Rapid turnover of 2-LTR HIV-1 DNA during early stage of highly active antiretroviral therapy.
Despite prolonged treatment with highly active antiretroviral therapy (HAART), the infectious HIV-1 continues to replicate and resides latently in the resting memory CD4+ T lymphocytes, which blocks the eradication of HIV-1. The viral persistence of HIV-1 is mainly caused by its proviral DNA being either linear nonintegrated, circular nonintegrated, or integrated. Previous reports have largely focused on the dynamics of HIV-1 DNA from the samples collected with relatively long time intervals during the process of disease and HAART treatment, which may have missed the intricate changes during the intervals in early treatment. In this study, we investigated the dynamics of HIV-1 DNA in patients during the early phase of HARRT treatment. Using optimized real time PCR, we observed significant changes in 2-LTR during the first 12-week of treatment, while total and integrated HIV-1 DNA remained stable. The doubling time and half-life of 2-LTR were not correlated with the baseline and the rate of changes in plasma viral load and various CD4+ T-cell populations. Longitudinal analyses on 2-LTR sequences and plasma lipopolysaccharide (LPS) levels did not reveal any significant changes in the same treatment period. Our study revealed the rapid changes in 2-LTR concentration in a relatively large number of patients during the early HAART treatment. The rapid changes indicate the rapid infusion and clearance of cells bearing 2-LTR in the peripheral blood. Those changes are not expected to be caused by the blocking of viral integration, as our study did not include the integrase inhibitor raltegravir. Our study helps better understand the dynamics of HIV-DNA and its potential role as a biomarker for the diseases and for the treatment efficacy of HAART.


Metal organic framework-mediated synthesis of highly active and stable Fischer-Tropsch catalysts.
Depletion of crude oil resources and environmental concerns have driven a worldwide research on alternative processes for the production of commodity chemicals. Fischer-Tropsch synthesis is a process for flexible production of key chemicals from synthesis gas originating from non-petroleum-based sources. Although the use of iron-based catalysts would be preferred over the widely used cobalt, manufacturing methods that prevent their fast deactivation because of sintering, carbon deposition and phase changes have proven challenging. Here we present a strategy to produce highly dispersed iron carbides embedded in a matrix of porous carbon. Very high iron loadings (>40 wt %) are achieved while maintaining an optimal dispersion of the active iron carbide phase when a metal organic framework is used as catalyst precursor. The unique iron spatial confinement and the absence of large iron particles in the obtained solids minimize catalyst deactivation, resulting in high active and stable operation.


Newer glycopeptide antibiotics for treatment of complicated skin and soft tissue infections: systematic review, network meta-analysis and cost analysis.
Skin and soft tissue infections (SSTIs) carry significant economic burden, as well as morbidity and mortality, especially when caused by methicillin-resistant Staphylococcus aureus (MRSA). Several new MRSA-active antibiotics have been developed, including semisynthetic glycopeptides (telavancin, dalbavancin and oritavancin). Of these, dalbavancin and oritavancin offer extended dosing intervals. We performed a systematic review, network meta-analysis and cost analysis to compare the newer glycopeptides to standard care and to each other for the treatment of complicated SSTIs (cSSTI). A search for randomized controlled trials (RCTs) was conducted in Medline, Embase and the Cochrane Central Register of Controlled Trials. We also developed a model to evaluate the costs associated with dalbavancin and oritavancin from the third-party payer perspective. Seven RCTs met the inclusion criteria. Network meta-analyses suggested that the clinical response to telavancin, dalbavancin and oritavancin was similar to standard care (odds ratio (OR) 1.09, 95% confidence interval (CI) 0.90-1.33; OR 0.78, 95% CI 0.52-1.18; and OR 1.06, 95% CI 0.85-1.33, respectively). Head-to-head comparisons showed no difference in clinical response between oritavancin and dalbavancin (OR 1.36; 95% CI 0.85-2.18), oritavancin and telavancin (OR 0.98; 95% CI 0.72-1.31) or dalbavancin and telavancin (OR 0.72; 95% CI 0.45-1.13). Telavancin had a higher incidence of overall adverse events compared to standard care (OR 1.33; 95% CI 1.10-1.61). Compared to telavancin, there were fewer overall adverse events with dalbavancin (OR 0.58; 95% CI 0.45-0.76) and oritavancin (OR 0.71; 95% CI 0.55-0.92). Studies were of high quality overall. Our cost analyses demonstrated that dalbavancin and oritavancin were less costly compared to standard care under baseline assumptions and many scenarios evaluated. The use of dalbavancin could save third-party payers $1442 to $4803 per cSSTI, while the use of oritavancin could save $3571 to $6932 per cSSTI. Dalbavancin and oritavancin demonstrate efficacy and safety comparable to standard care in well-designed RCTs and result in cost savings when standard care is treatment that covers MRSA.


Biosurfactant production by Mucor circinelloides on waste frying oil and possible uses in crude oil remediation.
Biosurfactants are biocompatible surface active agents which many microorganisms produce. This study investigated the production of biosurfactants by Mucor circinelloides. The effects of different factors on biosurfactant production, including carbon sources and concentrations, nitrogen sources, and iron (II) concentration, were studied and the optimum condition determined. Finally, the strain's ability to remove the crude oil and its relationship with biosurfactant production was evaluated. The results showed that M. circinelloides could reduce the surface tension of the culture medium to 26.6 mN/m and create a clear zone of 12.9 cm diameter in an oil-spreading test. The maximum surface tension reduction was recorded 3 days after incubation. The optimum condition for biosurfactant production was achieved in the presence of 8% waste frying oil as a carbon source, 2 g/L yeast extract as a nitrogen source, and 0.01 mM FeSO4. M. circinelloides could consume 8% waste frying oil in 5 days of incubation, and 87.6% crude oil in 12 days of incubation. A direct correlation was observed between oil degradation and surface tension reduction in the first 3 days of fungal growth. The results showed that the waste frying oil could be recommended as an inexpensive oily waste substance for biosurfactant production, and M. circinelloides could have the potential to treat waste frying oil. According to the results, the produced crude biosurfactant or fungal strain could be directly used for the mycoremediation of crude oil contamination in oil fields.


