Evaluation of Pringle maneuver during liver resection in a rat model of surgical obstructive jaundice.
Temporary portal triad clamping (Pringle maneuver) during liver resection reduces intraoperative blood loss. A normal liver can safely tolerate normothermic ischemia for up to 60 min. However, its safety in patients with surgical obstructive jaundice (SOJ) is not known. Therefore, we investigated the effect of hepatic ischemia in an experimental rat model of SOJ created by ligating the bile duct. Four groups of rats were created: Group I (sham operation, 10 days later, liver resection); Group II (sham operation, 10 days later, liver resection with 5 min of hepatic ischemia); Group III (bile duct ligation, 10 days later, liver resection); and Group IV (bile duct ligation, 10 days later, liver resection with 5 min of hepatic ischemia). The ischemic injury was assessed by the survival of rats, liver tissue malondialdehyde and total glutathione (markers of free radical injury), serum alanine aminotransferase, aspartate aminotransferase, and liver histology. The results showed decreased survival (47.6% vs. 90% [p = .046]), increased liver tissue malondialdehyde (161 +/- 35 vs. 129 +/- 33 microg/gm liver tissue [p = .05]), and decreased liver tissue total glutathione (565 +/- 169 vs. 1075 +/- 276 nmol/gm liver tissue [p = .05]) in rats with SOJ subjected to hepatic ischemia when compared to nonjaundiced rats. The changes in serum aspartate aminotransferase (AST), alanine aminotransferase (ALT), and alkaline phosphatase showed an increasing trend in the SOJ group but were not statistically significant. Ischemic changes in liver histology were seen more often in the SOJ group but were not statistically significant. These data suggest that temporary portal triad clamping in an experimental model of SOJ is detrimental to the outcome of liver resection.


Improvement of antibiotic therapy and ICU survival in severe non-pneumococcal community-acquired pneumonia: a matched case-control study.
We aimed to compare intensive care unit mortality due to non-pneumococcal severe community-acquired pneumonia between the periods 2000-2002 and 2008-2014, and the impact of the improvement in antibiotic strategies on outcomes. This was a matched case-control study enrolling 144 patients with non-pneumococcal severe pneumonia: 72 patients from the 2000-2002 database (CAPUCI I group) were paired with 72 from the 2008-2014 period (CAPUCI II group), matched by the following variables: microorganism, shock at admission, invasive mechanical ventilation, immunocompromise, chronic obstructive pulmonary disease, and age over 65 years. The most frequent microorganism was methicillin-susceptible Staphylococcus aureus (22.1%) followed by Legionella pneumophila and Haemophilus influenzae (each 20.7%); prevalence of shock was 59.7%, while 73.6% of patients needed invasive mechanical ventilation. Intensive care unit mortality was significantly lower in the CAPUCI II group (34.7% versus 16.7%; odds ratio (OR) 0.78, 95% confidence interval (CI) 0.64-0.95; p = 0.02). Appropriate therapy according to microorganism was 91.5% in CAPUCI I and 92.7% in CAPUCI II, while combined therapy and early antibiotic treatment were significantly higher in CAPUCI II (76.4 versus 90.3% and 37.5 versus 63.9%; p < 0.05). In the multivariate analysis, combined antibiotic therapy (OR 0.23, 95% CI 0.07-0.74) and early antibiotic treatment (OR 0.07, 95% CI 0.02-0.22) were independently associated with decreased intensive care unit mortality. In non-pneumococcal severe community-acquired pneumonia , early antibiotic administration and use of combined antibiotic therapy were both associated with increased intensive care unit survival during the study period.


Taxonomic revision of the Andean genus Xenophyllum (Compositae, Senecioneae).
The Andean genus Xenophyllum (Compositae, Senecioneae) is distributed along the high-Andes from northeastern Colombia to northern Chile and northwestern Argentina, mainly thriving in the paramo and puna ecoregions. It comprises suffruticose plants forming dense mats, hummocks, or clumps of erect stems. They are characterized by displaying involucral bracts fused at the base, supplementary bracts absent, and mostly radiate capitula with white ray corollas, seldom yellow or pink (disciform in one species). Traditionally, Xenophyllum species were treated as members of the genus Werneria, a morphologically close genus that includes rosettiform or scapiform perennial herbs. As currently circumscribed, Xenophyllum mostly differs from Werneria in having elongate stems. Herein, the first modern and comprehensive revision of the genus recognizing twenty-two species and two subspecies is presented. Werneria decumbens is synonymized with X. weddellii, as well as X. fontii with X. humile and X. oscartovarii with X. dactylophyllum. Likewise, four varietal names and two sectional names are proposed as new synonyms. Seven names are lectotypified, the name X. sotarense is epitypified, W. decumbens neotypified, and the supraspecific name W. sect. Integrifoliae Rockh. is typified. The combination X. crassum subsp. orientalecomb. nov. is made. Descriptions and distribution maps are provided for all accepted species, in addition to an identification key. Ten species are illustrated, three of them for the first time.


Green tea compound in chemoprevention of cervical cancer.
Human papillomavirus (HPV) infection is closely associated with the development of more than 95% of cervical cancer. Clinical trials using several chemopreventive agents are underway, but results are inconclusive. Most agents used in trials inhibited the growth of cancer cells in vitro, and about half of patients had some degree of clinical responses; however, the therapeutic effect was confounded by high rates of spontaneous regression and relapse. The selection of nontoxic agents especially food, beverage, and natural products that suppress oncogenic HPV, inhibit malignant transformation, and can additionally be used long term may be important for cervical cancer prevention. We evaluated green tea compound (epigallocatechin gallate and polyphenols E) effects on immortalized cervical epithelial and cervical cancer cells. HPV-immortalized cervical epithelial cells, TCL1, and HPV-positive cervical cancer cells, Me180 and HeLa, were used in the study. The effects of green tea compounds on cell growth, apoptosis, cell cycle, and gene expression were examined and characterized. Both epigallocatechin gallate and polyphenols E inhibited immortalized cervical epithelial and cancer cell growth. Apoptosis induction and cell cycle changes were observed in a dose-dependent manner. Western blot analysis of apoptosis-related proteins, p53 and p21, showed dose-dependent increase, whereas p27 was not affected. HPV-E7 protein expression was decreased by green tea compounds. This study provides information on the potential mechanisms of action of green tea compounds in suppression of HPV-related cervical cells, and it will enable us to assess the feasibility of using these agents.


Network-level analysis of metabolic regulation in the human red blood cell using random sampling and singular value decomposition.
Extreme pathways (ExPas) have been shown to be valuable for studying the functions and capabilities of metabolic networks through characterization of the null space of the stoichiometric matrix (S). Singular value decomposition (SVD) of the ExPa matrix P has previously been used to characterize the metabolic regulatory problem in the human red blood cell (hRBC) from a network perspective. The calculation of ExPas is NP-hard, and for genome-scale networks the computation of ExPas has proven to be infeasible. Therefore an alternative approach is needed to reveal regulatory properties of steady state solution spaces of genome-scale stoichiometric matrices. We show that the SVD of a matrix (W) formed of random samples from the steady-state solution space of the hRBC metabolic network gives similar insights into the regulatory properties of the network as was obtained with SVD of P. This new approach has two main advantages. First, it works with a direct representation of the shape of the metabolic solution space without the confounding factor of a non-uniform distribution of the extreme pathways and second, the SVD procedure can be applied to a very large number of samples, such as will be produced from genome-scale networks. These results show that we are now in a position to study the network aspects of the regulatory problem in genome-scale metabolic networks through the use of random sampling.


Management of refractory ascites.
Ascites that does not respond or recurs after high-dose diuresis and sodium restriction should be considered refractory ascites. As cirrhosis advances, the escaping fluid overwhelms the lymphatic return. Decrease in renal plasma flow leads to increased sodium reabsorption at the proximal tubule leading to decreased responsiveness to loop diuretics and mineralocorticoid antagonists, which work distally. These complex hemodynamic alterations lead to refractory ascites. In refractory ascites, high-dose diuresis (400 mg of spironolactone and 160 mg of furosemide) and sodium restriction (<90 mmol/d) result in inadequate weight loss and sub optimal sodium excretion (<78 mmol/d). Further use of diuretics is limited by complications such as encephalopathy, azotemia, renal insufficiency, hyponatremia, and hyperkalemia. Therapy for refractory ascites is limited. The available therapies are repeated large volume paracentesis (LVP), transjugular intrahepatic portosystemic shunts, peritoneovenous shunts, investigational medical therapies, and liver transplantation. LVP with concomitant volume expanders is the initial treatment of choice. Transjugular intrahepatic portosystemic seems to be superior to LVP in reducing the need for repeated paracentesis and improves the quality of life. Several treatments that act at different steps in the pathogenesis of ascites are investigational, and some show promising results. Splanchnic and peripheral vasoconstrictors (Octreotide, Midodrine, and Terlipressin) increase effective arterial volume and decrease activation of the renin-angiotensin system with resultant increase in renal sodium excretion. Clonidine when given with spironolactone has been shown to cause rapid mobilization of ascites by significantly decreasing the sympathetic activity and renin-aldosterone levels. Natural aquaretics and synthetic V2 receptor antagonists (satavaptan) are being evaluated for mobilization of ascites by increasing the excretion of solute-free water. Liver transplantation remains the only definitive therapy for refractory ascites. Because refractory ascites is a poor prognostic sign, liver transplantation should be considered and incorporated early in the treatment plan.


