (Mucosa-Submucosa, M-SM) and outer parietal layers (Muscular-Serosa, Ms-S). Disease activity was defined by CDAI > 150, serum CRP > 5 mg/dL and histologic results. About 9 patients showed a layered enhancement of bowel wall (8 active, 1 inactive), whereas inactive (7 cases) group presented a homogeneous pattern. In active patients we found a significant difference in parietal layered enhancement curves (M-SM vs. Ms-S, P < 0.03) not observed in inactive disease and controls (intra-group analysis). M-SM and Ms-S enhanced curves in clinically active patients were significantly different respect to those of patients with inactive CD (P < 0.001) (inter-group analysis). Parietal D-CE-MRI pattern well correlated with histologic features (r = 0.8; P < 0.001, Spearman test). D-CE-MRI can be a useful tool for clinical follow-up and in the treatment strategies in CD patients.


Links between anxiety and allergies: psychobiological reality or possible methodological bias?
The objective of the study was to examine the link between anxiety and allergies to establish whether it reflects a psychobiological reality or a possible methodological bias. A cohort of 1,037 children enrolled in the study. Anxiety disorders were assessed between 11 and 21 years. Anxious personality was assessed at 18 years. Allergies were examined at 21 years by (a) self reports, (b) skin pricks, and (c) serum total immunoglobulin E (IgE). Self-reported allergies were predicted by recurrent anxiety disorders (OR [95% CI]=1.56 [1.06-2.30], p=.023) and self-reports of anxious personality (OR [95% CI]=1.67 [1.17-2.37], p=.004): Objectively verified allergies were not. These results suggest that the link between anxiety and allergies may reflect a methodological artifact rather than a psychobiological reality.


Effects of imputation on correlation: implications for analysis of mass spectrometry data from multiple biological matrices.
With expanded access to, and decreased costs of, mass spectrometry, investigators are collecting and analyzing multiple biological matrices from the same subject such as serum, plasma, tissue and urine to enhance biomarker discoveries, understanding of disease processes and identification of therapeutic targets. Commonly, each biological matrix is analyzed separately, but multivariate methods such as MANOVAs that combine information from multiple biological matrices are potentially more powerful. However, mass spectrometric data typically contain large amounts of missing values, and imputation is often used to create complete data sets for analysis. The effects of imputation on multiple biological matrix analyses have not been studied. We investigated the effects of seven imputation methods (half minimum substitution, mean substitution, k-nearest neighbors, local least squares regression, Bayesian principal components analysis, singular value decomposition and random forest), on the within-subject correlation of compounds between biological matrices and its consequences on MANOVA results. Through analysis of three real omics data sets and simulation studies, we found the amount of missing data and imputation method to substantially change the between-matrix correlation structure. The magnitude of the correlations was generally reduced in imputed data sets, and this effect increased with the amount of missing data. Significant results from MANOVA testing also were substantially affected. In particular, the number of false positives increased with the level of missing data for all imputation methods. No one imputation method was universally the best, but the simple substitution methods (Half Minimum and Mean) consistently performed poorly.


The anaerobic bacterial flora of the mouse cecum.
Refinements of the anaerobic technique such as the utilization of prereduced media and the removal of traces of oxygen from the gases used by passage through a hot copper oven resulted in quantitative and qualitative improvements in the recovery of anaerobic bacteria from the cecum of SPF mice. The commonest morphologic types of bacteria in the mouse cecum were tapered rods. These characteristically gave "speckled" colonies on agar plates. Those identified were species of the genera Fusobacterium, Eubacterium, and Clostridium. Several of the morphologic types seen with phase microscopy still could not be cultivated. The implications of these findings for the study of the intestinal flora are discussed.


Core-shell designs of photoluminescent nanodiamonds with porous silica coatings for bioimaging and drug delivery I: fabrication.
A multifunctional core-shell nanocomposite platform consisting of a photoluminescent nanodiamond (ND) core with uniform porous silica coatings is presented. This design intended for drug delivery applications allows simultaneous stable fluorescent imaging with high loading capacity of bioactive molecules. Despite irregularly shaped starting cores, well-dispersed and uniformly shaped nanocomposite particles can be produced. Moreover, after optimization of the silica source-to-diamond ratio, the thickness of the porous layer can be tuned by adjusting the ethanol amount, allowing rational nanoparticle size control. The ND key property, photoluminescence, is not quenched regardless of coating with thick silica layers. The high loading capacity for incorporation of active agents, provided by the introduced porous layer, is demonstrated by adsorption of a hydrophobic model drug to the composite particles. The loading degree, as compared to a pure ND, increased by two orders of magnitude from 1 wt% for the ND to >100 wt% for the composite particles. Combining these two material classes, which both have well-documented excellent performance especially in biomedical applications, for the NDs with emphasis, but not exclusively, on imaging and mesoporous silica (MSN) on drug delivery, the advantages of both are shown here to be synergistically integrated into one multifunctional nanocomposite platform.