Plastic accumulation in the North Atlantic subtropical gyre.
Plastic marine pollution is a major environmental concern, yet a quantitative description of the scope of this problem in the open ocean is lacking. Here, we present a time series of plastic content at the surface of the western North Atlantic Ocean and Caribbean Sea from 1986 to 2008. More than 60% of 6136 surface plankton net tows collected buoyant plastic pieces, typically millimeters in size. The highest concentration of plastic debris was observed in subtropical latitudes and associated with the observed large-scale convergence in surface currents predicted by Ekman dynamics. Despite a rapid increase in plastic production and disposal during this time period, no trend in plastic concentration was observed in the region of highest accumulation.


A Multilevel Examination of Whether Child Welfare Worker Characteristics Predict the Substantiation Decision in Canada.
The decision to substantiate a report of child maltreatment represents a key decision point in the child welfare service decision-making continuum. This decision has various potential implications for children and their families, which may include more intensive child welfare involvement or the cessation of services. The substantiation decision is determined by whether there is enough evidence to suggest that maltreatment or the risk of maltreatment has occurred. To date, there has been minimal exploration of whether child welfare worker characteristics might influence this critical decision point. The Decision-Making Ecology would suggest that indeed, worker characteristics play a role in how they carry out their role. Given the importance of this decision point, this study uses secondary data to examine whether worker characteristics, such as education level and type, ethnoracial identity, caseload, and experience, predict substantiation in the Canadian child welfare context. Furthermore, this study utilizes multilevel modeling, a theoretically important and unique method of analyzing organizational data that considers differences in decisions among child welfare workers. The final model included 4,327 children and 567 workers from across Canada. Several case level factors (e.g., child age and functioning, caregiver risk factors) predicted the substantiation decision. Furthermore, and most importantly for this study, worker characteristics significantly predicted their substantiation decision. Workers with fewer years of experience, those in an Ongoing Services role, and with a lower caseload substantiated significantly more often than those with more work experience, in another role, and with higher caseloads. Lastly, caseload and years of experience, and training and caseload both interacted to predict the substantiation decision. Implications for policy and practice and future research areas are discussed.


Autophagy inhibition enhances therapy-induced apoptosis in a Myc-induced model of lymphoma.
Autophagy is a lysosome-dependent degradative pathway frequently activated in tumor cells treated with chemotherapy or radiation. Whether autophagy observed in treated cancer cells represents a mechanism that allows tumor cells to survive therapy or a mechanism for initiating a nonapoptotic form of programmed cell death remains controversial. To address this issue, the role of autophagy in a Myc-induced model of lymphoma generated from cells derived from p53ER(TAM)/p53ER(TAM) mice (with ER denoting estrogen receptor) was examined. Such tumors are resistant to apoptosis due to a lack of nuclear p53. Systemic administration of tamoxifen led to p53 activation and tumor regression followed by tumor recurrence. Activation of p53 was associated with the rapid appearance of apoptotic cells and the induction of autophagy in surviving cells. Inhibition of autophagy with either chloroquine or ATG5 short hairpin RNA (shRNA) enhanced the ability of either p53 activation or alkylating drug therapy to induce tumor cell death. These studies provide evidence that autophagy serves as a survival pathway in tumor cells treated with apoptosis activators and a rationale for the use of autophagy inhibitors such as chloroquine in combination with therapies designed to induce apoptosis in human cancers.


LIGHTCURVE PHOTOMETRY OPPORTUNITIES: 2019 APRIL-JUNE.
We present lists of asteroid photometry opportunities for objects reaching a favorable apparition and have no or poorly-defined lightcurve parameters. Additional data on these objects will help with shape and spin axis modeling via lightcurve inversion. We also include lists of objects that will or might be radar targets. Lightcurves for these objects can help constrain pole solutions and/or remove rotation period ambiguities that might not come from using radar data alone.


OCEANOGRAPHY: Tracer from the Sky.
The rate of marine photosynthesis is of great importance for the global carbon cycle but is difficult to measure from environmental properties or in vitro. In this Perspective, Bender highlights a study by Luz and Barkan, who exploit the anomalous isotopic composition of molecular oxygen to determine the gross rate of photosynthesis in seawater. The method allows much longer time scales and larger spatial scales to be covered than with traditional oceanographic techniques.


Screening and study enrolment in the Randomized Evaluation of Normal vs. Augmented Level (RENAL) Replacement Therapy Trial.
Aspects of trial design, screening and study efficiency can affect recruitment and the findings of the trial itself. A clear understanding of the screening and study inclusion process will assist clinicians in interpreting trial results. Prospective observational data collection on all patients screened for possible inclusion in a randomized controlled trial of normal vs. augmented renal replacement therapy in critically ill patients (the RENAL Trial). 35 hospitals in Australia and New Zealand. All patients screened for the RENAL Trial. We screened 4,551 patients. Of these patients, 767 were ineligible because of lack of inclusion criteria and 2,085 because of exclusion criteria. Of the remaining 1,699, 1,508 (88.7%) were enrolled. The three most common exclusion criteria which prevented recruitment of potentially eligible patients were that the patient had end-stage kidney failure and was already on chronic dialysis (484; 23.2%), the patient's body weight was either <60 or >120 kg (456; 21.8%), and the fact that the patient had already received renal replacement therapy during the index admission. Important modifiable impediments to recruitment were inability to obtain consent in 191 cases, unavailability of research staff in 124 cases, physician objection in 89 cases, and inability to deliver the trial protocol in 78 cases. The RENAL Trial's enrolment efficiency was high and compared favourably with previous large intensive care units trials and with that of trials in patients with acute renal failure. The high rate of enrolment suggests that the results can be applied with confidence to most patients with de novo acute renal failure. The loss of close to 1.5% of patients due to consent issues highlights a common problem in critical care trials. The low rate of physician objection suggests clinical equipoise.


Complete genome sequences of pooled genomic DNA from 10 marine bacteria using PacBio long-read sequencing.
High-quality, completed genomes are important to understand the functions of marine bacteria. PacBio sequencing technology provides a powerful way to obtain high-quality completed genomes. However individual library production is currently still costly, limiting the utility of the PacBio system for high-throughput genomics. Here we investigate how to generate high-quality genomes from pooled marine bacterial genomes. Pooled genomic DNA from 10 marine bacteria were subjected to a single library production and sequenced with eight SMRT cells on the PacBio RS II sequencing platform. In total, 7.35 Gbp of long-read data was generated, which is equivalent to an approximate 168× average coverage for the input genomes. Genome assembly showed that eight genomes with average nucleotide identities (ANI) lower than 91.4% can be assembled with high-quality and completion using standard assembly algorithms (e.g. HGAP or Canu). A reference-based reads phasing step was developed and incorporated to assemble the complete genomes of the remaining two marine bacteria that had an ANI > 97% and whose initial assemblies were highly fragmented. Ten complete high-quality genomes of marine bacteria were generated. The findings and developments made here, including the reference-based read phasing approach for the assembly of highly similar genomes, can be used in the future to design strategies to sequence pooled genomes using long-read sequencing.