Relative impacts of environmental variation and evolutionary history on the nestedness and modularity of tree-herbivore networks.
Nestedness and modularity are measures of ecological networks whose causative effects are little understood. We analyzed antagonistic plant-herbivore bipartite networks using common gardens in two contrasting environments comprised of aspen trees with differing evolutionary histories of defence against herbivores. These networks were tightly connected owing to a high level of specialization of arthropod herbivores that spend a large proportion of the life cycle on aspen. The gardens were separated by ten degrees of latitude with resultant differences in abiotic conditions. We evaluated network metrics and reported similar connectance between gardens but greater numbers of links per species in the northern common garden. Interaction matrices revealed clear nestedness, indicating subsetting of the bipartite interactions into specialist divisions, in both the environmental and evolutionary aspen groups, although nestedness values were only significant in the northern garden. Variation in plant vulnerability, measured as the frequency of herbivore specialization in the aspen population, was significantly partitioned by environment (common garden) but not by evolutionary origin of the aspens. Significant values of modularity were observed in all network matrices. Trait-matching indicated that growth traits, leaf morphology, and phenolic metabolites affected modular structure in both the garden and evolutionary groups, whereas extra-floral nectaries had little influence. Further examination of module configuration revealed that plant vulnerability explained considerable variance in web structure. The contrasting conditions between the two gardens resulted in bottom-up effects of the environment, which most strongly influenced the overall network architecture, however, the aspen groups with dissimilar evolutionary history also showed contrasting degrees of nestedness and modularity. Our research therefore shows that, while evolution does affect the structure of aspen-herbivore bipartite networks, the role of environmental variations is a dominant constraint.


A family of moderately repetitive sequences in mouse DNA.
When mouse DNA is digested to completion with restriction endonuclease Eco R1, a distinct band of 1.3 kb segments comprising about 0.5-3% of the genome is observed upon agarose gel electrophoresis. This DNA is not tandemly repeated in the genome and is not derived from mouse satellite DNA. Restriction endonuclease analysis suggested that the 1.3 kb segments are heterogeneous. Specific sequences were selected from the 1.3 kb segments and amplified by cloning in plasmid pBR322. Southern transfer experiments indicated that three separately cloned mouse DNA inserts hybridized predominantly to the Eco R1 1.3 kb band and to the conspicuous subsegments generated by secondary restriction endonuclease cleavage of the sucrose gradient purified 1.3 kb segments. Segments were also excised by Hha I (Hha I segments) from the chimeric plasmids containing mouse DNA inserts and subjected to restriction endonuclease and cross-hybridization analysis. It was found that the three Hha I segments were different, although two of them exhibited partial sequence homology. Cot analysis indicated that each of the Hha I segments are repeated about 10(4) times in the mouse genome. These findings indicate that a family of related but non-identical, moderately repetitive DNA sequences, rather than a single homogeneous repeat, is present in the 1.3 kb Eco R1 band.


Infant mortality in Costa Rica: explaining the recent decline.
Costa Rica has undergone a dramatic reduction in its infant mortality rate from 68 per 1,000 live births in 1970 to 20 per 1,000 in 1980. In the present study, changes during this century, mortality differentials, and causes of death are analyzed, and multiple regression techniques are used to identify the determinants of the decline in Costa Rica's 79 cantons (counties). Although socioeconomic development and greatly reduced fertility contributed to the infant mortality decline, as much as three-fourths of the decline is attributable to public health programs implemented during the 1970s. The extension of primary health care--especially rural and community programs--seems to be responsible for 40 percent of the reduction. In addition, health services produced a notable decrease in the socioeconomic differentials related to children's risk of death. The unique achievements of this developing country offer a new strategy for public health improvement.


Asthma caused by bromelain: an occupational allergy.
Bromelains consist of a group of proteolytic enzymes of Bromeliaceae. They are commonly used in pharmaceutical industries, food production and in diagnostic laboratories. Bromelains are known to cause IgE-mediated reactions of both the immediate type and the 'late phase reaction of immediate type reaction' with predominantly respiratory symptoms. We report four cases of occupational allergy to bromelain in workers of a blood grouping laboratory. These observations prompted us to investigate the sensitization rate to bromelain in all workers of the particular diagnostic laboratory who had contact with bromelain. These results were compared with those obtained from healthy, randomly selected individuals without evident bromelain exposure. Our findings indicate that (i) bromelain is a strong sensitizer, (ii) sensitization usually occurs due to inhalation and not to ingestion, (iii) bromelain allergy is occupationally acquired, and adequate precautions are necessary. We can further state that (iv) skin testing with relatively pure allergens such as isolated proteases like bromelain may induce systemic reactions, even at very high dilutions.


Longitudinal transcriptomics define the stages of myeloid activation in the living human brain after intracerebral hemorrhage.
Opportunities to interrogate the immune responses in the injured tissue of living patients suffering from acute sterile injuries such as stroke and heart attack are limited. We leveraged a clinical trial of minimally invasive neurosurgery for patients with intracerebral hemorrhage (ICH), a severely disabling subtype of stroke, to investigate the dynamics of inflammation at the site of brain injury over time. Longitudinal transcriptional profiling of CD14+ monocytes/macrophages and neutrophils from hematomas of patients with ICH revealed that the myeloid response to ICH within the hematoma is distinct from that in the blood and occurs in stages conserved across the patient cohort. Initially, hematoma myeloid cells expressed a robust anabolic proinflammatory profile characterized by activation of hypoxia-inducible factors (HIFs) and expression of genes encoding immune factors and glycolysis. Subsequently, inflammatory gene expression decreased over time, whereas anti-inflammatory circuits were maintained and phagocytic and antioxidative pathways up-regulated. During this transition to immune resolution, glycolysis gene expression and levels of the potent proresolution lipid mediator prostaglandin E2 remained elevated in the hematoma, and unexpectedly, these elevations correlated with positive patient outcomes. Ex vivo activation of human macrophages by ICH-associated stimuli highlighted an important role for HIFs in production of both inflammatory and anti-inflammatory factors, including PGE2, which, in turn, augmented VEGF production. Our findings define the time course of myeloid activation in the human brain after ICH, revealing a conserved progression of immune responses from proinflammatory to proresolution states in humans after brain injury and identifying transcriptional programs associated with neurological recovery.


Association of time to craniectomy with survival in patients with severe combat-related brain injury.
OBJECTIVEIn combat and austere environments, evacuation to a location with neurosurgery capability is challenging. A planning target in terms of time to neurosurgery is paramount to inform prepositioning of neurosurgical and transport resources to support a population at risk. This study sought to examine the association of wait time to craniectomy with mortality in patients with severe combat-related brain injury who received decompressive craniectomy.METHODSPatients with combat-related brain injury sustained between 2005 and 2015 who underwent craniectomy at deployed surgical facilities were identified from the Department of Defense Trauma Registry and Joint Trauma System Role 2 Registry. Eligible patients survived transport to a hospital capable of diagnosing the need for craniectomy and performing surgery. Statistical analyses included unadjusted comparisons of postoperative mortality by elapsed time from injury to start of craniectomy, and Cox proportional hazards modeling adjusting for potential confounders. Time from injury to craniectomy was divided into quintiles, and explored in Cox models as a binary variable comparing early versus delayed craniectomy with cutoffs determined by the maximum value of each quintile (quintile 1 vs 2-5, quintiles 1-2 vs 3-5, etc.). Covariates included location of the facility at which the craniectomy was performed (limited-resource role 2 facility vs neurosurgically capable role 3 facility), use of head CT scan, US military status, age, head Abbreviated Injury Scale score, Injury Severity Score, and injury year. To reduce immortal time bias, time from injury to hospital arrival was included as a covariate, entry into the survival analysis cohort was defined as hospital arrival time, and early versus delayed craniectomy was modeled as a time-dependent covariate. Follow-up for survival ended at death, hospital discharge, or hospital day 16, whichever occurred first.RESULTSOf 486 patients identified as having undergone craniectomy, 213 (44%) had complete date/time values. Unadjusted postoperative mortality was 23% for quintile 1 (n = 43, time from injury to start of craniectomy 30-152 minutes); 7% for quintile 2 (n = 42, 154-210 minutes); 7% for quintile 3 (n = 43, 212-320 minutes); 19% for quintile 4 (n = 42, 325-639 minutes); and 14% for quintile 5 (n = 43, 665-3885 minutes). In Cox models adjusted for potential confounders and immortal time bias, postoperative mortality was significantly lower when time to craniectomy was within 5.33 hours of injury (quintiles 1-3) relative to longer delays (quintiles 4-5), with an adjusted hazard ratio of 0.28, 95% CI 0.10-0.76 (p = 0.012).CONCLUSIONSPostoperative mortality was significantly lower when craniectomy was initiated within 5.33 hours of injury. Further research to optimize craniectomy timing and mitigate delays is needed. Functional outcomes should also be evaluated.


Voltage-dependent K+ channels improve the energy efficiency of signalling in blowfly photoreceptors.
Voltage-dependent conductances in many spiking neurons are tuned to reduce action potential energy consumption, so improving the energy efficiency of spike coding. However, the contribution of voltage-dependent conductances to the energy efficiency of analogue coding, by graded potentials in dendrites and non-spiking neurons, remains unclear. We investigate the contribution of voltage-dependent conductances to the energy efficiency of analogue coding by modelling blowfly R1-6 photoreceptor membrane. Two voltage-dependent delayed rectifier K+ conductances (DRs) shape the membrane's voltage response and contribute to light adaptation. They make two types of energy saving. By reducing membrane resistance upon depolarization they convert the cheap, low bandwidth membrane needed in dim light to the expensive high bandwidth membrane needed in bright light. This investment of energy in bandwidth according to functional requirements can halve daily energy consumption. Second, DRs produce negative feedback that reduces membrane impedance and increases bandwidth. This negative feedback allows an active membrane with DRs to consume at least 30% less energy than a passive membrane with the same capacitance and bandwidth. Voltage-dependent conductances in other non-spiking neurons, and in dendrites, might be organized to make similar savings.


Selenium-Containing Nanoparticles Combine the NK Cells Mediated Immunotherapy with Radiotherapy and Chemotherapy.
Considering the limited clinical benefits of individual approaches against malignancy, natural killer (NK) cell-mediated immunotherapy is increasingly utilized in combination with radiotherapy and target therapeutics. However, the interplay of targeted agents, immunotherapy, and radiotherapy is complex. An improved understanding of the effect of chemotherapy or radiotherapy on specific molecular pathways in immune cells would help to optimize the synergistic antitumor efficiency. In this study, the selenium-containing nanoparticles (NPs) could deliver the chemotherapeutic drug doxorubicin (DOX) to tumor sites by systemic administration. Radiation stimuli facilitate DOX release and enhance chemotherapy efficiency. Moreover, radiation could oxidize diselenide-containing NPs to seleninic acid, which have both synergistic antitumor effect and immunomodulatory activity through enhancing NK cells function. These results indicate that the selenium-containing NPs would be a potential approach to achieve simultaneous treatments of immunotherapy, chemotherapy, and radiotherapy by a simple but effective method.


