Sleep arrangements, parent-infant sleep during the first year, and family functioning.
The present longitudinal study addressed the ongoing debate regarding the benefits and risks of infant-parent cosleeping by examining associations between sleep arrangement patterns across the first year of life and infant and parent sleep, marital and family functioning, and quality of mothers' behavior with infants at bedtime. Patterns of infant sleep arrangements across the infants' first year were derived from information obtained from 139 families at 1, 3, 6, 9, and 12 months of infant age in a central Pennsylvania sample. Linkages between these patterns and parent-infant sleep, marital and coparenting stress, and maternal behavior at bedtime (from video-recordings) were assessed. Compared with families whose infants were solitary sleepers by 6 months, persistent cosleeping was associated with sleep disruption in mothers but not in infants, although mothers in persistent cosleeping arrangements reported that their infants had more frequent night awakenings. Persistent cosleeping was also associated with mother reports of marital and coparenting distress, and lower maternal emotional availability with infants at bedtime (from home observations). Persistent cosleeping appeared to be a marker of, though not necessarily a cause of, heightened family stress, although the present design did not enable strong tests of causal processes, and results may be particular to cultures that are not supportive of cosleeping. Findings are discussed in terms of cultural contexts of infant sleep and the need for further investigations into the role of the health of the family system in influencing how parents structure infant sleep. (PsycINFO Database Record


Inverse carbon isotope patterns of lipids and kerogen record heterogeneous primary biomass.
Throughout the Proterozoic δ(13)C values for preserved n-alkyl lipids are more positive than for syngenetic kerogen. This pattern is the inverse of biosynthetic expectations. It has been suggested that this isotopic inversion results from selective preservation of lipids from (13)C-enriched heterotrophic populations, while the bulk of kerogen derives from primary producers. Here, we formulate a degradation model to calculate the (13)C content of sedimentary total organic carbon and lipid. The model addresses two scenarios. The first scenario explores preferential preservation of heterotrophic lipid, thereby quantifying the existing hypothesis. In the second, we suggest that an inverse signature could be the result of prokaryotic phytoplankton contributing the majority of the total ecosystem biomass. Photosynthetic prokaryotes bearing a relative (13)C enrichment would contribute much of the resulting preserved lipids, while primary eukaryotic biomass would dominate the total organic carbon. We find that our hypothesis of a mixed primary producer community generates inverse isotopic patterns while placing far fewer requirements on specific degradation conditions. It also provides a possible explanation as to why there are large variations in the (13)C content of the isoprenoid lipids pristane and phytane relative to n-alkyl lipid, while the difference between n-alkyl lipid and kerogen is more constant. Our results suggest that the disappearance of the inverse (13)C signature in the late Ediacaran is a natural consequence of the fundamental shift to oceans in which export production has a higher ratio of eukaryotic biomass.


New Raffaelea species (Ophiostomatales) from the USA and Taiwan associated with ambrosia beetles and plant hosts.
Raffaelea (Ophiostomatales) is a genus of more than 20 ophiostomatoid fungi commonly occurring in symbioses with wood-boring ambrosia beetles. We examined ambrosia beetles and plant hosts in the USA and Taiwan for the presence of these mycosymbionts and found 22 isolates representing known and undescribed lineages in Raffaelea. From 28S rDNA and β-tubulin sequences, we generated a molecular phylogeny of Ophiostomatales and observed morphological features of seven cultures representing undescribed lineages in Raffaelea s. lat. From these analyses, we describe five new species in Raffaelea s. lat.: R. aguacate, R. campbellii, R. crossotarsa, R. cyclorhipidia, and R. xyleborina spp. nov. Our analyses also identified two plant-pathogenic species of Raffaelea associated with previously undocumented beetle hosts: (1) R. quercivora, the causative agent of Japanese oak wilt, from Cyclorhipidion ohnoi and Crossotarsus emancipatus in Taiwan, and (2) R. lauricola, the pathogen responsible for laurel wilt, from Ambrosiodmus lecontei in Florida. The results of this study show that Raffaelea and associated ophiostomatoid fungi have been poorly sampled and that future investigations on ambrosia beetle mycosymbionts should reveal a substantially increased diversity.


Developing attributes for a generic quality of life measure for older people: preferences or capabilities?
Current UK policy with respect to the provision of health and social care for older people suggests that greater integration is required. Economists' attempts to assist resource allocation decisions, however, are very health focused, with concentration on the use of health-related quality of life measures. This paper reports an attempt to determine attributes for a new index clearly focusing on quality of life for older people rather than health or other influences on quality of life. In-depth interviews were conducted with 40 purposively selected informants aged 65 and over in private households to explore their views about what is important to them in terms of quality of life. Data were analysed using Framework qualitative analysis. Initial discussions tended to concentrate upon factors influencing quality of life including activities, relationships, health, wealth and surroundings. Further probing and analysis suggested five conceptual attributes: attachment, role, enjoyment, security and control. The data also suggested that the quality of informants' lives was limited by the loss of ability to pursue these attributes. So, for example, it is not poor health in itself, which reduces quality of life, but the influence of that poor health upon each informant's ability to, say, be independent, that is important. Amartya Sen's work on functioning and capability is particularly pertinent here. Using this work, it is possible to interpret the five conceptual attributes as a set of functionings-important for older people in the UK in the 21st century-but noting that it is the capacity to achieve these functionings that appears to be of importance. This suggests that further development of this measure should focus on an index of capability rather than preference-based utility.


Molecular regulation of cholesterol biosynthesis: implications in carcinogenesis.
Cholesterol synthesis was demonstrated to be mandatory for cellular growth and serves to supply one of the necessary building blocks for new membranes demanded by dividing cells during growth. The mevalonate pathway, which is regulated through a finely tuned mechanism, is responsible mainly for cholesterol enrichment to cells. Among the various steps, the production of mevalonate from 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) is the most critically regulated step catalyzed by HMG-CoA reductase. The ability of sterols to regulate both the transcriptional rates of the reductase gene and the degradative machinery for the reductase protein provides a multilevel system for controlling the expression of this enzyme. Much convincing evidence indicates that cells manifest a higher flux through the mevalonate pathway when proliferating than when they are in the cell cycle arrest condition; furthermore, tumors undergo deregulated cholesterogenesis mainly at the critical rate-controlling juncture (i.e., the reaction catalyzed by HMG-CoA reductase). The mevalonate component of the cholesterol biosynthesis plays a key role in controlling cell proliferation by generating prenyl intermediates, particularly farnesyl and geranyl-geranyl moieties. These isoprenoids covalently modify and thus modulate the biological activity of signal transducing proteins, such as that of the Ras superfamily. The prenylated Ras-mediated signal transduction pathway provides much of the molecular information needed to trigger cell proliferation. Therefore, depletion of mevalonate can block both the processing and the transforming activities of Ras, indicating that drugs such as lovastatin and compactin, which had previously been exploited for lowering cholesterol levels, may be useful chemotherapeutic agents for treating tumors harboring oncogenic Ras mutation. In addition, Ras prenylation, which provides much of the molecular information needed to trigger cell proliferation, represents an inviting target for the design of chemotherapeutic drugs that would interrupt such signaling events and arrest tumor cell proliferation.


Further insights into the syndrome of prolapsing non-coronary aortic cusp and ventricular septal defect.
Ventricular septal defect (VSD) with prolapse of the right coronary cusp and aortic regurgitation can be managed surgically with the anatomical correction technique. However when the VSD is located underneath the non coronary cusp surgical management differs due to anatomical constraints and secondary pathological changes seen in the non coronary cusp. It is therefore important that the location of the VSD and the morphology of prolapsing cusp be characterised preoperatively in order to plan appropriate surgical repair. We present a case study in which we discuss the salient differences in the surgical management of the prolapsing right and the prolapsing non coronary cusps.


Characterising the STR locus D6S1043 and examination of its effect on stutter rates.
The forensic analysis of DNA is most often undertaken by the amplification of short tandem repeats (STR) using the polymerase chain reaction (PCR). DNA amplification can result in production of the target allele amplicon and a by-product called stutter. Stutter is the result of the miscopy of the target allele and is typically one repeat smaller. Stutter is traditionally described as a ratio of stutter and allele height; stutter ratio (SR). The challenge to DNA profile interpretation is most serious whenever stutter products are of a similar height to the minor allelic peaks in a mixed DNA profile. An accurate assignment of peaks and the prediction of their height is important when objectively interpreting forensic DNA profiles. The longest uninterrupted stretch (LUS) of tandem repeats within the allele has previously been shown to be a good predictor of stutter ratio. LUS is determined by sequencing a range of observed alleles at a locus. The locus D6S1043 is a relatively new locus to appear in commercial forensic DNA testing kits. To date however, there has been no comprehensive report of sequencing of this locus. In this work, we sequence a sample of D6S1043 alleles to determine LUS values and investigate allele repeat number and LUS as explanatory variables for SR.


Molecular mechanisms of drug resistance and its reversal in cancer.
Chemotherapy is the main strategy for the treatment of cancer. However, the main problem limiting the success of chemotherapy is the development of multidrug resistance. The resistance can be intrinsic or acquired. The resistance phenotype is associated with the tumor cells that gain a cross-resistance to a large range of drugs that are structurally and functionally different. Multidrug resistance arises via many unrelated mechanisms, such as overexpression of energy-dependent efflux proteins, decrease in uptake of the agents, increase or alteration in drug targets, modification of cell cycle checkpoints, inactivation of the agents, compartmentalization of the agents, inhibition of apoptosis and aberrant bioactive sphingolipid metabolism. Exact elucidation of resistance mechanisms and molecular and biochemical approaches to overcome multidrug resistance have been a major goal in cancer research. This review comprises the mechanisms guiding multidrug resistance in cancer chemotherapy and also touches on approaches for reversing the resistance.


Laser irradiated nano-architectured undoped tin oxide arrays: mechanism of ultrasensitive room temperature hydrogen sensing.
Undoped nanostructured tin oxide (SnO(2)) arrays were prepared on oxidized Si substrates by nanosecond pulsed laser interference irradiation for hydrogen gas sensing applications. Scanning electron microscopy (SEM), in combination with Atomic Force Microscopy (AFM), showed that the SnO(2) surface consisted of periodic features of ∼130 nm width, ∼228 nm spacing, an average height of ∼8 nm along the periodicity and tens of microns length. The SnO(2) nanostructured arrays and precursor thin films were tested by cyclic exposure under dynamic conditions of hydrogen in the concentration range of 300-9000 ppm. The observed electrical response of SnO(2) towards hydrogen at low concentrations and room temperature drastically improved in the nanostructured array as compared to the thin film. The results suggest that this method to fabricate SnO(2) nanostructured arrays has the potential to produce nanodevices that have ultra-low detection limits, and fast response and recovery times, which are suited for practical hydrogen sensing applications.