Spatial attention affects brain activity in human primary visual cortex.
Functional MRI was used to test whether instructing subjects to attend to one or another location in a visual scene would affect neural activity in human primary visual cortex. Stimuli were moving gratings restricted to a pair of peripheral, circular apertures, positioned to the right and to the left of a central fixation point. Subjects were trained to perform a motion discrimination task, attending (without moving their eyes) at any moment to one of the two stimulus apertures. Functional MRI responses were recorded while subjects were cued to alternate their attention between the two apertures. Primary visual cortex responses in each hemisphere modulated with the alternation of the cue; responses were greater when the subject attended to the stimuli in the contralateral hemifield. The attentional modulation of the brain activity was about 25% of that evoked by alternating the stimulus with a uniform field.


Production of unusual dispiro metabolites in Pestalotiopsis virgatula endophyte cultures: HPLC-SPE-NMR, electronic circular dichroism, and time-dependent density-functional computation study.
The endophytic fungus Pestalotiopsis virgatula, derived from the plant Terminalia chebula and previously found to produce a large excess of a single metabolite when grown in the minimal M1D medium, was induced to produce a variety of unusual metabolites by growing in potato dextrose broth medium. Analysis of the fermentation medium extract was performed using an HPLC-PDA-MS-SPE-NMR hyphenated system, which led to the identification of a total of eight metabolites (1-8), six of which are new. Most of the metabolites are structurally related and are derivatives of benzo[c]oxepin, rare among natural products. This includes dispiro derivatives 7 and 8 (pestalospiranes A and B), having a novel 1,9,11,18-tetraoxadispiro[22.214.171.124]octadecane skeleton. Relative and absolute configurations of the latter were determined by a combination of NOESY spectroscopy and electronic circular dichroism spectroscopy supported by time-dependent density-functional theory calculations (B3LYP/TZVP level). This work demonstrates that a largely complete structure elucidation of numerous metabolites present in a raw fermentation medium extract can be performed by the HPLC-SPE-NMR technique using only a small amount of the extract, even with unstable metabolites that are difficult to isolate by traditional methods.


Re-thinking resuscitation: leaving blood pressure cosmetics behind and moving forward to permissive hypotension and a tissue perfusion-based approach.
Definitions of shock and resuscitation endpoints traditionally focus on blood pressures and cardiac output. This carries a high risk of overemphasizing systemic hemodynamics at the cost of tissue perfusion. In line with novel shock definitions and evidence of the lack of a correlation between macro- and microcirculation in shock, we recommend that macrocirculatory resuscitation endpoints, particularly arterial and central venous pressure as well as cardiac output, be reconsidered. In this viewpoint article, we propose a three-step approach of resuscitation endpoints in shock of all origins. This approach targets only a minimum individual and context-sensitive mean arterial blood pressure (for example, 45 to 50 mm Hg) to preserve heart and brain perfusion. Further resuscitation is exclusively guided by endpoints of tissue perfusion irrespectively of the presence of arterial hypotension ('permissive hypotension'). Finally, optimization of individual tissue (for example, renal) perfusion is targeted. Prospective clinical studies are necessary to confirm the postulated benefits of targeting these resuscitation endpoints.


Dental implications of bisphophonate-related osteonecrosis.
The aim is to explore the current theories about clinical , pathological and dental management of bisphosphonate related osteonecrosis of the jaws. Also discussed are the actions of bisphosphonates, pathogenesis related to the susceptibility of jaws, the predisposing risk factors for the development of bisphosphonate-related osteonecrosis of the jaws (BRONJ) and diagnostic criteria based on the literature review. Osteoporosis is a disease that generally affects the mineral status of both cortical and trabecular bone in post menopausal women. Bisphosphonates are a group of drugs that preserve and increase bone mass. Bisphosphonate drugs are classified according to use and method of delivery. The bisphosphonates used for the treatment of osteoporosis are taken orally. Little is known about the side effects and dangers of the long-term use of therapeutic doses of Bisphosphonates. A recent complication reported is osteonecrosis of jaws. The use of IV bisphosphonates for multiple myeloma and metastatic bone diseases suggests that dosage, length of treatment, and route of administration, as well as cofactors such as use of glucocorticoids and immunosuppressive agents, and dental surgery, could all be related to the incidence of BRONJ. This review provides an update on current knowledge about clinical, pathological and management aspects of BRONJ. Little evidence exists to direct the prosthodontic management of patients with a history of bisphosphonate use. Patients with active osteonecrosis related to bisphosphonate use have reduced tissue tolerance to function with removable prostheses and decreased potential for osseointegration of dental implants. Decisions should be based on clinical judgment tempered by the presenting conditions, medical profile, and patient needs. A better understanding would help in a dental setting to prevent any complication and help to improve the prognosis for those being treated for osteoradionecrosis.Until further evidence emerges regarding management of patients with active bisphosphonate- related osteonecrosis, conservative prosthodontic treatment is reasonable and prudent.


The well-being of relatives of patients with atrial fibrillation: a critical incident technique analysis.
The well-being of relatives of patients having chronic heart diseases (CHD) has been found to be negatively affected by the patient's condition. Studies examining relatives of patients with atrial fibrillation (AF) indicate that their well-being may be affected in a similar manner, but further research is needed. To explore and describe critical incidents in which relatives of patients experience how AF affects their well-being and what actions they take to handle these situations. An explorative, descriptive design based on the critical incident technique (CIT) was used. Interviews were conducted with 19 relatives (14 women and five men) of patients hospitalised in southern Sweden due to acute symptoms of the AF. The well-being of relatives was found to be affected by their worries (patient-related health), as well as the sacri-ficing of their own needs (self-related health). In handling their own well-being, these relatives adjusted to and supported the patient (practical involvement), along with adjusting their own feelings and responding to the mood of the patients (emotional involvement). The well-being of relatives of patients with AF was affected depending on the patients' well-being. In their attempt to handle their own well-being, the relatives adjusted to and supported the patients. Further research is needed in order to evaluate the effects of support to relatives and patients respectively and together.