The "con" of concept analysis A discussion paper which explores and critiques the ontological focus, reliability and antecedents of concept analysis frameworks.
This paper draws on the work of Paley and Duncan et al in order to extend and engender debate regarding the use of Concept Analysis frameworks. Despite the apparent plethora of Concept Analysis frameworks used in nursing studies we found that over half of those used were derived from the work of one author. This paper explores the suitability and use of these frameworks and is set at a time when the numbers of published concept analysis papers are increasing. For the purpose of this study thirteen commonly used frameworks, identified from the nursing journals 1993 to 2005, were explored to reveal their origins, ontological and philosophical stance, and any common elements. The frameworks were critiqued and links made between their antecedents. It was noted if the articles contained discussion of any possible tensions between the ontological perspective of the framework used, the process of analysis, praxis and possible nursing theory developments. It was found that the thirteen identified frameworks are mainly based on hermeneutic propositions regarding understandings and are interpretive procedures founded on self-reflective modes of discovery. Six frameworks rely on or include the use of casuistry. Seven of the frameworks identified are predicated on, or adapt the work of Wilson, a school master writing for his pupils. Wilson's framework has a simplistic eleven step, binary and reductionist structure. Other frameworks identified include Morse et al's framework which this article suggests employs a contestable theory of concept maturity. Based on the findings revealed through our exploration of the use of concept analysis frameworks in the nursing literature, concerns were raised regarding the unjustified adaptation and alterations and the uncritical use of the frameworks. There is little evidence that these frameworks provide the necessary depth, rigor or replicability to enable the development in nursing theory which they underpin.


Factors affecting reliability and validity of self-directed automatic in situ audiometry: implications for self-fitting hearing AIDS.
A reliable and valid method for the automatic in situ measurement of hearing thresholds is a prerequisite for the feasibility of a self-fitting hearing aid, whether such a device becomes an automated component of an audiological management program or is fitted by the user independently of a clinician. Issues that must be addressed before implementation of the procedure into a self-fitting hearing aid include the role of real-ear-to-dial difference correction factors in ensuring accurate results and the ability of potential users to successfully self-direct the procedure. The purpose of this study was to evaluate the reliability and validity of an automatic audiometry algorithm that is fully implemented in a wearable hearing aid, to determine to what extent reliability and validity are affected when the procedure is self-directed by the user, and to investigate contributors to a successful outcome. Design was a two-phase correlational study. A total of 60 adults with mild to moderately severe hearing loss participated in both studies: 20 in Study 1 and 40 in Study 2. Twenty-seven participants in Study 2 attended with a partner. Participants in both phases were selected for inclusion if their thresholds were within the output limitations of the test device. In both phases, participants performed automatic audiometry through a receiver-in-canal, behind-the-ear hearing aid coupled to an open dome. In Study 1, the experimenter directed the task. In Study 2, participants followed a set of written, illustrated instructions to perform automatic audiometry independently of the experimenter, with optional assistance from a lay partner. Standardized measures of hearing aid self-efficacy, locus of control, cognitive function, health literacy, and manual dexterity were administered. Statistical analysis examined the repeatability of automatic audiometry; the match between automatically and manually measured thresholds; and contributors to successful, independent completion of the automatic audiometry procedure. When the procedure was directed by an audiologist, automatic audiometry yielded reliable and valid thresholds. Reliability and validity were negatively affected when the procedure was self-directed by the user, but the results were still clinically acceptable: test-retest correspondence was 10 dB or lower in 97% of cases, and 91% of automatic thresholds were within 10 dB of their manual counterparts. However, only 58% of participants were able to achieve a complete audiogram in both ears. Cognitive function significantly influenced accurate and independent performance of the automatic audiometry procedure; accuracy was further affected by locus of control and level of education. Several characteristics of the automatic audiometry algorithm played an additional role in the outcome. Average transducer- and coupling-specific correction factors are sufficient for a self-directed in situ audiometry procedure to yield clinically reliable and valid hearing thresholds. Before implementation in a self-fitting hearing aid, however, the algorithm and test instructions should be refined in an effort to increase the proportion of users who are able to achieve complete audiometric results. Further evaluation of the procedure, particularly among populations likely to form the primary audience of a self-fitting hearing aid, should be undertaken.


Organic aerosols and inorganic species from post-harvest agricultural-waste burning emissions over northern India: impact on mass absorption efficiency of elemental carbon.
Atmospheric PM2.5 (particulate matter with aerodynamic diameter of ≤ 2.5 μm), collected from a source region [Patiala: 30.2 °N; 76.3 °E; 250 m above mean sea level] of emissions from post-harvest agricultural-waste (paddy-residue) burning in the Indo-Gangetic Plain (IGP), North India, has been studied for its chemical composition and impact on regional atmospheric radiative forcing. On average, organic aerosol mass accounts for 63% of PM2.5, whereas the contribution of elemental carbon (EC) is ∼3.5%. Sulphate, nitrate and ammonium contribute up to ∼85% of the total water-soluble inorganic species (WSIS), which constitutes ∼23% of PM2.5. The potassium-to-organic carbon ratio from paddy-residue burning emissions (KBB(+)/OC: 0.05 ± 0.01) is quite similar to that reported from Amazonian and Savanna forest-fires; whereas non-sea-salt-sulphate-to-OC ratio (nss-SO4(2-)/OC: 0.21) and nss-SO4(2-)/EC ratio of 2.6 are significantly higher (by factor of 5 to 8). The mass absorption efficiency of EC (3.8 ± 1.3 m(2) g(-1)) shows significant decrease with a parallel increase in the concentrations of organic aerosols and scattering species (sulphate and nitrate). A cross plot of OC/EC and nss-SO4(2-)/EC ratios show distinct differences for post-harvest burning emissions from paddy-residue as compared to those from fossil-fuel combustion sources in south-east Asia.


PAIN OUT: an international acute pain registry supporting clinicians in decision making and in quality improvement activities.
Management of post-operative pain is unsatisfactory worldwide. An estimated 240 million patients undergo surgery each year. Forty to 60% of these patients report clinically significant pain. Discrepancy exists between availability of evidence-based medicine (EBM)-derived knowledge about management of perioperative pain and increased implementation of related practices versus lack of improvement in patient-reported outcomes (PROs). We aimed to assist health care providers to optimize perioperative pain management by developing and validating a medical registry that measures variability in care, identifies best pain management practices and assists clinicians in decision making. PAIN OUT was established from 2009 to 2012 with funding from the European Commission. It now continues as a self-sustaining, not-for-profit project, targeting health care professionals caring for patients undergoing surgery. The growing registry includes data from 40 898 patients, 60 hospitals and 17 countries. Collaborators upload data (demographics, clinical, PROs) from patients undergoing surgery in their hospital/ward into an Internet-based portal. Two modules make use of the data: (1) online, immediate feedback and benchmarking compares PROs across sites while offline analysis permits in-depth analysis; and (2) the case-based clinical decision support system offers practice-based treatment recommendations for individual patients; it is available now as a prototype. The Electronic Knowledge Library provides succinct summaries on perioperative pain management, supporting knowledge transfer and application of EBM. PAIN OUT, a large, growing international registry, allows use of 'real-life' data related to management of perioperative pain. Ultimately, comparative analysis through audit, feedback and benchmarking will improve quality of care.


Finding the direction of lowest resilience in multivariate complex systems.
The dynamics of complex systems, such as ecosystems, financial markets and the human brain, emerge from the interactions of numerous components. We often lack the knowledge to build reliable models for the behaviour of such network systems. This makes it difficult to predict potential instabilities. We show that one could use the natural fluctuations in multivariate time series to reveal network regions with particularly slow dynamics. The multidimensional slowness points to the direction of minimal resilience, in the sense that simultaneous perturbations on this set of nodes will take longest to recover. We compare an autocorrelation-based method with a variance-based method for different time-series lengths, data resolution and different noise regimes. We show that the autocorrelation-based method is less robust for short time series or time series with a low resolution but more robust for varying noise levels. This novel approach may help to identify unstable regions of multivariate systems or to distinguish safe from unsafe perturbations.


Complaint-based surveillance for foodborne illness in the United States: a survey of local health departments.
Foodborne illnesses are an important public health problem in the United States in terms of both the burden of illness and cost to the health care system. Strengthening foodborne illness surveillance helps address the growing issues of food safety in the United States. Very little is known about the use of consumer complaint surveillance systems for foodborne illness. This study evaluates the use of these surveillance systems by local health departments (LHDs) in the United States and their practices and policies for investigating complaints. Data for this study were collected through two Web-based surveys based on a representative sample of LHDs in the United States; 81% of LHDs use complaint-based surveillance. Of those that did not have a complaint system, 64% reported that the state health department or another agency ran their complaint system. Health departments collect a wide variety of information from callers through their complaint systems, including food intake history. Most of the LHDs, however, do not store the information in an electronic database. Outbreak rates and complaint rates were found to be positively correlated, with a Pearson's correlation coefficient of 0.38. Complaints were the most common outbreak detection mechanism reported by respondents, with a median of 69% of outbreaks during the previous year found through complaints. Complaint systems are commonly used in the United States. Increasing the rate at which illnesses are reported by the public and improving investigation practices could help increase the number of outbreaks detected through complaint surveillance.


