The Drosophila copper transporter Ctr1C functions in male fertility.
Living organisms have evolved intricate systems to harvest trace elements from the environment, to control their intracellular levels, and to ensure adequate delivery to the various organs and cellular compartments. Copper is one of these trace elements. It is at the same time essential for life but also highly toxic, not least because it facilitates the generation of reactive oxygen species. In mammals, copper uptake in the intestine and copper delivery into other organs are mediated by the copper importer Ctr1. Drosophila has three Ctr1 homologs: Ctr1A, Ctr1B, and Ctr1C. Earlier work has shown that Ctr1A is an essential gene that is ubiquitously expressed throughout development, whereas Ctr1B is responsible for efficient copper uptake in the intestine. Here, we characterize the function of Ctr1C and show that it functions as a copper importer in the male germline, specifically in maturing spermatocytes and mature sperm. We further demonstrate that loss of Ctr1C in a Ctr1B mutant background results in progressive loss of male fertility that can be rescued by copper supplementation to the food. These findings hint at a link between copper and male fertility, which might also explain the high Ctr1 expression in mature mammalian spermatozoa. In both mammals and Drosophila, the X chromosome is known to be inactivated in the male germline. In accordance with such a scenario, we provide evidence that in Drosophila, the autosomal Ctr1C gene originated as a retrogene copy of the X-linked Ctr1A, thus maintaining copper delivery during male spermatogenesis.


Alpha-synuclein dysfunction in Lewy body diseases.
alpha-Synuclein belongs to a small group of natively unfolded proteins that can transiently bind to lipid membranes and acquire a partial alpha-helical conformation. Its relevance to Parkinson's disease (PD) is based on mutations found in familial cases of the disease and its presence in filaments of Lewy bodies (LB) and Lewy neurites (LN) in sporadic cases where it is packed in a beta-sheet configuration. This structural plasticity of alpha-synuclein has raised the possibility that neurodegeneration may be a consequence of abnormal protein folding. The extent to which abnormal folding and aggregation of neuronal proteins is directly toxic to the cell, an inert biochemical marker of an underlying harmful metabolic defect, or a protective reaction remains to be seen. We review the function of alpha-synuclein and recent studies that have shed light on the mechanisms by which it aggregates.


Nurses' work environments, care rationing, job outcomes, and quality of care on neonatal units.
This paper is a report of a study of the relationship between work environment characteristics and neonatal intensive care unit nurses' perceptions of care rationing, job outcomes, and quality of care. International evidence suggests that attention to work environments might improve nurse recruitment and retention, and the quality of care. However, comparatively little attention has been given to neonatal care, a specialty where patient and nurse outcomes are potentially quite sensitive to problems with staffing and work environments. Over a 6-month period in 2007-2008, a questionnaire containing measures of work environment characteristics, nursing care rationing, job satisfaction, burnout and quality of care was distributed to 553 nurses in all neonatal intensive care units in the province of Quebec (Canada). A total of 339 nurses (61.3%) completed questionnaires. Overall, 18.6% were dissatisfied with their job, 35.7% showed high emotional exhaustion, and 19.2% rated the quality of care on their unit as fair or poor. Care activities most frequently rationed because of insufficient time were discharge planning, parental support and teaching, and comfort care. In multivariate analyses, higher work environment ratings were related to lower likelihood of reporting rationing and burnout, and better ratings of quality of care and job satisfaction. Additional research on the determinants of nurse outcomes, the quality of patient care, and the impact of rationing of nursing care on patient outcomes in neonatal intensive care units is required. The Neonatal Extent of Work Rationing Instrument appears to be a useful tool for monitoring the extent of rationing of nursing care in neonatal units.


Experimental and modeling studies of desensitization of P2X3 receptors.
The function of ATP-activated P2X3 receptors involved in pain sensation is modulated by desensitization, a phenomenon poorly understood. The present study used patch-clamp recording from cultured rat or mouse sensory neurons and kinetic modeling to clarify the properties of P2X3 receptor desensitization. Two types of desensitization were observed, a fast process (t1/2 = 50 ms; 10 microM ATP) following the inward current evoked by micromolar agonist concentrations, and a slow process (t1/2 = 35 s; 10 nM ATP) that inhibited receptors without activating them. We termed the latter high-affinity desensitization (HAD). Recovery from fast desensitization or HAD was slow and agonist-dependent. When comparing several agonists, there was analogous ranking order for agonist potency, rate of desensitization and HAD effectiveness, with 2-methylthioadenosine triphosphate the strongest and beta,gamma-methylene-ATP the weakest. HAD was less developed with recombinant (ATP IC50 = 390 nM) than native P2X3 receptors (IC50 = 2.3 nM). HAD could also be induced by nanomolar ATP when receptors seemed to be nondesensitized, indicating that resting receptors could express high-affinity binding sites. Desensitization properties were well accounted for by a cyclic model in which receptors could be desensitized from either open or closed states. Recovery was assumed to be a multistate process with distinct kinetics dependent on the agonist-dependent dissociation rate from desensitized receptors. Thus, the combination of agonist-specific mechanisms such as desensitization onset, HAD, and resensitization could shape responsiveness of sensory neurons to P2X3 receptor agonists. By using subthreshold concentrations of an HAD-potent agonist, it might be possible to generate sustained inhibition of P2X3 receptors for controlling chronic pain.


A novel recursive Bayesian learning-based method for the efficient and accurate segmentation of video with dynamic background.
Segmentation of video with dynamic background is an important research topic in image analysis and computer vision domains. In this paper, we present a novel recursive Bayesian learning-based method for the efficient and accurate segmentation of video with dynamic background. In the algorithm, each frame pixel is represented as the layered normal distributions which correspond to different background contents in the scene. The layers are associated with a confident term and only the layers satisfy the given confidence which will be updated via the recursive Bayesian estimation. This makes learning of background motion trajectories more accurate and efficient. To improve the segmentation quality, the coarse foreground is obtained via simple background subtraction first. Then, a local texture correlation operator is introduced to fill the vacancies and remove the fractional false foreground regions. Extensive experiments on a variety of public video datasets and comparisons with some classical and recent algorithms are used to demonstrate its improvements in both segmentation accuracy and efficiency.


The Mid- to Long-Term Outcomes of the Lateral Domed Oxford Unicompartmental Knee Replacement: An Analysis From the National Joint Registry for England, Wales, Northern Ireland, and the Isle of Man.
Lateral unicompartmental knee replacement (UKR) is an alternative to total knee replacement for isolated lateral unicompartmental knee arthritis. The geometry and mechanics of the lateral compartment differ to the medial compartment with the Lateral Domed Oxford UKR designed to address this. We used the National Joint Registry (NJR) data to report the mid- to long-term outcomes of this device. We performed a retrospective observational study using NJR data on 992 Lateral Domed Oxford UKRs implanted between 1st January 2005 and 31st December 2017. Outcomes of interest were implant survival and revision indications. The 10 year cumulative implant survival rates were 88.6% (CI 85.3-91.2). When compared with <55 year age group, the 55-64, 65-74 and ≥75 groups had significantly lower revision rates (hazard ratio (HR) = 0.56 (CI 0.32-0.98, P = .04), HR 0.40 (CI 0.22-0.72, P = .003), and HR 0.27 (CI 0.12-0.58, P = .001), respectively). The obese group had significantly (P = .04) increased revision risk compared with normal BMI (HR 2.33, CI 1.06-5.12). The commonest reasons for revision surgery were dislocation (n = 23, 2.3%), pain (n = 15, 1.5%), and aseptic loosening (n = 14, 1.4%). The Lateral Domed Oxford UKR provides a good option for isolated lateral compartment osteoarthritis. However, dislocation of the mobile bearing remains a problem, occurring in 2.3% of the patients and accounting for 30% of the revisions. To help prevent dislocation, it is now possible to assess bearing stability intraoperatively and if very unstable to implant a compatible fixed bearing tibial component, without the need for further bone preparation.


Establishment of a real-time, quantitative, and reproducible mouse model of Staphylococcus osteomyelitis using bioluminescence imaging.
Osteomyelitis remains a serious problem in the orthopedic field. There are only a few animal models in which the quantity and distribution of bacteria can be reproducibly traced. Here, we established a real-time quantitative mouse model of osteomyelitis using bioluminescence imaging (BLI) without sacrificing the animals. A bioluminescent strain of Staphylococcus aureus was inoculated into the femurs of mice. The bacterial photon intensity (PI) was then sequentially measured by BLI. Serological and histological analyses of the mice were performed. The mean PI peaked at 3 days, and stable signals were maintained for over 3 months after inoculation. The serum levels of interleukin-6, interleukin-1β, and C-reactive protein were significantly higher in the infected mice than in the control mice on day 7. The serum monocyte chemotactic protein 1 level was also significantly higher in the infected group at 12 h than in the control group. A significantly higher proportion of granulocytes was detected in the peripheral blood of the infected group after day 7. Additionally, both acute and chronic histological manifestations were observed in the infected group. This model is useful for elucidating the pathophysiology of both acute and chronic osteomyelitis and to assess the effects of novel antibiotics or antibacterial implants.


CLV3 is localized to the extracellular space, where it activates the Arabidopsis CLAVATA stem cell signaling pathway.
Plant growth and development depends on the activity of a continuously replenished pool of stem cells within the shoot apical meristem to supply cells for organogenesis. In Arabidopsis, the stem cell-specific protein CLAVATA3 (CLV3) acts cell nonautonomously to restrict the size of the stem cell population, but the hypothesis that CLV3 acts as an extracellular signaling molecule has not been tested. We used genetic and immunological assays to show that CLV3 localizes to the apoplast and that export to the extracellular space is required for its function in activating the CLV1/CLV2 receptor complex. Apoplastic localization allows CLV3 to signal from the stem cell population to the organizing center in the underlying cells.


Long lives for homozygous trembler mutant mice despite virtual absence of peripheral nerve myelin.
Nervous system functions are dependent on point-to-point communication of signals along neuronal axons, and axonal insulation by myelin is thought to speed such conduction. Loss of previously formed myelin or lack of myelin formation can have serious, even fatal, consequences. Mice homozygous for the trembler mutation make virtually no peripheral nervous system myelin, yet have long and functional lives. This result calls into question the view that peripheral nervous system myelin plays a vital role, at least in this species.