Roles of replicative and specialized DNA polymerases in frameshift mutagenesis: mutability of Salmonella typhimurium strains lacking one or all of SOS-inducible DNA polymerases to 26 chemicals.
Progression of DNA replication is occasionally blocked by endogenous and exogenous DNA damage. To circumvent the stalling of DNA replication, cells possess a variety of specialized DNA polymerases that replicate through DNA damage. Salmonella typhimurium strain TA1538 has six DNA polymerases and four of them are encoded by damage-inducible SOS genes, i.e. polB(ST) (pol II), dinB(ST) (pol IV), umuDC(ST) (pol V) and samAB. The strain has been used for the detection of a variety of chemical mutagens because of the high sensitivity to -2 frameshift occurring in CGCGCGCG sequence. To assign the role of each DNA polymerase in the frameshift mutagenesis, we have constructed the derivatives lacking one or all of SOS-inducible DNA polymerases and examined the mutability to 26 chemical mutagens. Interestingly, the chemicals could be categorized into four classes: class I whose mutagenicity was reduced by the deletion of dinB(ST) (1-aminoanthracene and other four chemicals); class II whose mutagenicity was reduced by the deletion of either dinB(ST) or umuDC(ST) plus samAB (7,12-dimethylbenz[a]anthracene and other three chemicals); class III whose mutagenicity largely depended on the presence of umuDC(ST) plus samAB (1-N-6-azabenzo[a]pyrene and other three chemicals) and class IV whose mutagenicity was not reduced by deletion of any of the genes encoding SOS-inducible DNA polymerases (Glu-P-1 and other 12 chemicals). Deletion of polB(ST) reduced by 30-60% the mutagenicity of six chemicals of classes II and III. These results suggest that multiple DNA polymerases including the replicative DNA polymerase, i.e. DNA polymerase III holoenzyme, play important roles in chemically induced -2 frameshift and also that different sets of DNA polymerases are engaged in the translesion bypass of different DNA lesions.


Thyroglossal Duct Cyst Occupying Posterior Hyoid Space with Endolaryngeal Extension Presenting After Neck Trauma.
Thyroglossal duct cysts are the most common congenital neck mass. They typically present as a painless midline neck mass in a child or young adult, but may also present later in adulthood when the cyst becomes symptomatic. Thyroglossal duct cysts are most commonly located inferior to the hyoid bone in close relation with the thyrohyoid membrane. Very rarely, they may extend intralaryngeal, occupy the posterior hyoid space, and present with dysphonia and/or dysphagia. To our knowledge, this is the 24th reported case in the literature. Case report with a comprehensive review of the literature. The patient was a 43-year-old male experiencing dysphonia and dysphagia following a motor vehicle accident. He was subsequently found to have a large thyroglossal duct cyst with endolaryngeal extension that was previously asymptomatic and undiagnosed. He underwent successful surgical excision which resulted in resolution of symptoms. This is the first reported case of a thyroglossal duct cyst in the posterior hyoid space with endolaryngeal extension being diagnosed following a traumatic event. This case illustrates the need to consider thyroglossal duct cyst in the differential diagnosis when working up a post-traumatic intralaryngeal neck mass. A secondary educational objective in this case is to be diligent to consider and rule out laryngeal fracture in the case of a neck mass presenting after trauma as they can easily be missed and present with many overlapping symptoms.


CASK participates in alternative tripartite complexes in which Mint 1 competes for binding with caskin 1, a novel CASK-binding protein.
CASK, an adaptor protein of the plasma membrane, is composed of an N-terminal calcium/calmodulin-dependent protein (CaM) kinase domain, central PSD-95, Dlg, and ZO-1/2 domain (PDZ) and Src homology 3 (SH3) domains, and a C-terminal guanylate kinase sequence. The CaM kinase domain of CASK binds to Mint 1, and the region between the CaM kinase and PDZ domains interacts with Velis, resulting in a tight tripartite complex. CASK, Velis, and Mint 1 are evolutionarily conserved in Caenorhabditis elegans, in which homologous genes (called lin-2, lin-7, and lin-10) are required for vulva development. We now demonstrate that the N-terminal CaM kinase domain of CASK binds to a novel brain-specific adaptor protein called Caskin 1. Caskin 1 and a closely related isoform, Caskin 2, are multidomain proteins containing six N-terminal ankyrin repeats, a single SH3 domain, and two sterile alpha motif domains followed by a long proline-rich sequence and a short conserved C-terminal domain. Unlike CASK and Mint 1, no Caskin homolog was detected in C. elegans. Immunoprecipitations showed that Caskin 1, like Mint 1, is stably bound to CASK in the brain. Affinity chromatography experiments demonstrated that Caskin 1 coassembles with CASK on the immobilized cytoplasmic tail of neurexin 1, suggesting that CASK and Caskin 1 coat the cytoplasmic tails of neurexins and other cell-surface proteins. Detailed mapping studies revealed that Caskin 1 and Mint 1 bind to the same site on the N-terminal CaM kinase domain of CASK and compete with each other for CASK binding. Our data suggest that in the vertebrate brain, CASK and Velis form alternative tripartite complexes with either Mint 1 or Caskin 1 that may couple CASK to distinct downstream effectors.


Clinical characteristics of echovirus 11 and coxsackievirus B5 infections in Taiwanese children requiring hospitalization.
Severe illness can occur in young children infected with certain types of enteroviruses including echovirus 11 (Echo11) and coxsackievirus B5 (CoxB5). The manifestations and outcomes of Echo11 and CoxB5 diseases across all ages of children remained not comprehensively characterized in Taiwan. Culture-confirmed Echo11 (60 patients) or CoxB5 (65 patients) infections were identified in a hospital from 2010 to 2018. The demographics, clinical presentations, laboratory data and outcomes were abstracted and compared between the two viruses infections. Echo11 and CoxB5 was respectively identified in 7 (77.8%) and 2 (22.2%) of 9 calendar years. The median age of all patients was 15 months (range, 1 day-14.5 years). For infants ≤3 months old, Echo11 (23 cases) was associated with higher incidence of aseptic meningitis (35% versus 0%, P = 0.003), and a lower rate of upper respiratory tract infections (URI) (22% versus 65%, P = 0.004) compared to CoxB5 (20 cases) infections. For patients >3 months old, URI was the cardinal diagnosis (60%) for both viruses. Aseptic meningitis was also more commonly identified in elder children with Echo11 infections (27% versus 11%), though with marginal significance (P = 0.07). Acute liver failure was identified in four young infants with Echo11 infections including one neonate dying of severe sepsis and myocarditis. All patients with CoxB5 infections recovered uneventfully. Aseptic meningitis, sepsis-like illness and acute liver failure were more commonly identified in children with Echo11 than those with CoxB5 infections, suggesting greater neurological tropism and virulence toward Echo11.