Selective killing of transformed cells by methotrexate with histidine deprivation or with alpha-amino alcohols.
The effects of amino acid deprivation and treatment with amino alcohols upon the growth, viability, and susceptibility to methotrexate (MTX) cytotoxicity were examined in BALB/3T3 cells and their virally transformed counterparts, SV-T2 cells. Cells were deprived of either histidine or tyrosine plus phenylalanine, or they were treated with amino alcohol analogues of histidine and tyrosine (histidinol and tyrosinol). When incubated in medium lacking histidine and supplemented with dialyzed serum (histidine-deficient medium), the BALB/3T3 cells remained viable for at least 3 days, but they ceased logarithmic growth, and the cell number reached an early plateau. In contrast, the SV-T2 cells continued to divide in histidine-deficient medium. Neither cell line ceased division in medium deficient in both phenylalanine and tyrosine. Incubation of the BALB/3T3 cells with 1.5 mM histidinol or 1.0 mM tyrosinol caused an early plateau similar to the effect of histidine deprivation. SV-T2 cells were not affected by these concentrations of histidinol or tyrosinol, but growth was arrested at higher concentrations. Any of the conditions used which caused a plateau in the number of BALB/3T3 cells substantially protected the treated cells from cell death caused by MTX. Therefore, pretreatment of BALB/3T3 cells with histidine deprivation, 1.5 mM histidinol, or 1.0 mM tyrosinol protected this cell line against MTX-induced cell death, while the same pretreatment conditions failed to protect SV-T2 cells. (SV-T2 cells were protected by 4.0 mM histidinol.) Thus, the ability of MTX to kill cells in vitro can be selectively modified by conditions which protect cells which retain normal growth control characteristics, but which do not protect virally transformed cells.


Fruits, vegetables, and bladder cancer risk: a systematic review and meta-analysis.
Smoking is estimated to cause about half of all bladder cancer cases. Case-control studies have provided evidence of an inverse association between fruit and vegetable intake and bladder cancer risk. As part of the World Cancer Research/American Institute for Cancer Research Continuous Update Project, we conducted a systematic review and meta-analysis of prospective studies to assess the dose-response relationship between fruit and vegetables and incidence and mortality of bladder cancer. We searched PubMed up to December 2013 for relevant prospective studies. We conducted highest compared with lowest meta-analyses and dose-response meta-analyses using random effects models to estimate summary relative risks (RRs) and 95% confidence intervals (CIs), and used restricted cubic splines to examine possible nonlinear associations. Fifteen prospective studies were included in the review. The summary RR for an increase of 1 serving/day (80 g) were 0.97 (95% CI: 0.95-0.99) I(2) = 0%, eight studies for fruits and vegetables, 0.97 (95% CI: 0.94-1.00, I(2) = 10%, 10 studies) for vegetables and 0.98 (95% CI: 0.96-1.00, I(2) = 0%, 12 studies) for fruits. Results were similar in men and women and in current, former and nonsmokers. Amongst fruits and vegetables subgroups, for citrus fruits the summary RR for the highest compared with the lowest intake was 0.87 (95% CI: 0.76-0.99, I(2) = 0%, eight studies) and for cruciferous vegetables there was evidence of a nonlinear relationship (P = 0.001). The current evidence from cohort studies is not consistent with a role for fruits and vegetables in preventing bladder cancer.


Universal health coverage necessitates a system approach: an analysis of Community-based Health Planning and Services (CHPS) initiative in Ghana.
The 9th Global conference on health promotion (Shanghai 2016) reaffirmed the role of primary health care (PHC) in achieving the 2030 Sustainable Development Agenda. Gaining much international recognition, the community-based health planning and services (CHPS) initiative is considered one of the pragmatic strategy in delivering on the promise of universal health coverage (UHC) through the PHC strategy, in Ghana. Yet, certain key factors threaten its successes - renewing the relevance of this study to present the barriers to and facilitators of the initiative. According to our findings, CHPS contribution particularly in bridging geographical access to health cannot be demeaned. Nevertheless, the full functioning of the initiative is limited by factors centered on the following themes: health governance and leadership, provision of services of quality, financial risk protection strategies targeting public health, information and care continuity, and the right mix of trained health professionals of even distribution across communities. Addressing the challenges of CHPS demand a system-approach. Substantial progress is more likely to emerge with improved leadership especially on the part of Governments to take bold political step to provide adequate financial and material resources. However, much will be achieved when stakeholders including the community work in synergies, to manage competing priorities by focusing on the core values and goals of CHPS.


Total and methylmercury concentrations in ground and surface waters in natural and restored freshwater wetlands in northern New York.
Nearly half of freshwater wetlands have been lost due to human disturbance. In response, wetlands are being restored to retain their ecosystem services. A potentially adverse consequence of wetland function is the production of methylmercury (MeHg). We measured concentrations of mercury (Hg) species and ancillary parameters in groundwaters and surface waters from four natural and 16 restored wetlands in northern New York State, USA to investigate differences in concentrations of Hg species among wetlands. We found no obvious differences in concentrations of total mercury (THg) and methylmercury in pond waters between natural and restored wetlands. High values of %methylmercury were evident in both ground (38.8 ± 27.6%) and surface waters (43.4 ± 25.6%) suggesting these wetland complexes are highly efficient in converting ionic Hg to methylmercury, regardless if restored or natural. High methylation efficiency may be due to observed drying and rewetting cycles. Hg in pond waters is likely derived from direct atmospheric deposition or by mobilization from near-wetland shallow sediments, in addition to groundwater inflows. Water flow of groundwaters from the associated watershed into pond waters resulted in increases in concentrations of THg and methylmercury. Dissolved organic matter likely plays an important role in the supply of Hg to pond waters. Relationships between methylmercury and %methylmercury with sulfate and nitrate in groundwaters may suggest some chemical limitation on Hg methylation at higher concentrations of these anions. Because of the similarity in Hg dynamics for natural and restored wetlands, the most effective strategy to mitigate methylmercury production would be to decrease atmospheric Hg deposition.


Baseline characteristics associated with different BMI trajectories in weight loss trials: a case for better targeting of interventions.
Dietary weight loss interventions have heterogeneous outcomes in long-term studies, with many participants regaining part or all of the lost weight. Growth mixture modelling is a novel analytic approach that can be used to identify different trajectories of weight change during a trial rather than focussing on the total amount of weight lost. Data were pooled from two 12-month dietary weight loss studies where no significant difference was detected between the treatment and control arms, thus allowing analysis independent of treatment. The data set included 231 subjects (74.5% female), with a mean weight loss of 6.40 kg (4.96). Growth mixture models were used to identify participants with similar trajectories of change in body mass index (BMI). Three subgroups were identified. A rapid and continuing BMI loss over the study period (rapid, n=53), a rapid initial weight loss in the first 3 months with a slowing rate over the remaining 9 months (maintainers, n=146) and those with an initial loss trajectory, which slowed and began to increase at 9 months (recidivists, n=53). Age (s.d.) and BMI (s.d.) were significantly different between the three groups (rapid 53 years (7), 28.99 kg/m(2) (3.30); maintainers 47 years (9), 30.90 kg/m(2) (2.95); recidivists 44 years (7), 34.84 kg/m(2) (1.92), both P<0.001). Older subjects with lower BMIs were more likely to have a rapid and continuing weight loss in a 1-year dietary-based weight loss intervention. Different interventional approaches may be necessary for different ages and baseline BMIs and stratification prior to randomisation may be necessary to prevent confounding in weight loss trials.


Tracking implementation and (un)intended consequences: a process evaluation of an innovative peripheral health facility financing mechanism in Kenya.
In many African countries, user fees have failed to achieve intended access and quality of care improvements. Subsequent user fee reduction or elimination policies have often been poorly planned, without alternative sources of income for facilities. We describe early implementation of an innovative national health financing intervention in Kenya; the health sector services fund (HSSF). In HSSF, central funds are credited directly into a facility's bank account quarterly, and facility funds are managed by health facility management committees (HFMCs) including community representatives. HSSF is therefore a finance mechanism with potential to increase access to funds for peripheral facilities, support user fee reduction and improve equity in access. We conducted a process evaluation of HSSF implementation based on a theory of change underpinning the intervention. Methods included interviews at national, district and facility levels, facility record reviews, a structured exit survey and a document review. We found impressive achievements: HSSF funds were reaching facilities; funds were being overseen and used in a way that strengthened transparency and community involvement; and health workers' motivation and patient satisfaction improved. Challenges or unintended outcomes included: complex and centralized accounting requirements undermining efficiency; interactions between HSSF and user fees leading to difficulties in accessing crucial user fee funds; and some relationship problems between key players. Although user fees charged had not increased, national reduction policies were still not being adhered to. Finance mechanisms can have a strong positive impact on peripheral facilities, and HFMCs can play a valuable role in managing facilities. Although fiduciary oversight is essential, mechanisms should allow for local decision-making and ensure that unmanageable paperwork is avoided. There are also limits to what can be achieved with relatively small funds in contexts of enormous need. Process evaluations tracking (un)intended consequences of interventions can contribute to regional financing and decentralization debates.


Imperfect centered miRNA binding sites are common and can mediate repression of target mRNAs.
MicroRNAs (miRNAs) bind to mRNAs and target them for translational inhibition or transcriptional degradation. It is thought that most miRNA-mRNA interactions involve the seed region at the 5' end of the miRNA. The importance of seed sites is supported by experimental evidence, although there is growing interest in interactions mediated by the central region of the miRNA, termed centered sites. To investigate the prevalence of these interactions, we apply a biotin pull-down method to determine the direct targets of ten human miRNAs, including four isomiRs that share centered sites, but not seeds, with their canonical partner miRNAs. We confirm that miRNAs and their isomiRs can interact with hundreds of mRNAs, and that imperfect centered sites are common mediators of miRNA-mRNA interactions. We experimentally demonstrate that these sites can repress mRNA activity, typically through translational repression, and are enriched in regions of the transcriptome bound by AGO. Finally, we show that the identification of imperfect centered sites is unlikely to be an artifact of our protocol caused by the biotinylation of the miRNA. However, the fact that there was a slight bias against seed sites in our protocol may have inflated the apparent prevalence of centered site-mediated interactions. Our results suggest that centered site-mediated interactions are much more frequent than previously thought. This may explain the evolutionary conservation of the central region of miRNAs, and has significant implications for decoding miRNA-regulated genetic networks, and for predicting the functional effect of variants that do not alter protein sequence.