Atypical Chikungunya virus infections: clinical manifestations, mortality and risk factors for severe disease during the 2005-2006 outbreak on Réunion.
In April 2005, an outbreak of Chikungunya fever occurred on the island of Réunion in the Indian Ocean. During winter 2005, six patients developed meningoencephalitis and acute hepatitis due to Chikungunya virus. Our objectives were to determine the incidence and mortality of atypical Chikungunya viral infections and to identify risk factors for severe disease. A hospital-based surveillance system was established to collect data on atypical Chikungunya cases. Case reports, medical records and laboratory results were reviewed and analysed. We defined an atypical case as one in which a patient with laboratory-confirmed Chikungunya virus infection developed symptoms other than fever and arthralgia. We defined a severe atypical case as one which required maintenance of at least one vital function. We recorded 610 atypical cases of Chikungunya fever: 222 were severe cases, 65 affected patients died. Five hundred and forty-six cases had underlying medical conditions (of which 226 suffered from cardiovascular, 147 from neurological and 150 from respiratory disorders). Clinical features that had never been associated with Chikungunya fever were recorded, such as bullous dermatosis, pneumonia, and diabetes mellitus. Hypertension, and underlying respiratory or cardiological conditions were independent risk factors for disease severity. The overall mortality rate was 10.6% and it increased with age. This is the first time that severe cases and deaths due to Chikungunya fever have been documented. The information presented in this article may assist clinicians in identifying the disease, selecting the treatment strategy, and anticipating the course of illness.


First Report of Persian Buttercup Downy Mildew Caused by Peronospora sp. in Korea.
Persian buttercup (Ranunculus asiaticus L.) is an ornamental plant cultivated mainly in the countries surrounding the Mediterranean Sea, and has recently become popular in Korea. During March and April 2012, Persian buttercups 'Elegance' showing symptoms of downy mildew were found in plastic greenhouses in Hwaseong City of Korea. Infection resulted in chlorotic leaves with a dark greyish and dense fungal-like growth on the lower surfaces, and finally led to necrosis of the lesions. A sample was deposited in the Korea University herbarium (KUS-F26431). Conidiophores emerging from stomata were hyaline, 250 to 550 × 7 to 15 μm, straight, and dichotomously branched in 6 to 8 orders. Ultimate branchlets were mostly in pairs, slightly curved, 5 to 15 μm long, and had obtuse tips. Conidia were brown, broadly ellipsoidal to subglobose or ellipsoidal, often pedicellated, and measured 24 to 33 × 20 to 27 μm with a length/width ratio of 1.15 to 1.30. Fourteen species of Peronospora have previously been described on the genus Ranunculus (2), of which P. ficariae was mostly considered the causal agent of downy mildew on Persian buttercup (1,3). The present Korean accession is morphologically distinct from P. ficariae on R. ficaria (a synonym of Ficaria verna) by somewhat larger conidia with often pedicel-like ends. The nuclear ribosomal LSU and ITS regions were PCR-amplified and sequenced as described in Göker et al. (4), and the resulting sequences deposited in GenBank (Accession Nos. KC111207 and JX465737, respectively). A comparison with the GenBank sequences revealed that the present Korean pathogen differed from P. ficariae on R. ficaria at 10 of 688 characters (about 1.5%) in LSU (AF119600) and 11 of 802 characters (about 1.4%) in ITS sequences (unpublished sequence). In addition, the ITS sequence exhibits a dissimilarity of 1.5 to 2.0% from three species of Peronospora parasitic on Ranunculus; P. alpicola on R. aconitifolius (AY198271), P. illyrica on R. illyricus (AY198268), and P. ranunculi on R. acris (AY198267) and R. recurvatus (AY198269). Based on morphological and molecular distinction between P. ficariae and the Korean pathogen, we provisionally indicate this pathogen as an undetermined species of Peronospora. Pathogenicity was demonstrated by shaking diseased leaves onto the leaves of healthy Persian buttercup 'Elegance', incubating the plants in a dew chamber at 20°C for 24 h, and then maintaining them in a greenhouse (20 to 24°C and relative humidity 60 to 80%). After 3 to 4 days, inoculated plants developed downy mildew symptoms, from which an identical fungus was observed, thus fulfilling Koch's postulates. Control plants treated with sterile water did not develop any symptoms of downy mildew. To our knowledge, this is the first report of a downy mildew on Persian buttercup in Asia, although this disease has been found in other continental countries, such as Italy (1), New Zealand, South Africa, and the United States (3). The presence of a downy mildew on Persian buttercup in Asia can be considered as a potentially new and serious threat to commercial production of this ornamental plant. References: (1) E. Buonocore and R. Areddia. Informatore Fitopatologico 49:25, 1999. (2) O. Constantinescu. Thunbergia 15:1, 1991. (3) D. F. Farr and A. Y. Rossman. Fungal Databases, Syst. Mycol. Microbiol. Lab., Online publication, ARS, USDA, Retrieved August 4, 2012. (4) M. Göker et al. Mycol. Res. 113:308, 2009.


Genes affected by mouse mammary tumor virus (MMTV) proviral insertions in mouse mammary tumors are deregulated or mutated in primary human mammary tumors.
The accumulation of mutations is a contributing factor in the initiation of premalignant mammary lesions and their progression to malignancy and metastasis. We have used a mouse model in which the carcinogen is the mouse mammary tumor virus (MMTV) which induces clonal premalignant mammary lesions and malignant mammary tumors by insertional mutagenesis. Identification of the genes and signaling pathways affected in MMTV-induced mouse mammary lesions provides a rationale for determining whether genetic alteration of the human orthologues of these genes/pathways may contribute to human breast carcinogenesis. A high-throughput platform for inverse PCR to identify MMTV-host junction fragments and their nucleotide sequences in a large panel of MMTV-induced lesions was developed. Validation of the genes affected by MMTV-insertion was carried out by microarray analysis. Common integration site (CIS) means that the gene was altered by an MMTV proviral insertion in at least two independent lesions arising in different hosts. Three of the new genes identified as CIS for MMTV were assayed for their capability to confer on HC11 mouse mammary epithelial cells the ability for invasion, anchorage independent growth and tumor development in nude mice. Analysis of MMTV induced mammary premalignant hyperplastic outgrowth (HOG) lines and mammary tumors led to the identification of CIS restricted to 35 loci. Within these loci members of the Wnt, Fgf and Rspo gene families plus two linked genes (Npm3 and Ddn) were frequently activated in tumors induced by MMTV. A second group of 15 CIS occur at a low frequency (2-5 observations) in mammary HOGs or tumors. In this latter group the expression of either Phf19 or Sdc2 was shown to increase HC11 cells invasion capability. Foxl1 expression conferred on HC11 cells the capability for anchorage-independent colony formation in soft agar and tumor development in nude mice. The published transcriptome and nucleotide sequence analysis of gene expression in primary human breast tumors was interrogated. Twenty of the human orthologues of MMTV CIS associated genes are deregulated and/or mutated in human breast tumors.


Three new cases of late-onset cblC defect and review of the literature illustrating when to consider inborn errors of metabolism beyond infancy.
The cblC defect is a rare inborn error of intracellular cobalamin metabolism. Biochemical hallmarks are elevated homocysteine and low methionine in plasma accompanied by methylmalonic aciduria. Due to the heterogeneous clinical picture, patients with the late-onset form of the disease (onset >12 months) come to the attention of diverse medical specialists, e.g. paediatricians, neurologists, nephrologists, psychiatrists or haematologists. The report reviews the published clinical data and adds three new cases to raise awareness for this severe but often treatable disease. The Pubmed and the Cochrane databases were searched for clinical reports on cblC patients and three unreported cases are presented to illustrate the clinical spectrum. Reports on 58 cases (30 females, 22 males, 6 = no information) and the three new cases underlined the clinical heterogeneity of the disease. Time between first symptoms and diagnosis ranged from three months to more than 20 years. Haemolytic uraemic syndrome and pulmonary hypertension were main presenting symptoms in preschool children. In older children/adolescents, psychiatric symptoms, cognitive impairment, ataxia and myelopathy were frequently observed while thromboembolic events and glomerulopathies were almost exclusively seen in adults. Brain atrophy, white matter lesions and myelopathy were frequently encountered. The majority of patients showed marked biochemical and clinical response to treatment with parenteral hydroxocobalamin combined with oral betaine, folate, carnitine and rarely methionine. The course was less favourable in late treated or untreated patients. The late-onset cblC defect is a rare disease and unfortunately, diagnosis is often delayed. Raising awareness for this disorder can significantly improve patients' outcome and perspective by timely initiation of targeted treatment. Newborn screening (NBS) for the cblC defect might be of benefit especially for late-onset patients since treatment seems efficient when initiated before irreversible organ damage. In general, inborn errors of metabolisms should be considered in unexplained medical cases at any age, especially in patients with multisystemic disease. More specifically, total homocysteine in plasma and methylmalonic acid in urine/plasma should be measured in unexplained neurologic, psychiatric, renal, haematologic and thromboembolic disease.


Disrupted prefrontal interhemispheric structural coupling in schizophrenia related to working memory performance.
Prominent regional cortical thickness reductions have been shown in schizophrenia. In contrast, little is known regarding alterations of structural coupling between regions in schizophrenia and how these alterations may be related to cognitive impairments in this disorder. T1-weighted magnetic resonance images were acquired in 54 patients with schizophrenia and 68 healthy control subjects aged 18-55 years. Cortical thickness was compared between groups using a vertex-wise approach. To assess structural coupling, seeds were selected within regions of reduced thickness, and brain-wide cortical thickness correlations were compared between groups. The relationships between identified patterns of circuit structure disruption and cognitive task performance were then explored. Prominent cortical thickness reductions were found in patients compared with controls at a 5% false discovery rate in a predominantly frontal and temporal pattern. Correlations of the left dorsolateral prefrontal cortex (DLPFC) with right prefrontal regions were significantly different in patients and controls. The difference remained significant in a subset of 20 first-episode patients. Participants with stronger frontal interhemispheric thickness correlations had poorer working memory performance. We identified structural impairment in a left-right DLPFC circuit in patients with schizophrenia independent of illness stage or medication exposure. The relationship between left-right DLPFC thickness correlations and working memory performance implicates prefrontal interhemispheric circuit impairment as a vulnerability pathway for poor working memory performance. Our findings could guide the development of novel therapeutic interventions aimed at improving working memory performance in patients with schizophrenia.