Effect of supervised exercise training during pregnancy on neonatal and maternal outcomes among overweight and obese women. Secondary analyses of the ETIP trial: A randomised controlled trial.
Maternal obesity associates with complications during pregnancy and childbirth. Our aim was to investigate if exercise during pregnancy in overweight/obese women could influence birth weight or other neonatal and maternal outcomes at delivery. This is a secondary analysis of a randomised controlled trial of exercise training in pregnancy for women with body mass index (BMI) ≥ 28 kg/m2. Ninety-one women (31.3 ± 4.3 years, BMI 34.5 ± 4.2 kg/m2) were allocated 1:1 to supervised exercise during pregnancy or to standard care. The exercise group was offered three weekly training sessions consisting of 35 minutes of moderate intensity walking/running followed by 25 minutes of strength training. Data from 74 women (exercise 38, control 36) were analysed at delivery. Birth weight was 3719 ± 695 g in the exercise group and 3912 ± 413 g in the control group (CI -460.96, 74.89, p = 0.16). Birth weight > 4000 g was 35% in the exercise group and 52% in the control group (p = 0.16). Mean gestational age at delivery was 39.1 weeks in the exercise group and 39.5 weeks in the control group (CI -1.33, 0.43, p = 0.31). No significant between-group differences were found in neonatal body size, skinfold thickness, placental weight ratio, or Apgar score. The prevalence of caesarean section was 24% in the exercise group and 17% in the control group (CI 0.20, 2.05, p = 0.57). Mean length of hospital stay was 4.8 days in the exercise group and 4.5 days in the control group (CI -0.45, 1.00, p = 0.45). Offering supervised exercise during pregnancy for overweight and obese women did not influence birth weight or other neonatal and maternal outcomes at delivery. However our trial was limited by low sample size and poor adherence to the exercise protocol, and further research is needed. ClinicalTrials.gov NCT01243554.


Impaired intracellular trafficking defines early Parkinson's disease.
Parkinson's disease (PD) is an insidious and incurable neurodegenerative disease, and represents a significant cost to individuals, carers, and ageing societies. It is defined at post-mortem by the loss of dopamine neurons in the substantia nigra together with the presence of Lewy bodies and Lewy neurites. We examine here the role of α-synuclein and other cellular transport proteins implicated in PD and how their aberrant activity may be compounded by the unique anatomy of the dopaminergic neuron. This review uses multiple lines of evidence from genetic studies, human tissue, induced pluripotent stem cells, and refined animal models to argue that prodromal PD can be defined as a disease of impaired intracellular trafficking. Dysfunction of the dopaminergic synapse heralds trafficking impairment.


The zebrafish homeobox gene hox[zf-114]: primary structure, expression pattern and evolutionary aspects.
It is gradually becoming accepted that vertebrate homeobox genes, like their counterparts in Drosophila, are crucial for normal development of the embryo. Most vertebrate homeoboxes reported so far are related to the Drosophila Antennapedia (Antp) sequence, and here we describe hox[zf-114], a novel Antp-like homeobox gene from the zebrafish. The sequence of the hox[zf-114] homeodomain indicates that this gene could be a member of a subfamily defined by the mouse Hox-1.5/-2.7/-4.1 genes. However, the evolutionary origin of hox[zf-114] is unclear and, based on the putative protein sequence, we conclude that it is not directly homologous to Hox-1.5, Hox-2.7 or Hox-4.1, or to other known mammalian homeobox genes. Nevertheless, as revealed by in situ hybridization, hox[zf-114] exhibits a spatial expression pattern typical for vertebrate Antp-like homeobox genes. Transcripts are detected in the posterior hindbrain, where a sharp anterior border of expression is observed, and throughout the spinal cord. The hox[zf-114] gene is also active in a region that gives rise to the pectoral fins. These findings suggest a role for hox[zf-114] in anteroposterior patterning of the neural tube and in pectoral fin development.


Antiprotozoal Effect of Saponins in the Rumen Can Be Enhanced by Chemical Modifications in Their Structure.
The antiprotozoal effect of saponins is transitory, as when saponins are deglycosylated to the sapogenin by rumen microorganisms they become inactive. We postulated that the substitution of the sugar moiety of the saponin with small polar residues would produce sapogen-like analogs which might be resistant to degradation in the rumen as they would not be enzymatically cleaved, allowing the antiprotozoal effect to persist over time. In this study, we used an acute assay based on the ability of protozoa to break down [14C] leucine-labeled Streptococcus bovis and a longer term assay based on protozoal motility over 24 h to evaluate both the antiprotozoal effect and the stability of this effect with fifteen hederagenin bis-esters esterified with two identical groups, and five cholesterol and cholic acid based derivatives carrying one to three succinate residues. The acute antiprotozoal effect of hederagenin derivatives was more pronounced than that of cholesterol and cholic acid derivatives. Modifications in the structure of hederagenin, cholesterol, and cholic acid derivatives resulted in compounds with different biological activities in terms of acute effect and stability, although those which were highly toxic to protozoa were not always the most stable over time. Most of the hederagenin bis-esters, and in particular hederagenin bis-succinate (TSB24), hederagenin bis-betainate dichloride (TSB37) and hederagenin bis-adipate (TSB47) had a persistent effect against rumen protozoa in vitro, shifting the fermentation pattern toward higher propionate and lower butyrate. These chemically modified triterpenes could potentially be used in ruminant diets as an effective defaunation agent to, ultimately, increase nitrogen utilization, decrease methane emissions, and enhance animal production. Further trials in vivo or in long term rumen simulators are now needed to confirm the in vitro observations presented.