Familial idiopathic basal ganglia calcification with a heterozygous missense variant (c.902C>T/p.P307L) in SLC20A2 showing widespread cerebrovascular lesions.
We describe a postmortem case of familial idiopathic basal ganglia calcification (FIBGC) in a 72-year-old Japanese man. The patient showed progressive cognitive impairment with a seven-year clinical course and calcification of the basal ganglia, thalami, and cerebellar dentate nuclei. A novel heterozygous missense variant in SLC20A2 (c.920C>T/p.P307L), a type III sodium-dependent phosphate transporter (PiT-2), was subsequently identified, in addition to typical neuropathological findings of FIBGC, such as capillary calcification of the occipital gray matter, confluent calcification of the basal ganglia and cerebellar white matter, widespread occurrence of vasculopathic changes, cerebrovascular lesions, and vascular smooth muscle cell depletion. Immunohistochemistry for PiT-2 protein revealed no apparent staining in endothelial cells in the basal ganglia and insular cortex; however, the immunoreactivity in endothelial cells of the cerebellum was preserved. Moreover, Western blot analysis identified preserved PiT-2 immunoreactivity signals in the frontal cortex and cerebellum. The variant identified in the present patient could be associated with development of FIBGC and is known to be located at the large intracytoplasmic part of the PiT-2 protein, which has potential phosphorylation sites with importance in the regulation of inorganic phosphate transport activity. The present case is an important example to prove that FIGBC could stem from a missense variant in the large intracytoplasmic loop of the PiT-2 protein. Abnormal clearance of inorganic phosphate in the brain could be related to the development of vascular smooth muscle damage, the formation of cerebrovascular lesions, and subsequent brain calcification in patients with FIBGC with SLC20A2 variants.


Health status of the residents in occidental mindoro, Philippines: a way to make a healthy community.
Even though Philippines is widely known as exporters of health workers in the world, the Occidental Mindoro province suffers from a lack of health workers compared with the total population of each municipality. The aim of this study was to observe, identify, and understand the persisting health status, knowledge, and practices among the three selected communities in Occidental Mindoro, Philippines. The study applied a survey using basic questions with three key topics, with relevance to the health condition of the villagers, such as demographics (social capital and regional characteristics), lifestyle (healthy living, and healthy lifestyle and behavior), and status or position in the society (general demographics, and personal behavior and attitudes), with a random sample of 256 adult respondents. Only about 54.3% rated themselves as fair/moderately healthy, and a total of 17.2% suffered from chronic diseases such as diabetes mellitus, cancers, chronic obstructive pulmonary disease, and cardiovascular disease, while 9% have been diagnosed with tuberculosis in the past 6 months. Respondents mostly have low income and low education. The majority of the respondents have only primary and secondary education, and a very low average income; these suggest that respondents were afflicted with poverty and low educational attainment. Respondents who are deprived of their rights to obtain a higher education also have a higher chance of having less knowledge on their well-being. Health programs do not guarantee a healthy individual and a healthy society, but a combination of health programs and socioeconomic support can help in creating a healthy community.


Novel protocol to establish the myocardial infarction model in rats using a combination of medetomidine-midazolam-butorphanol (MMB) and atipamezole.
Myocardial infarction (MI) is one of the most common cardiac problems causing deaths in humans. Previously validated anesthetic agents used in MI model establishment are currently controversial with severe restrictions because of ethical concerns. The combination between medetomidine, midazolam, and butorphanol (MMB) is commonly used in different animal models. The possibility of MMB combination to establish the MI model in rats did not study yet which is difficult because of severe respiratory depression and delayed recovery post-surgery, resulting in significant deaths. Atipamezole is used to counter the cardiopulmonary suppressive effect of MMB. The aim of the present study is to establish MI model in rats using a novel anesthetic combination between MMB and Atipamezole. Twenty-five Sprague Dawley (SD) rats were included. Rats were prepared for induction of the Myocardial infarction (MI) model through thoracotomy. Anesthesia was initially induced with a mixture of MMB (0.3/5.0/5.0 mg/kg/SC), respectively. After endotracheal intubation, rats were maintained with isoflurane 1% which gradually reduced after chest closing. MI was induced through the left anterior descending (LAD) artery ligation technique. Atipamezole was administered after finishing all surgical procedures at a dose rate of 1.0 mg/kg/SC. Cardiac function parameters were evaluated using ECG (before and after atipamezole administration) and transthoracic echocardiography (before and 1 month after MI induction) to confirm the successful model. The induction time, operation time, and recovery time were calculated. The success rate of the MI model was also calculated. MI was successfully established with the mentioned anesthetic protocol through the LAD ligation technique and confirmed through changes in ECG and echocardiographic parameters after MI. ECG data was improved after atipamezole administration through a significant increase in heart rate (HR), PR Interval, QRS Interval, and QT correction (QTc) and a significant reduction in RR Interval. Atipamezole enables rats to recover voluntary respiratory movement (VRM), wakefulness, movement, and posture within a very short time after administration. Echocardiographic ally, MI rats showed a significant decrease in the left ventricular wall thickness, EF, FS, and increased left ventricular diastolic and systolic internal diameter. In addition, induction time (3.440 ± 1.044), operation time (29.40 ± 3.663), partial recovery time (10.84 ± 3.313), and complete recovery time (12.36 ± 4.847) were relatively short. Moreover, the success rate of the anesthetic protocol was 100%, and all rats were maintained for 1 month after surgery with a survival rate of 88%. Our protocol produced a more easy anesthetic effect and time-saving procedures with a highly successful rate in MI rats. Subcutaneous injection of Atipamezole efficiently counters the cardiopulmonary side effect of MMB which is necessary for rapid recovery and subsequently enhancing the survival rate during the creation of the MI model in rats.