Peripheral blood stem cell mobilisation by granulocyte-colony stimulating factor in patients with acute and old myocardial infarction for intracoronary cell infusion.
Peripheral blood stem cells (PBSC) are one of the most promising stem cell sources for treatment of ischaemic heart disease. However, the experience of mobilisation and collection of PBSC using granulocyte-colony stimulating factor (G-CSF) in patients with myocardial infarction (MI) is still limited. We report our experiences with the feasibility and safety of collection of mobilised PBSC with G-CSF in MI patients, and the influence of acute ischaemia on efficacy of PBSC collection. 74 patients with acute or old myocardial infarction (AMI vs OMI, n = 46 and n = 28) underwent PBSC collection after administration of G-CSF twice a day at a dose of 5 microg/kg for 3 days. Flow cytometric analysis of cell surface markers was performed. No evidence of inflammation or ischaemia was observed during G-CSF mobilisation and PBSC collection. The yield of CD34(+) cells was 12.9 (SD 15.92) x10(9)/l (5.04% (5.30%) of total cells) with a product volume of 37.9 (8.4) ml after 5650 (987) ml of blood were processed during PBSC collection. Stem cell mobilisation and collection by G-CSF is more efficient in AMI than in OMI, and proportions of cells positive for VE-cadherin or KDR/CD34 are significantly greater in AMI than in OMI (p<0.01). We could obtain sufficient numbers of PBSC for intracoronary infusion with the G-CSF-based mobilisation strategy without complications even in patients with MI. PBSC collection after mobilisation with G-CSF is a safe and feasible method of stem cell collection for therapeutic purpose in patients with MI.


In vivo matrix metalloproteinase-7 substrates identified in the left ventricle post-myocardial infarction using proteomics.
Matrix metalloproteinase-7 (MMP-7) deletion has been shown to improve survival after myocardial infarction (MI). MMP-7 has a large array of in vitro substrates, but in vivo substrates for MMP-7 following MI have not been fully identified. Accordingly, we evaluated the infarct regions of wild-type (WT; n = 12) and MMP-7 null (null; n = 10) mice using a proteomic strategy. Seven days post-MI, infarct regions of the left ventricles were excised, homogenized, and protein extracts were analyzed by two-dimensional gel electrophoresis and mass spectrometry. Of 13 spots that showed intensity differences between WT and null, the intensities of eight spots were higher and those of five spots were lower in the null group (p < 0.05). Fibronectin and tenascin-C, known in vitro substrates of MMP-7, were identified in spots that showed lower intensity in the null. Immunoblotting and in vitro cleavage assays confirmed reduced fibronectin and tenascin-C fragment generation in the null, and this effect was restored by exogenous administration of MMP-7. Lower levels of full-length peroxiredoxin-1 and -2 and higher levels of the full-length peroxiredoxin-3 were detected in the null group, suggesting MMP-7 deletion may also indirectly regulate protein levels through nonenzymatic mechanisms. In conclusion, this is the first study to identify fibronectin and tenascin-C as in vivo MMP-7 substrates in the infarcted left ventricle using a proteomic approach.


Physician assisted suicide perceptions among patients presenting to family physicians at a teaching hospital in Karachi, Pakistan.
To study the perceptions on physician assisted suicide, among patients presenting to family physicians, at a teaching hospital in Karachi, Pakistan. The study was carried out at the Community Health Center of The Aga Khan University Hospital, Karachi, between December 1999 and May 2000. The principal and co-investigators filled a pre-coded and pretested questionnaire consisting of important demographic characteristics and bio-medical ethics issues. A system of convenience sampling was used and a written consent was taken from respondents over the age of 16 years, who agreed to participate. The data were managed by using the Epi Info (version 6.0) program. Four hundred twenty respondents were interviewed against an estimated sample size of 385. Majority (88%) were males between 25 and 34 years of age, and were self employed or in private service. Overall 9% respondents were advocated of physician assisted suicide, advocates was 9%. Those who advocated the Physician assisted suicide were more likely to be female, elderly, married and educated amongst the total respondents. Those who support Physician assisted suicide were less likely to attach divine qualities to physicians and were more bold, courageous and tolerant towards broader biomedical ethics issues. We have found a substantial acceptability to the idea of Physician assisted suicide in a Muslim society and have identified characteristics of those who support it.


Design and optimization of a series of 1-sulfonylpyrazolo[4,3-b]pyridines as selective c-Met inhibitors.
c-Met has emerged as an attractive target for targeted cancer therapy because of its abnormal activation in many cancer cells. To identify high potent and selective c-Met inhibitors, we started with profiling the potency and in vitro metabolic stability of a reported hit 7. By rational design, a novel sulfonylpyrazolo[4,3-b]pyridine 9 with improved DMPK properties was discovered. Further elaboration of π-π stacking interactions and solvent accessible polar moieties led to a series of highly potent and selective type I c-Met inhibitors. On the basis of in vitro and in vivo pharmacological and pharmacokinetics studies, compound 46 was selected as a preclinical candidate for further anticancer drug development.


Concurrent advanced HIV disease and viral load suppression in a high-burden setting: Findings from the 2015-6 ZIMPHIA survey.
As Zimbabwe approaches epidemic control of HIV, programs now prioritize viral load over CD4 monitoring, making it difficult to identify persons living with HIV (PLHIV) suffering from advanced disease (AD). We present an analysis of cross-sectional ZIMPHIA data, highlighting PLHIV with AD and concurrent viral load suppression (VLS). ZIMPHIA collected blood specimens for HIV testing from 22,501 consenting adults (ages 15 years and older); 3,466 PLHIV had CD4 and VL results. Household HIV testing used the national serial algorithm, and those testing positive then received point-of-care CD4 enumeration with subsequent VL testing. We used logistic regression analysis to explore factors associated with concurrent AD and VLS (<1000 copies/mL). All analyses were weighted to account for complex survey design. Of the 3,466 PLHIV in the survey with CD4 and VL results, 17% were found to have AD (CD4<200cells/mm3). Of all AD patients, 30% had VLS. Concurrent AD and VLS was associated with male sex (aOR 2.45 95%CI 1.61-3.72), older age (35-49 years [aOR 2.46 95%CI 1.03-5.91] and 50+ years [aOR 4.82 95%CI 2.02-11.46] vs 15-24 years), and ART duration (<6 months [aOR 0.46 95%CI 0.29-0.76] and 6-24 months [aOR 2.07 95%CI 1.35-3.17] vs more than 2 years). The relationship between sex and AD is driven by age with significant associations among men aged 25-34, (aOR 3.37 95%CI 1.35-8.41), 35-49 (aOR 5.13 95%CI 2.16-12.18), and 50+ (aOR 12.56 95%CI 4.82-32.72) versus men aged 15-24. The percentage of PLHIV with AD and VLS illustrates the conundrum of decreased support for CD4 monitoring, as these patients may not receive appropriate clinical services for advanced HIV disease. In high-prevalence settings such as Zimbabwe, CD4 monitoring support warrants further consideration to differentiate care appropriately for the most vulnerable PLHIV. Males may need to be prioritized, given their over-representation in this sub-population.


The Salmonella effector SifA initiates a kinesin-1 and kinesin-3 recruitment process mirroring that mediated by Arl8a and Arl8b.
When intracellular, pathogenic Salmonella reside in a membrane compartment composed of interconnected vacuoles and tubules, the formation of which depends on the translocation of bacterial effectors into the host cell. Cytoskeletons and their molecular motors are prime targets for these effectors. In this study, we show that the microtubule molecular motor KIF1Bβ (a splice variant of KIF1B), a member of the kinesin-3 family, is a key element for the establishment of the Salmonella replication niche as its absence is detrimental to the stability of bacterial vacuoles and the formation of associated tubules. Kinesin-3 interacts with the Salmonella effector SifA but also with SKIP (also known as PLEKHM2), a host protein complexed to SifA. The interaction with SifA is essential for the recruitment of kinesin-3 on Salmonella vacuoles whereas that with SKIP is incidental. In the non-infectious context, however, the interaction with SKIP is essential for the recruitment and activity of kinesin-3 only on a fraction of the lysosomes. Finally, our results show that, in infected cells, the presence of SifA establishes a kinesin-1 and kinesin-3 recruitment pathway that is analogous to and functions independently of that mediated by the Arl8a and Arl8b GTPases. This article has an associated First Person interview with the first author of the paper.


High and low pressure pulsatile lavage of contaminated tibial fractures: an in vitro study of bacterial adherence and bone damage.
This study was designed to examine the effect of pulsatile irrigation on microscopic bone architecture and its time-dependent efficacy in removing adherent slime-producing bacteria from cortical bone. Using an in vitro model, ten-millimeter transverse cut sections from five human tibiae were contaminated with Staphylococcus aureus and subjected to either high pressure pulsatile lavage (HPPL; seventy pounds per square inch, normal saline) or low pressure pulsatile lavage (LPPL; fourteen pounds per square inch, normal saline) or served as controls. Alteration of bony architecture was quantified by using a previously described ordinal scale and histomorphometric analysis of each transverse cut section of tibia. To assess the time-dependent effectiveness of pulsatile lavage in removing adherent bacteria from bone, ten-millimeter transverse cut sections from ten canine tibiae were contaminated with S. aureus and subjected to high or low pressure pulsatile lavage immediately or after one, three, or six hours. Scanning electron microscopy and bacterial cultures were used to assess the removal of adherent bacteria. HPPL resulted in significantly greater macroscopic damage than was seen with LPPL or in controls (ANOVA, p < 0.001). Histomorphometry revealed that HPPL was associated with significantly larger and more numerous fissures or defects in the cortical bone when compared with low pressure irrigation (p < 0.001). However, high and low pressure lavage were associated with similar degrees of periosteal separation from the cortical bone surface (p = 0.87). Both high and low pressure lavage were effective in removing adherent bacteria from bone at three hours irrigation delay, but only high pressure lavage removed adherent bacteria from bone at six hours delay. In this in vitro study, compared with HPPL, LPPL led to less structural damage and was equally effective in removing bacteria within three hours debridement delay; however, the efficacy of LPPL at six hours debridement delay is questionable. This finding may have clinical significance in the development of infection following open tibial fractures.