Microsphere passage through intestinal circulation: via shunts or capillaries?
Significant quantities of 9-micron microspheres (20-30%) are not trapped in the intestine following intracardiac or intra-arterial injection, but reach venous blood. Some investigators propose that the passage of 9-micron spheres measures blood flow through noncapillary connections. Because frequency distributions of intestinal capillary diameters and 9-micron spheres overlap, microspheres could simply pass through capillaries. Therefore, we developed simple probabilistic models to predict both the size distribution and the percentage of injected spheres [9 +/- 1 (SD) micron] that should appear in venous blood. Chief assumptions in models are that microsphere delivery and sphere diameter are independent and that microspheres pass through capillaries of equal or larger size. The passage predicted by the models was consistent with values in canine intestinal circulation, demonstrating that passage through capillaries [7.38 +/- 1.4 (SD) micron] adequately accounts for spheres in venous blood. Because the diameters of nominal 9-micron spheres were distributed too narrowly to show a marked sieving effect on passage through the intestinal circulation, we also injected microspheres varying from 5 to 20 micron in diameter. This mixture demonstrated a marked sieving effect. The predicted frequency distribution for microsphere diameters in venous blood agreed with the observed distribution. Our models demonstrate that the passage of 9-micron spheres through capillaries, rather than through "shunts," adequately accounts for the appearance of spheres in venous blood and suggests that the frequency distribution of venous microspheres can provide an in vivo method for estimating the frequency distribution of intestinal capillary diameters.


Natural history of chronic Chagas' heart disease: prognosis factors.
Chronic Chagas' disease shows several progression modes. Usually, the different clinical syndromes manifest themselves together, however, isolated forms can occur. Cardiac arrhythmias, which are very frequent, are present in about 50% of patients. The cardiac damage manifests itself later, with the emergence of heart failure. Thromboembolism can occur in both pulmonary and systemic circulation. Pulmonary embolism is the most frequent, appearing in more advanced phases of heart disease. Sudden death is the fatal outcome of these patients. It predominates in males and generally occurs in a disease stage when patients have their highest productivity. The presence of serious ventricular arrhythmias, conduction disturbances in the electrocardiogram, and heart failure, provide an unfavorable prognosis.


Purpose and pleasure in late life: Conceptualising older women's participation in art and craft activities.
The fourth age, as the last stage of life, represents a final challenge to find personal meaning in the face of changing capacities, illness and disability. Participation in valued activities is important for sustaining interest in life and has been associated with enhanced health and well-being. Art and craft activities are a popular form of participation amongst women in late life with growing international interest in the potential for these types of activities to maintain health and well-being and address problems of social isolation. Drawing on open text comments from 114 women enrolled in the Australian Longitudinal Study on Women's Health and in-depth interviews with 23 women all aged in their eighties, this paper explores the nature of older women's participation in art and craft activities and conceptualises links between participation in these activities and health and well-being in late life. Participation in art and craft activities is complex and dynamic, comprising cognitive and physical processes infused with emotion and occurs in the context of social relationships, physical spaces, physical ailments and beliefs about the value of the activities. By participating in art and craft activities, older women find purpose in their lives, contributing to their subjective well-being whilst helping and being appreciated by others. They develop a self view as enabled and as such take on new art and craft challenges, continue to learn and develop as art and craft makers and remain open to new possibilities.


Aquaglyceroporin PbAQP during intraerythrocytic development of the malaria parasite Plasmodium berghei.
The malaria parasite can use host plasma glycerol for lipid biosynthesis and membrane biogenesis during the asexual intraerythrocytic development. The molecular basis for glycerol uptake into the parasite is undefined. We hypothesize that the Plasmodium aquaglyceroporin provides the pathway for glycerol uptake into the malaria parasite. To test this hypothesis, we identified the orthologue of Plasmodium falciparum aquaglyceroporin (PfAQP) in the rodent malaria parasite, Plasmodium berghei (PbAQP), and examined the biological role of PbAQP by performing a targeted deletion of the PbAQP gene. PbAQP and PfAQP are 62% identical in sequence. In contrast to the canonical NPA (Asn-Pro-Ala) motifs in most aquaporins, the PbAQP has NLA (Asn-Leu-Ala) and NPS (Asn-Leu-Ser) in those positions. PbAQP expressed in Xenopus oocytes was permeable to water and glycerol, suggesting that PbAQP is an aquaglyceroporin. In P. berghei, PbAQP was localized to the parasite plasma membrane. The PbAQP-null parasites were viable; however, they were highly deficient in glycerol transport. In addition, they proliferated more slowly compared with the WT parasites, and mice infected with PbAQP-null parasites survived longer. Taken together, these findings suggest that PbAQP provides the pathway for the entry of glycerol into P. berghei and contributes to the growth of the parasite during the asexual intraerythrocytic stages of infection. In conclusion, we demonstrate here that PbAQP plays an important role in the blood-stage development of the rodent malaria parasite during infection in mice and could be added to the list of targets for the design of antimalarial drugs.


Ideas of caring in nursing practice.
In nursing practice, awareness of ethical inner values and a common understanding of nursing and caring are needed. It is therefore important to highlight ideas of caring in nursing practice. The aim of this paper was to illuminate nursing, caring and ethical inner values in caring and caring in nursing practice. By being attentive, open, respectful and treating the patient as a person, nurses can enhance both their own and the patient's sense of personal meaning in the caring relationship. Nurses can use self-reflection to create an awareness of nursing, caring and ethical inner values in caring.


A randomized controlled trial on Stroke telerehabilitation: The effects on falls self-efficacy and satisfaction with care.
We determined the effect of a multifaceted stroke telerehabilitation (STeleR) intervention on falls-related self-efficacy and satisfaction with care. We conducted a prospective, randomized, multisite, single-blinded trial in 52 veterans from three Veterans Affairs Medical Centers. Participants who experienced a stroke in the past 24 months were randomized to the STeleR intervention or usual care. Participants in the intervention arm were administered an exit interview to gather specific patient satisfaction data three months after their final outcome measure. The STeleR intervention consisted of three home visits, five telephone calls, and an in-home messaging device provided over three months to instruct patients in functionally based exercises and adaptive strategies. The outcome measures included Falls Efficacy Scale to measure fall-related self-efficacy and a Stroke-Specific Patient Satisfaction with Care (SSPSC) scale, a measure separated into two subscales (satisfaction with home care and satisfaction with hospital care) was employed to measure the participants' satisfaction. At six months, compared with the usual care group, the STeleR group showed statistically significant improvements in one of the two SSPSC scales (satisfaction with hospital care, p = .029) and approached significance in the second SSPSC scale (satisfaction with home care, p = .077). There were no improvements in fall-related self-efficacy. Core concepts identified were: (a) beneficial impact of the trained assistant; (b) exercises helpful; (c) home use of technology. The STeleR intervention improved satisfaction with care, especially as it relates to care following their experience from the hospital. With the limited resources available for in-home rehabilitation for stroke survivors, STeleR (and especially its exercise components) can be a useful complement to traditional post-stroke rehabilitation.


A comparison of questionnaire, accelerometer, and pedometer: measures in older people.
To compare (i) the convergent validity of the self-report Zutphen Physical Activity Questionnaire with the 7-d objective physical activity (PA) measurement by accelerometers and pedometers and (ii) the construct validity of these measures by examining their associations with physical health and psychological and anthropometric variables. Five hundred and sixty community-dwelling people aged > or =65 yr were invited from a UK primary care practice and 238 (43%) participated (mean age = 74, 53% male). PA was assessed subjectively by the Zutphen questionnaire (modified to include housework questions) and objectively by the 7-d accelerometer monitoring: a random half also had a pedometer. A questionnaire assessed health, disability, and psychological factors, and anthropometric assessment was performed. Mean daily PA levels were as follows: Zutphen = 9.1 kcal x kg(-1) x d(-1) (SD = 6.6 kcal x kg(-1) x d(-1)); accelerometer activity count = 226,648 (SD = 121,966); accelerometer step count = 6495 (SD = 3212); and pedometer step count = 6712 (SD = 3526). Zutphen score was moderately correlated with accelerometer activity count (R = 0.34, P < 0.001) and pedometer step count (R = 0.36, P < 0.001). Pedometer step count was highly correlated with accelerometer activity count (R = 0.82, P< 0.001) and accelerometer step count (R = 0.86, P < 0.001). Objective PA measures showed strong associations with health and anthropometric and psychological variables. Zutphen score was not significantly related to most health or anthropometric measures but was associated with psychological variables and provided information about activity type. Convergent validity was strong between accelerometers and pedometers but weaker between these and self-report Zutphen. Pedometers may be preferred to accelerometers for simple studies due to their lower cost. Objective measures had better construct validity, being more strongly associated with established PA determinants, and thus offered better value to researchers than the questionnaire, but the latter provided useful detail on activity type, so a combined approach to PA assessment may be preferable.


Low Serum Testosterone is Present in Nearly Half of Men Undergoing Artificial Urinary Sphincter Placement.
To report the prevalence of low serum testosterone (LST) in men undergoing artificial urinary sphincter (AUS) placement at a single high-volume institution. We retrospectively reviewed all men undergoing AUS procedures by a single surgeon from January 2015 to January 2018 to identify men with pretreatment total serum testosterone levels. LST was defined as less than 280 ng/dL. Patients with only posttreatment testosterone levels were excluded. Demographic characteristics and clinical outcomes were compared between men with and without LST. Among 113 patients who underwent AUS with pretreatment serum testosterone levels drawn an average of 2.2 months before AUS surgery, 45.1% (51 of 113) met criteria for LST, including 18 patients on androgen deprivation therapy. The rate of primary LST was 34.7% (33 of 95). The median total serum testosterone level among men with LST was 118 ng/dL (interquartile range 6-211), and 413 mg/dL (interquartile range 333-550) in the normal serum testosterone group. There were no differences in patient age, history of radiation, erectile dysfunction, or other comorbidities between the groups. Body mass index was higher in the LST group compared to normal serum testosterone (30 vs 27 kg/m2, P = .001). Cuff size and rates of transcorporal cuff placement were similar between groups. Nearly one-half of men with stress urinary incontinence undergoing AUS placement present with LST. While AUS cuff erosion appears to be more common in men with LST, further study is needed to determine if treating LST will reduce cuff erosion rates.