Chronic alcohol feeding inhibits physiological and molecular parameters of intestinal and renal riboflavin transport.
Vitamin B2 (riboflavin, RF) is essential for normal human health. Mammals obtain RF from exogenous sources via intestinal absorption and prevent its urinary loss by reabsorption in the kidneys. Both of these absorptive events are carrier-mediated and involve specific RF transporters (RFVTs). Chronic alcohol consumption in humans is associated with a high prevalence of RF deficiency and suboptimal levels, but little is known about the effect of chronic alcohol exposure on physiological and molecular parameters of the intestinal and renal RF transport events. We addressed these issues using rats chronically fed an alcohol liquid diet and pair-fed controls as a model. The results showed that chronic alcohol feeding significantly inhibits carrier-mediated RF transport across the intestinal brush-border and basolateral membrane domains of the polarized enterocytes. This inhibition was associated with a parallel reduction in the expression of the rat RFVT-1 and -3 at the protein, mRNA, and heterogeneous nuclear RNA (hnRNA) levels. Chronic alcohol feeding also caused a significant inhibition in RF uptake in the colon. Similarly, a significant inhibition in carrier-mediated RF transport across the renal brush-border and basolateral membrane domains was observed, which again was associated with a significant reduction in the level of expression of RFVT-1 and -3 at the protein, mRNA, and hnRNA levels. These findings demonstrate that chronic alcohol exposure impairs both intestinal absorption and renal reabsorption processes of RF and that these effects are, at least in part, mediated via transcriptional mechanism(s) involving the slc52a1 and slc52a3 genes.


Obesity with insulin resistance increase thrombosis in wild-type and bone marrow-transplanted Zucker Fatty rats.
Obesity induces metabolic and inflammatory alterations that contribute to the presentation of cardiovascular events. Although obesity is a risk factor for atherosclerosis and vascular disease, its role on thrombosis has not been directly explored. Therefore, we aimed to investigate the mechanisms by which obesity affects thrombosis. Thrombus formation was monitored by real-time intravital microscopy in Zucker Fatty rats (ZF) and lean controls (ZC). Crossed bone marrow (BM) transplants between ZF and ZC were performed. Intravital microscopy showed that ZF had significantly shorter occlusion times (OTs) than ZC, reflecting a three-fold higher thrombotic risk. Transplantation of ZC-BM to ZF recipients significantly reduced thrombosis, reducing their thrombotic risk to one third of that observed in non-transplanted ZF. Wild-type ZF showed increased platelet counts and increased platelet size compared to wild-type ZC and platelet number remained unaltered after transplantation. However, ZF-BM produced a significant increase in platelet size in ZC recipients. Thrombotic risk was found to be inversely correlated with both weight and insulin levels and directly correlated to HOMA-IR, while platelet number and size were directly correlated with weight. Thus, our data shows that obesity with insulin resistance significantly increases thrombosis and that alterations in BM-derived cells significantly contribute to this prothrombotic behaviour. Importantly, the reduction of insulin resistance was associated with reduced thrombotic risk even in the presence of obesity.


Pleural effusion in breast cancer. Thoracoscopy for hormone receptor determination.
Metastatic breast cancer frequently presents as a malignant pleural effusion. Knowledge of the estrogen and progesterone receptor status of the tumor predicts response to hormonal therapy, but breast cancer tissue in the pleural space is not readily accessible for hormone receptor determination. Thoracoscopy was used in six breast cancer patients with pleural effusions; all but one had concurrent sites of metastases. In five of six women recurrent breast cancer in the pleural cavity was diagnosed by thoracoscopy, and in four sufficient tissue was obtained for receptor assay. All patients achieved excellent control of their pleural effusions through a combination of local sclerotic measures and systemic therapy. Thoracoscopy is a safe procedure that can be performed under local anesthesia and is useful to visualize the pleural space, not only for diagnosis but also for obtaining breast cancer tissue for hormone receptor determination.


Results of a community-based antiretroviral treatment program for HIV-1 infection in Western Uganda.
To compare the treatment outcomes and mortality in a rural community-based ART (CBART) program with a hospital-based ART program in the same district. The study design was a non-randomized cohort study consisting of 185 persons living with HIV (PLWHIV) in the CBART cohort and 200 PLWHIV in the hospital cohort. Eligibility for both cohorts was: being HIV-infected and eligible for ART, being treatment naïve, age 18 years or older, and being a resident of Rwimi sub-county. The intervention consisted of a community-based program which included weekly home visits to patients by trained volunteers who delivered antiretroviral drugs (ARVs), monitored and supported adherence to treatment, and identified and reported adverse reactions and other clinical symptoms. Outcome variables were compared to patients in a hospital-based cohort who received the standard care delivered to all other HIV patients in the hospital. The main outcome measures were HIV-1 RNA viral load (VL), CD4 cell count and mortality after six months of treatment. Successful ART treatment outcome as measured by virological suppression (VL<400 copies/ml) in the CBART cohort were similar to those in the hospital-based cohort (90.1% vs 89.3%, p=0.47). The median CD4 cell count increased significantly in both cohorts (community-based cohort 159 cells/microl vs 145 cells/microl in the hospital-based cohort). Mortality was not significantly different in both cohorts (community-based cohort 11.9%, hospital-based cohort 9.0%). The findings show that outcomes of a CBART intervention in a rural area compare favorably to outcomes of hospital-based care. If the study results are sustainable over a longer time period, this model could be considered for ART roll-out to impoverished rural/remote populations in Uganda and elsewhere.