In a randomized trial, outcomes were not affected by intensive follow-up over 1 year.
This study tested the impact on response and psychological measures of different follow-up schedules for participants in a study screening for type 2 diabetes risk. Participants were randomly allocated either to limited follow-up (n=213), with a single questionnaire at 1 year, or to intensive follow-up (n=218), with questionnaires sent 1, 6, and 12 months after screening. There were no statistically significant differences between the groups at 1 year in either response rate (P=.08) or change from baseline to 1-year follow-up in scores on the short form of the Spielberger State Anxiety Inventory (SSAI-SF; P=.13), although there was a statistically significant but small difference between the groups in the change score of the 12-item Well-Being Questionnaire (WBQ-12; P=.003). The proportion of participants completing a WBQ-12 measure at 1 year was significantly greater in the intensive follow-up group (P=.03). This study shows no important adverse effect of repeated questionnaire use on response rates or psychological outcomes and suggests that some measures may be more fully completed with intensive follow-up.


Occupational respiratory allergy in bakery workers: a review of the literature.
Baker's asthma has long been recognized as a serious disease among workers in the bakery industry and the number of cases with baker's asthma is steadily increasing. This paper presents a review of the available literature on baker's allergy with a special focus on the allergens involved, the epidemiologic research and issues on exposure assessment, evidence of exposure-response-relationships, and possible prevention strategies. A large number of potential allergens have been identified and are described here. At present little is known about the incidence of baker's allergy. On the other hand, a large number of cross-sectional studies have been performed, showing that sensitization and work-related symptoms are common among bakery workers. Only atopy and exposure level have consistently been reported as determinants of this occupational disease. Age, gender, and smoking habits do not seem to be associated with sensitization or work-related respiratory symptoms. Recently, immunochemical methods have been developed to measure specific allergens in the bakery industry, which have been used to unravel the role of allergen exposure in the development of baker's asthma. Clear exposure-response-relationships have been found. The implications of these recent findings for prevention strategies and standard setting are discussed.


Molecular markers of angiogenesis and metastasis in lines of oral carcinoma after treatment with melatonin.
Oral cancer is the most common type of head and neck cancer and its high rate of mortality and morbidity is closely related to the processes of angiogenesis and tumor metastasis. The overexpression of the pro-angiogenic genes, HIF-1α and VEGF, and pro-metastatic gene, ROCK-1, are associated with unfavorable prognosis in oral carcinoma. Melatonin has oncostatic, antiangiogenic and antimetastatic properties in several types of neoplasms, although its relationship with oral cancer has been little explored. This study aims to analyze the expression of the genes HIF-1α, VEGF and ROCK-1 in cell lines of squamous cell carcinoma of the tongue, after treatment with melatonin. SCC9 and SCC25 cells were cultured and cell viability was assessed by MTT assay, after treatment with 100 μM of CoCl2 to induce hypoxia and with melatonin at different concentrations. The analysis of quantitative RT-PCR and the immunocytochemical analysis were performed to verify the action of melatonin under conditions of normoxia and hypoxia, on gene and protein expression of HIF-1α, VEGF and ROCK-1. The MTT assay showed a decrease in cell viability in both cell lines, after the treatment with melatonin. The analysis of quantitative RT-PCR indicated an inhibition of the expression of the pro-angiogenic genes HIF-1α (P < 0.001) and VEGF (P < 0.001) under hypoxic conditions, and of the pro-metastatic gene ROCK-1 (P < 0.0001) in the cell line SCC9, after treatment with 1 mM of melatonin. In the immunocytochemical analysis, there was a positive correlation with gene expression data, validating the quantitative RT-PCR results for cell line SCC9. Treatment with melatonin did not demonstrate inhibition of the expression of genes HIF-1α, VEGF and ROCK-1 in line SCC25, which has different molecular characteristics and greater degree of malignancy when compared to the line SCC9. Melatonin affects cell viability in the SCC9 and SCC25 lines and inhibits the expression of the genes HIF-1α, VEGF and ROCK-1 in SCC9 line. Additional studies may confirm the potential therapeutic effect of melatonin in some subtypes of oral carcinoma.


Translational health economics: The key to accountable adoption of in vitro diagnostic technologies.
Adoption of new technologies, including diagnostic tests, is often considered not to deliver the expected return on investment. The reasons for this poor link between expectation and outcome include lack of evidence, variation in use of the technology, and an inability of the health system to manage the balance between investment and disinvestment associated with the change in care pathway. The challenges lie in the complex nature of healthcare provision where the investment is likely to be made in the jurisdiction of one stakeholder while the benefits (as well as dis-benefits) accrue to the other stakeholders. A prime example is found in the field of laboratory medicine and the use of diagnostic tests. The current economic tools employed in healthcare are primarily used to make policy and strategic decisions, particularly across health systems, and in purchaser and provider domains. These tools primarily involve cost effectiveness and budget impact analyses, both of which have been applied in health technology assessment of diagnostic technologies. However, they lack the granularity to translate findings down to the financial management and operational decision making at the provider department level. We propose an approach to translational health economics based on information derived from service line management and time-driven activity-based costing, identifying the resource utilisation for each of the units involved in the delivery of a care pathway, before and after adoption of new technology. This will inform investment and disinvestment decisions, along with identifying where the benefits, and dis-benefits, can be achieved for all stakeholders.


Mycobacterium tuberculosis requires phosphate-responsive gene regulation to resist host immunity.
Mycobacterium tuberculosis persists in the tissues of mammalian hosts despite inducing a robust immune response dominated by the macrophage-activating cytokine gamma interferon (IFN-γ). We identified the M. tuberculosis phosphate-specific transport (Pst) system component PstA1 as a factor required to resist IFN-γ-dependent immunity. A ΔpstA1 mutant was fully virulent in IFN-γ(-/-) mice but attenuated in wild-type (WT) mice and mice lacking specific IFN-γ-inducible immune mechanisms: nitric oxide synthase (NOS2), phagosome-associated p47 GTPase (Irgm1), or phagocyte oxidase (phox). These phenotypes suggest that ΔpstA1 bacteria are sensitized to an IFN-γ-dependent immune mechanism(s) other than NOS2, Irgm1, or phox. In other species, the Pst system has a secondary role as a negative regulator of phosphate starvation-responsive gene expression through an interaction with a two-component signal transduction system. In M. tuberculosis, we found that ΔpstA1 bacteria exhibited dysregulated gene expression during growth in phosphate-rich medium that was mediated by the two-component sensor kinase/response regulator system SenX3-RegX3. Remarkably, deletion of the regX3 gene suppressed the replication and virulence defects of ΔpstA1 bacteria in NOS2(-/-) mice, suggesting that M. tuberculosis requires the Pst system to negatively regulate activity of RegX3 in response to available phosphate in vivo. We therefore speculate that inorganic phosphate is readily available during replication in the lung and is an important signal controlling M. tuberculosis gene expression via the Pst-SenX3-RegX3 signal transduction system. Inability to sense this environmental signal, due to Pst deficiency, results in dysregulation of gene expression and sensitization of the bacteria to the host immune response.


[Non-fatal work-related accidents in adolescents].
This community-based survey estimates the annual incidence of non-fatal work-related injuries in adolescents and describes their characteristics. Using a random area sampling design, 2,513 households were selected from the city of Salvador, capital of Bahia State, Northeast Brazil. The study population included 361 young people 10 to 20 years of age who answered individual questionnaires on socio-demographic, occupational, and health data. Annual incidence of non-fatal work-related injuries was 6.4%. Among females, most injuries were mild and were related to household chores; some 38.5% required medical care, and 36.4% caused permanent non-disabling lesions. Among males, work-related injuries took place mainly in public byways, and the majority (60.0%) were treated in emergency-care facilities. These results show that work-related injuries need to be considered in adolescent protection policies.


The sll1951 gene encodes the surface layer protein of Synechocystis sp. strain PCC 6803.
Sll1951 is the surface layer (S-layer) protein of the cyanobacterium Synechocystis sp. strain PCC 6803. This large, hemolysin-like protein was found in the supernatant of a strain that was deficient in S-layer attachment. An sll1951 deletion mutation was introduced into Synechocystis and was easily segregated to homozygosity under laboratory conditions. By thin-section and negative-stain transmission electron microscopy, a ~30-nm-wide S-layer lattice covering the cell surface was readily visible in wild-type cells but was absent in the Δsll1951 strain. Instead, the Δsll1951 strain displayed a smooth lipopolysaccharide surface as its most peripheral layer. In the presence of chaotropic agents, the wild type released a large (>150-kDa) protein into the medium that was identified as Sll1951 by mass spectrometry of trypsin fragments; this protein was missing in the Δsll1951 strain. In addition, Sll1951 was prominent in crude extracts of the wild type, indicating that it is an abundant protein. The carotenoid composition of the cell wall fraction of the Δsll1951 strain was similar to that of the wild type, suggesting that the S-layer does not contribute to carotenoid binding. Although the photoautotrophic growth rate of the Δsll1951 strain was similar to that of the wild-type strain, the viability of the Δsll1951 strain was reduced upon exposure to lysozyme treatment and hypo-osmotic stress, indicating a contribution of the S-layer to the integrity of the Synechocystis cell wall. This work identifies the S-layer protein in Synechocystis and shows that, at least under laboratory conditions, this very abundant, large protein has a supportive but not a critical role in the function of the cyanobacterium.


Diagnosis of dementia in a heterogeneous population. Development of a neuropsychological paradigm-based diagnosis of dementia and quantified correction for the effects of education.
A brief diagnostic battery of neuropsychological tests was developed for a large-scale epidemiological study of dementia. We operationally defined dementia as defective memory and defective performance in at least two other areas, including orientation, abstract reasoning, construction, and language. Criterion scores for defining defective performance on each test were developed. In a pilot study that used 51 different subjects with a working diagnosis based on physicians' assessment (ie, 32 demented and 29 nondemented subjects), the test-based diagnosis agreed with the working diagnosis in all but two cases. The test battery was then applied to 430 healthy elderly subjects. Eighteen percent of those with 8 or less years of education met criteria for dementia compared with 5% of those with more than 8 years of education. We computed education-corrected scores for each test with the use of residuals from the regression of each test score on education. Based on corrected scores, 12 subjects were reclassified as nondemented and 11 as demented. Subjects who were reclassified as demented were significantly more impaired in activities of daily living than nondemented subjects who were not reclassified. Activities of daily living in subjects who were reclassified as nondemented did not differ from those in demented subjects who were not reclassified. These findings suggest that the neuropsychological battery may have utility in the diagnosis of dementia. However, neuropsychological performance may be influenced by education, and some form of adjustment, such as correction for activities of daily living, may be required in epidemiological studies.