SHARAKU: an algorithm for aligning and clustering read mapping profiles of deep sequencing in non-coding RNA processing.
Deep sequencing of the transcripts of regulatory non-coding RNA generates footprints of post-transcriptional processes. After obtaining sequence reads, the short reads are mapped to a reference genome, and specific mapping patterns can be detected called read mapping profiles, which are distinct from random non-functional degradation patterns. These patterns reflect the maturation processes that lead to the production of shorter RNA sequences. Recent next-generation sequencing studies have revealed not only the typical maturation process of miRNAs but also the various processing mechanisms of small RNAs derived from tRNAs and snoRNAs. We developed an algorithm termed SHARAKU to align two read mapping profiles of next-generation sequencing outputs for non-coding RNAs. In contrast with previous work, SHARAKU incorporates the primary and secondary sequence structures into an alignment of read mapping profiles to allow for the detection of common processing patterns. Using a benchmark simulated dataset, SHARAKU exhibited superior performance to previous methods for correctly clustering the read mapping profiles with respect to 5'-end processing and 3'-end processing from degradation patterns and in detecting similar processing patterns in deriving the shorter RNAs. Further, using experimental data of small RNA sequencing for the common marmoset brain, SHARAKU succeeded in identifying the significant clusters of read mapping profiles for similar processing patterns of small derived RNA families expressed in the brain. The source code of our program SHARAKU is available at http://www.dna.bio.keio.ac.jp/sharaku/, and the simulated dataset used in this work is available at the same link. Accession code: The sequence data from the whole RNA transcripts in the hippocampus of the left brain used in this work is available from the DNA DataBank of Japan (DDBJ) Sequence Read Archive (DRA) under the accession number DRA004502. email@example.com Supplementary data are available at Bioinformatics online.


Computed tomography features of middle ear cholesteatoma in dogs.
We describe the computed tomography (CT) findings in 11 dogs with middle ear cholesteatoma. The cholesteatoma appeared as an expansile tympanic cavity mass with a mean attenuation value of 55.8 +/- 4.2 Hounsfield units. There was no appreciable contrast enhancement of the tympanic bulla contense but ring enhancement was seen in four dogs. Due to the slow progressive growth, the lesion causes severe bone changes at the contour of the tympanic bulla, including osteolysis, osteoproliferation and osteosclerosis, expansion of the tympanic cavity, and sclerosis or osteoproliferation of the ipsilateral temporomandibular joint and paracondylar process. Cholesteatoma can cause lysis of the petrosal part of the temporal bone, leading to intracranial complications. Although not definitive, CT provides useful information for distinguishing a middle ear cholesteatoma from otitis media and neoplasia. In otitis media, enlargement of the tympanic cavity is not routinely observed. In tumors that primarily affect the middle or inner ear, the predominant signs are lysis of the contour of the tympanic bulla or the petrosal part of the temporal bone, soft tissue swelling around the middle ear and marked contrast enhancement. In tumors that arise from the external ear, a soft tissue mass is visible within the external acusticus meatus, and the middle ear is only involved secondarily.


A redox-switchable alpha-cyclodextrin-based [2]rotaxane.
A bistable [2]rotaxane comprising an alpha-cyclodextrin (alpha-CD) ring and a dumbbell component containing a redox-active tetrathiafulvalene (TTF) ring system within its rod section has been synthesized using the Cu(I)-catalyzed azide-alkyne cycloaddition, and the redox-driven movements of the alpha-CD ring between the TTF and newly formed triazole ring systems have been elucidated. Microcalorimetric titrations on model complexes suggested that the alpha-CD ring prefers to reside on the TTF rather than on the triazole ring system by at least an order of magnitude. The fact that this situation does pertain in the bistable [2]rotaxane has not only been established quantitatively by electrochemical experiments and backed up by spectroscopic and chiroptical measurements but also been confirmed semiquantitatively by the recording of numerous cyclic voltammograms which point, along with the use of redox-active chemical reagents, to a mechanism of switching that involves the oxidation of the neutral TTF ring system to either its radical cationic (TTF*+) or dicationic (TTF2+) counterparts, whereupon the alpha-CD ring, moves along the dumbbell to encircle the triazole ring system. Since redox control by both chemical and electrochemical means is reversible, the switching by the bistable [2]rotaxane can be reversed on reduction of the TTF*+ or TTF2+ back to being a neutral TTF.


[The effects of estrogen-like products in milk on prostate and testes].
To study the effects of estrogen-like products in milk on the male reproductive system. The incidence rates of testicular and prostatic cancers in 42 countries (regions), collected from the cancer incidence in five continent database (1988-1992), were correlated with the relative food consumption in these countries, which was provided by FAO database (1961-1990); two-generation reproduction test was taken by using Wistar rats according to FDA standard. Among the food items examined, milk was most closely correlated (r = 0.711) with prostatic cancer incidence, followed by meat and coffee. As for testicular cancer, cheese was most closely correlated (r = 0.804) with the incidences in ages of 20-39, followed by animal fat and milk. No significant decreases of reproductive organ weight were found after the rats had been fed a lot of milk for long time. The results suggest that further studies should be taken on the effects of estrogen-like products in milk on the male reproductive health.


Poisoning by Datura leaves used as edible wild vegetables.
The causes of Datura intoxication include medication overdose, misuse of edible vegetables, deliberate abuse as a hallucinogen, homicidal or robbery and accidental intoxication from contaminated food. We report an incident of 14 people with Datura intoxication caused by ingesting wild Datura suaveolans for food. The incubation period was 15 to 30 min. The symptoms/signs were dizziness, dry mouth, flushed skin, palpitation, nausea, drowsiness, tachycardia, blurred vision, mydriasis, hyperthermia, disorientation, vomiting, agitation, delirium, urine retention, hypertension and coma. Three patients were hospitalized for 2-3 days. Thirteen persons received supportive fluid therapy. One patient did not receive medical therapy, he induced vomiting and drank a lot of water. Four patients presented with delirium/coma and 3 received physostigmine therapy with good response. One patient was intubated because of coma and respiratory depression. Three persons needed Foley catheterization for urine retention or coma status. One patient had a complication of urinary tract infection and antibiotic management. All patients recovered with no sequelae.


Sleep Deprivation and Neurological Disorders.
Sleep plays an important role in maintaining neuronal circuitry, signalling and helps maintain overall health and wellbeing. Sleep deprivation (SD) disturbs the circadian physiology and exerts a negative impact on brain and behavioural functions. SD impairs the cellular clearance of misfolded neurotoxin proteins like α-synuclein, amyloid-β, and tau which are involved in major neurodegenerative diseases like Alzheimer's disease and Parkinson's disease. In addition, SD is also shown to affect the glymphatic system, a glial-dependent metabolic waste clearance pathway, causing accumulation of misfolded faulty proteins in synaptic compartments resulting in cognitive decline. Also, SD affects the immunological and redox system resulting in neuroinflammation and oxidative stress. Hence, it is important to understand the molecular and biochemical alterations that are the causative factors leading to these pathophysiological effects on the neuronal system. This review is an attempt in this direction. It provides up-to-date information on the alterations in the key processes, pathways, and proteins that are negatively affected by SD and become reasons for neurological disorders over a prolonged period of time, if left unattended.


Patterns of population genetic structure for springtails and mites in southern Victoria Land, Antarctica.
We sequenced the mitochondrial (mt) DNA cytochrome c oxidase subunit I gene to examine comparative phylogeographic patterns for the springtail Gomphiocephalus hodgsoni and the mite Stereotydeus mollis throughout their ranges in southern Victoria Land, Antarctica. Our aim was to extend previous genetic work to encompass a large ice-free area in the Dry Valleys. In particular, we sought to determine if this new region harboured high levels of genetic diversity and if patterns of genetic structure were congruent across taxa. Phylogenetic and nested clade analyses for G. hodgsoni and S. mollis showed similar patterns of population sub-structuring among locations and highlighted several potential refugia that may have existed during glacial maxima. We identified greater levels of genetic divergence in S. mollis and suggest that there is a nucleotide substitution (mutation) rate difference between S. mollis and G. hodgsoni, and/or that S. mollis has had a longer association with the Antarctic landscape.


Effects of noradrenaline depletion in the brain on response on novelty in isolation-reared rats.
Social isolation from weaning in the rat produces a variety of neurochemical and behavioural effects in the adult that in part parallel changes seen in human schizophrenia. The study investigated the effects of central noradrenaline (NA) depletion by the selective neurotoxin, N-(2-chloroethyl)-N-ethyl-2-bromobenzylamine (DSP-4), on the behaviour of isolation-reared rats. Male Lister hooded rats were reared singly or in groups after weaning. During week 2, the rats were tested in photocell activity cages and were then injected with DSP-4 (25 mg/kg, IP). During week 4, rats were tested in the open field under the following conditions: open field alone, with two novel stimuli (T1), and with a familiar and a novel object (T2), and in the activity cages. DSP-4 significantly reduced cortical and hippocampal NA levels with no effect on the hypothalamus. Isolation-reared rats exhibited locomotor hyperactivity and reduced habituation to the testing arena, although their exploration of the novel objects in T1 was not significantly different from group-reared rats. DSP-4 treatment in group-reared rats increased inner zone activity in the open field but did not significantly affect the exploration of novel objects. DSP-4 treatment in isolates reduced exploration of objects at T2 while increasing exploration of the general environment. Isolation rearing influences the behavioural effects of central NA depletion. The results suggest isolation-induced changes in the central noradrenergic system in the isolated rat, supporting the view that early environmental factors can have long-term effects on central noradrenergic function as well as other neurotransmitter systems.


Residuals and Diagnostics for Ordinal Regression Models: A Surrogate Approach.
Ordinal outcomes are common in scientific research and everyday practice, and we often rely on regression models to make inference. A long-standing problem with such regression analyses is the lack of effective diagnostic tools for validating model assumptions. The difficulty arises from the fact that an ordinal variable has discrete values that are labeled with, but not, numerical values. The values merely represent ordered categories. In this paper, we propose a surrogate approach to defining residuals for an ordinal outcome Y. The idea is to define a continuous variable S as a "surrogate" of Y and then obtain residuals based on S. For the general class of cumulative link regression models, we study the residual's theoretical and graphical properties. We show that the residual has null properties similar to those of the common residuals for continuous outcomes. Our numerical studies demonstrate that the residual has power to detect misspecification with respect to 1) mean structures; 2) link functions; 3) heteroscedasticity; 4) proportionality; and 5) mixed populations. The proposed residual also enables us to develop numeric measures for goodness-of-fit using classical distance notions. Our results suggest that compared to a previously defined residual, our residual can reveal deeper insights into model diagnostics. We stress that this work focuses on residual analysis, rather than hypothesis testing. The latter has limited utility as it only provides a single p-value, whereas our residual can reveal what components of the model are misspecified and advise how to make improvements.