Context-dependent functions of specific microRNAs in neuronal development.
MicroRNAs (miRNAs) are small noncoding RNAs that regulate multiple developmental processes at the post-transcriptional level. Recent rapid progresses have demonstrated critical roles for a number of miRNAs in neuronal development and function. In particular, miR-9 and miR-124 are specifically expressed in the mammalian nervous system, and their respective nucleotide sequences are 100% identical among many species. Yet, their expression patterns and mRNA targets are less conserved throughout evolution. As a consequence, these miRNAs exhibit diverse context-dependent functions in different aspects of neuronal development, ranging from early neurogenesis and neuronal differentiation to dendritic morphogenesis and synaptic plasticity. Some other neuronal miRNAs also exhibit context-dependent functions in development. Thus, post-transcriptional regulation of spatial and temporal expression levels of protein-coding genes by miRNAs contributes uniquely to the proper development and evolution of the complex nervous system.


Influence of the dietary carotenoids lutein and zeaxanthin on visual performance: application to baseball.
Macular pigment (MP) is composed of the yellow, blue-absorbing carotenoids lutein and zeaxanthin. Although distributed throughout the visual system, MP is heavily concentrated in the central retinal area (eg, screening the foveal cones). Because light must pass through MP before reaching the receptors, it filters significant amounts of short-wave energy. Individual variation in peak absorbance is large and ranges from 0.0 to 1.6 optical density units depending largely on dietary intake. Several important functions of MP have been proposed. MP may serve to protect the retina from damage by absorbing actinic short-wave light (analogous to internal sunglasses) or by inactivating highly reactive free radicals and oxygen triplicates that are the by-product of light-driven cellular activity. MP may also serve, as proposed more than a century ago, to improve the retinal image through optical mechanisms. Recent data suggest that the MP carotenoids reduce glare discomfort and disability, shorten photostress recovery times, enhance chromatic contrast, and increase visual range (how far one can see in the distance). Lutein and zeaxanthin within the brain might also increase temporal processing speeds. This article reviews the influences of MP on visual function by exploring the implications of these visual improvements for baseball players.


Automated cross-sectional and longitudinal hippocampal volume measurement in mild cognitive impairment and Alzheimer's disease.
Volume and change in volume of the hippocampus are both important markers of Alzheimer's disease (AD). Delineation of the structure on MRI is time-consuming and therefore reliable automated methods are required. We describe an improvement (multiple-atlas propagation and segmentation (MAPS)) to our template library-based segmentation technique. The improved technique uses non-linear registration of the best-matched templates from our manually segmented library to generate multiple segmentations and combines them using the simultaneous truth and performance level estimation (STAPLE) algorithm. Change in volume over 12months (MAPS-HBSI) was measured by applying the boundary shift integral using MAPS regions. Methods were developed and validated against manual measures using subsets from Alzheimer's Disease Neuroimaging Initiative (ADNI). The best method was applied to 682 ADNI subjects, at baseline and 12-month follow-up, enabling assessment of volumes and atrophy rates in control, mild cognitive impairment (MCI) and AD groups, and within MCI subgroups classified by subsequent clinical outcome. We compared our measures with those generated by Surgical Navigation Technologies (SNT) available from ADNI. The accuracy of our volumes was one of the highest reported (mean(SD) Jaccard Index 0.80(0.04) (N=30)). Both MAPS baseline volume and MAPS-HBSI atrophy rate distinguished between control, MCI and AD groups. Comparing MCI subgroups (reverters, stable and converters): volumes were lower and rates higher in converters compared with stable and reverter groups (p< or =0.03). MAPS-HBSI required the lowest sample sizes (78 subjects) for a hypothetical trial. In conclusion, the MAPS and MAPS-HBSI methods give accurate and reliable volumes and atrophy rates across the clinical spectrum from healthy aging to AD.


The protective effects of parental monitoring and internet restriction on adolescents' risk of online harassment.
With many adolescents using the internet to communicate with their peers, online harassment is on the rise among youth. The purpose of this study was to understand how parental monitoring and strategies parents use to regulate children's internet use (i.e., internet restriction) can help reduce online harassment among adolescents. Online survey data were collected from a nationally representative sample of parents and their 12-17 year old adolescents (n = 629; 49 % female). Structural equation modeling was used to test direct and indirect effects of parental monitoring and internet restriction on being a victim of online harassment. Potential mediators included adolescents' frequency of use of social networking websites, time spent on computers outside of school, and internet access in the adolescent's bedroom. Age and gender differences were also explored. Adolescents' reports of parental monitoring and efforts to regulate specific forms of internet use were associated with reduced rates of online harassment. Specifically, the effect of parental monitoring was largely direct and 26 times greater than parental internet restriction. The latter was associated with lower rates of harassment only indirectly by limiting internet access in the adolescent's bedroom. These effects operated similarly for younger and older adolescents and for males and females. Adolescents' perceptions of parental monitoring and awareness can be protective against online harassment. Specific restriction strategies such as regulating internet time and content can also help reduce the risk of online harassment.


Impact of Peer Support on Acute Care Visits and Hospitalizations for Individuals With Diabetes and Depressive Symptoms: A Cluster-Randomized Controlled Trial.
Comorbid depression is associated with increased health care utilization and cost. We examined the effects of peer support on acute care (AC) and hospital utilization in individuals with diabetes with or without depressive symptoms. This was a cluster-randomized controlled trial conducted in 2010-2012, with the clusters being practices and their surrounding communities. Adults with type 2 diabetes who wanted help with self-management were eligible to participate. Those without a doctor, with limited life expectancy, with plans to move within the next year, and with an unwillingness to work with a peer advisor were excluded. Intervention participants received 1 year of peer support. Control participants received usual care. The Patient Health Questionnaire (PHQ-8) (range 0-24; 5 indicates mild and 10 indicates moderate depressive symptoms) assessed depressive symptoms. AC and hospital utilization were measured by self-report. Data were collected at baseline, 6 months, and 12 months. Quasi-Poisson regression using generalized estimating equations examined differences in utilization per year attributable to the intervention for those with and without mild depressive symptoms (and separately, moderate depressive symptoms), controlling for imbalance across treatment arms. At baseline, half of the sample reported mild depressive symptoms (52% intervention and 48% control, P = 0.37), a quarter reported moderate depressive symptoms (25% intervention and 26% control, P = 1.0), and there were no significant differences in utilization. A total of 168 intervention (six clusters) and 187 control (five clusters) participants had follow-up data. In individuals with mild depressive symptoms, the incident rate ratio (IRR) for hospitalization among intervention compared with control was 0.26 (95% CI 0.08-0.84) per 10 patient-years. The IRR for AC was 0.55 (95% CI 0.28-1.07) per 10 person-years. Findings were similar for individuals with moderate depressive symptoms. Peer support lowered AC visits and hospitalizations for individuals with depressive symptoms but not for those without depressive symptoms; these findings can guide resource allocation for population health management.


Stability and Change of the Personality Traits Languidity and Flexibility in a Sample of Nurses: A 7-8 Years Follow-Up Study.
The traits languidity (tendency to become tired/sleepy upon losing sleep) and flexibility (ability to sleep and work at odd times) have been implicated in shift work tolerance. However, there is a dearth of knowledge about their temporal stability. The aim of the present study was to explore these traits during a long follow-up (FU) period and identify factors related to potential changes in trait scores over time. In all, 1,652 nurses completed the Circadian Type Inventory-revised (CTI-r), which measures languidity and flexibility, at both 2008/2009 (baseline, BL) and again in 2016 (FU). The latent scores of these two constructs at BL, in addition to age, sex, childcare responsibility, marital status, night work status, and insomnia status, were regressed on the corresponding latent scores at follow-up using a structural equation modeling (SEM) approach. Stability was found for both languidity (rho = 0.59) and flexibility (rho = 0.58). Both composite scores declined significantly from baseline (20.62 and 12.48) to follow-up (19.96 and 11.77). Languidity at baseline was positively associated with languidity at follow-up (β = 0.89, p < 0.009). Undertaking childcare responsibility between baseline and follow-up was inversely related to languidity at follow-up (β = -0.09, p < 0.05). Starting night work was positively related to languidity at follow-up (β = 0.06, p < 0.05). Developing insomnia between baseline and follow-up (β = 0.15, p < 0.05) was positively, whereas remitting from insomnia during the same period was negatively (β = -0.11, p < 0.01) associated with languidity at follow-up. Flexibility at baseline was positively associated with flexibility at follow-up (β = 0.64, p < 0.05). Having childcare responsibility at baseline, but not at follow-up was inversely related to flexibility at follow-up (β = -0.05 p < 0.05). Becoming cohabitant with a partner between baseline and follow-up (β = -0.07, p < 0.05) was negatively associated with flexibility at follow-up. Starting night work between baseline and follow-up (β = 0.17, p < 0.01) and reporting night work at both baseline and follow-up (β = 0.18, p < 0.01) were both positively associated with flexibility at follow-up, whereas stopping working nights was negatively (β = -0.09, p < 0.05), associated with flexibility at follow-up. The personality traits languidity and flexibility show fairly high stability, albeit the mean scores were significantly reduced during the 7-8 years follow-up period. The results suggest that these personality traits are partly modifiable.


Gene expression in cell lines from propionic acidemia patients, carrier parents, and controls.
Propionic acidemia (PA) is an inborn of metabolism which usually presents with metabolic acidosis and accumulation of 3-hydroxypropionate among other toxins. Examining the gene expression in lymphoblastoid cell lines (LCLs) from PA patients, their carrier parents and age/sex-matched controls at normal glucose and low glucose growth conditions demonstrated differences among and between these groups. Using three-way ANOVA analysis, four DAVID clusters of response were identified of which three of the four clusters showed that LCLs from carrier parents had an intermediate response between healthy controls and PA patients. These differences included changes in expression of cell cycle regulatory genes, mitochondrial related genes, and transcriptional regulation. In addition, differences also were observed in expression of genes involved in transendothelial migration and focal adhesion at normal growth conditions when comparing the LCLs from PA patients and controls. These studies demonstrate transcriptional differences between LCLs from PA patients, their parents and biochemically normal controls.