Nasal osteotomies: a clinical comparison of the perforating methods versus the continuous technique.
Do perforating lateral osteotomies cause less ecchymosis and edema compared with the popular continuous method? Many studies have demonstrated that perforated osteotomies cause less trauma and periosteal disruption. Numerous investigators have subjectively perceived less postoperative ecchymosis and edema, but no clinical study has compared the perforated methods versus the continuous technique in the same patient. This prospective, randomized, partially blinded study was designed to test the hypothesis that the perforating method causes less postoperative ecchymosis and edema compared with the continuous lateral osteotomy technique. The questions remain: does the type of perforating osteotomy affect the results? Does a percutaneous approach cause more ecchymosis and edema by the access maneuver of piercing the skin? The two perforating lateral nasal osteotomy techniques require the same 2-mm straight osteotome, so any genuine difference in postoperative ecchymosis or edema could only be attributed to the differing surgical approaches. Accordingly, this study also tests whether the external percutaneous perforating osteotomy causes more ecchymosis and edema than the internal transnasal perforating method. Twenty-five consecutive rhinoplasty patients (group A) requiring bilateral osteotomies (50 total lateral osteotomies) were randomized so that each patient received an internal/transnasal perforating lateral osteotomy (2-mm straight chisel) on one side and an internal/transnasal continuous osteotomy (4-mm curved, guarded osteotome) on the other. The next 25 patients studied (group B) received an external/percutaneous perforating lateral osteotomy (same 2-mm straight chisel as used in group A) on one side and the same internal/transnasal continuous osteotomy on the other. The final 25 consecutive rhinoplasty patients (group C) received an external percutaneous perforating lateral osteotomy on one side and an internal transnasal perforating lateral osteotomy on the other. The entry sites for the perforating osteotomies were either external (groups B and C) with a percutaneous skin puncture or intranasal (groups A and C) at the pyriform aperture. All 75 patients (150 total lateral osteotomies) initialed the surgical plan on the Gunter rhinoplasty worksheet, which has been approved by the Institutional Review Board of Abbott-Northwestern Hospital, Minneapolis, Minnesota (study no. 1341-1 M). All patients were evaluated for ecchymosis and edema on the left versus the right side of the face at 2 to 3, 7, and 21 days after the operation. The clinical evaluation was performed by two blinded examiners (clinic registered nurse and the patient with his or her family) and a partially blinded examiner (the surgeon, who did not refresh his memory about the randomization). To compare the two methods in each study (groups A, B, and C) for the six outcomes (edema and ecchymosis at 2 to 3, 7, and 21 days), the authors used an exact binomial test of the null hypothesis that the treatments do not differ. To compare the two methods in each study (groups A, B, and C) using all six outcomes simultaneously, the authors used a permutation test. By both testing methods, the perforating internal method was superior to the continuous technique (group A; p < 0.01 in both tests). Although the perforating external method gave better results than the continuous technique (group B) and the perforating internal method gave better results than the perforating external method (group C), neither of these differences was significant by either testing method. A lateral osteotomy technique should be precise, reproducible, and safe, and it should minimize ecchymosis and edema. Since edema and ecchymosis are comparable regardless of osteotome size, this prospective randomized study confirms the subjective clinical impression that perforating lateral osteotomies with a 2-mm straight osteotome reduce postoperative ecchymosis and edema in rhinoplasty patients compared with the continuous osteotomy (4-mm curved, guarded osteotome). These findings should encourage te the use of perforating osteotomies rather than continuous osteotomies.


Evaluation of intermittent preventive treatment of malaria against group B Streptococcus colonization in pregnant women: a nested analysis of a randomized controlled clinical trial of sulfadoxine/pyrimethamine versus mefloquine.
Streptococcus agalactiae constitutes an important cause of neonatal infections in sub-Saharan Africa. Sulfadoxine/pyrimethamine-the current intermittent preventive treatment of malaria in pregnancy (IPTp)-has proven in vitro activity against group B Streptococcus (GBS). Because of specific drug resistance to sulfadoxine/pyrimethamine, mefloquine-an antimalarial without in vitro activity against GBS-was evaluated as a potential alternative. This study assessed the potential of sulfadoxine/pyrimethamine-IPTp to reduce the prevalence of GBS colonization in pregnant women in Gabon when compared with the inactive control mefloquine-IPTp. Pregnant women participating in a randomized controlled clinical trial evaluating mefloquine-IPTp versus sulfadoxine/pyrimethamine-IPTp were invited to participate and recto-vaginal swabs were collected at delivery for detection of GBS colonization. Prevalence of recto-vaginal GBS colonization was compared between IPTp regimens and risk factor and birth outcome analyses were computed. Among 549 participants, 106 were positive for GBS colonization at delivery (19%; 95% CI = 16%-23%). Prevalence of maternal GBS colonization showed no significant difference between the two IPTp regimens (mefloquine-IPTp: 67 of 366 women = 18%; 95% CI = 14%-22%; sulfadoxine/pyrimethamine-IPTp: 39 of 183 women = 21%; 95% CI = 15%-27%). Risk factor analysis for GBS colonization demonstrated a significant association with illiteracy (adjusted OR = 2.03; 95% CI = 1.25-3.30). GBS colonization had no impact on birth outcome, anaemia at delivery, gestational age and birth weight. Sulfadoxine/pyrimethamine did not reduce colonization rates when used as the IPTp drug during pregnancy. Illiteracy was associated with GBS colonization.


The E6 and E7 genes of human papillomavirus type 6 have weak immortalizing activity in human epithelial cells.
Previous studies have shown that the E7 gene of human papillomavirus (HPV) type 16 or 18 alone was sufficient for immortalization of human foreskin epithelial cells (HFE) and that the efficiency was increased in cooperation with the respective E6 gene, whereas the HPV6 E6 or E7 gene was not active in HFE. To detect weak immortalizing activities of the HPV6 genes, cells were infected with recombinant retroviruses containing HPV genes, alone and in homologous and heterologous combinations. The HPV6 genes, alone or together (HPV6 E6 plus HPV6 E7), were not able to immortalize cells. However the HPV6 E6 gene, in concert with HPV16 E7, increased the frequency of immortalization threefold over that obtained with HPV16 E7 alone. Interestingly, 6 of 20 clones containing the HPV16 E6 gene and the HPV6 E7 gene were immortalized, whereas neither gene alone was sufficient. Thus, the HPV6 E6 and E7 genes have weak immortalizing activities which can be detected in cooperation with the more active transforming genes of HPV16. Acute expression of the HPV6 and HPV16 E6 and E7 genes revealed that only HPV16 E7 was able to stimulate the proliferation of cells in organotypic culture, resulting in increased expression of the proliferative cell nuclear antigen and the formation of a disorganized epithelial layer. Additionally, combinations of genes that immortalized HFE cells (HPV16 E6 plus HPV16 E7, HPV16 E6 plus HPV6 E7, and HPV6 E6 plus HPV16 E7) also stimulated proliferation.