Individual differences and self-regulatory fatigue: optimism, conscientiousness, and self-consciousness.
Ability to self-regulate varies and self-regulatory strength is a limited source that can be depleted or fatigued. Research on the impact of individual differences on self-regulatory capacity is still scarce, and this study aimed to examine whether personality factors such as dispositional optimism, conscientiousness, and self-consciousness can impact or buffer self-regulatory fatigue. Participants were patients diagnosed with chronic multi-symptom illnesses (N = 50), or pain free matched controls (N = 50), randomly assigned to either a high or low self-regulation task, followed by a persistence task. Higher optimism predicted longer persistence (p = .04), and there was a trend towards the same effect for conscientiousness (p = .08). The optimism by self-regulation interaction was significant (p = .01), but rather than persisting despite self-regulatory effort, optimists persisted longer only when not experiencing self-regulatory fatigue. The effects of optimism were stronger for controls than patients. There was also a trend towards a similar conscientiousness by self-regulation interaction (p = .06). These results suggest that the well-established positive impact of optimism and conscientiousness on engagement and persistence may be diminished or reversed in the presence of self-regulatory effort or fatigue, adding an important new chapter to the self-regulation, personality, and pain literature.


MR measurement of visceral fat: assessment of metabolic syndrome.
One diagnostic criterion for metabolic syndrome is obesity from the accumulation of visceral fat; others include abdominal circumference and area of visceral fat as measured by computed tomography (CT) at the umbilical level. We evaluated visceral fat using frequency-selective excitation magnetic resonance (MR) imaging SPAIR (spectral attenuation with inversion recovery) water suppression THRIVE (3D T1-high resolution isotropic volume examination). Fifty of 70 slices with 2-mm interval were used to render and measure volume of visceral fat ranging within 10 cm of the umbilicus; the area of visceral fat at the umbilical level was also measured. Imaging was completed using breath hold within 14 s. Image processing was easier than using CT.


Serum total cholesterol and long-term coronary heart disease mortality in different cultures. Twenty-five-year follow-up of the seven countries study.
To compare the relationship between serum total cholesterol and long-term mortality from coronary heart disease (CHD) in different cultures. Total cholesterol was measured at baseline (1958 through 1964) and at 5- and 10-year follow-up in 12,467 men aged 40 through 59 years in 16 cohorts located in seven countries: five European countries, the United States, and Japan. To increase statistical power six cohorts were formed, based on similarities in culture and cholesterol changes during the first 10 years of follow-up. Relative risks (RRs), estimated with Cox proportional hazards (survival) analysis, for 25-year CHD mortality for cholesterol quartiles and per 0.50-mmol/L (20-mg/dL) cholesterol increase. Adjustment was made for age, smoking, and systolic blood pressure. The age-standardized CHD mortality rates in the six cohorts ranged from 3% to 20%. The RRs for the highest compared with the lowest cholesterol quartile ranged from 1.5 to 2.3, except for Japan's RR of 1.1. For a cholesterol level of around 5.45 mmol/L (210 mg/dL), CHD mortality rates varied from 4% to 5% in Japan and Mediterranean Southern Europe to about 15% in Northern Europe. However, the relative increase in CHD mortality due to a given cholesterol increase was similar in all cultures except Japan. Using a linear approximation, a 0.50-mmol/L (20-mg/dL) increase in total cholesterol corresponded to an increase in CHD mortality risk of 12%, which became an increase in mortality risk of 17% when adjusted for regression dilution bias. Across cultures, cholesterol is linearly related to CHD mortality, and the relative increase in CHD mortality rates with a given cholesterol increase is the same. The large difference in absolute CHD mortality rates at a given cholesterol level, however, indicates that other factors, such as diet, that are typical for cultures with a low CHD risk are also important with respect to primary prevention.


Lunasin is prevalent in barley and is bioavailable and bioactive in in vivo and in vitro studies.
Lunasin, a unique 43-amino acid peptide found in a number of seeds, has been shown to be chemopreventive in mammalian cells and in a skin cancer mouse model. To elucidate the role of cereals in cancer prevention, we report here the prevalence, bioavailability, and bioactivity of lunasin from barley. Lunasin is present in all cultivars of barley analyzed. The liver and kidney of rats fed with lunasin-enriched barley (LEB) show the presence of lunasin in Western blot. Lunasin extracted from the kidney and liver inhibits the activities of HATs (histone acetyl transferases), yGCN5 by 20% and 18% at 100 nM, and PCAF activity by 25% and 24% at 100 nM, confirming that the peptide is intact and bioactive. Purified barley lunasin localizes in the nuclei of NIH 3T3 cells. Barley lunasin added to NIH 3T3 cells in the presence of the chemical carcinogen MCA activates the expression of tumor suppressors p21 and p15 by 45% and 47%, decreases cyclin D1 by 98%, and inhibits Rb hyperphosphorylation by 45% compared with the MCA treatment alone. We conclude that lunasin is prevalent in barley, bioavailable, and bioactive and that consumption of barley could play an important role of cancer prevention in barley-consuming populations.


Androgens and Androgen Derivatives: Science, Myths, and Theories: Explored From a Special Operations Perspective.
Androgen use outside of legitimate medical therapy is a perceived concern that is drawing attention across military and specifically Special Operations Forces (SOF) communities. For leadership and the medical community to properly address the issue and relate to those individuals who are using or considering use, it will be crucial to understand the scope of the problem. Limited data suggest that the prevalence of androgen use may be increasing, and inferences made from the scientific literature suggest that SOF may be a population of concern. While risks of androgen use are well known, there are little data specific to military performance that can be applied to a rigorous risk:benefit analysis, allowing myths and poorly supported theories to perpetuate within the community. Further efforts to define the potential benefits balanced against the short- and long-term risks should be undertaken. Providers within the SOF community should arm themselves with information to engage androgen users and leadership in meaningful discussion regarding androgen use.


The sleeper effect: Artifact or phenomenon-A brief comment on Bell et al. (2013).
Bell, Marcus, and Goodlad (2013) recently conducted a meta-analysis of randomized controlled additive trials and found that adding an additional component to an existing treatment vis-à-vis the existing treatment produced larger effect sizes on targeted outcomes at 6-months follow-up than at termination, an effect they labeled as a sleeper effect. One of the limitations with Bell et al.'s detection of the sleeper effect was that they did not conduct a statistical test of the size of the effect at follow-up versus termination. To statistically test if the differences of effect sizes between the additive conditions and the control conditions at follow-up differed from those at termination, we used a restricted maximum-likelihood random-effect model with known variances to conduct a multilevel longitudinal meta-analysis (k = 30). Although the small effects at termination detected by Bell et al. were replicated (ds = 0.17-0.23), none of the analyses of growth from termination to follow-up produced statistically significant effects (ds < 0.08; p > .20), and when asymmetry was considered using trim-and-fill procedure or the studies after 2000 were analyzed, magnitude of the sleeper effect was negligible (d = 0.00). There is no empirical evidence to support the sleeper effect. (PsycINFO Database Record


Cold-water immersion and iced-slush ingestion are effective at cooling firefighters following a simulated search and rescue task in a hot environment.
Firefighters are exposed to hot environments, which results in elevated core temperatures. Rapidly reducing core temperatures will likely increase safety as firefighters are redeployed to subsequent operational tasks. This study investigated the effectiveness of cold-water immersion (CWI) and iced-slush ingestion (SLUSH) to cool firefighters post-incident. Seventy-four Australian firefighters (mean ± SD age: 38.9 ± 9.0 years) undertook a simulated search and rescue task in a heat chamber (105 ± 5 °C). Testing involved two 20-min work cycles separated by a 10-min rest period. Ambient temperature during recovery periods was 19.3 ± 2.7 °C. Participants were randomly assigned one of three 15-min cooling protocols: (i) CWI, 15 °C to umbilicus; (ii) SLUSH, 7 g·kg(-1) body weight; or (iii) seated rest (CONT). Core temperature and strength were measured pre- and postsimulation and directly after cooling. Mean temperatures for all groups reached 38.9 ± 0.9 °C at the conclusion of the second work task. Both CWI and SLUSH delivered cooling rates in excess of CONT (0.093 and 0.092 compared with 0.058 °C·min(-1)) and reduced temperatures to baseline measurements within the 15-min cooling period. Grip strength was not negatively impacted by either SLUSH or CONT. CWI and SLUSH provide evidence-based alternatives to passive recovery and forearm immersion protocols currently adopted by many fire services. To maximise the likelihood of adoption, we recommend SLUSH ingestion as a practical and effective cooling strategy for post-incident cooling of firefighters in temperate regions.


α7 and β2 nicotinic receptors control monoamine-mediated locomotor response.
Earlier studies reported exacerbated locomotor response to stress and tranylcypromine in β2 nicotinic acetylcholine receptor (nAChR) knockout (KO) mice. This study aimed to further assess the role of β2 and coexpressed nAChR subunits in the brain (α4, α6 and α7) to control monoamine-mediated locomotor response, that is, response to novelty, saline, nicotine with tranylcypromine pretreatment, cocaine, d-amphetamine and morphine treatments. Results show that β2 KO mice were hyperreactive to novelty, cocaine and morphine. In contrast, α7 KO mice were hyporeactive to tranylcypromine and cocaine. These results suggest that endogenous nAChR stimulation may exert a tonic control on monoamine-mediated locomotor responses. β2 and α7-containing nAChR may contribute, respectively, to the inhibitory and permissive pathways of this tonic control.


Nonthrombogenic plastic surfaces.
Heparin has been chemically combined with a number of plastic surfaces rendering them nonthrombogenic as judged by Lee-White coagulation tests in vitro with human blood. Addition of quaternary ammonium groups to the plastic permitted formation of insoluble complexes with heparin. These heparinized surfaces were essentially nonthrombogenic and adsorb blood proteins to a significantly smaller degree from dilute solution than do the unmodified plastic surfaces. The affinity of the formed blood elements for these modified surfaces is much less than for the unmodified surfaces.