Hemoglobinuria-related acute kidney injury is driven by intrarenal oxidative reactions triggering a heme toxicity response.
Intravascular hemolysis can result in hemoglobinuria with acute kidney injury. In this study we systematically explored two in vivo animal models and a related cell culture system to identify hemoglobinuria-triggered damage pathways. In models of stored blood transfusion and hemoglobin (Hb) exposure in guinea pigs and beagle dogs we found that hemoglobinuria led to intrarenal conversion of ferrous Hb(Fe(2+)) to ferric Hb(Fe(3+)), accumulation of free heme and Hb-cross-linking products, enhanced 4-hydroxynonenal reactivity in renal tissue, and acute tubule injury. These changes were associated in guinea pigs with activation of a renal cortex gene expression signature indicative of oxidative stress and activation of the unfolded protein response (UPR). Tubule cells of hemolytic animals demonstrated enhanced protein expression of heme oxygenase and heat shock protein and enhanced expression of acute kidney injury-related neutrophil gelatinase-associated lipocalin. These adverse changes were completely prevented by haptoglobin treatment. The in vivo findings were extrapolated to a MS-based proteome analysis of SILAC-labeled renal epithelial cells that were exposed to free heme within a concentration range estimate of renal tubule heme exposure. These experiments confirmed that free heme is a likely trigger of tubule barrier deregulation and oxidative cell damage and reinforced the hypothesis that uncontrolled free heme could trigger the UPR as an important pathway of renal injury during hemoglobinuria.


Angiotensin Type 1 Receptor Antagonists Protect Against Alpha-Synuclein-Induced Neuroinflammation and Dopaminergic Neuron Death.
The loss of dopaminergic neurons and α-synuclein accumulation are major hallmarks of Parkinson's disease (PD), and it has been suggested that a major mechanism of α-synuclein toxicity is microglial activation. The lack of animal models that properly reproduce PD, and particularly the underlying synucleinopathy, has hampered the clarification of PD mechanisms and the development of effective therapies. Here, we used neurospecific adeno-associated viral vectors serotype 9 coding for either the wild-type or mutated forms of human alpha-synuclein (WT and SynA53T, respectively) under the control of a synapsin promoter to further induce a marked dopaminergic neuron loss together with an important microglial neuroinflammatory response. Overexpression of neuronal alpha-synuclein led to increased expression of angiotensin type 1 receptors and NADPH oxidase activity, together with a marked increase in the number of OX-6-positive microglial cells and expression of markers of phagocytic activity (CD68) and classical pro-inflammatory/M1 microglial phenotype markers such as inducible nitric oxide synthase, tumor necrosis factor alpha, interleukin-1β, and IL-6. Moreover, a significant decrease in the expression of markers of immunoregulatory/M2 microglial phenotype such as the enzyme arginase-1 was constantly observed. Interestingly, alpha-synuclein-induced changes in microglial phenotype markers and dopaminergic neuron death were inhibited by simultaneous treatment with the angiotensin type 1 blockers candesartan or telmisartan. Our results suggest the repurposing of candesartan and telmisartan as a neuroprotective strategy for PD.


Functional adult acetylcholine receptor develops independently of motor innervation in Sol 8 mouse muscle cell line.
We have defined culture conditions, using a feeder layer of cells from the embryonic mesenchymal cell line, 10T1/2 and a serum-free medium, which allow cells from the mouse myogenic cell line Sol 8 to form contracting myotubes for two weeks. Under these culture conditions, Sol 8 myotubes undergo a maturation process characterized by a sequential expression of two phenotypes. An early phenotype is typified by the expression of the nicotinic acetylcholine receptor (AChR) gamma-subunit transcripts and the presence of low conductance ACh-activated channels, typical of embryonic AChR. A late phenotype is characterized by the expression of AChR epsilon-subunit transcripts, the decreased accumulation of gamma-subunit transcripts and the appearance of high conductance ACh-activated channels, typical of adult AChR. These results indicate that the expression of functional adult type AChR does not require the presence of the motor nerve and therefore represents an intrinsic feature of the Sol 8 muscle cells. Chronic exposure of the cells to the voltage-sensitive Na+ channel blocking agent tetrodotoxin does not affect the appearance of the AChR epsilon-subunit transcripts but prevents the reduction of the steady-state level of the AChR gamma-subunit transcripts and yields a reduced proportion of the adult type channels. Thus, activity seems to facilitate the switch from the embryonic to the adult phenotype of the AChR protein. The Sol 8 cell system might be useful to analyse further the genetic and epigenetic regulation of muscle fibre maturation in mammals.


Are screening guidelines for abdominal aortic aneurysms being implemented within a large VA primary health care system?
Screening men aged 65 to 75 years who have ever smoked for abdominal aortic aneurysm (AAA) has been recommended to reduce AAA-related mortality. However, it is unknown whether the evidence-based recommendation has been implemented within primary care. The aim of this study was to determine whether screening for AAA is being performed within a large Veterans Affairs (VA) primary health care system. This was a retrospective cohort study examining AAA screening practices within the VA Connecticut Healthcare System. Any of the following imaging procedures were considered screening tests for AAA: abdominal ultrasound, computed tomography (CT) of the abdomen, CT colography, or magnetic resonance imaging of the abdomen. A total of 279 patients were included in the cohort: 83 (30%) were offered screening for AAA or had recent imaging performed that would have allowed for detection of an AAA. Seventy-three patients (26%) underwent AAA screening or had recent imaging of their abdomens, while 10 patients either refused imaging or were awaiting ultrasonographic screening at the time of this study. Of the 73 patients who had undergone screening or other abdominal imaging evaluations, 9 (12.3%) were found to have AAAs. There appears to be a low rate of screening for AAA within 1 primary care setting in a large VA health care system. If this finding is replicated within other VA primary health care settings, then the VA health care system should consider implementing a performance metric within primary care to improve AAA screening rates.


Benefits and risks of using gelatin solution as a plasma expander for perioperative and critically ill patients: a meta-analysis.
This meta-analysis aimed to evaluate the benefits and risks of gelatin solutions compared to other intravenous fluids for patients in perioperative and critical care settings. Of the 66 studies identified from MEDLINE and EMBASE databases, 30 randomised controlled trials involving 2709 patients met the inclusion criteria and were subject to meta-analysis. The risk of mortality (odds ratio 1.03, 95% confidence interval 0.80 to 1.32) and amount of blood loss (weighted-mean-difference 7.56 ml, 95% confidence interval 18.75 to 33.87) were not significantly different between patients who were treated with gelatin solutions and other types of intravenous fluids. When compared to starches, gelatin solutions were associated with a lower risk of acute renal failure (odds ratio 0.43, 95% confidence interval 0.20 to 0.92; P=0.03). When gelatin solutions were compared to isotonic albumin, patients who were treated with gelatin solutions required a small, but significantly greater amount of blood transfusion (weighted-mean-difference 180 ml, 95% confidence interval 8.1 to 353.6; P=0.04). These findings suggest that using gelatin solutions is associated with a lower risk of acute renal failure compared to older starches. Using gelatin as a plasma expander appears to have no significant advantages over crystalloids or isotonic albumin on mortality and may have a slightly higher risk of requiring allogeneic blood transfusion in perioperative and critically ill patients. An adequately powered randomised controlled trial with economic analysis is needed before gelatin solution can be recommended as a routine plasma expander for patients undergoing major surgery or who are critically ill.