Neglected zoonotic helminthiases in wild canids: new insights from South America.
The global threat of neglected tropical diseases (NTDs) constitutes a public health issue in underdeveloped countries. Zoonotic helminthiases are the most common human NTD agents in developing countries in sub-Saharan Africa, Asia, and the Americas, causing a global burden of disease that exceeds that of more recognized infectious diseases such as malaria and tuberculosis. Wild canids are well-known mammals that act as natural reservoirs of zoonotic-relevant helminthiasis worldwide, thus playing a pivotal role in their epidemiology and transmission to humans. Here we evaluate the occurrence of zoonotic gastrointestinal helminths in two Neotropical wild canid species from the Amazonian and Andean regions of Colombia, i.e., the bush dog (Speothos venaticus) and the crab-eating fox (Cerdocyon thous). We recovered tapeworm proglottids from bush dog fecal samples and identified them molecularly as the canine-specific lineage of Dipylidium caninum by using cytochrome c oxidase subunit I (cox1) gene sequences. Moreover, examination of a crab-eating fox during necropsy revealed the presence of non-embryonated eggs of the neglected nematode Lagochilascaris cf. minor, in addition to eggs and gravid proglottids of the cestode Spirometra mansoni. These findings represent the first report of zoonotic-relevant cestodes, i.e., D. caninum ("canine genotype"), S. mansoni, and the nematode L. cf. minor, in bush dogs and crab-eating foxes as final hosts. The occurrence of these zoonotic helminthiases in wild canid species calls for regular monitoring programs to better understand the epidemiology and transmission routes of neglected dipylidiasis, lagochilascariosis, and sparganosis in South America.


State mandatory overtime regulations and newly licensed nurses' mandatory and voluntary overtime and total work hours.
Nurse overtime has been used to handle normal variations in patient census and to control chronic understaffing. By 2010, 16 states had regulations to limit nurse overtime. We examined mandatory overtime regulations and their association with mandatory and voluntary overtime and total hours worked by newly licensed registered nurses (NLRNs). For this secondary data analysis, we used a panel survey of NLRNs; the final dataset consisted of 1,706 NLRNs. Nurses working in states that instituted overtime regulations after 2003 or in states that restricted any type of mandatory overtime had a lower probability of experiencing mandatory overtime than those nurses working in states without regulations. Nurses who worked in states with mandatory overtime regulations reported fewer total hours worked per week. The findings of this study provided insight into how mandatory overtime regulations were related to nurse mandatory and voluntary overtime and the total number of hours worked. Future research should investigate institutions' compliance with regulations and the impact of regulations on nurse and patient outcomes.


Detection of swine caliciviruses by indirect immunofluorescence.
The indirect immunofluorescence test is a rapid method for detecting the presence of vesicular exanthema of swine virus or San Miguel sea lion virus in cell culture. A serological relationship exists between vesicular exanthema of swine virus and San Miguel sea lion virus, as shown by the fluorescence-positive reactions between swine antisera to vesicular exanthema of swine virus A48 and San Miguel sea lion virus type 5 and cell cultures infected with San Miguel sea lion virus types 1, 2, 3, 4 and 5 as well as vesicular exanthema of swine virus B51, C52, D53, E54, F55, G55, H55, I55, J56 and K55. The indirect immunofluorescence test detects group-specific antibody to caliciviruses in swine sera.


Strong differentiation between amphibian communities on two adjacent mountains in the Upper Rio Pastaza watershed of Ecuador, with descriptions of two new species of terrestrial frogs.
We present the results of herpetological surveys in two adjacent mountains where the EcoMinga Foundation protects the cloud forest in the Upper Rio Pastaza watershed, in the Llanganates Sangay Ecological Corridor in Ecuador. A rapid assessment of the amphibian communities of the study sites reveals a diverse and heterogeneous composition, dominated by terrestrial frogs from the genus Pristimantis. We also identify a cryptic diversity with a significant number of candidate new species. We describe two new species of terrestrial frogs of the genus Pristimantis. Pristimantismaryanneae sp. nov. is characterised by not having tympanum externally visible and having 2-3 subconical tubercles in the upper eyelid; and Pristimantisburtoniorum sp. nov. is characterised by the presence of red colouration in hidden surfaces of the hind-limbs, tubercles on the upper eyelid, interorbital tubercle and a row of rounded tubercles along the snout to the tip and a pale red venter with dark brown mottled pattern. Our samples from the two Reserves do not share species between them, so the proportion of shared species seems to be relatively low. In addition, we highlight the importance of updating the knowledge of amphibians that are restricted to this important conservation region and comment about the threats and composition of the amphibian communities on the eastern slopes of the Upper Rio Pastaza watershed.


Hepatobiliary-specific MR contrast agents: role in imaging the liver and biliary tree.
Hepatobiliary-specific contrast agents are one of several classes of contrast agents available for magnetic resonance (MR) imaging of the liver. These agents are taken up by functioning hepatocytes and excreted in the bile, and their paramagnetic properties cause shortening of the longitudinal relaxation time (T1) of the liver and biliary tree. The three contrast agents that have been developed are mangafodipir trisodium (Mn-DPDP), gadobenate dimeglumine (Gd-BOPTA), and gadoxetic acid (Gd-EOB-DTPA). These three MR contrast agents vary in mode of administration and dose, mechanism of cellular uptake, degree of excretion through the biliary pathway, and imaging characteristics. In the liver, hepatobiliary-specific agents can be used to improve lesion detection, to characterize lesions as hepatocellular or nonhepatocellular, and to specifically characterize some hepatocellular lesions, notably focal nodular hyperplasia. Biliary excretion of these agents can be used to evaluate the anatomic structure and function of the biliary tree. In the future, hepatobiliary-specific contrast agents may have wider applications, such as grading of cirrhosis and quantification of liver function.