Two-photon absorption and self-phase modulation measurements with shaped femtosecond laser pulses.
We show that phase-sensitive detection of spectral hole refilling can yield information about self-phase modulation and two-photon absorption coefficients. We expect that, when applied to tissue microscopy, this technique will allow the study of endogenous molecular markers beneath the surface, even if those markers are nonfluorescent.


Ursolic Acid Activates Intracellular Killing Effect of Macrophages During Mycobacterium tuberculosis Infection.
Tuberculosis is one of the most threatening infectious diseases to public health all over the world, for which Mycobacterium tuberculosis (MTB) is the etiological agent of pathogenesis. Ursolic acid (UA) has immunomodulatory function and exhibits antimycobacterial activity. However, the intracellular killing effect of UA has yet to be elucidated. The aim of this study was to evaluate the intracellular killing effect of UA during mycobacterial infection. The intracellular killing activity of UA was evaluated in the macrophage cell line THP-1 by the MGIT 960 system as well as by CFU count. The production of reactive oxygen species (ROS) and the level of nitric oxide (NO) were measured using DCF-DA and Griess reagent, respectively. Phagocytosis was observed by a fluorescence-based staining method, and the colony forming units were enumerated on 7H11 agar medium following infection. In addition, MRP8 mRNA expression was measured by qRT-PCR. UA significantly decreased the number of intracellular Mycobacterium through generation of ROS and NO. In addition, it profoundly activated the phagocytosis process of THP-1 cells during MTB-infection. Furthermore, our data demonstrated that UA activated the phagocytosis process in human monocyte cells through MRP8 induction. These data suggest that UA firmly contributes to the intracellular killing effect of macrophages during mycobacterial infection.


Upper eyelid skin grafting: an effective treatment for lagophthalmos following blepharoplasty.
Patients who have undergone upper blepharoplasty occasionally develop anterior lamellar insufficiency, which can result in lagophthalmos, corneal decompensation, and even blindness. Historically, skin grafts in the upper eyelid have been considered a last-resort procedure because of poor cosmetic outcomes. Poor cosmetic outcomes result from the traditional practice of placing the skin graft above the eyelid crease. This article describes a surgical technique for upper eyelid skin grafting in which the graft is placed in a supraciliary position. Presented are results of a retrospective study of 20 patients (31 eyelids) who underwent supraciliary upper eyelid skin grafting. The postoperative results were evaluated by examining the improvement in lagophthalmos, the improvement of keratopathy and comfort of the patient, and the cosmetic appearance of the graft. Upper eyelid skin grafting using this surgical technique is an effective and cosmetically acceptable method to improve corneal integrity and comfort in patients who have corneal exposure from insufficient anterior lamella after upper eyelid or eyebrow surgery.


Increased proximal sodium reabsorption is associated with increased cardiovascular risk in men.
To explore the relationship between fractional lithium excretion (taken as an index of proximal sodium excretion), blood pressure and metabolic risk factors for coronary heart disease in a sample of a general white male population. A cross-sectional survey of a sample of a white male working population carried out as part of the Olivetti Heart Study. The Olivetti factory in Pozzuoli, a suburb of Naples, Italy. Five hundred and sixty-seven untreated white male workers aged 21-68 years. Anthropometry, blood pressure, a blood test to determine the lipid profile, electrolytes, glucose and uric acid levels, a detailed questionnaire and urinary measurements on a fasting timed urine sample after a 300 mg lithium carbonate capsule taken the night before the investigation. The fractional excretion of lithium was inversely associated with body weight, body mass index, diastolic blood pressure, serum triglycerides level and serum uric acid level whereas it was positively associated with serum high-density lipoprotein cholesterol level. The greater the percentage of filtered sodium reabsorbed at the proximal tubule the greater the overall metabolic cardiovascular risk. The association was consistent across tertiles of fractional excretion of lithium, was independent of age, smoking, alcohol consumption and sodium excretion (taken as an index of sodium intake) and persisted in multiple regression analysis. Men with high serum triglycerides, high serum uric acid and low serum high-density lipoprotein cholesterol levels and higher blood pressure had significantly lower fractional excretion of lithium (and thus enhanced proximal sodium reabsorption) than did men with a normal metabolic profile (21.8 +/- 5.6 versus 26.5 +/- 5.1%, p = 0.002). The independent association between a cluster of risk factors for cardiovascular disease suggestive of insulin resistance and increased proximal sodium reabsorption at the renal tubule indicates a new feature of the metabolic syndrome that, if causal, could be amenable to intervention.


Intratumoral Agreement of High-Resolution Magic Angle Spinning Magnetic Resonance Spectroscopic Profiles in the Metabolic Characterization of Breast Cancer.
High-resolution magic angle spinning (HR-MAS) magnetic resonance (MR) spectroscopy data may serve as a biomarker for breast cancer, with only a small volume of tissue sample required for assessment. However, previous studies utilized only a single tissue sample from each patient. The aim of this study was to investigate whether intratumoral location and biospecimen type affected the metabolic characterization of breast cancer assessed by HR-MAS MR spectroscopy. This prospective study was approved by the institutional review board and informed consent was obtained. Preoperative core-needle biopsies (CNBs), central, and peripheral surgical tumor specimens were prospectively collected under ultrasound (US) guidance in 31 patients with invasive breast cancer. Specimens were assessed with HR-MAS MR spectroscopy. The reliability of metabolite concentrations was evaluated and multivariate analysis was performed according to intratumoral location and biospecimen type. There was a moderate or higher agreement between the relative concentrations of 94.3% (33 of 35) of metabolites in the center and periphery, 80.0% (28 of 35) of metabolites in the CNB and central surgical specimens, and 82.9% (29 of 35) of metabolites between all 3 specimen types. However, there was no significant agreement between the concentrations of phosphocholine (PC) and phosphoethanolamine (PE) in the center and periphery. The concentrations of several metabolites (adipate, arginine, fumarate, glutamate, PC, and PE) had no significant agreement between the CNB and central surgical specimens. In conclusion, most HR-MAS MR spectroscopic data do not differ based on intratumoral location or biospecimen type. However, some metabolites may be affected by specimen-related variables, and caution is recommended in decision-making based solely on metabolite concentrations, particularly PC and PE. Further validation through future studies is needed for the clinical implementation of these biomarkers based on data from a single tissue sample.


A straightforward approach to antibodies recognising cancer specific glycopeptidic neoepitopes.
Aberrantly truncated immature O-glycosylation in proteins occurs in essentially all types of epithelial cancer cells, which was demonstrated to be a common feature of most adenocarcinomas and strongly associated with cancer proliferation and metastasis. Although extensive efforts have been made toward the development of anticancer antibodies targeting MUC1, one of the most studied mucins having cancer-relevant immature O-glycans, no anti-MUC1 antibody recognises carbohydrates and the proximal MUC1 peptide region, concurrently. Here we present a general strategy that allows for the creation of antibodies interacting specifically with glycopeptidic neoepitopes by using homogeneous synthetic MUC1 glycopeptides designed for the streamlined process of immunization, antibody screening, three-dimensional structure analysis, epitope mapping and biochemical analysis. The X-ray crystal structure of the anti-MUC1 monoclonal antibody SN-101 complexed with the antigenic glycopeptide provides for the first time evidence that SN-101 recognises specifically the essential epitope by forming multiple hydrogen bonds both with the proximal peptide and GalNAc linked to the threonine residue, concurrently. Remarkably, the structure of the MUC1 glycopeptide in complex with SN-101 is identical to its solution NMR structure, an extended conformation induced by site-specific glycosylation. We demonstrate that this method accelerates dramatically the development of a new class of designated antibodies targeting a variety of "dynamic neoepitopes" elaborated by disease-specific O-glycosylation in the immunodominant mucin domains and mucin-like sequences found in intrinsically disordered regions of many proteins.


The Saccharomyces cerevisiae Start-specific transcription factor Swi4 interacts through the ankyrin repeats with the mitotic Clb2/Cdc28 kinase and through its conserved carboxy terminus with Swi6.
At a point in late G1 termed Start, yeast cells enter S phase, duplicate their spindle pole bodies, and form buds. These events require activation of Cdc28 kinase by G1 cyclins. Swi4 associates with Swi6 to form the SCB-binding factor complex which activates G1 cyclin genes CLN1 and CLN2 in late G1. In G2 and M phases, the transcriptional activity of SCB-binding factor is repressed by the mitotic Clb2/Cdc28 kinase. Mbp1, a transcription factor related to Swi4, forms the MCB-binding factor complex with Swi6, which activates DNA synthesis genes and S-phase cyclin genes CLB5 and CLB6 in late G1. Clb2/Cdc28 kinase is not required for the repression of MCB-binding factor transcriptional activity in G2 and M phase. We show here that the Swi4 carboxy terminus is sufficient for interaction with Swi6 in vitro. A carboxy-terminal domain of Swi6 is required and sufficient for interaction with Swi4. The carboxy terminus of Mbp1 is sufficient for interaction with Swi6, and the carboxy terminus of Swi6 is required for interaction with Mbp1. By coimmunoprecipitation, we show that Swi4 but not Mbp1 interacts with Clb2/Cdc28 kinase in vivo during the G2 and M phases of the cell cycle. We demonstrate that the ankyrin repeats of Swi4 mediate the interaction with Clb2/Cdc28 kinase. The ankyrin repeats constitute a domain by which a cell cycle-specific transcription factor can interact with cyclin-dependent kinase complexes, thus enabling it to link its transcriptional activity to cell cycle progression.


Structural anomalies of fluids: origins in second and higher coordination shells.
Compressing or cooling a fluid typically enhances its static interparticle correlations. However, there are notable exceptions. Isothermal compression can reduce the translational order of fluids that exhibit anomalous waterlike trends in their thermodynamic and transport properties, while isochoric cooling (or strengthening of attractive interactions) can have a similar effect on fluids of particles with short-range attractions. Recent simulation studies by Yan [Phys. Rev. E 76, 051201 (2007)] on the former type of system and Krekelberg [J. Chem. Phys. 127, 044502 (2007)] on the latter provide examples where such structural anomalies can be related to specific changes in second and more distant coordination shells of the radial distribution function. Here, we confirm the generality of this microscopic picture through analysis, via molecular simulation and integral equation theory, of coordination shell contributions to the two-body excess entropy for several related model fluids which incorporate different levels of molecular resolution. The results suggest that integral equation theory can be an effective and computationally inexpensive tool for assessing, based on the pair potential alone, whether new model systems are good candidates for exhibiting structural (and hence thermodynamic and transport) anomalies.