Multidisciplinary approach for the treatment of extensive external cervical resorption after dental trauma.
External cervical resorption (ECR) is a sequela of dentoalveolar trauma that may cause functional, esthetic, and psychologic alterations. The aim of this study was to report a successful multidisciplinary treatment approach performed in a 12-year-old patient who presented with posttraumatic ECR associated with extensive opened cavity, pulp necrosis, and periapical lesion of tooth number 9, with an initial unfavorable prognosis. Crown lengthening was done to enable restoration of vestibular surface with resin composite, forming a barrier that allowed endodontic treatment. Afterwards, a prefabricated fiberglass post was cemented and esthetic restoration was performed using the adhesive technique and direct composite veneer. Reconstructive periodontal surgery was performed to correct irregular gingival contour. After treatment and successive follow-up sessions, it was concluded that although the tooth had been indicated for extraction, low invasive direct techniques were effective to recover function and esthetics and to maintain the tooth in the oral cavity.


Amended safety assessment of Sesamum indicum (sesame) seed oil, hydrogenated sesame seed oil, Sesamum indicum (sesame) oil unsaponifiables, and sodium sesameseedate.
Sesamum indicum (sesame) seed oil and related cosmetic ingredients are derived from Sesamum indicum. Sesamum indicum (sesame) seed oil, sesamum indicum (sesame) oil unsaponifiables, and hydrogenated sesame seed oil function as conditioning agents. Sodium sesameseedate functions as a cleansing agent, emulsifying agent, and a nonaqueous viscosity increasing agent. These ingredients are neither skin irritants, sensitizers, teratogens, nor carcinogens at exposures that would result from cosmetic use. Both animal and human data relevant to the cosmetic use of these ingredients were reviewed. The CIR Expert Panel concluded that these ingredients are safe in the present practices of use and concentration as described in this safety assessment.


Design of computational retrobiosynthesis tools for the design of de novo synthetic pathways.
Designing putative metabolic pathways is of great interest in synthetic biology. Retrobiosynthesis is a discipline that involves the design, evaluation, and optimization of de novo biosynthetic pathways for the production of high-value compounds and drugs from renewable resources and natural or engineered enzymes. The best candidate pathways are then engineered within a metabolic network of microorganisms that serve as synthetic platforms for synthetic biology. The complexity of biological chemistry and metabolism requires computational approaches to explore the full possibilities of engineering synthetic pathways towards target compounds. Herein, we discuss recent developments in the design of computational tools for retrosynthetic biochemistry and outline the workflow and design elements for such tools.


A side-by-side comparison of dust collection methods for sampling lead-contaminated house dust.
The Environmental Protection Agency is required to set a standard for lead-contaminated house dust, but whether dust lead loading (micrograms/ft2) or concentration (micrograms/g) is more predictive of children's blood lead levels, which dust collection method should be used, and which surfaces should be sampled are unknown. Using a random sample of sequential births, we enrolled 205 urban children, 12 to 30 months of age, who had lived in the same house since at least 6 months of age. Samples of dust were obtained from predetermined surfaces in each child's residence using a wipe method and two vacuum methods, the Baltimore repair and maintenance method (BRM) and the dust vacuum method (DVM). Other potential sources of environmental exposure also were analyzed for lead, including soil, water, and paint. In general, dust lead loading is more predictive of children's blood lead levels than is dust lead concentration. Dust lead loading as measured with the BRM sampler explained more of the variation in children's blood lead levels than did wipe loading and DVM loading (13.7, 10.1, and 5.9%, respectively, adjusted for other significant predictors). The partial correlation between BRM lead loading and children's blood lead was significantly different than that for DVM lead loading, but it was not significantly different than that for wipe lead loading. Of the four surfaces measured, noncarpeted floors and interior window sills or wells were significantly associated with children's blood lead levels in multiple regression models. These data indicate that dust lead loading is more predictive of children's blood lead levels than is dust lead concentration and that, to determine if a housing unit is safe for children, noncarpeted floors and interior window sills or window wells should be measured using either the BRM or wipe sampling method.


Neural network of cognitive emotion regulation--an ALE meta-analysis and MACM analysis.
Cognitive regulation of emotions is a fundamental prerequisite for intact social functioning which impacts on both well being and psychopathology. The neural underpinnings of this process have been studied intensively in recent years, without, however, a general consensus. We here quantitatively summarize the published literature on cognitive emotion regulation using activation likelihood estimation in fMRI and PET (23 studies/479 subjects). In addition, we assessed the particular functional contribution of identified regions and their interactions using quantitative functional inference and meta-analytic connectivity modeling, respectively. In doing so, we developed a model for the core brain network involved in emotion regulation of emotional reactivity. According to this, the superior temporal gyrus, angular gyrus and (pre) supplementary motor area should be involved in execution of regulation initiated by frontal areas. The dorsolateral prefrontal cortex may be related to regulation of cognitive processes such as attention, while the ventrolateral prefrontal cortex may not necessarily reflect the regulatory process per se, but signals salience and therefore the need to regulate. We also identified a cluster in the anterior middle cingulate cortex as a region, which is anatomically and functionally in an ideal position to influence behavior and subcortical structures related to affect generation. Hence this area may play a central, integrative role in emotion regulation. By focusing on regions commonly active across multiple studies, this proposed model should provide important a priori information for the assessment of dysregulated emotion regulation in psychiatric disorders.