[Monoclonal antibodies in the diagnosis and therapy of breast carcinoma: current status and future prospects].
Hybridoma technology has now advanced to the point where hundreds of monoclonal antibodies have been produced and are reported to bind to normal and malignant cell surfaces or intracellular organelles. The original hope for tumor-specific antigens has not materialized, but certain antibodies with rather restricted ranges of binding activities have been identified. For the first time, one now has tools other than morphology or the presence of hormone receptors to distinguish between two different tumors, or among different cell populations within a given tumor mass. New techniques including gene cloning and in situ hybridization will undoubtedly provide insights in the field of tumor biology and immunology, and hopefully, will foster developments toward the mechanisms of oncogenesis as well as toward more immediate clinical applications.


The action logic of the older adults about health-seeking in South Rural China.
The Chinese government has invested significant resources to build many rural healthcare stations. However, in the face of convenient medical paths and accessible medical resources, the utilization rate of health services for older adults in rural areas is surprisingly low. This study explored why health-seeking behavior among older adults in rural China was not active. Data were collected through participatory rural appraisal (PRA) with 108 participants in 12 villages in southern China. Daily schedule and social and resource mapping were employed to outline the range of activities and the routine of the older adults, as well as in-depth interviews to understand the logic of their healthcare choices. Data collected were analyzed by content analysis. Three themes were generated: (1) perceptions of health status (being healthy or sick): the rural older adults used the ability to handle routine chores as a measure of health status; (2) prioritization of solving symptoms over curing diseases: the older adults preferred the informal self-medication to cope with diseases, as long as there were no symptoms and no pain; (3) 'unpredictable' troubles: they tended to favor the 'optimal' solution of keeping their lives in order rather than the best medical treatment options. This study showed that the medical practices of the rural elderly were profoundly influenced by their perceptions of health and their life experiences. In the face of diseases, they tended to keep their lives in order, preferring self-treatment practices that address symptoms or selectively following medical advice rather than medical and science-based clinical solutions. In the future, the construction of rural health care should focus on changing the 'inaccessibility' of healthcare resources at the subjective level of the rural elderly and develop culturally adaptable health education.


Effects of room temperature on reproduction, body and organ weights, food and water intake, and hematology in rats.
Two generations of rats were raised at animal room temperature ranging from 12 to 32 degrees C at steps of 2 degrees C. The body weight of rats born in 22 degrees C environment and exposed to each temperature did not significantly differ within the range of 16-28 degrees C. The delivery rate, litter size, and weaning rate decreased at 30 degrees C and 32 degrees C, and only the weaning rate had a tendency to decrease at 12 degrees C. No significant difference was demonstrated within the 14-28 degrees C range for any reproduction parameter observed. The body weight of sucklings did not differ at birth within the 12-32 degrees C range; and at 3 weeks of age, there were no differences within the range of 18-28 degrees C. The body weight gain in both sexes after weaning was generally small when the temperature was below 18 degrees C or above 30 degrees C. There were no significant differences in food intake within the 20-26 degrees C range, in water intake within the 12-26 degrees C range, in hematological and serum biochemical values within the 20-26 degrees C range, and in organ weights within the 18-28 degrees C range. Therefore, the range of 20-26 degrees C (68-78 degrees F) was the optimum temperature range in rat rooms.


[Corrective effect of olmesartan on parameters of the local rigidity using the echo-tracking technology].
Purpose of this study was to determine clinical value of intima-media thickness (IMT) and indicators of local stiffness obtained by ultrasonography of the carotid arteries by echo-tracking technology in patients with hypertension and coronary artery disease (CAD), and to assess vasoprotective effect of 24-week treatment with olmesartan. We included into the study 22 patients with 1-2 degree hypertension, 25 patients with CAD combined with hypertension, and 44 healthy subjects. In patients with CAD and hypertension local stiffness was reassessed after 24-week treatment with olmesartan (20-40 mg/day). Patients with-hypertension compared with healthy subjects had higher values of loc P sys and loc P dia, P (T1), pulse wave propagation velocity (PWPV), stiffness index, as well as lower compliance and distensibility coefficients (CC and DC). In patients with CAD and hypertension all parameters of local stiffness and IMT were significantly higher, while CC and DC were significantly lower than in control group. Therapy with olmesartan for 24-weeks was associated with reductions of IMT (-4.8%, p < 0.05), loc P sys and loc P dia (-13.5 and -11%, respectively, p < 0.05), stiffness index β (-19.1%, p < 0.05), loc PWPV (-17%) and P (T1) (-7%, p < 0.05), and increase of CC (3.3%, p < 0.05). Conclusion. Combination of CAD and hypertension worsened the local stiffness and increased IMT. Therapy with olmesartan in this group of patients was associated with indications of vasoprotective effect.