Superficial radially resolved fluorescence and 3D photochemical time-dependent model for photodynamic therapy.
Photodynamic therapy (PDT) dosimetric tools are crucial for treatment planning and noninvasive monitoring by means of fluorescence. Present approaches consider usually a 1D problem, a simple photochemical process, or a spatially homogeneous photosensitizer. In this work, a radially resolved superficial photosensitizer fluorescence and 3D photochemical time-dependent PDT model are presented. The model provides a time-dependent estimation of tissue fluorescence and the photosensitizer and singlet oxygen 3D concentrations. The model is applied to a basal cell carcinoma treated by Metvix topical photosensitizer protocol. The analysis shows the potentiality in treatment planning and monitoring. The fluorescence results are in agreement with previous measurements.


Non-damaging retinal phototherapy: dynamic range of heat shock protein expression.
Subthreshold retinal phototherapy demonstrated clinical efficacy for the treatment of diabetic macular edema without visible signs of retinal damage. To assess the range of cellular responses to sublethal hyperthermia, expression of the gene encoding a 70 kDa heat shock protein (HSP70) was evaluated after laser irradiation using a transgenic reporter mouse. One hundred millisecond, 532 nm laser exposures with 400 μm beam diameter were applied to the retina surrounding the optic nerve in 32 mice. Transcription from the HSP70 promoter was assessed relative to the control eye using a bioluminescence assay at 7 hours after laser application. The retinal pigmented epithelium (RPE) viability threshold was determined with a fluorescence assay. A computational model was developed to estimate temperature and the extent of cell damage. A significant increase in HSP70 transcription was found at exposures over 20 mW, half the threshold power for RPE cell death. Computational modeling estimated peak temperature T = 49°C at HSP70 expression threshold. At RPE viability threshold, T = 57°C. Similar temperatures and damage indices were calculated for clinical subvisible retinal treatment parameters. Beneficial effects of laser therapy have been previously shown to extend beyond those resulting from destruction of tissue. One hundred millisecond laser exposures at approximately half the threshold power of RPE damage induced transcription of HSP70, an indication of cellular response to sublethal thermal stress. A computational model of retinal hyperthermia can guide further optimization of laser parameters for nondamaging phototherapy.


Astrocytes as a predominant cellular site of (99m)Tc-HMPAO retention.
Technetium-99m-d,l-hexamethylpropylene amine oxime ((99m)Tc-HMPAO) retention in the brain monitored by single photon emission computed tomography (SPECT) is currently used as a marker of cerebral blood flow. The purported mechanism by which (99m)Tc-HMPAO accumulates in the brain is through its intracellular conversion from a lipophilic form to more hydrophilic derivatives within the brain parenchyma. The issue of the contribution of different cell types on (99m)Tc-HMPAO retention was investigated in vitro by studying the accumulation of (99m)Tc-HMPAO in primary cultures of mouse cortical astrocytes and neurons. Results show that (99m)Tc-HMPAO retention predominates in astrocytes over neurons by a factor of approximately 2.5 (0.26 +/- 0.05 vs. 0.095 +/- 0.042 fmol/mg protein after 120 minutes, respectively). Diethyl maleate (60 micromol/L), ethacrynic acid (1 mmol/L) and buthionine sulfoximine (1 mmol/L), 3 agents which significantly reduced glutathione levels also decreased (99m)Tc-HMPAO retention in both astrocytes (29%, 3%, and 46% of control, respectively) and neurons (69%, 11% and 63% of control). Decrease did not always correlate with glutathione levels, however, which suggests that other factors could be involved. The possibility that cell energy status determines (99m)Tc-HMPAO retention was also assessed. Agents that activate (glutamate, azide) or inhibit (cytochalasin B) glucose utilization in astrocytes, as measured by the (3)H-2-deoxyglucose method, were without effect on (99m)Tc-HMPAO retention. In conclusion, the data presented indicate that astrocytes may constitute a prominent site of (99m)Tc-HMPAO retention and most likely contribute significantly to the SPECT signal. In addition, the data also suggest that specific alterations in glial cell metabolism could explain flow-independent changes in (99m)Tc-HMPAO retention in the brain as observed by SPECT in some pathologies.


The GTEx Consortium atlas of genetic regulatory effects across human tissues.
The Genotype-Tissue Expression (GTEx) project was established to characterize genetic effects on the transcriptome across human tissues and to link these regulatory mechanisms to trait and disease associations. Here, we present analyses of the version 8 data, examining 15,201 RNA-sequencing samples from 49 tissues of 838 postmortem donors. We comprehensively characterize genetic associations for gene expression and splicing in cis and trans, showing that regulatory associations are found for almost all genes, and describe the underlying molecular mechanisms and their contribution to allelic heterogeneity and pleiotropy of complex traits. Leveraging the large diversity of tissues, we provide insights into the tissue specificity of genetic effects and show that cell type composition is a key factor in understanding gene regulatory mechanisms in human tissues.


TMS over the intraparietal sulcus induces perceptual fading.
During prolonged fixation, visual objects presented in the periphery of the visual field often fade from awareness. This phenomenon, known as the Troxler effect, has been largely attributed to adaptation of neurons responding to peripheral targets. Here, we hypothesized that perceptual disappearance might result from degeneration of feedback from attention-related cortical areas to early visual areas and that visual transients disrupt the feedback loop sustaining low-level signals and thereby trigger perceptual fading. We examined this hypothesis by briefly disrupting the functions of attention-related regions in the intraparietal sulcus (IPS) using transcranial magnetic stimulation (TMS). The hypothesis predicted that temporary disruption of IPS would trigger perceptual disappearances. We measured perceptual disappearance using a task in which participants were asked to discriminate the presence or absence (fading) of a peripheral green target immediately after a TMS pulse. On one half of the trials, the target remained on the screen until the end of a trial, and on the other half of the trials, it gradually faded. The results of this experiment show that brief disruption of the IPS with a single pulse TMS is sufficient to trigger perceptual disappearance. TMS over the IPS resulted in a fewer reports of continuous percepts (i.e., more fading) without changes in the perception of physically fading stimuli. Our control experiment shows the loss of sensitivity is not caused by suppression of microsaccades in response to the sound produced by TMS. This study supports the idea that conscious visual perception requires a coupling between the early visual areas representing sensory data and the parietal areas subserving spatial localization functions.


Acute effects of cocaine on spontaneous and discriminative motor functions: relation to route of administration and pharmacokinetics.
Rats administered cocaine i.p. and p.o. (7.5-30 mg/kg) showed dose-related increases in locomotor (LM) and small-movement activities, with LM rates decreasing over the 2-hr session, except at the largest i.p. dose, for which rates were greater in the 2nd hr. Lidocaine p.o. (15-30 mg/kg) did not increase activity. Relating the area under the curve measures for serum cocaine (concentration-time) and LM activity (LM activity-time) for 2 hr postadministration indicated that cocaine was about twice as potent i.p., compared to p.o., in increasing LM activity. Cocaine (i.p. and p.o.) produced dose-related decrements in both discriminative motor control performance and in task work rate, whereas lidocaine p.o. did not. The motor control decrements produced by cocaine were approximately comparable by i.p. and p.o. routes, whereas effects on LM rates were much greater by i.p. than by p.o. administration. The effects of cocaine by both routes on LM rates were proportionally much greater than its effect on motor control performance. Changes in LM rates and motor control performance over the postadministration period were related to the pharmacokinetic features (maximum serum concentration, time to maximum serum concentration and elimination half-life) of cocaine observed for the routes explored (i.p., p.o. and s.c.). Tail-tip serum samples, although yielding conservative estimates of cocaine concentration, correlated well with trunk serum and brain cocaine levels.


Nano-channels in the spider fang for the transport of Zn ions to cross-link His-rich proteins pre-deposited in the cuticle matrix.
We identify the presence of multiple vascular channels within the spider fang. These channels seem to serve the transport of zinc to the tip of the fang to cross-link the protein matrix by binding to histidine residues. According to amino acid and elemental analysis of fangs extracted shortly after ecdysis, His-rich proteins are deposited before Zn is incorporated into the cuticle. Microscopic and spectroscopic investigations in the electron microscope and synchrotron radiation experiments suggest that Zn ions are transported through these channels in a liable (yet unidentified) form, and then form stable complexes upon His binding. The resulting cross-linking through the Zn-His complexes is conferring hardness to the fang. Our observations of nano-channels serving the Zn-transport within the His-rich protein matrix of the fibre reinforced spider fang may also support recent bio-inspired attempts to design artificial polymeric vascular materials for self-healing and in-situ curing.


A spatial pattern analysis of beta-amyloid (Abeta) deposition in the temporal lobe in Alzheimer's disease.
To determine the factors influencing the distribution of -amyloid (Abeta) deposits in Alzheimer's disease (AD), the spatial patterns of the diffuse, primitive, and classic A deposits were studied from the superior temporal gyrus (STG) to sector CA4 of the hippocampus in six sporadic cases of the disease. In cortical gyri and in the CA sectors of the hippocampus, the Abeta deposits were distributed either in clusters 200-6400 microm in diameter that were regularly distributed parallel to the tissue boundary or in larger clusters greater than 6400 microm in diameter. In some regions, smaller clusters of Abeta deposits were aggregated into larger 'superclusters'. In many cortical gyri, the density of Abeta deposits was positively correlated with distance below the gyral crest. In the majority of regions, clusters of the diffuse, primitive, and classic deposits were not spatially correlated with each other. In two cases, double immunolabelled to reveal the Abeta deposits and blood vessels, the classic Abeta deposits were clustered around the larger diameter vessels. These results suggest a complex pattern of Abeta deposition in the temporal lobe in sporadic AD. A regular distribution of Abeta deposit clusters may reflect the degeneration of specific cortico-cortical and cortico-hippocampal pathways and the influence of the cerebral blood vessels. Large-scale clustering may reflect the aggregation of deposits in the depths of the sulci and the coalescence of smaller clusters.