Relationship between microbiology of throat swab and clinical course among primary care patients with acute cough: a prospective cohort study.
Acute lower respiratory tract infections (ALRTIs) account for most antibiotics prescribed in primary care despite lack of efficacy, partly due to clinician uncertainty about aetiology and patient concerns about illness course. Nucleic acid amplification tests could assist antibiotic targeting. In this prospective cohort study, 645 patients presenting to primary care with acute cough and suspected ALRTI, provided throat swabs at baseline. These were tested for respiratory pathogens by real-time polymerase chain reaction and classified as having a respiratory virus, bacteria, both or neither. Three hundred fifty-four participants scored the symptoms severity daily for 1 week in a diary (0 = absent to 4 = severe problem). Organisms were identified in 346/645 (53.6%) participants. There were differences in the prevalence of seven symptoms between the organism groups at baseline. Those with a virus alone, and those with both virus and bacteria, had higher average severity scores of all symptoms combined during the week of follow-up than those in whom no organisms were detected [adjusted mean differences 0.204 (95% confidence interval 0.010 to 0.398) and 0.348 (0.098 to 0.598), respectively]. There were no differences in the duration of symptoms rated as moderate or severe between organism groups. Differences in presenting symptoms and symptoms severity can be identified between patients with viruses and bacteria identified on throat swabs. The magnitude of these differences is unlikely to influence management. Most patients had mild symptoms at 7 days regardless of aetiology, which could inform patients about likely symptom duration.


Associations of genetic variants in/near body mass index-associated genes with type 2 diabetes: a systematic meta-analysis.
Genome-wide association studies have identified many obesity/body mass index (BMI)-associated loci in Europeans and East Asians. Since then, a large number of studies have investigated the role of BMI-associated loci in the development of type 2 diabetes (T2D). However, the results have been inconsistent. The objective of this study was to investigate the associations of eleven obesity/BMI loci with T2D risk and explore how BMI influences this risk. We retrieved published literature from PubMed and Embase. The pooled odds ratios (OR) with 95% confidence intervals (CI) were calculated using fixed- or random-effect models. In the meta-analysis of 42 studies for 11 obesity/BMI-associated loci, we observed a statistically significant association of the FTO rs9939609 polymorphism (66 425 T2D cases/239 689 normoglycaemic subjects; P = 1·00 × 10(-41) ) and six other variants with T2D risk (17 915 T2D cases/27 531 normoglycaemic individuals: n = 40 629-130 001; all P < 0·001 for SH2B1 rs7498665, FAIM2 rs7138803, TMEM18 rs7561317, GNPDA2 rs10938397, BDNF rs925946 and NEGR1 rs2568958). After adjustment for BMI, the association remained statistically significant for four of the seven variants (all P < 0·05 for FTO rs9939609, SH2B1 rs7498665, FAIM2 rs7138803, GNPDA2 rs10938397). Subgroup analysis by ethnicity demonstrated similar results. This meta-analysis indicates that several BMI-associated variants are significantly associated with T2D risk. Some variants increase the T2D risk independent of obesity, while others mediate this risk through obesity.


Use of ultrafiltration to isolate viruses from seawater which are pathogens of marine phytoplankton.
Viruses may be major structuring elements of phytoplankton communities and hence important regulators of nutrient and energy fluxes in aquatic environments. In order to ascertain whether viruses are potentially important in dictating phytoplankton community structure, it is essential to determine the extent to which representative phytoplankton taxa are susceptible to viral infection. We used a spiral ultrafiltration cartridge (30,000-molecular-weight cutoff) to concentrate viruses from seawater at efficiencies approaching 100%. Natural virus communities were concentrated from stations in the Gulf of Mexico, a barrier island pass, and a hypersaline lagoon (Laguna Madre) and added to cultures of potential phytoplankton hosts. By following changes in in vivo fluorescence over time, it was possible to isolate several viruses that were pathogens to a variety of marine phytoplankton, including a prasinophyte (Micromonas pusilla), a pennate diatom (likely a Navicula sp.), a centric diatom (of unknown taxa), and a chroococcoid cyanobacterium (a Synechococcus sp.). As well, we observed changes in fluorescence in cultures of a cryptophyte (a Rhodomonas sp.) and a chlorophyte (Nannochloropsis oculata) which were consistent with the presence of viral pathogens. Although pathogens were isolated from all stations, all the pathogens were not isolated from every station. Filterability studies on the viruses infecting M. pusilla and the Navicula sp. showed that the viruses were consistently infective after filtration through polycarbonate and glass-fiber filters but were affected by most other filter types. Establishment of phytoplankton-pathogen systems will be important in elucidating the effect that viruses have on primary producers in aquatic systems.


Seasonal changes in the genetic structure of an aphid-ant mutualism as revealed using microsatellite analysis of the aphid Tuberculatus quercicola and the ant Formica yessensis.
The present study examined whether the mutualistic relationship between the aphid Tuberculatus quercicola (Matsumura) (Homoptera: Aphididae) and the attending ant Formica yessensis Forel (Hymenoptera: Formicidae) has had any mutual effects on the microgeographical genetic population structure of both partner species. The aphids and the attending ants were collected in June, August, and October 2004 from six trees of the Daimyo oak Quercus dentata Thunberg (Fagales: Fagaceae) and were genotyped using microsatellite loci. Significant genetic differentiation was detected among T. quercicola populations on the respective trees across seasons (an average of pairwise F(ST) = 0.183). Similarly, significant genetic differentiation was found among populations of F. yessensis that attended aphid colonies on the respective host trees, though the averages of pairwise F(ST) were lower (an average of pairwise F(ST) = 0.070). An analysis of molecular variance and two-way ANOVA detected a significantly large genetic difference between spring and summer samples in F. yessensis but not in T. quercicola, indicating that changes in genetic composition occurred in the F. yessensis colony. In spite of a drastic seasonal change in the genetic difference in F. yessensis, principle coordinate analysis showed that the relative position among the six populations was maintained from spring to summer, suggesting that the tree where honeydew was available for a long time was occupied by F. yessensis over the same period and that the honeydew sources were inherited at the level of the ant colony. It is hypothesized that the suitability of host trees for the aphid T. quercicola may have an affect on the genetic structure of the attending ant F. yessensis. Within a colony of aphids, clonal diversity decreased significantly as the season progressed. The reduction in clonal diversity may be due to an increase in identical genotypes by parthenogenesis or selective pressure from host plant deterioration.


Genome-wide association study of major depressive disorder: new results, meta-analysis, and lessons learned.
Major depressive disorder (MDD) is a common complex disorder with a partly genetic etiology. We conducted a genome-wide association study of the MDD2000+ sample (2431 cases, 3673 screened controls and >1 M imputed single-nucleotide polymorphisms (SNPs)). No SNPs achieved genome-wide significance either in the MDD2000+ study, or in meta-analysis with two other studies totaling 5763 cases and 6901 controls. These results imply that common variants of intermediate or large effect do not have main effects in the genetic architecture of MDD. Suggestive but notable results were (a) gene-based tests suggesting roles for adenylate cyclase 3 (ADCY3, 2p23.3) and galanin (GAL, 11q13.3); published functional evidence relates both of these to MDD and serotonergic signaling; (b) support for the bipolar disorder risk variant SNP rs1006737 in CACNA1C (P=0.020, odds ratio=1.10); and (c) lack of support for rs2251219, a SNP identified in a meta-analysis of affective disorder studies (P=0.51). We estimate that sample sizes 1.8- to 2.4-fold greater are needed for association studies of MDD compared with those for schizophrenia to detect variants that explain the same proportion of total variance in liability. Larger study cohorts characterized for genetic and environmental risk factors accumulated prospectively are likely to be needed to dissect more fully the etiology of MDD.


Serum growth hormone and glucose levels in acute exercise and in the recovery period in athletes.
Growth hormone exerts several metabolic effects, including effects on proteins, fats and carbohydrates. Among the many metabolic activities of GH, two contradictory actions were described: acute and early insulin-like activity and chronic and late anti-insulin like activity also called diabetogenic activity. A dramatic increase in plasma concentration of GH was found during endurance exercise, but its role during exercise is not well known. According to its metabolic effects a possible role of growth hormone may be in maintenance of glucose level during exercise. The aim of this study was to analyze dynamics of changes in GH and glucose levels during acute workload and in the recovery period, in a group of well trained athletes. All the subjects exercised for 30 minutes on cycle ergometer in sitting position (work intensity 50% of VO2 max, RPM 60/min). Serum GH concentrations were measured by IRMA (immunoradiometric assays) method in blood samples obtained at rest and 6-min intervals during exercise, and 15-min intervals during recovery period. Serum glucose levels were determined by standard enzymatic method glucose oxidase (GOD PAP) at the same intervals. There were no correlations between serum GH and glucose levels either during exercise or in the recovery period. There were no differences between glucose levels during exercise, so we can not exclude possible role of GH in glucose concentration maintenance.


Predicted philogeny, secondary conformational structure, and epitope antigenicity of immunological sequences in poultry.
Poultry production is faced with different types of stresses that are responsible for issues of animal welfare as well as for economic losses. Moreover, the immunity decreases when animals are stressed. In silico analyses are important in reducing the cost and in increasing the accuracy of scientific results. A bioinformatics tool was used to perform ontology studies on 15 different immunological sequences of poultry. The mRNA structures and sequences with maximum antigenic residues were also predicted. No homology was found between the sequences of poultry and mammals. These results helped in the prediction of new potential molecular markers. Of the 15 sequences that were analyzed, predictions could not be made for five because they were longer than 2500 nucleotides; for the remaining 10 sequences, 20 conformational structures per sequence were predicted and the most stable sequences were identified by their minimum free energy values. The highest antigenic epitopes were accepted by the maximum scores; 15 of the total 8934 epitopes that were predicted were analyzed. These results would aid future studies that use synthetic peptides or recombinants as markers or immunomodulators and would expand our understanding on how stress can modulate the immune system. These would also help in developing rapid diagnostic tools, in increasing animal welfare, biosecurity, and productivity, and also in developing of food additives and environmental enrichment for stress control, thereby, making animal production more sustainable.