The influence of smoking, age and stage at diagnosis on the survival after larynx, hypopharynx and oral cavity cancers in Europe: The ARCAGE study.
Head and neck cancer (HNC) is a preventable malignancy that continues to cause substantial morbidity and mortality worldwide. Using data from the ARCAGE and Rome studies, we investigated the main predictors of survival after larynx, hypopharynx and oral cavity (OC) cancers. We used the Kaplan-Meier method to estimate overall survival, and Cox proportional models to examine the relationship between survival and sociodemographic and clinical characteristics. 604 larynx, 146 hypopharynx and 460 OC cancer cases were included in this study. Over a median follow-up time of 4.6 years, nearly 50% (n = 586) of patients died. Five-year survival was 65% for larynx, 55% for OC and 35% for hypopharynx cancers. In a multivariable analysis, we observed an increased mortality risk among older (≥71 years) versus younger (≤50 years) patients with larynx/hypopharynx combined (LH) and OC cancers [HR = 1.61, 95% CI 1.09-2.38 (LH) and HR = 2.12, 95% CI 1.35-3.33 (OC)], current versus never smokers [HR = 2.67, 95% CI 1.40-5.08 (LH) and HR = 2.16, 95% CI 1.32-3.54 (OC)] and advanced versus early stage disease at diagnosis [IV versus I, HR = 2.60, 95% CI 1.78-3.79 (LH) and HR = 3.17, 95% CI 2.05-4.89 (OC)]. Survival was not associated with sex, alcohol consumption, education, oral health, p16 expression, presence of HPV infection or body mass index 2 years before cancer diagnosis. Despite advances in diagnosis and therapeutic modalities, survival after HNC remains low in Europe. In addition to the recognized prognostic effect of stage at diagnosis, smoking history and older age at diagnosis are important prognostic indicators for HNC.


Incidental Memory Encoding Assessed with Signal Detection Theory and Functional Magnetic Resonance Imaging (fMRI).
In functional magnetic resonance imaging (fMRI) studies that apply a "subsequent memory" approach, successful encoding is indicated by increased fMRI activity during the encoding phase for hits vs. misses, in areas underlying memory encoding such as the hippocampal formation. Signal-detection theory (SDT) can be used to analyze memory-related fMRI activity as a function of the participant's memory trace strength (d(')). The goal of the present study was to use SDT to examine the relationship between fMRI activity during incidental encoding and participants' recognition performance. To implement a new approach, post-experimental group assignment into High- or Low Performers (HP or LP) was based on 29 healthy participants' recognition performance, assessed with SDT. The analyses focused on the interaction between the factors group (HP vs. LP) and recognition performance (hits vs. misses). A whole-brain analysis revealed increased activation for HP vs. LP during incidental encoding for remembered vs. forgotten items (hits > misses) in the insula/temporo-parietal junction (TPJ) and the fusiform gyrus (FFG). Parameter estimates in these regions exhibited a significant positive correlation with d('). As these brain regions are highly relevant for salience detection (insula), stimulus-driven attention (TPJ), and content-specific processing of mnemonic stimuli (FFG), we suggest that HPs' elevated memory performance was associated with enhanced attentional and content-specific sensory processing during the encoding phase. We provide first correlative evidence that encoding-related activity in content-specific sensory areas and content-independent attention and salience detection areas influences memory performance in a task with incidental encoding of facial stimuli. Based on our findings, we discuss whether the aforementioned group differences in brain activity during incidental encoding might constitute the basis of general differences in memory performance between HP and LP.


The structure of a GH149 β-(1 → 3) glucan phosphorylase reveals a new surface oligosaccharide binding site and additional domains that are absent in the disaccharide-specific GH94 glucose-β-(1 → 3)-glucose (laminaribiose) phosphorylase.
Glycoside phosphorylases (GPs) with specificity for β-(1 → 3)-gluco-oligosaccharides are potential candidate biocatalysts for oligosaccharide synthesis. GPs with this linkage specificity are found in two families thus far-glycoside hydrolase family 94 (GH94) and the recently discovered glycoside hydrolase family 149 (GH149). Previously, we reported a crystallographic study of a GH94 laminaribiose phosphorylase with specificity for disaccharides, providing insight into the enzyme's ability to recognize its' sugar substrate/product. In contrast to GH94, characterized GH149 enzymes were shown to have more flexible chain length specificity, with preference for substrate/product with higher degree of polymerization. In order to advance understanding of the specificity of GH149 enzymes, we herein solved X-ray crystallographic structures of GH149 enzyme Pro_7066 in the absence of substrate and in complex with laminarihexaose (G6). The overall domain organization of Pro_7066 is very similar to that of GH94 family enzymes. However, two additional domains flanking its catalytic domain were found only in the GH149 enzyme. Unexpectedly, the G6 complex structure revealed an oligosaccharide surface binding site remote from the catalytic site, which, we suggest, may be associated with substrate targeting. As such, this study reports the first structure of a GH149 phosphorylase enzyme acting on β-(1 → 3)-gluco-oligosaccharides and identifies structural elements that may be involved in defining the specificity of the GH149 enzymes.


Evaluation of Aspergillus niger as host for virus-like particle production, using the hepatitis B surface antigen as a model.
The filamentous fungus Aspergillus niger was transformed with the hepatitis B virus S gene encoding the major viral envelope protein under control of the constitutive A. nidulans glyceraldehyde-3-phosphate dehydrogenase ( gpdA) promoter. Approximately seven copies of the expression cassette were integrated on the genome, resulting in high-level transcription of the S gene. Production of the 24-kDa S protein and a 48-kDa S protein dimer in the membrane-associated protein fraction of the recombinant A. niger strain was shown through Western analysis. Electron microscopy of partially purified recombinant S protein revealed the formation of spherical pseudoviral particles with a diameter of 22 nm. The production level of hepatitis B pseudoviral particles was estimated to be 0.4 mg/l culture, which compares favourably with the reported levels initially obtained in yeast, indicating the potential of the Aspergillus expression system as an alternative, cost-effective vaccine production system.


Genetic ablation of Drosophila phagocytes reveals their contribution to both development and resistance to bacterial infection.
Drosophila phagocytes participate in development and immune responses through their abilities to perform phagocytosis and/or secrete extra-cellular matrix components, antimicrobial peptides, clotting factors and signalling molecules. However, our knowledge of their functional impact on development and host resistance to infection is limited. To address this, we have used a genetic cell ablation strategy to generate Drosophila individuals lacking functional phagocytes. Our results highlight the essential contribution of phagocytes to embryonic development including central nervous system morphogenesis. Phagocytes also ensure optimal viability during post-embryonic development through immune functions. The use of phagocyte-depleted flies reveals the contribution of phagocytes in the resistance of Drosophila adults upon systemic infections with specific bacteria. Phagocytes were not involved in the expression of antimicrobial peptides by the fat body indicating a clear separation between cellular and humoral immune responses at this stage. Finally, we confirm that phagocytosis is a critical effector mechanism of the cellular arm by demonstrating that phagocytosis contributes to resistance to infection with Staphylococcus aureus in adults. Our results highlight the power of this cell ablation strategy to reveal the contribution of phagocytes to specific biological processes. We now provide a blueprint of phagocyte importance during both development and innate immune responses in Drosophila.


Quantification of transcription factor-DNA binding affinity in a living cell.
The apparent dissociation constant (Kd) for specific binding of glucocorticoid receptor (GR) and androgen receptor (AR) to DNA was determined in vivo in Xenopus oocytes. The total nuclear receptor concentration was quantified as specifically retained [(3)H]-hormone in manually isolated oocyte nuclei. DNA was introduced by nuclear microinjection of single stranded phagemid DNA, chromatin is then formed during second strand synthesis. The fraction of DNA sites occupied by the expressed receptor was determined by dimethylsulphate in vivo footprinting and used for calculation of the receptor-DNA binding affinity. The forkhead transcription factor FoxA1 enhanced the DNA binding by GR with an apparent Kd of ∼1 μM and dramatically stimulated DNA binding by AR with an apparent Kd of ∼0.13 μM at a composite androgen responsive DNA element containing one FoxA1 binding site and one palindromic hormone receptor binding site known to bind one receptor homodimer. FoxA1 exerted a weak constitutive- and strongly cooperative DNA binding together with AR but had a less prominent effect with GR, the difference reflecting the licensing function of FoxA1 at this androgen responsive DNA element.


Production of perfluoroalkylated nanospheres from buckminsterfullerene.
Perfluoroalkylated nanospheres have been prepared by reaction of fullerenes with a variety of fluoroalkyl radicals. The latter are generated by thermal or photochemical decomposition of fluoroalkyl iodides or fluorodiacyl peroxides. Up to 16 radicals add to C(60) to afford easily isolable fluoroalkylated derivatives. The monosubstituted radical adducts were detected by electron spin resonance in the early stages of the fluoroalkylation reactions. These spheroidal molecules are thermally quite stable, soluble in fluoroorganic solvents, chemically resistant to corrosive aqueous solutions, and more volatile than the parent fullerenes. Films of the sublimed material display properties typical for a perfluoroalkylated material.


Inhibition of TBK1/IKKε Promotes Regeneration of Pancreatic β-cells.
β-cell proliferation induction is a promising therapeutic strategy to restore β-cell mass. By screening small molecules in a transgenic zebrafish model of type 1 diabetes, we identified inhibitors of non-canonical IκB kinases (IKKs), TANK-binding kinase 1 (TBK1) and IκB kinase ε (IKKε), as enhancers of β-cell regeneration. The most potent β-cell regeneration enhancer was a cinnamic acid derivative (E)-3-(3-phenylbenzo[c]isoxazol-5-yl)acrylic acid (PIAA), which, acting through the cAMP-dependent protein kinase A (PKA), stimulated β-cell-specific proliferation by increasing cyclic AMP (cAMP) levels and mechanistic target of rapamycin (mTOR) activity. A combination of PIAA and cilostamide, an inhibitor of β-cell-enriched cAMP hydrolyzing enzyme phosphodiesterase (PDE) 3, enhanced β-cell proliferation, whereas overexpression of PDE3 blunted the mitogenic effect of PIAA in zebrafish. PIAA augmented proliferation of INS-1β-cells and β-cells in mammalian islets including human islets with elevation in cAMP levels and insulin secretion. PIAA improved glycemic control in streptozotocin (STZ)-induced diabetic mice with increases in β-cell proliferation, β-cell area, and insulin content in the pancreas. Collectively, these data reveal an evolutionarily conserved and critical role of TBK1/IKKε suppression in expanding functional β-cell mass.