The Genetic Information Nondiscrimination Act (GINA): public policy and medical practice in the age of personalized medicine.
Survey data suggest that many people fear genetic discrimination by health insurers or employers. In fact, such discrimination has not yet been a significant problem. This article examines the fear and reality of genetic discrimination in the United States, describes how Congress sought to prohibit such discrimination by passing the Genetic Information Nondiscrimination Act of 2008 (GINA), and explores the implications of GINA for general internists and their institutions. It concludes that medical providers and health care institutions must be familiar with the general intent and specific terms of GINA, and should continue to collect genetic information that can contribute to the high quality provision of medical treatment. Not doing so violates their medical mission and diminishes the quality of care patients deserve.


Peer support: healthcare professionals supporting each other after adverse medical events.
The patient safety movement in healthcare is beginning to openly acknowledge the need to support the human side of adverse medical events in conjunction with evidence-based improvement initiatives. While medical literature has sporadically reported on the emotional impact of adverse events on healthcare professionals, little has been documented on the implementation of support services following these events. This article describes an adverse medical event where open communication and apology catalysed the development and implementation of a structured peer support service for care providers at the Brigham and Women's Hospital following adverse events. The Peer Support Service bypasses the stigmas that limit the utilisation of formal support services and offers care providers a safe environment to share the emotional impact of adverse events while serving as a foundation for open communication and a renewal of compassion in the workplace. As the breadth of stressors impacting healthcare professionals is revealed, the Peer Support Service is being recognised as a vital hospital-wide service. It also appears to offer an important leap forward in the critical areas of patient safety and quality of care.


Noradrenergic denervation alters serotonin2-mediated behavior but not serotonin2 receptor number in rats: modulatory role of beta adrenergic receptors.
Recent behavioral evidence suggests that enhancement of noradrenergic neurotransmission may alter the functional sensitivity of serotonin2 (5-HT2) receptors in the central nervous system. The present studies have examined the effects of two types of noradrenergic denervation [neurotoxic: via N-(2-chloroethyl)-N-ethyl-2-bromobenzylamine (DSP4) treatment; and pharmacologic: via chronic beta adrenergic receptor blockade] on the 5-HT2-mediated head shake response and cortical beta adrenergic and 5-HT2 receptor number in the rat. No changes in quipazine-induced head shakes were observed 3 days after DSP4 lesion. However, the frequency of head shakes was significantly enhanced 10 days after DSP4 treatment in the presence of a 39% up-regulation of beta adrenergic receptors. Pretreatment with propranolol 10 days after DSP4 lesion selectively antagonized the enhancement of the behavioral response to quipazine without altering base-line response rate, whereas pretreatment with the 5-HT2 antagonist ketanserin totally blocked head shakes in both control and DSP4-treated rats. Pharmacologic denervation achieved by continuous (14 day) administration of the beta adrenergic antagonist propranolol also resulted in a potentiation of the head shake response (274% of control) and an upregulation of beta adrenergic receptors (44%). Conversely, continuous treatment with the beta adrenergic agonist clenbuterol resulted in a marked reduction in head shakes (36% of control) with a concomitant 29% down-regulation of beta adrenergic receptors. 5-HT2 receptor binding was not modified by either DSP4 lesion or continuous administration of beta adrenergic agonists or antagonists. These studies demonstrate that changes in cortical beta adrenergic receptor density may modify 5-HT2-mediated behavior in a manner that is independent of changes in 5-HT2 receptor number.


Intracellular trafficking and subcellular distribution of a large array of HPMA copolymers.
The basic physicochemical properties that determine the distribution and fate of synthetic macromolecules in living cells were characterized using fluorescently labeled HPMA (N-(2-hydroxypropyl)methacrylamide) copolymers. Twelve different classes of water-soluble copolymers were created by incorporating eight different functionalized comonomers. These comonomers possessed functional groups with positive or negative charges or contained short hydrophobic peptides. The copolymers were fractionated to create parallel "ladders" consisting of 10 fractions of narrow polydispersity with molecular weights ranging from 10 to 200 kDa. The intracellular distributions were characterized for copolymer solutions microinjected into the cytoplasm of cultured ovarian carcinoma cells. Even the highest molecular weight HPMA copolymers were shown to quickly and evenly diffuse throughout the cytoplasm and remain excluded from membrane-bound organelles, regardless of composition. The exceptions were the strongly cationic copolymers, which demonstrated a pronounced localization to microtubules. For all copolymers, nuclear entry was consistent with passive transport through the nuclear pore complex (NPC). Nuclear uptake was shown to be largely dictated by the molecular weight of the copolymers, however, detailed kinetic analyses showed that nuclear import rates were moderately, but significantly, affected by differences in comonomer composition. HPMA copolymers containing amide-terminated phenylalanine-glycine (FG) sequences, analogous to those found in the NPC channel protein, demonstrated a potential to regulate import to the nuclear compartment. Kinetic analyses showed that 15 kDa copolymers containing GGFG, but not those containing GGLFG, peptide pendant groups altered the size-exclusion characteristics of NPC-mediated nuclear import.


Baltic Sea sediment records: unlikely near-future declines in PCBs and HCB.
We present a comprehensive study on PCBs and HCB in dated sediment cores from the Baltic Sea covering the 20th century, and compare their spatiotemporal trends with those of PCDD/Fs from the same areas. PCB concentrations in coastal impacted sediment followed the temporal trend of estimated global emissions of PCBs and thus responded quickly to changes in global industrial use, whereas concentrations in offshore sediment needed 10-20 years longer to respond. Differences in spatiotemporal trends of PCDD/Fs and PCBs were smaller than expected based on documented differences in key sources and source areas. Sediment concentrations of HCB varied little over time and space, but concentrations are increasing in recent years. The steep PCB concentration reduction over time observed for the late 20th century levelled off during the last 20 years, and levels of PCBs appear to be at or near a steady-state condition. PCB concentrations in Baltic Sea sediments appear to be at or near steady-state, and no significant concentration decreases are to be expected in the near future.


Louis K. Dahl Memorial Lecture. Renal and cardiovascular mechanisms of hypertension in obesity.
In all forms of hypertension, including human essential hypertension, pressure natriuresis is reset to higher blood pressures. Because human essential hypertension is a heterogeneous disease, it is likely that there are multiple neurohumoral and intrarenal causes of abnormal pressure natriuresis and increased blood pressure. Weight gain is recognized to be an important contributor to essential hypertension, although the mechanisms that link obesity with altered renal function and high blood pressure have not been fully elucidated. In obese dogs and humans, the shift of pressure natriuresis to higher blood pressures appears to be due mainly to increased tubular reabsorption, as glomerular filtration rate and renal plasma flow are increased compared with normal. Multiple causes of increased tubular reabsorption and hypertension in obesity have been postulated, including insulin resistance and hyperinsulinemia, activation of the sympathetic nervous and renin-angiotensin systems, and physical changes within the kidney itself. Support for the insulin resistance-hyperinsulinemia link between obesity and hypertension has been inferred mainly from acute and epidemiologic studies showing a correlation between insulin and blood pressure. Recent studies suggest that chronic hyperinsulinemia, comparable to that found in obesity, cannot account for obesity hypertension in dogs or humans. Activation of the sympathetic nervous system may play a role in obesity-induced hypertension, and there is evidence for a role of altered intrarenal physical forces caused by histological changes within the renal medulla. The quantitative importance of each of these abnormalities in altering renal function and raising blood pressure in obesity remains to be determined but is an important area of research for understanding human essential hypertension.


No longer diseases of the wealthy: prevalence and health-seeking for self-reported chronic conditions among urban poor in Southern India.
The burden of chronic conditions is high in low- and middle-income countries and poses a significant challenge to already weak healthcare delivery systems in these countries. Studies investigating chronic conditions among the urban poor remain few and focused on specific chronic conditions rather than providing overall profile of chronic conditions in a given community, which is critical for planning and managing services within local health systems. We aimed to assess the prevalence and health- seeking behaviour for self-reported chronic conditions in a poor neighbourhood of a metropolitan city in India. We conducted a house-to-house survey covering 9299 households (44514 individuals) using a structured questionnaire. We relied on self-report by respondents to assess presence of any chronic conditions, including diabetes and hypertension. Multivariable logistic regression was used to analyse the prevalence and health-seeking behaviour for self-reported chronic conditions in general as well as for diabetes and hypertension in particular. The predictor variables included age, sex, income, religion, household poverty status, presence of comorbid chronic conditions, and tiers in the local health care system. Overall, the prevalence of self-reported chronic conditions was 13.8% (95% CI = 13.4, 14.2) among adults, with hypertension (10%) and diabetes (6.4%) being the most commonly reported conditions. Older people and women were more likely to report chronic conditions. We found reversal of socioeconomic gradient with people living below the poverty line at significantly greater odds of reporting chronic conditions than people living above the poverty line (OR = 3, 95% CI = 1.5, 5.8). Private healthcare providers managed over 80% of patients. A majority of patients were managed at the clinic/health centre level (42.9%), followed by the referral hospital (38.9%) and the super-specialty hospital (18.2%) level. An increase in income was positively associated with the use of private facilities. However, elderly people, people below the poverty line, and those seeking care from hospitals were more likely to use government services. Our findings provide further evidence of the urgent need to improve care for chronic conditions for urban poor, with a preferential focus on improving service delivery in government health facilities.


Against COVID-19 vaccination of healthy children.
COVID-19 vaccination of children has begun in a number of countries with provisional regulatory approval and public support. This article provides an ethical analysis of COVID-19 vaccination of healthy children. Specifically, we present three of the strongest arguments that might justify COVID-19 vaccination of children: (a) an argument from paternalism, (b) an argument from indirect protection and altruism, and (c) an argument from global eradication. We offer a series of objections to each of these arguments to show that none of them is currently tenable. Given the minimal direct benefit of COVID-19 vaccination for healthy children, the potential for rare risks to outweigh these benefits and to undermine vaccine confidence, the substantial evidence that COVID-19 vaccination confers adequate protection to risk groups whether or not healthy children are vaccinated and that current vaccines do not provide sterilizing immunity, and given that eradication of the virus is neither feasible nor a high priority for global health, we argue that routine COVID-19 vaccination of healthy children is currently ethically unjustified. Since mandates for children have already been implemented in some places (e.g., California) and may be considered elsewhere, we also present two additional arguments explicitly against making COVID-19 vaccination mandatory for children.