[Factors for voiding dysfunction and cystocele].
To evaluate the clinical and urodynamic features of a series of women with post void residual urine (disbalanced voiding) and various degrees of associated cystocele. 119 female patients were studied by clinical evaluation, urodynamics, and imaging tests (VCUG). All patients underwent history and genitourological examination (evaluating cystoceles from grade o to 3), neuro-urological examination, and complete urodynamic study. Fifty patients (42%) underwent radiological studies of the upper urinary tract. Disbalanced voiding was defined as existence of post void residual greater than 20% of the voided volume. Urethral resistance was measured by URA. Structural obstruction was characterized by PURR (CHESS classification). Functional obstruction was studied by DURR and perineal EMG (associated with flowmetry). Detrusor contractile power was evaluated by W max, W 80-20, and duration of contraction. Urodynamic terminology and measurements complied with the International Continence Society (ICS) standards. Statistical significance was established at 0.05. Statistical analysis was done by Student's t for quantitative variables, and Pearson's chi-square for non parametric variables. 119 patients were enrolled. Mean age was 55.84 yr. (range 15-87). Regarding post void residual (114 valid uroflowmetry studies), 25 patients were classified as voiding disbalance (21.9%) and 89 as balanced (74.8%). Regarding clinical data, there were only significant differences between groups in voiding difficulty. For uroflowmetry, only the percentile of the Maximal flow (Qmax) showed significant differences (35 vs. 22 for balanced/disbalanced voiding respectively, p = 0.02). Pressure/volume studies demonstrated bladder hyperactivity in 16 cases (64%) in the group of disbalanced voiding and 31 cases (34.8%) in the normal voiding group (p = 0.008), which presented associated with increased urethral resistance (URA)(p = 0.01) . In the pressure/flow study, there were significant differences in the URA (14.7 vs. 25.3, p = 0.001). There were statistically significant differences in the degree of constrictive (0.5 vs. 1.1, p = 0.009) and compressive (0.5 vs. 1.1; p = 0.04) obstruction (Chess classification). There were not significant differences in the analysis of isometric contractility (Wmax), but there were in the isotonic contractility (W80-20) and detrusor contraction duration. These latter differences presented significant association with the degree of cystocele. DURR and perineal EMG data did not show differences between groups. Radiological abnormalities of urethral morphology were statistically different between groups, presenting in 10% of the patients with normal voiding and 50% of the disbalanced voiding group, although there was not statistical association with obstruction (p = 0.64). The existence of cystocele did not show a statistical association with these variables either. Disbalanced voiding appeared with organic obstruction of the lower urinary tract (constrictive most significantly), as well as detrusor abnormal contractility, but whereas the first was not significantly associated with presence and grade of cystocele, the second showed such association.


Demystifying biotrophs: FISHing for mRNAs to decipher plant and algal pathogen-host interaction at the single cell level.
Plant-pathogen interactions follow spatial and temporal developmental dynamics where gene expression in pathogen and host undergo crucial changes. Therefore, it is of great interest to detect, quantify and localise where and when key genes are active to understand these processes. Many pathosystems are not accessible for genetic amendments or other spatially-resolved gene expression monitoring methods. Here, we adapt single molecule FISH techniques to demonstrate the presence and activity of mRNAs at the single-cell level using phytomyxids in their plant and algal host in lab and field material. This allowed us to monitor and quantify the expression of genes from the clubroot pathogen Plasmodiophora brassicae, several species of its Brassica hosts, and of several brown algae, including the genome model Ectocarpus siliculosus, infected with the phytomyxid Maullinia ectocarpii. We show that mRNAs are localised along a spatiotemporal gradient, thus providing a proof-of-concept of the usefulness of single-molecule FISH to increase knowledge about the interactions between plants, algae and phytomyxids. The methods used are easily applicable to any interaction between microbes and their algal or plant host, and have therefore the potential to rapidly increase our understanding of key, spatially- and temporally-resolved processes underpinning complex plant-microbe interactions.


Microwave-assisted synthesis of water-dispersed CdTe/CdSe core/shell type II quantum dots.
A facile synthesis of mercaptanacid-capped CdTe/CdSe (core/shell) type II quantum dots in aqueous solution by means of a microwave-assisted approach is reported. The results of X-ray diffraction and high-resolution transmission electron microscopy revealed that the as-prepared CdTe/CdSe quantum dots had a core/shell structure with high crystallinity. The core/shell quantum dots exhibit tunable fluorescence emissions by controlling the thickness of the CdSe shell. The photoluminescent properties were dramatically improved through UV-illuminated treatment, and the time-resolved fluorescence spectra showed that there is a gradual increase of decay lifetime with the thickness of CdSe shell.


Metallization of vanadium dioxide driven by large phonon entropy.
Phase competition underlies many remarkable and technologically important phenomena in transition metal oxides. Vanadium dioxide (VO2) exhibits a first-order metal-insulator transition (MIT) near room temperature, where conductivity is suppressed and the lattice changes from tetragonal to monoclinic on cooling. Ongoing attempts to explain this coupled structural and electronic transition begin with two alternative starting points: a Peierls MIT driven by instabilities in electron-lattice dynamics and a Mott MIT where strong electron-electron correlations drive charge localization. A key missing piece of the VO2 puzzle is the role of lattice vibrations. Moreover, a comprehensive thermodynamic treatment must integrate both entropic and energetic aspects of the transition. Here we report that the entropy driving the MIT in VO2 is dominated by strongly anharmonic phonons rather than electronic contributions, and provide a direct determination of phonon dispersions. Our ab initio calculations identify softer bonding in the tetragonal phase, relative to the monoclinic phase, as the origin of the large vibrational entropy stabilizing the metallic rutile phase. They further reveal how a balance between higher entropy in the metal and orbital-driven lower energy in the insulator fully describes the thermodynamic forces controlling the MIT. Our study illustrates the critical role of anharmonic lattice dynamics in metal oxide phase competition, and provides guidance for the predictive design of new materials.


Differences in inducibility of the latent HIV reservoir in perinatal and adult infection.
The HIV latent reservoir in resting memory CD4+ T cells precludes cure. Therapeutics to reactivate and eliminate this reservoir are in clinical trials in adults, but not yet in pediatric populations. We determined, ex vivo, the inducibility of the latent reservoir in perinatal infection as compared with adult infections using the Tat/rev induced limiting dilution assay (TILDA), in which a single round (12 hours) of CD4+ T cell stimulation with PMA/ionomycin maximally activates T cells and leads to proviral expression with multiply spliced HIV RNA production. Markers of immune activation and exhaustion were measured to assess interactions with inducibility. Although rates of T cell activation with PMA/ionomycin were similar, the latent reservoir in perinatal infection was slower to reactivate and of lower magnitude compared with adult infection, independent of proviral load. An enhanced TILDA with the addition of phytohemagglutin and a duration of 18 hours augmented proviral expression in perinatal but not adult infection. The baseline HLA-DR+CD4+ T cell level was significantly lower in perinatal compared with adult infections, but not correlated with induced reservoir size. These data support the hypothesis that there are differences in kinetics of latency reversal and baseline immune activation in perinatal compared with adult infections, with implications for latency reversal strategies toward reservoir clearance and remission.


Perturbation of single hematopoietic stem cell fates in artificial niches.
Hematopoietic stem cells (HSCs) are capable of extensive self-renewal in vivo and are successfully employed clinically to treat hematopoietic malignancies, yet are in limited supply as in culture this self-renewal capacity is lost. Using an approach at the interface of stem cell biology and bioengineering, here we describe a novel platform of hydrogel microwell arrays for assessing the effects of either secreted or tethered proteins characteristic of the in vivo microenvironment, or niche, on HSC fate in vitro. Time-lapse microscopic analyses of single cells were crucial to overcoming inevitable heterogeneity of FACS-enriched HSCs. A reduction in proliferation kinetics or an increase in asynchronous division of single HSCs in microwells in response to specific proteins (Wnt3a and N-Cadherin) correlated well with subsequent serial long-term blood reconstitution in mice in vivo. Single cells that divided once in the presence of a given protein were capable of in vivo reconstitution, providing evidence of self-renewal divisions of HSCs in vitro. These results validate the hydrogel microwell platform as a broadly applicable paradigm for dissecting the regulatory role of specific signals within a complex stem cell niche.


Separation between the different fluxes scattered by art glazes: explanation of the special color saturation.
In a previous paper, the special visual appearance of art glazes was explained using the auxiliary function method (AFM) for solving the radiative transfer equation. Glazes are made of low concentrated colored scattering centers embedded in a transparent medium and the artist modulates the color by varying the number of glaze layers. A simple model of glazes and the new solving method have both been validated by comparison between flux measurements and modeling. The color of art glazes is analyzed here, and the study shows a spectacular maximum of saturation (purity) of the color that is never reached, to the best of our knowledge, with other techniques, such as pigment mixtures. This phenomenon is explained once more using the AFM that allows separation of the different contributions to the scattered fluxes. It is then shown that, on the one hand, single scattering never induces a maximum of saturation. On the other hand, multiple scattering has a typical increasing and decreasing behavior with an increasing number of glaze layers and thus participates to the maximum of saturation, just as the scattering by the diffuse base layer. A comparison between glazes and pigment mixtures, where the proportion of colored pigments with white pigments varies instead of the number of layers, shows that this maximum of saturation is much smaller with the second technique. To the best of our knowledge, we present a new development of the AFM that allows separation of the different origins of light scattering. We also show that it is possible to determine the optical properties of the scattering centers and of the base layer to create the required visual effect of a scattering medium.


Sibling interaction of children with learning disabilities: a comparison of autism and Down's syndrome.
Two potentially contrasting hypotheses can be generated about sibling interactions involving a child with Down's syndrome or autism. Research on siblings would predict that learning disabled children adopt responsive roles. Studies of children with autism would predict impoverished interactions. Home observations were conducted on 30 sibling pairs involving children with autism or Down's syndrome. Both hypotheses were partially supported. All learning disabled children engaged in frequent bouts of interaction, usually directed by their sibling. While children with autism engaged in fewer bouts and imitated less, they did reciprocate their siblings' initiations. Sibling encounters provide a unique opportunity for such children to learn about social relationships.