Transcranial direct current stimulation alleviates seizure severity in kainic acid-induced status epilepticus rats.
Status epilepticus (SE) is a state of prolonged and repeated seizures that can lead to permanent brain damage or life-threatening conditions. Transcranial direct current stimulation (tDCS) non-invasively provides a polarity-specific electric current to modulate brain excitability. Little is known about the therapeutic potential of tDCS in SE. Here, we aim to determine the tDCS effects on seizure severity, EEG and post-SE consequences in rats with kainic acid (KA)-induced SE. Rats were subjected to cathodal tDCS or sham stimulation over the dorsal hippocampus for 5 days. KA was intraperitoneally injected to induce SE. We used continuous video-EEG recording to monitor seizure activity, immunostaining and Timm staining to evaluate neuron counts and mossy fiber sprouting, and ELISA for Brain-derived neurotrophic factor (BDNF) protein measurement. Two featured EEG patterns, gamma ranged high-frequency polyspikes and low-frequency spike-and-wave complexes, were identified in the hippocampal CA1 of KA-induced SE rats. tDCS elicited a significant decrease in severe seizures of Racine stages 4-5 in KA-induced SE rats. tDCS-treated rats manifested diminished high-frequency oscillation during SE, decreased chronic spontaneous spike activities and mossy fiber sproutings compared to sham. tDCS-treated rats also exhibited significantly lower hippocampal BDNF protein levels than sham immediately and 4 weeks after SE. A positive correlation between the hippocampal BDNF level and the seizure severity of SE was found. Altogether, our results show that repeated cathodal tDCS can mitigate seizure severity, alter ictal EEG pattern and reduce the chronic adverse consequences in KA-induced SE rats, supporting the therapeutic potential of tDCS in severe prolonged epileptic seizures.


Specificity of transfer in basic and applied perceptual-motor tasks.
We conducted research on transfer of skills using basic stimulus-response compatibility tasks and applied tasks requiring control of a hydraulic excavator simulator. The basic tasks show rapid acquisition of practiced spatial mappings, for which transfer is specific to the procedures used in training. The applied tasks show transfer across alternative control configurations that maintain practiced spatial mappings, as well as from part to whole practice. Transfer from simulated to real equipment also seems to occur; however, studies involving cooperation of academia and industry are needed to provide more definitive evidence on this question.


Curative surgery for adenocarcinoma of the pancreas/ampulla of Vater: the role of adjuvant pre or postoperative radiation therapy.
From 1972 to 1981, 7 patients received adjuvant external beam radiation therapy before (5 patients) or after (2 patients) a curative Whipple operation for adenocarcinoma of the pancreas (5 patients) or ampulla of Vater (2 patients). Via supervoltage, 4000-4520 rad/20-25 fractions/4-5 1/2 weeks were delivered to the tumor bed and draining lymphatics. Two patients (of 4 at risk) are 5-year survivors; 3 are alive-without-disease at 1-2 years follow-up. Distant metastases have developed to date in 3 patients. A marginal miss occurred at the edge of radiation portal in 1 patient, and no true in-field failures have occurred. Complications occurred in 2 patients. Because of the high incidence of local-regional failures after curative surgery alone for adenocarcinoma of the pancreas or ampulla of Vater, the above results suggest that adjuvant radiation therapy should be considered as a modality to potentially improve treatment results.


Male partner participation in maternity care and social support for childbearing women: a discussion paper.
Male partners/fathers are key support persons for many childbearing women and their involvement in pregnancy, childbirth and the postpartum/postnatal period has beneficial effects on a wide range of outcomes related to maternal and child health and family wellbeing. Social support is implicated in the relevant causal pathway, but has received largely tangential attention in the public health literature. This discussion paper aims to reframe men's participation in maternity care as an opportunity to enhance their readiness and ability to provide social support to women, contributing to the debate on the definition and rationale for male partner involvement, and paving the way for further empirical work. I begin by presenting a theory of change illustrating the causal pathway leading from male partner participation, through the key intermediate step of social support, to improved health and wellbeing for women and children. I proceed by arguing that many people desire male partner participation in maternity care; however, in practice, this is often limited owing to cultural, social and institutional barriers. I use examples from the intervention literature to demonstrate how participation in care can boost men's motivation to support women and enhance their ability to do so by increasing their knowledge and skills. Finally, I draw up general implications for further male partner involvement programmes, suggesting that in order to achieve meaningful and sustainable gains, attention to design is crucial in order to avoid reinforcing patriarchal gender norms. Programmes should be implemented alongside other efforts to improve quality and promote woman-centred care. This article is part of the theme issue 'Multidisciplinary perspectives on social support and maternal-child health'.


Onset of immune senescence defined by unbiased pyrosequencing of human immunoglobulin mRNA repertoires.
The immune system protects us from foreign substances or pathogens by generating specific antibodies. The variety of immunoglobulin (Ig) paratopes for antigen recognition is a result of the V(D)J rearrangement mechanism, while a fast and efficient immune response is mediated by specific immunoglobulin isotypes obtained through class switch recombination (CSR). To get a better understanding on how antibody-based immune protection works and how it changes with age, the interdependency between these two parameters need to be addressed. Here, we have performed an in depth analysis of antibody repertoires of 14 healthy donors representing different gender and age groups. For this task, we developed a unique pyrosequencing approach, which is able to monitor the expression levels of all immunoglobulin V(D)J recombinations of all isotypes including subtypes in an unbiased and quantitative manner. Our results show that donors have individual immunoglobulin repertoires and cannot be clustered according to V(D)J recombination patterns, neither by age nor gender. However, after incorporating isotype-specific analysis and considering CSR information into hierarchical clustering the situation changes. For the first time the donors cluster according to age and separate into young adults and elderly donors (>50). As a direct consequence, this clustering defines the onset of immune senescence at the age of fifty and beyond. The observed age-dependent reduction of CSR ability proposes a feasible explanation why reduced efficacy of vaccination is seen in the elderly and implies that novel vaccine strategies for the elderly should include the "Golden Agers".