Could clinical audit improve the diagnosis of pulmonary tuberculosis in Cuba, Peru and Bolivia?
To assess the effectiveness of clinical audit in improving the quality of diagnostic care provided to patients suspected of tuberculosis; and to understand the contextual factors which impede or facilitate its success. Twenty-six health centres in Cuba, Peru and Bolivia were recruited. Clinical audit was introduced to improve the diagnostic care for patients attending with suspected TB. Standards were based on the WHO and TB programme guidelines relating to the appropriate use of microscopy, culture and radiological investigations. At least two audit cycles were completed over 2 years. Improvement was determined by comparing the performance between two six-month periods pre- and post-intervention. Qualitative methods were used to ascertain facilitating and limiting contextual factors influencing change among healthcare professionals' clinical behaviour after the introduction of clinical audit. We found a significant improvement in 11 of 13 criteria in Cuba, in 2 of 6 criteria in Bolivia and in 2 of 5 criteria in Peru. Twelve out of 24 of the audit criteria in all three countries reached the agreed standards. Barriers to quality improvement included conflicting objectives for clinicians and TB programmes, poor coordination within the health system and patients' attitudes towards illness. Clinical audit may drive improvements in the quality of clinical care in resource-poor settings. It is likely to be more effective if integrated within and supported by the local TB programmes. We recommend developing and evaluating an integrated model of quality improvement including clinical audit.


Combining electron microscopy with single molecule DNA fiber approaches to study DNA replication dynamics.
Replication stress is a crucial driver of genomic instability. Understanding the mechanisms of replication stress response is instrumental to improve diagnosis and treatment of human disease. Electron microscopy (EM) is currently the technique of choice to directly visualize a high number of replication intermediates and to monitor their remodeling upon stress. At the same time, DNA fiber analysis is useful to gain mechanistic insight on how genotoxic agents perturb replication fork dynamics genome-wide at single-molecule resolution. Combining these techniques has proven invaluable to achieve a comprehensive view of the mechanisms that ensure error-free processing of damaged replication forks. Here, we review how EM and single-molecule DNA fiber approaches can be used together to shed light into the mechanisms of replication stress response and discuss important cautions to be taken into account when comparing results obtained by EM and DNA fiber.


Use of verbal autopsy to determine mortality patterns in an urban slum in Kolkata, India.
To define mortality patterns in an urban slum in Kolkata, India, in the context of a cholera and typhoid fever project. In a well-defined population that was under surveillance for 18 months, we followed a dynamic cohort of 63 788 residents whose households were visited monthly by community health workers to identify deaths. Trained physicians performed verbal autopsies and experienced senior physicians assigned the primary cause of death according to the International classification of diseases, 10th edition. We tabulated causes of death in accordance with Global Burden of Disease 2000 categories and assessed overall and cause-specific mortality rates per age group and gender. During 87 921 person-years of follow-up, we recorded 544 deaths. This gave an overall mortality rate of 6.2 per 1000 person-years. We assigned a cause to 89% (482/544) of the deaths. The leading causes of death, in descending order, were cardiovascular diseases (especially among adults aged over 40 years), cancer, respiratory ailments and digestive disorders. Most deaths in children under 5 years of age were caused by tuberculosis, respiratory infections and diarrhoeal diseases. Although the most common causes of death in children were infectious, non-communicable diseases were predominant among adults. There is a need for continuing interventions against infectious diseases in addition to new and innovative strategies to combat non-infectious conditions.


Early Mobilization of Infants Intubated for Acute Respiratory Failure.
Early mobility in the intensive care unit is being promoted as a way to improve outcomes but has not been studied in young infants. To determine the safety of a holding intervention for infants intubated for respiratory failure. Infants less than 6 months of age intubated for respiratory failure underwent a holding intervention in which a caretaker held the infant at least twice a day once the infant was medically stabilized. Rates of adverse events were compared with historical controls matched by age and Pediatric Index of Mortality 2 score. Vital signs before and during holding were recorded to assess for physiologic tolerance. Twenty-four infants with a total of 158 holding interventions were studied. Mean holding duration was 99 minutes. Twenty holding interventions were terminated early, most commonly because of agitation. The 24 historical controls had 1 unplanned extubation and 2 arterial catheter removals. The intervention group had no unplanned extubations, inadvertent central catheter removals, or urinary catheter removals and had 1 arterial catheter removal that was not associated with holding. Vital signs before and during holding were similar. There were no differences between the control and intervention groups in duration of intubation (7.3 vs 6.0 days, P = .98), length of intensive care unit stay (9.1 vs 9.0 days, P = .52), or length of hospital stay (11.5 vs 12.5 days, P = .28). Holding intubated infants in the intensive care unit was well tolerated, without an increase in adverse events.


The Radial Artery Pedicle Perforator Flap: Vascular Analysis and Clinical Implications.
The purpose of this study was to determine the location, size, and vascular territory of the radial artery cutaneous perforators. Twenty-six human cadaveric forearms were dissected. All cutaneous radial artery perforators were analyzed for total number, orientation, location, and external diameter. A cluster analysis was performed to determine the overall distribution of perforators. The cutaneous territory of a distally based pedicled perforator flap was determined using methylene blue injection and three-dimensional computed tomographic angiography in five flaps. Six hundred thirty-nine perforators (399 perforators smaller than 0.5 mm compared with 240 perforators 0.5 mm or larger) were dissected in 26 forearms. Of the 639 radial artery perforators dissected, 328 (51 percent) were radially distributed and 311 (49 percent) were ulnarly distributed. There are two main clusters of clinically relevant perforators at a relative distance of 17.6 percent and 61.7 percent along the radial styloid-to-lateral epicondyle interval. In all cases, two or more perforators were found within 2 cm proximal to the styloid. Dye injection of the most dominant distally based perforators revealed a cutaneous territory ranging from 104 cm2 to 333 cm2. The case presented is of a patient with a dorsal hand defect, which was resurfaced with a pedicled perforator flap based on a distal perforator proximal to the radial styloid. There are two main clusters of clinically significant radial artery perforators. Increased knowledge of size, location, and cutaneous territory of the radial artery perforators can lead to expanded use of the radial artery forearmflap based on cutaneous perforators alone, without sacrificing the radial artery.


Predator functional response and prey survival: direct and indirect interactions affecting a marked prey population.
1. Predation plays an integral role in many community interactions, with the number of predators and the rate at which they consume prey (i.e. their functional response) determining interaction strengths. Owing to the difficulty of directly observing predation events, attempts to determine the functional response of predators in natural systems are limited. Determining the forms that predator functional responses take in complex systems is important in advancing understanding of community interactions. 2. Prey survival has a direct relationship to the functional response of their predators. We employed this relationship to estimate the functional response for bald eagle Haliaeetus leucocepalus predation of Canada goose Branta canadensis nests. We compared models that incorporated eagle abundance, nest abundance and alternative prey presence to determine the form of the functional response that best predicted intra-annual variation in survival of goose nests. 3. Eagle abundance, nest abundance and the availability of alternative prey were all related to predation rates of goose nests by eagles. There was a sigmoidal relationship between predation rate and prey abundance and prey switching occurred when alternative prey was present. In addition, predation by individual eagles increased as eagle abundance increased. 4. A complex set of interactions among the three species examined in this study determined survival rates of goose nests. Results show that eagle predation had both prey- and predator-dependent components with no support for ratio dependence. In addition, indirect interactions resulting from the availability of alternative prey had an important role in mediating the rate at which eagles depredated nests. As a result, much of the within-season variation in nest survival was due to changing availability of alternative prey consumed by eagles. 5. Empirical relationships drawn from ecological theory can be directly integrated into the estimation process to determine the mechanisms responsible for variation in observed survival rates. The relationship between predator functional response and prey survival offers a flexible and robust method to advance our understanding of predator-prey interactions in many complex natural systems where prey populations are marked and regularly visited.


Substrates of the human oligopeptide transporter hPEPT2.
Oligopeptide transporters serve important functions in nutrition and pharmacology. In particular, these transporters help maintain the homeostasis of peptides. The peptide-transporter PEPT2 is a high-affinity and low-capacity type oligopeptide transporter from the proton-coupled oligopeptide transporter family. PEPT2 has recently received attention because of its potential application in targeted drug delivery. PEPT2 is widely distributed in kidney, central nervous system, and lung of organisms. In general, all dipeptides, tripeptides, and peptide-like drugs such as β-lactam antibiotics and angiotensin-converting enzyme inhibitors could be mediated and transported as a substrate of PEPT2. The design of many extant drugs and prodrugs is based on the substrate structure of PEPT2 to accelerate absorption via peptide transporters. Thus, this paper summarizes the substrate features of PEPT2 to promote the rational design of drugs and prodrugs that target peptide transporters.


Long-term survival in patients with coronary artery disease: importance of peripheral vascular disease. The Coronary Artery Surgery Study (CASS) Investigators.
The purpose of this study was to determine the importance of peripheral arterial disease in predicting long-term survival in patients with clinically evident coronary artery disease. Patients in the Coronary Artery Surgery Study (CASS) Registry were followed up for > 10 years. Survival in 2,296 patients with peripheral arterial disease was compared with that of 13,953 patients without peripheral arterial disease using Kaplan-Meier survival curves. All patients had known stable coronary artery disease. Clinical, electrocardiographic (ECG), chest X-ray film and catheterization variables of the two groups were compared using the chi-square statistic or the two-sample t test. The independent effect of peripheral arterial disease (as well as other variables) on mortality was determined utilizing a Cox proportional hazards model. Patients with peripheral vascular disease were more likely to have hypertension, diabetes, family history of coronary artery disease, previous angina or myocardial infarction, previous coronary bypass surgery or to have smoked. They also had a higher incidence of congestive heart failure, ECG abnormality and modestly increased frequency of three-vessel disease. Independent correlates of long-term mortality for the entire cohort included age, smoking, diabetes, number of diseased coronary vessels, left ventricular function, hypertension, pulmonary disease, anginal class, previous myocardial infarction and peripheral vascular disease (all p < 0.001). At any point in time, patients with peripheral vascular disease had a 25% greater likelihood of mortality than patients without peripheral vascular disease (multivariate chi-square 25.83, hazard ratio 1.25, 95% confidence interval 1.15 to 1.36, p < 0.001). Peripheral vascular disease is a strong, independent predictor of long-term mortality in patients with stable coronary artery disease. Aggressive attempts at secondary disease prevention are warranted in this high risk group.