Five year outcomes of Boston type I keratoprosthesis as primary versus secondary penetrating corneal procedure in a matched case control study.
Despite improved retention and reduced complication rates paving the way for the current expansion of applications and surge in prevalence for the Boston type I Keratoprosthesis (KPro), the most frequent indication for its implantation today remains prior graft failure. The purpose of this study is to evaluate the long-term outcomes of primary KPro and compare to secondary implantation in a matched cohort study. This study included patients who underwent KPro implantation in a single center by two surgeons between July 2008 and October 2014. All eyes with KPro implantation as the primary procedure with a minimum follow up of 12 months were matched with eyes with same preoperative diagnoses that underwent secondary KPro implantation. Main outcomes included visual acuity and device retention. A total of 56 eyes were included with 28 eyes in each group. Mean follow up was 5.0 years for both groups. Twenty-nine percent (8) of the eyes in the primary group had a diagnosis of chemical or thermal injuries, 25% (7) aniridia, 18% (5) autoimmune disease, 4% (1) infectious keratitis/neurotrophic cornea, 7% (2) gelatinous corneal dystrophy, 7% (2) ectrodactyly ectodermal dysplasia/limbal stem cell deficiency, and 11% (3) uveitis/hypotony. Sixty-one percent (17) of the eyes in the primary group and 39% (11) in the secondary group maintained a final best-corrected visual acuity of 20/200 or better at a mean follow up of 5.0 years; the probability of maintaining best-corrected vision is 0.83 and 0.49 for primary and secondary groups at 5.0 years (p = 0.02). There is no statistically significant difference between groups in device retention (p = 0.22) or postoperative complication rates (p >0.05). This study demonstrates that Boston KPro implantation may be successful as a primary procedure in patients at high risk of failure with traditional penetrating keratoplasty. The device has a good long-term retention rate and visual outcomes are promising however a larger study is needed for more definitive results.


Msp1 Is a Membrane Protein Dislocase for Tail-Anchored Proteins.
Mislocalized tail-anchored (TA) proteins of the outer mitochondrial membrane are cleared by a newly identified quality control pathway involving the conserved eukaryotic protein Msp1 (ATAD1 in humans). Msp1 is a transmembrane AAA-ATPase, but its role in TA protein clearance is not known. Here, using purified components reconstituted into proteoliposomes, we show that Msp1 is both necessary and sufficient to drive the ATP-dependent extraction of TA proteins from the membrane. A crystal structure of the Msp1 cytosolic region modeled into a ring hexamer suggests that active Msp1 contains a conserved membrane-facing surface adjacent to a central pore. Structure-guided mutagenesis of the pore residues shows that they are critical for TA protein extraction in vitro and for functional complementation of an msp1 deletion in yeast. Together, these data provide a molecular framework for Msp1-dependent extraction of mislocalized TA proteins from the outer mitochondrial membrane.


Zinc prevention and treatment of alcoholic liver disease.
Alcoholic liver disease (ALD) is associated with decreases in zinc (Zn) and its major binding protein, metallothionein (MT), in the liver. Studies using animal models have shown that Zn supplementation prevents alcohol-induced liver injury under both acute and chronic alcohol exposure conditions. There are hepatic and extrahepatic actions of Zn in the prevention of alcoholic liver injury. Zn supplementation attenuates ethanol-induced hepatic Zn depletion and suppresses ethanol-elevated cytochrome P450 2E1 (CYP2E1) activity, but increases the activity of alcohol dehydrogenase in the liver; an action that is likely responsible for Zn suppression of alcohol-induced oxidative stress. Zn also enhances glutathione-related antioxidant capacity in the liver. At the cellular level, Zn inhibits alcohol-induced hepatic apoptosis partially through suppression of the Fas/FasL-mediated pathway. Zn supplementation preserves intestinal integrity and prevents endotoxemia, leading to inhibition of endotoxin-induced tumor necrosis factor-alpha (TNF-alpha) production in the liver. Zn also directly inhibits the signaling pathway involved in endotoxin-induced TNF-alpha production. These hepatic and extrahepatic effects of Zn are independent of MT. However, low levels of MT in the liver sensitize the organ to alcohol-induced injury, and elevation of MT enhances the endogenous Zn reservoir and makes Zn available when oxidative stress is imposed. Zn has a high potential to be developed as an effective agent in the prevention and treatment of ALD.


Contribution of the novel inflammatory bowel disease gene IL23R to disease susceptibility and phenotype.
A North American genome-wide single nucleotide polymorphism (SNP) association study identified IL23R as a novel inflammatory bowel disease (IBD) susceptibility gene. Association was reported with multiple risk variants in the centromeric portion of IL23R in 3 large independent cohorts. The aims of this study were to replicate the association of IL23R with Crohn's disease (CD), examine subphenotype relationships, and look for evidence of epistasis with the known CD susceptibility gene CARD15 and susceptibility haplotype IBD5 in a large collection of CD patients. We further investigated the relationship between IL23R and ulcerative colitis (UC). In all, 604 CD and 647 UC patients who had been rigorously phenotyped and who had been recruited from a single UK center were used in this study. Controls were either spouses of patients (141) or were recruited from well-person clinics (993). Eight SNPs were genotyped using MassArray (Sequenom). All 8 SNPs genotyped were significantly associated with CD. The association with the nonsynonymous SNP rs11209026 was confirmed (P=6.65x10(-6), odds ratio [OR], 0.43, 95% confidence interval [CI]: 0.29-0.64). The most significant SNP in our study was rs7517847 (P=4.9x10(-9), OR 0.65, 0.56-0.75), which is statistically independent of rs11209026. Preliminary evidence suggests an epistatic interaction with the IBD5 risk haplotype. The effects of mutations in this IL23R appear weaker in UC (P=0.008, OR 0.63, 0.45-0.89 and 0.005 OR, 0.81, 0.71-0.94, respectively). No subphenotype associations were identified. We confirmed the findings that IL23R is a susceptibility gene for IBD with suggestive epistasis with the IBD5 locus in the CD population.


Remote ischemic preconditioning with--but not without--metabolic support protects the neonatal porcine heart against ischemia-reperfusion injury.
While remote ischemic preconditioning (rIPC) protects the mature heart against ischemia-reperfusion (IR) injury, the effect on the neonatal heart is not known. The neonatal heart relies almost solely on carbohydrate metabolism, which is modified by rIPC in the mature heart. We hypothesized that rIPC combined with metabolic support with glucose-insulin (GI) infusion improves cardiac function and reduces infarct size after IR injury in neonatal piglets in-vivo. 32 newborn piglets were randomized into 4 groups: control, GI, GI+rIPC and rIPC. GI and GI+rIPC groups received GI infusion continuously from 40 min prior to ischemia. rIPC and GI+rIPC groups underwent four cycles of 5 min limb ischemia. Myocardial IR injury was induced by 40 min occlusion of the left anterior descending artery followed by 2 h reperfusion. Myocardial lactate concentrations were assessed in microdialysis samples analyzed by mass spectrometry. Infarct size was measured using triphenyltetrazolium chloride staining. Systolic recovery (dP/dt(max) as % of baseline) after 2 h reperfusion was 68.5±13.8% in control, 53.7±11.2% in rIPC (p<0.05), and improved in GI (83.6±18.8%, p<0.05) and GI+rIPC (87.0±15.7%, p<0.01). rIPC+GI protects the neonatal porcine heart against IR injury in-vivo. rIPC alone has detrimental metabolic and functional effects that are abrogated by simultaneous GI infusion.


Chromosomally integrated HHV-6: impact on virus, cell and organismal biology.
HHV-6 integrates its genome into telomeres of human chromosomes. Integration can occur in somatic cells or gametes, the latter leading to individuals harboring the HHV-6 genome in every cell. This condition is transmitted to descendants and referred to as inherited chromosomally integrated human herpesvirus 6 (iciHHV-6). Although integration can occur in different chromosomes, it invariably takes place in the telomere region. This integration mechanism represents a way to maintain the virus genome during latency, which is so far unique amongst human herpesviruses. Recent work provides evidence that the integrated HHV-6 genome can be mobilized from the host chromosome, resulting in the onset of disease. Details on required structural determinants, putative integration mechanisms and biological and medical consequences of iciHHV-6 are discussed.


The Prevalence and Risk Factors for S2 Alar-Iliac Screw Loosening with a Minimum 2-Year Follow-up.
A retrospective cohort study. The purpose of this study was to investigate the prevalence and risk factors for S2 alar-iliac (SAI) screw loosening following lumbosacral fixation, with a minimum 2-year follow-up. Although SAI screws allow surgeons to perform lumbosacral fixation with a low profile and enhanced biomechanical strength, screw loosening following surgery can occur in some cases. However, few studies have investigated the prevalence and risk factors for SAI screw loosening. This retrospective study included 35 patients (mean age, 72.8±8.0 years; male, 10; female, 25) who underwent lumbosacral fixation using SAI screws with at least 2 years of follow-up. SAI screw loosening and L5-S bony fusion were assessed using computed tomography. The period for which the screws appeared loose and the risk factors for SAI screw loosening were investigated 2 years after surgery. A total of 70 SAI screws and 70 S1 pedicle screws were inserted. Loosening was observed 0.5, 1, and 2 years after surgery in 17 (24.3%), 35 (50.0%), and 35 (50.0%) SAI screws, respectively. Bony fusion rate at L5-S was significantly lower in patients with SAI screw loosening than in those without screw loosening (65.0% vs. 93.3%, p =0.048). The score for SAI screw contact with the iliac cortical bone and the bony fusion rate at L5-S were significantly lower in the loosening group than in the non-loosening group (1.8±0.5 vs. 2.2±0.3, p <0.001, respectively). Postoperative pelvic incidence-lumbar lordosis was significantly higher in the loosening group than in the non-loosening group (7.9°±15.4° vs. 0.5°±8.7°, p =0.02, respectively). SAI screw loosening is closely correlated with pseudoarthrosis at L5-S. Appropriate screw insertion and optimal lumbar lordosis restoration are important to prevent postoperative complications related to SAI screws.


Clinical evaluation of USCOM ultrasonic cardiac output monitor in cardiac surgical patients in intensive care unit.
The USCOM ultrasonic cardiac output monitor (USCOM Pty Ltd, Coffs Harbour, NSW, Australia) is a non-invasive device that determines cardiac output by continuous-wave Doppler ultrasound. The aim of this study was to evaluate the accuracy of the USCOM device compared with the thermodilution technique in intensive care patients who had just undergone cardiac surgery. We conducted a prospective study in the 18-bed intensive care unit of a 600-bed tertiary referral hospital. Twenty-four mechanically ventilated patients were studied immediately following cardiac surgery. We evaluated the USCOM monitor by comparing its output with paired measurements obtained by the standard thermodilution technique using a pulmonary artery catheter. Forty paired measurements were obtained in 22 patients. We were unable to obtain an acceptable signal in the remaining two patients. Comparison of the two techniques showed a bias of 0.18 and limits of agreement of -1.43 to 1.78. The agreement may not be as good between techniques at higher cardiac output values. The USCOM monitor has a place in intensive care monitoring. It is accurate, rapid, safe, well-tolerated, non-invasive and cost-effective. The learning curve for skill acquisition is very short. However, during the learning phase the USCOM monitor measurements are rather 'operator dependent'. Its suitability for use in high and low cardiac output states requires further validation.