VDAC1 is a transplasma membrane NADH-ferricyanide reductase.
Porin isoform 1 or VDAC (voltage-dependent anion-selective channel) 1 is the predominant protein in the outer mitochondrial membrane. We demonstrated previously that a plasma membrane NADH-ferricyanide reductase activity becomes up-regulated upon mitochondrial perturbation, and therefore suggested that it functions as a cellular redox sensor. VDAC1 is known to be expressed in the plasma membrane; however, its function there remained a mystery. Here we show that VDAC1, when expressed in the plasma membrane, functions as a NADH-ferricyanide reductase. VDAC1 preparations purified from both plasma membrane and mitochondria fractions exhibit NADH-ferricyanide reductase activity, which can be immunoprecipitated with poly- and monoclonal antibodies directed against VDAC(1). Transfecting cells with pl-VDAC1-GFP, which carries an N-terminal signal peptide, directs VDAC1 to the plasma membrane, as shown by confocal microscopy and FACS analysis, and significantly increases the plasma membrane NADH-ferricyanide reductase activity of the transfected cells. This novel enzymatic activity of the well known VDAC1 molecule may provide an explanation for its role in the plasma membrane. Our data suggest that a major function of VDAC1 in the plasma membrane is that of a NADH(-ferricyanide) reductase that may be involved in the maintenance of cellular redox homeostasis.


Association between urine osmolality and specific gravity in dogs and the effect of commonly measured urine solutes on that association.
To determine the association between urine osmolality and specific gravity (USG) in dogs and to evaluate the effect of commonly measured urine solutes on that association. 60 dogs evaluated by an internal medicine service. From each dog, urine was obtained by cystocentesis and USG was determined with a refractometer. The sample was divided, and one aliquot was sent to a diagnostic laboratory for urinalysis and the other was frozen at -80°C until osmolality was determined. Urine samples were thawed and osmolality was measured in duplicate with a freezing-point depression osmometer. The correlation between mean urine osmolality and USG was determined; the effect of pH, proteinuria, glucosuria, ketonuria, bilirubinuria, and hemoglobinuria on this relationship was investigated with multiple regression analysis. The Pearson correlation coefficient between urine osmolality and USG was 0.87. The final multivariable regression model for urine osmolality included USG and the presence of ketones; ketonuria had a small negative association with urine osmolality. Results indicated a strong linear correlation between osmolality and USG in urine samples obtained from dogs with various pathological conditions, and ketonuria had a small negative effect on that correlation.


Associations of neighborhood characteristics with sleep timing and quality: the Multi-Ethnic Study Of Atherosclerosis.
To investigate the associations of specific neighborhood features (disorder, safety, social cohesion, physical environment, and socioeconomic status) with sleep duration and quality. Cross-sectional. One wave of a population-based study (Multi-Ethnic Study of Atherosclerosis). Community-dwelling participants in New York, NY and Los Angeles, CA. There were 1,406 participants (636 males, 770 females). NA. Sleep was assessed using reported hours of sleep, the Epworth Sleepiness Scale, and insomnia symptoms. Neighborhood characteristics were assessed via questionnaires administered to neighbors of study participants and were aggregated to the neighborhood (census tract) level using empirical Bayes estimation. An adverse social environment (characterized by high disorder, and low safety and social cohesion) was associated with shorter sleep duration after adjustment for the physical environment, neighborhood and individual-level socioeconomic status (SES), and other short sleep risk factors (mean difference per standard deviation increase in summary social environment scale 0.24 h 95% confidence interval 0.08, 0.43). Adverse neighborhood social and physical environments, and neighborhood SES were associated with greater sleepiness, but associations with physical environments were no longer statistically significant after adjustment for sociodemographic characteristics. Neighborhood SES was a weaker and less consistent predictor of specific measures of neighborhood social and physical environments. Neighborhood characteristics were not associated with insomnia. Shortened sleep related to adverse social environments represents one potential pathway through which neighborhoods may influence health.


Tallimustine lesions in cellular DNA are AT sequence-specific but not region-specific.
Tallimustine (FCE 24517) is an AT-specific alkylating antitumor derivative of distamycin. This study examined levels of tallimustine lesions in intracellular DNA, their sequence- and region-specificity, and the long-range distribution of the drug binding motif. Tallimustine adducts in DNA converted to strand breaks by heating allowed the quantitation of drug lesions. In bulk DNA of intact human leukemia CEM cells, tallimustine formed 0.15 +/- 0.04 and 0.64 +/- 0.18 lesions/kbp at 5 and 50 microM, respectively. These lesions represent monoadducts as no interstrand cross-links or DNA-protein cross-links were detected. Tallimustine adducts in intracellularly treated DNA showed a general preference for sequences with T-tracts, suggesting a propensity for intrinsically bent motifs. Major drug-adducted sites identified by repetitive primer extension, included 5'-TTTTGPu-3' and 5'-TTTTGC-3' motif. Despite the high specificity at the nucleotide level, tallimustine did not differentiate among bulk DNA and three discrete AT-rich regions of genomic DNA examined by quantitative PCR stop assay with lesion frequencies ranging from 0.23 to 0.39 lesions/kbp at 25 microM drug. In comparisons of lesion frequencies and cytotoxicity, tallimustine adducts are approximately 50 times more lethal than relatively nonsequence specific cisplatin adducts but are >100 times less lethal than lesions by an unrelated AT-specific drug, bizelesin. However, the 5'-TTTTGPu-3' motifs targeted by tallimustine are relatively infrequent and scattered throughout the genome. In contrast, the motifs 5'-T(A/T)(4)A-3' motifs targeted by bizelesin, while also infrequent, cluster in defined AT-rich islands. The lack of region-specificity may be the reason tallimustine adducts, despite high AT-specificity at the nucleotide level, are less lethal than region-specific bizelesin adducts.