Human breast development.
This review presents an atlas of the histology of the normal physiological states of the human breast including prenatal, prepubertal, and pubertal development, adult resting gland, pregnancy, lactation, and postinvolution. The aim is to produce a pictorial overview of the main stages in development and the common findings in the adult that are considered to be within the range of normality. Unlike inbred strains of animals, in humans it is clear that the chronology of ductal and lobular development is not predictable, either in the fetus, the infant, the peripubertal breast, or the adult. This is probably due to the individual variation in hormone levels both in utero and after birth. For many of the developmental time points there are very little data available. In this review we indicate the current state of knowledge of human breast development and some of the main similarities and differences with the rodent, the main animal model. The major phases of growth and development are described and accompanied by photographs that are representative of each stage. Stress is placed on terminology as there is confusion in the literature. This article is written as an accessory to the companion review on breast cancer.


Transition metal catalysis in confined spaces.
Transition metal catalysis plays an important role in both industry and in academia where selectivity, activity and stability are crucial parameters to control. Next to changing the structure of the ligand, introducing a confined space as a second coordination sphere around a metal catalyst has recently been shown to be a viable method to induce new selectivity and activity in transition metal catalysis. In this review we focus on supramolecular strategies to encapsulate transition metal complexes with the aim of controlling the selectivity via the second coordination sphere. As we will discuss, catalyst confinement can result in selective processes that are impossible or difficult to achieve by traditional methods. We will describe the template-ligand approach as well as the host-guest approach to arrive at such supramolecular systems and discuss how the performance of the catalyst is enhanced by confining it in a molecular container.


BMP signaling induces astrocytic differentiation of clinically derived oligodendroglioma propagating cells.
Oligodendrogliomas are a type of glioma that lack detailed investigation because of an inability to cultivate oligodendroglioma cells that faithfully recapitulate their salient qualities. We have successfully isolated and propagated glioma stem-like cells from multiple clinical oligodendroglioma specimens. These oligodendroglioma-propagating cells (OligPC) are multipotent and form xenografts with oligodendroglioma features. Bone morphogenetic proteins (BMP) are considered potent inhibitors of oligodendrogliogenesis during development; therefore, the effects of BMP signaling in OligPCs were characterized. BMP pathway components are expressed by OligPCs and canonical signaling via Smad proteins is intact. This signaling potently depletes CD133-positive OligPCs, decreasing proliferation, and inducing astrocytic differentiation. Furthermore, analyses revealed that cytoplasmic sequestration of the oligodendrocyte differentiation factors OLIG1/2 by the BMP signaling effectors ID2 and ID4 is a plausible underlying mechanism. These findings elucidate the molecular pathways that underlie the effects of BMP signaling on oligodendroglioma stem-like cells. Stem-like cells are capable of propagating oligodendrogliomas, and BMP signaling potently diminishes their stemness by inducing astrocytic differentiation, suggesting that BMP activation may be effective as a cancer stem cell-targeted therapy.


A new method for studying problem solving and tool use in stingrays (Potamotrygon castexi).
Testing the cognitive abilities of cartilaginous fishes is important in understanding the evolutionary origins of cognitive functions in higher vertebrates. We used five South American fresh water stingrays (Potamotrygon castexi) in a learning and problem-solving task. A tube test apparatus was developed to provide a simple but sophisticated procedure for testing cognitive abilities of aquatic animals. All five subjects quickly learned to use water as a tool to extract food from the testing apparatus. The experimental protocol, which gave the animals the opportunity of correcting a wrong visual cue decision, resulted in four out of five subjects correcting an error rather than making an initial right choice. One of five subjects reached 100% correct trials in the visual discrimination task. The ability to use water as an agent to extract food from the testing apparatus is a first indication of tool use in batoid fishes. Performance in the instrumental task of retrieving food from a novel testing apparatus and the rapid learning in the subsequent discrimination/error correction task shows that cartilaginous fish can be used to study the origins of cognitive functions in the vertebrate lineage.


Isolation and characterization of microsatellite markers and analysis of genetic diversity in Chinese jujube (Ziziphus jujuba Mill.).
Chinese jujube (Ziziphus jujuba Mill, 2n = 2× = 24, Rhamnaceae) is an economically important Chinese native species. It has high nutritional value, and its medicinal properties have led to extensive use in traditional oriental medicine. The characterization of genotypes using molecular markers is important for genetic studies and plant breeding. However, few simple sequence repeat (SSR) markers are available for this species. In this study, 1,488 unique SSR clones were isolated from Z. jujuba 'Dongzao' using enriched genomic libraries coupled with a three-primer colony PCR screening strategy, yielding a high enrichment rate of 73.3%. Finally, 1,188 (80.87%) primer pairs were amplified successfully in the size expected for 'Dongzao'. A total of 350 primer pairs were further selected and evaluated for their ability to detect polymorphisms across a panel of six diverse cultivars; among these, 301 primer pairs detected polymorphisms, and the polymorphism information content (PIC) value across all loci ranged from 0.15 to 0.82, with an average of 0.52. An analysis of 76 major cultivars employed in Chinese jujube production using 31 primer pairs revealed comparatively high genetic diversity among these cultivars. Within-population differences among individuals accounted for 98.2% of the observed genetic variation. Neighbor-joining clustering divided the cultivars into three main groups, none of which correspond to major geographic regions, suggesting that the genetics and geographical origin of modern Chinese jujube cultivars might not be linked. The current work firstly reports the large-scale development of Chinese jujube SSR markers. The development of these markers and their polymorphic information represent a significant improvement in the available Chinese jujube genomic resources and will facilitate both genetic and breeding applications, further accelerating the development of new cultivars.


Effect of body weight and caloric restriction on serum complement proteins, including Factor D/adipsin: studies in anorexia nervosa and obesity.
Complement plays important roles in host immune defences, and recent studies suggest that adipose tissue is an important site of production for some complement proteins. Starvation has been associated with low complement levels, but studied populations have usually had concomitant opportunistic infections or other conditions which might affect complement levels. To determine the impact of body weight and changes in body weight on serum complement, we investigated levels of complement proteins in otherwise healthy patients with a wide range of body weights, including patients with anorexia nervosa before and after treatment, obese dieters before and after weight loss, and normal weight controls. We found that complement proteins of the alternative pathway (C3, B, and D), alternative pathway haemolytic activity (AP50) and the inhibitors H and I were low in starving anorectics and normalized with weight gain. C3a levels were comparable in anorectics at low weight and after weight gain, indicating that low serum complement levels were attributable to hypoproduction and not complement cascade activation with consumption. Further, levels of C3, B, AP50, H and I, but not D, were higher than controls in obese patients and decreased toward normal after weight loss. Overall, percentage of ideal body weight, changes in body weight, and serum transferrin were each highly correlated with serum levels of complement proteins. We conclude that levels of alternative pathway complement components are determined in part by factors that influence body weight and by weight changes, possibly due to changes in production in adipose tissue or at other sites.


Sex and estrogens and responsiveness of terminal arterioles to neurohypophyseal hormones and catecholamines.
It has been generally assumed that reactivity (or responsiveness) of mammalian microcirculatory blood vessels to vasoactive hormones is not influenced by sex. In view of the paucity of knowledge in this area, the present in vivo studies were initiated to determine the quantitative dose-response relationships (by means of a high magnification image-splitting television microscope recording system) of catecholamines, selected sympathomimetics, neurohypophyseal hormones (NHPH), serotonin and angiotensin on mesenteric arterioles of unpretreated male and female rats and male rats pretreated with single low doses of 17 beta-estradiol (2 and 10 mug/100 g s.c.). The results indicate: 1) dose-response curves for the constrictor catecholamines (epinephrine and norepinephrine) and NHPH (vasopressin, oxytocin, vasotocin) in female rats are significantly shifted in parallel manner to the left of those obtained in male rats; 2) pretreatment of male rats with estrogen results in an enhancement of the constrictor actions of catecholamines, NHPH and phenylephrine; 3) the maximal arteriolar contractile responses (i.e., lumen narrowings) to norepinephrine and phenylphrine were enhanced by pretreatment with estrogen; and 4) dose-response curves for arteriolar constrictions induced by dopamine, serotonin or angiotensin were not affected by sex or estrogen pretreatment. The sex-linked differences, observed in this study, thus appear to be specific for NHPH and certain alpha adrenergic agonists. Overall, these findings reinforce the concept that sex hormones may play a role in control of peripheral blood flow and reactivity of arteriolar smooth muscle.


An open-label trial of acamprosate in the treatment of pathological gambling.
The efficacy and tolerability of acamprosate has been tested in the treatment of pathological gambling (PG). Acamprosate is known to reduce alcohol craving and use in persons with alcohol dependence, and it has been hypothesized that the drug would have a similar effect in individuals with PG. Participants with DSM-IV criteria for PG received acamprosate in an 8-week, open-label trial following a 2-week observation. The primary efficacy measure was the Yale-Brown Obsessive Compulsive Scale modified for PG (Y-BOCS-PG). Secondary efficacy measures included the Gambling Severity Assessment Scale (GSAS), the Clinical Global Impression (CGI) Improvement and Severity Scales, a patient self-rated global rating, the Hamilton Depression Rating Scale (HDRS), the Sheehan Disability Scale (SDS), and the Timeline Follow Back (TLFB). The study was conducted at 2 sites. Twenty-six participants (11 men, 15 women) had at least 1 post-baseline visit and were included in the analysis. Twenty participants (77%) completed the protocol. Significant improvement was observed in Y-BOCS-PG and GSAS scores, both CGI scales, a patient self-rated global scale, all 3 SDS subscales, and number of gambling episodes. Seventeen participants (65%) were considered responders (ie, achieved "much" or "very much" improvement). Improvements on the HDRS, in money wagered, and in time spent gambling were not significant. Few adverse events were reported. The results suggest that acamprosate is well tolerated and may be effective in the treatment of PG.