Digital microfluidics for time-resolved cytotoxicity studies on single non-adherent yeast cells.
Single cell analysis (SCA) has gained increased popularity for elucidating cellular heterogeneity at genomic, proteomic and cellular levels. Flow cytometry is considered as one of the most widely used techniques to characterize single cell responses; however, its inability to analyse cells with spatio-temporal resolution poses a major drawback. Here, we introduce a digital microfluidic (DMF) platform as a useful tool for conducting studies on isolated yeast cells in a high-throughput fashion. The reported system exhibits (i) a microwell array for trapping single non-adherent cells by shuttling a cell-containing droplet over the array, and allows (ii) implementation of high-throughput cytotoxicity assays with enhanced spatio-temporal resolution. The system was tested for five different concentrations of the antifungal drug Amphotericin B, and the cell responses were monitored over time by time lapse fluorescence microscopy. The DMF platform was validated by bulk experiments, which mimicked the DMF experimental design. A correlation analysis revealed that the results obtained on the DMF platform are not significantly different from those obtained in bulk; hence, the DMF platform can be used as a tool to perform SCA on non-adherent cells, with spatio-temporal resolution. In addition, no external forces, other than the physical forces generated by moving the droplet, were used to capture single cells, thereby avoiding cell damage. As such, the information on cellular behaviour during treatment could be obtained for every single cell over time making this platform noteworthy in the field of SCA.


Dasatinib inhibits the growth of molecularly heterogeneous myeloid leukemias.
Dasatinib is a dual Src/Abl inhibitor recently approved for Bcr-Abl+ leukemias with resistance or intolerance to prior therapy. Because Src kinases contribute to multiple blood cell functions by triggering a variety of signaling pathways, we hypothesized that their molecular targeting might lead to growth inhibition in acute myeloid leukemia (AML). We studied growth factor-dependent and growth factor-independent leukemic cell lines, including three cell lines expressing mutants of receptor tyrosine kinases (Flt3 or c-Kit) as well as primary AML blasts for responsiveness to dasatinib. Dasatinib resulted in the inhibition of Src family kinases in all cell lines and blast cells at approximately 1 x 10(-9) mol/L. It also inhibited mutant Flt3 or Kit tyrosine phosphorylation at approximately 1 x 10(-6) mol/L. Mo7e cells expressing the activating mutation (codon 816) of c-Kit were most sensitive to growth inhibition with a GI(50) of 5 x 10(-9) mol/L. Primary AML blast cells exhibited a growth inhibition of <1 x 10(-6) mol/L. Cell lines that showed growth inhibition at approximately 1 x 10(-6) mol/L showed a G(1) cell cycle arrest and correlated with accumulation of p21 and p27 protein. The addition of rapamycin or cytotoxic agents enhanced growth inhibition. Dasatinib also caused the apoptosis of Mo7e cells expressing oncogenic Kit. Although all of the precise targets for dasatinib are not known, this multikinase inhibitor causes either growth arrest or apoptosis in molecularly heterogeneous AML. The addition of cytotoxic or targeted agents can enhance its effects.


Observational study of dronedarone in Taiwanese patients with atrial fibrillation.
Currently, data on the real-world use of dronedarone, an antiarrhythmic drug for atrial fibrillation (AF), are contradictory and often based on patient populations comprised of Caucasians. We prospectively investigated the efficacy and safety of dronedarone and risk factors related to treatment outcomes in a real-world use setting. The prospective, observational, single-arm, multi-center study included a total of 824 Taiwanese patients with a diagnosis of paroxysmal or persistent AF and receiving dronedarone treatment. Risk factors analysis, efficacy, and safety of dronedarone were assessed with a follow-up of six months. Of the 824 patients enrolled (mean age, 75.3 ± 7.2 years), 95.2% had at least one cardiovascular risk factor. An increase in the proportion of patients with sinus rhythm following treatment was seen (52.1% at baseline vs. 67.4% at 6 months). A decrease in the mean duration of AF episodes (388.4 min vs. 62.3 min) and an increase in total AFEQT (65.4 ± 16.2 vs. 74.0 ± 11.8) were also observed after 6 months of treatment. Females, those under the age of 75, and those with symptomatic AF had higher odds of treatment success. At 6 months, 10.5% of patients reported treatment-related AEs. However, only 0.2% of the AEs were both severe in nature and causally related to dronedarone. This six-month study showed dronedarone to be relatively safe and efficacious and to improve quality-of-life in Taiwanese patients with atrial fibrillation. Odds of treatment success were related to the patient's gender, age, and AF type.


In-vitro evaluation of the tolerance of surgical instruments in templates for computer-assisted guided implantology produced by 3-D printing.
The aim of this in-vitro study was to compare the tolerance of surgical instruments in surgical guides produced by 3-D printing, without metal sleeves to a surgical guide with conventional metal sleeves from two different manufacturers. Lateral movements of drill tips caused by tolerance between the sleeve and drill key and between the drill key and the drill were recorded after application of a standardized force to the surgical instruments. Four groups were tested: Control 1 (C1): metal sleeve from commercially available surgical system 1; Test 1 (T1): 3-D-printed sleeve for surgical system 1; Control 2 (C2): metal sleeve from commercially available surgical system 2. Test 2 (T2): 3-D-printed sleeve for surgical system 2. The mean total lateral movement was 0.75 mm (0.5-1.04 mm) in the C1 group and 0.91 mm (0.54-1.34 mm) in the C2 group. The mean amount of movement from tolerance between sleeve and drill-guiding key was 0.31 mm (range 0.22-0.41 mm) in C1 and 0.42 mm (range 0.29-0.56 mm) in C2. This lateral movement was in mean reduced by 0.24 mm (32%) in T1 and by 0.39 mm (43%) in T2 group. This reduction was statistically significant in both groups (P < 0.001). The tolerance of surgical instruments and the lateral movements of the drills were significantly reduced by the use of 3-D printing with reduced sleeve diameter. This reduction could improve the overall accuracy in computer-assisted template-guided implant dentistry. The lateral movement of the drill can be further reduced by using a shorter drill and a higher drill key. This can be considered during implant planning and CAD/CAM of surgical guides.


Expression of human MRP6, a homologue of the multidrug resistance protein gene MRP1, in tissues and cancer cells.
The human multidrug resistance protein (MRP) family contains at least six members: MRP1, the gene encoding the multidrug resistance protein; cMOAT or MRP2, encoding the canalicular multispecific organic anion transporter; and four homologues, called MRP3, MRP4, MRP5, and MRP6. The most recently discovered member of the family, MRP6, is analyzed in this report. The MRP6 gene is located on chromosome 16, immediately next to MRP1, and encodes a protein of 1503 amino acids with a predicted molecular weight of Mr 165,000. The 3' end of the MRP6 protein was found to be almost identical with the anthracycline resistance associated (ARA) protein identified previously in epirubicin-selected leukemia cells. Using both 3'- and 5'-derived MRP6 probes, we found that MRP6 is highly expressed in liver and kidney and to a low or very low extent in a few other tissues. No evidence was obtained for an independent expression of the ARA part of the MRP6 gene in normal tissues. To assess a possible role for MRP6 in multidrug resistance, we examined a large panel of resistant cell lines for the (over)expression of MRP6. We found overexpression of the complete MRP6 gene or part of it only in those cell lines with high overexpression and amplification of the MRP1 gene. DNA blot (Southern) analysis showed that MRP6 or a part of it is also amplified in these cell lines. Our results suggest that MRP6 does not play a role in the resistance of the resistant cells analyzed, and that MRP6/ARA is only coamplified with MRP1 because of its location immediately next to it on the same chromosome.


Legalization of marijuana: potential impact on youth.
This technical report provides historical perspectives and comparisons of various approaches to the legal status of marijuana to aid in forming public policy. Information on the impact that decriminalization and legalization of marijuana could have on adolescents, in addition to concerns surrounding medicinal use of marijuana, are also addressed in this report. Recommendations are included in the accompanying policy statement.


What core primary health care services should be available to Australians living in rural and remote communities?
Australians living in rural and remote areas experience poorer access to primary health care (PHC) and poorer health outcomes compared to metropolitan populations. Current health reform in Australia aims to ensure all Australians, regardless of where they live, have access to essential PHC services. However, at a national level policy makers and health planners lack an evidence-based set of core PHC services to assist in implementing this goal. A Delphi method was used to reach consensus on an evidence-based list of core PHC services to which all Australians should have access and their necessary support functions. Experts in rural and remote and/or Indigenous PHC, including policy-makers, academics, clinicians and consumers, were invited to consider a list of core services derived from the literature. Thirty nine experts agreed to participate. After three survey rounds there was a strong consensus (≥80% agreement) on core PHC services namely; 'care of the sick and injured', 'mental health', 'maternal/child health', 'allied health', 'sexual/reproductive health', 'rehabilitation', 'oral/dental health' and 'public health/illness prevention'; and on the PHC support functions of; 'management/governance/leadership', 'coordination', 'health infrastructure', 'quality systems', 'data systems', 'professional development' and 'community participation'. Themes emerging from qualitative data included challenges in providing equitable PHC in rural and remote areas, the importance of service coordination and diverse strategies to overcome access barriers. This study identifies a basket of PHC services that consumers in rural and remote communities can expect to access. It provides rigorously derived evidence that will contribute to a more systematic approach to PHC service planning and availability and will assist policy makers in the allocation of scarce resources necessary to improve the health outcomes of residents of rural and remote areas.


Task- and time-dependent memory enhancement by dehydroepiandosterone in day-old chicks.
We have previously reported the presence of dehydroepiandosterone (DHEA) in the day-old-chick brain, and a role for it in enhanced memory formation. Here we confirm that intracerebral injections of DHEA 5 min before training on the weak passive avoidance task enhanced recall 24 hours after training. Recall per se on an appetitive visual categorization task was not altered by administration of DHEA 5 min before training. However administration of DHEA 5 min before limited or very limited training on a visual categorization task (20 or 10 pecks only) appeared to enhance consolidation of this task at test 24 h after training; reducing the latency and total time taken to complete the test (60 pecks), while not detrimentally altering accuracy. Moreover, DHEA is unlikely to induce this effect via possible anxiolytic effects because it did not alter behavior in the open field test. We also examined diffusion of DHEA throughout the brain at various stages following intracerebral injection.


Adaptive changes in harvested populations: plasticity and evolution of age and size at maturation.
We investigate harvest-induced adaptive changes in age and size at maturation by modelling both plastic variation and evolutionary trajectories. Harvesting mature individuals displaces the reaction norm for age and size at maturation toward older ages and larger sizes and rotates it clockwise, whereas harvesting immature individuals has the reverse qualitative effect. If both immature and mature individuals are harvested, the net effect has approximately the same trend as when harvesting immature individuals only. This stems from the sensitivity of the evolutionary response, which depends on the maturity state of harvested individuals, but also on the type of harvest mortality (negatively or positively density dependent, density independent) and the value of three life-history parameters (natural mortality, growth rate and the trade-off between growth and reproduction). Evolutionary changes in the maturation reaction norm have strong repercussions for the mean size and the density of harvested individuals that, in most cases, result in the reduction of biomass--a response that population dynamical models would overlook. These results highlight the importance of accounting for evolutionary trends in the long-term management of exploited living resources and give qualitative insights into how to minimize the detrimental consequences of harvest-induced evolutionary changes in maturation reaction norms.


Aberrant function and differentiation of monocytes in end stage renal disease.
Patients with end stage renal disease (ESRD) suffer from many disturbances of the immune system. These immunopathologies are related to the higher failure of vaccination, and increased prevalence of infections and neoplasms. In the presented article, we review the current data regarding the role of monocytes in immune dysfunctions which are observed in terminal renal failure. As monocytes play a pivotal role in regulating the function of the immune system, their dysfunction can have a profound effect on the immune system and may lead to accelerated arteriosclerosis and deteriorating overall health conditions. More specifically, we suggest that peripheral blood monocytes in patients with ESRD are chronically activated, and their functional and phenotypical features resemble those of inflammatory macrophages. This state of chronic inflammation is unfavorable for dendritic cells and consequently, the prevalence of dendritic cells is reduced. As these effects are consistent across different modes of dialysis, they are probably mediated by the uremia itself.


Causes of death among lead smelters in relation to the glucose-6-phosphate dehydrogenase polymorphism.
To assess, by updating a follow-up mortality study of a lead smelters cohort in Sardinia, Italy, the adverse health effects following occupational lead exposure in relation to the glucose-6-phosphate dehydrogenase (G6PD) polymorphism. The 1973-2003 mortality of 1017 male lead smelters were followed-up, divided into two subcohorts according to the G6PD phenotype: whether G6PD deficient (G6PD-) or wild-type (wtG6PD). Deaths observed in the overall cohort and the two subcohorts were compared with those expected, on the basis of the age-, sex- and calendar year-specific mortality in the general male population of the island. Directly standardised mortality rates (sr) in the two subcohorts were also compared. Cardiovascular mortality was strongly reduced among production and maintenance workers, which is most related to the healthy worker effect. However, the sr for cardiovascular diseases was substantially lower among the G6PD- subcohort (5.0x10(-4)) than among the wtG6PD subcohort (33.6x10(-4); chi2 = 1.10; p = NS). Neoplasms of the haemopoietic system exceeded the expectation in the G6PD- subcohort (SMR = 388; 95% CI 111 to 1108). No other cancer sites showed any excess in the overall cohort or in the two subcohorts. No death from haemolytic anaemia occurred in the G6PD- subcohort. With due consideration of the limited statistical power of our study, previous results suggesting that in workplaces where exposure is under careful control, expressing the G6PD- phenotype does not convey increased susceptibility to lead toxicity are confirmed. The observed excess risk of haematopoietic malignancies seems to have most likely resulted from chance.


Polyclonal origin of pancreatic islets in aggregation mouse chimaeras.
In the present study, we have examined the origin and growth pattern of the beta cells in pancreatic islets, to determine whether a single progenitor cell gave rise to all the precursors of the islets, or if each of a few progenitor cells is the founder of a different islet, or if each islet is a mixture of cells originating from a pool of progenitor cells. Aggregation mouse chimaeras where the pancreatic beta cells derived from each embryo can be identified in the islets on histological sections were analyzed. In two chimaeras, all the islets contained cells from both the aggregated embryo. This clearly demonstrates that each islet resulted from several independent cells. In addition, the beta cells derived from either embryo component were in very small clusters in the islets, suggesting that in situ cell division did not account significantly for islet growth.


Formulated Chinese Medicine Shaoyao Gancao Tang Reduces Tau Aggregation and Exerts Neuroprotection through Anti-Oxidation and Anti-Inflammation.
Misfolded tau proteins induce accumulation of free radicals and promote neuroinflammation by activating microglia-releasing proinflammatory cytokines, leading to neuronal cell death. Traditional Chinese herbal medicines (CHMs) have been widely used in clinical practice to treat neurodegenerative diseases associated with oxidative stress and neuroinflammation. This study examined the neuroprotection effects of formulated CHMs Bai-Shao (made of Paeonia lactiflora), Gan-Cao (made of Glycyrrhiza uralensis), and Shaoyao Gancao Tang (SG-Tang, made of P. lactiflora and G. uralensis at 1 : 1 ratio) in cell model of tauopathy. Our results showed that SG-Tang displayed a greater antioxidative and antiaggregation effect than Bai-Shao and Gan-Cao and a stronger anti-inflammatory activity than Bai-Shao but similar to Gan-Cao. In inducible 293/SH-SY5Y cells expressing proaggregant human tau repeat domain (ΔK280 tauRD), SG-Tang reduced tau misfolding and reactive oxygen species (ROS) level in ΔK280 tauRD 293 cells and promoted neurite outgrowth in ΔK280 tauRD SH-SY5Y cells. Furthermore, SG-Tang displayed anti-inflammatory effects by reducing nitric oxide (NO) production in mouse BV-2 microglia and increased cell viability of ΔK280 tauRD-expressing SH-SY5Y cells inflamed by BV-2 conditioned medium. To uncover the neuroprotective mechanisms of SG-Tang, apoptosis protein array analysis of inflamed tau expressing SH-SY5Y cells was conducted and the suppression of proapoptotic proteins was confirmed. In conclusion, SG-Tang displays neuroprotection by exerting antioxidative and anti-inflammatory activities to suppress neuronal apoptosis in human tau cell models. The study results lay the base for future applications of SG-Tang on tau animal models to validate its effect of reducing tau misfolding and potential disease modification.


Methanomethylophilus alvus Mx1201 Provides Basis for Mutual Orthogonal Pyrrolysyl tRNA/Aminoacyl-tRNA Synthetase Pairs in Mammalian Cells.
Genetic code expansion via stop codon suppression is a powerful technique for engineering proteins in mammalian cells with site-specifically encoded noncanonical amino acids (ncAAs). Current methods rely on very few available tRNA/aminoacyl-tRNA synthetase pairs orthogonal in mammalian cells, the pyrrolysyl tRNA/aminoacyl-tRNA synthetase pair from Methanosarcina mazei ( Mma PylRS/PylT) being the most active and versatile to date. We found a pyrrolysyl tRNA/aminoacyl-tRNA synthetase pair from the human gut archaeon Methanomethylophilus alvus Mx1201 (Mx1201 PylRS/PylT) to be active and orthogonal in mammalian cells. We show that this PylRS enzyme can be engineered to expand its ncAA substrate spectrum. We find that due to the large evolutionary distance of the two pairs, Mx1201 PylRS/PylT is partially orthogonal to Mma PylRS/PylT. Through rational mutation of Mx1201 PylT, we abolish its noncognate interaction with Mma PylRS, creating two mutually orthogonal PylRS/PylT pairs. Combined in the same cell, we show that the two pairs can site-selectively introduce two different ncAAs in response to two distinct stop codons. Our work expands the repertoire of mutually orthogonal tools for genetic code expansion in mammalian cells and provides the basis for advanced in vivo protein engineering applications for cell biology and protein production.


International Humanitarian Award.
The International Humanitarian Award recognizes extraordinary humanitarian services and activism by psychologists, including professional and volunteer work conducted primarily in the field with underserved populations. Award recipients are psychologists who, by their extraordinary service at a difficult time, improve the lives and contribute to the well-being of people in a large or small geographic area anywhere in the world. The 2020 recipient of the APA International Humanitarian Award was selected by the Board of Directors, as recommended by the Committee on International Relations in Psychology (CIRP). (PsycInfo Database Record (c) 2020 APA, all rights reserved).


Generation of HER2-specific antibody immunity during trastuzumab adjuvant therapy associates with reduced relapse in resected HER2 breast cancer.
Resected HER2 breast cancer patients treated with adjuvant trastuzumab and chemotherapy have superior survival compared to patients treated with chemotherapy alone. We previously showed that trastuzumab and chemotherapy induce HER2-specific antibodies which correlate with improved survival in HER2 metastatic breast cancer patients. It remains unclear whether the generation of immunity required trastuzumab and whether endogenous antibody immunity is associated with improved disease-free survival in the adjuvant setting. In this study, we addressed this question by analyzing serum anti-HER2 antibodies from a subset of patients enrolled in the NCCTG trial N9831, which includes an arm (Arm A) in which trastuzumab was not used. Arms B and C received trastuzumab sequentially or concurrently to chemotherapy, respectively. Pre-and post-treatment initiation sera were obtained from 50 women enrolled in N9831. Lambda IgG antibodies (to avoid detection of trastuzumab) to HER2 were measured and compared between arms and with disease-free survival. Prior to therapy, across all three arms, N9831 patients had similar mean anti-HER2 IgG levels. Following treatment, the mean levels of antibodies increased in the trastuzumab arms but not the chemotherapy-only arm. The proportion of patients who demonstrated antibodies increased by 4% in Arm A and by 43% in the Arms B and C combined (p = 0.003). Cox modeling demonstrated that larger increases in antibodies were associated with improved disease-free survival in all patients (HR = 0.23; p = 0.04). These results show that the increased endogenous antibody immunity observed in adjuvant patients treated with combination trastuzumab and chemotherapy is clinically significant, in view of its correlation with improved disease-free survival. The findings may have important implications for predicting treatment outcomes in patients treated with trastuzumab in the adjuvant setting. ClinicalTrials.gov, NCT00005970 . Registered on July 5, 2000.