Changing the paradigm of 1-stage total callosotomy for the treatment of pediatric generalized epilepsy.
The authors propose that anterior callosotomy -- thought to have some advantages over total callosotomy -- is not superior to total callosotomy for prevention of seizure propagation or other complications. The study comprised 34 patients in whom generalized epilepsy syndrome or frontal lobe seizures with a secondary generalization were diagnosed. Preoperatively, all patients suffered from disabling drop attacks or intense head-drop seizures, and some patients also experienced other types of seizures. The male/female ratio was 22:12, and patients ranged in age between 1 to 19 years (mean 8.7 years). The follow-up period ranged from 1.08 to 5.0 years (mean 2.58 years). Seizure outcome, parental assessment of daily function, and parental satisfaction with the outcome was assessed postoperatively. After undergoing 1-stage total callosotomy, drop attacks disappeared completely in 25 patients during the follow-up period, and in 6 patients the frequency of drop attacks decreased to < 10% of baseline. With regard to other types of seizures, seizures resolved completely in 12 patients, and in another 18 seizure frequency decreased. Two patients experienced a transient disconnection syndrome for 2 and 4 weeks. One patient experienced ataxic hemiparesis for 3 weeks before it completely abated. The overall daily function of the patients improved, and all parents were satisfied with the surgical outcome. For pediatric generalized epilepsy syndrome, 1-stage total callosotomy will be the first choice in treatment for controlling generalized seizures.


Body-image distress is increased in women with polycystic ovary syndrome and mediates depression and anxiety.
To evaluate differences in body-image distress (BID) scores between women with polycystic ovary syndrome (PCOS) and controls and whether BID mediates anxiety and depression. Cross-sectional study. Academic institution. Reproductive-aged women with PCOS (n = 189) and controls (n = 225). Administering the Multidimensional Body-Self Relations-Appearance Subscale (MBSRQ-AS), Stunkard Figure Rating Scale (FRS), Hospital Anxiety and Depression Scale (HADS), and PCOS quality of life survey (MPCOS-Q). BID, depression, and anxiety scores and mediation of depression and anxiety scores by BID. Women with PCOS had worse BID scores on all five MBSRQ-AS subscales adjusted for age, body mass index, race, pregnancy history, income, and employment, and larger differences on the FRS compared with the control women. In multivariable regression models, the prevalence of depressive (28% vs. 19.2%) and anxiety (76.5% vs. 56.5%) symptoms were also statistically significantly higher in women with PCOS compared with the controls. Most MBSRQ-AS subscale scores statistically significantly correlated with depression, anxiety, and quality of life scores. The association between PCOS/control status and higher anxiety and depression scores was completely mediated by the appearance evaluation and body areas satisfaction subscales and partially mediated by overweight preoccupation, appearance orientation, and self-classified weight. Women with PCOS have increased BID and depressive and anxiety symptoms. In our study different aspects of BID either fully or partially mediated the association between PCOS/control status and depression and anxiety scores, suggesting that therapeutic interventions targeted at improving body image may decrease depressive and anxiety symptoms.


Efficacy of curcumin, and a saffron/curcumin combination for the treatment of major depression: A randomised, double-blind, placebo-controlled study.
Several studies have supported the antidepressant effects of curcumin (from the spice turmeric) and saffron for people with major depressive disorder. However, these studies have been hampered by poor designs, small sample sizes, short treatment duration, and similar intervention dosages. Furthermore, the antidepressant effects of combined curcumin and saffron administration are unknown. In a randomised, double-blind, placebo-controlled study, 123 individuals with major depressive disorder were allocated to one of four treatment conditions, comprising placebo, low-dose curcumin extract (250mg b.i.d.), high-dose curcumin extract (500mg b.i.d.), or combined low-dose curcumin extract plus saffron (15mg b.i.d.) for 12 weeks. The outcome measures were the Inventory of Depressive Symptomatology self-rated version (IDS-SR30) and Spielberger State-Trait Anxiety Inventory (STAI). The active drug treatments (combined) were associated with significantly greater improvements in depressive symptoms compared to placebo (p=.031), and superior improvements in STAI-state (p<.001) and STAI-trait scores (p=.001). Active drug treatments also had greater efficacy in people with atypical depression compared to the remainder of patients (response rates of 65% versus 35% respectively, p=.012). No differences were found between the differing doses of curcumin or the curcumin/saffron combination. Investigations with larger sample sizes are required to examine the efficacy of differing doses of curcumin and saffron/curcumin combination. Its effects in people with atypical depression also require examination in larger scale studies. Active drug treatments comprising differing doses of curcumin and combined curcumin/saffron were effective in reducing depressive and anxiolytic symptoms in people with major depressive disorder.


Effect of mitotane on pituitary corticotrophs in clinically normal dogs.
To evaluate the effects of mitotane administration on the function and morphology of pituitary corticotrophs in clinically normal dogs. 12 clinically normal adult Beagles. Dogs were randomly assigned to the control group or the mitotane treatment group. In mitotane treatment group dogs, mitotane was administered for 1 month. In both groups, ACTH stimulation testing and corticotrophin-releasing hormone (CRH) stimulation testing were performed. Magnetic resonance imaging (MRI) of the pituitary gland and brain was performed in mitotane treatment group dogs before and after administration of mitotane. After CRH stimulation testing and MRI, dogs were euthanatized and the pituitary gland and adrenal glands were excised for gross and histologic examination. ACTH concentrations in mitotane treatment group dogs were significantly higher than in the control group dogs following CRH stimulation. Magnetic resonance imaging revealed that pituitary glands were significantly larger in treatment group dogs after administration of mitotane, compared with before administration. On gross and histologic examinations, the adrenal cortex was markedly atrophied. Immunohistochemistry revealed hypertrophy of corticotrophs in pituitary glands of mitotane treatment group dogs. These findings indicate that inhibition of the adrenal cortex by continuous administration of mitotane leads to functional amplification and morphologic enhancement of corticotrophs in clinically normal dogs. In instances of corticotroph adenoma, hypertrophy of individual corticotrophs induced by mitotane may greatly facilitate enlargement of the pituitary gland and increases in ACTH secretion.


Proprotein-processing endoprotease furin controls growth of pancreatic beta-cells.
We have previously reported that in the well-differentiated beta-cell line MIN6 cells, the beta-cell-specific differentiated characteristics, such as insulin content, expression of prohormone convertases PC2 and PC3, and glucose-regulated insulin secretion, diminished when the proprotein-processing endoprotease furin was highly expressed. Since furin converts many growth-related protein precursors to their bioactive forms, we compared the four pancreatic islet cell lines RINm5F, betaTC3, betaHC9, and MIN6 with respect to cell growth rate, furin expression, endoprotease activity, and insulin content. RINm5F cells exhibited the strongest furin expression, higher furin-type endoprotease activity, and the fastest cell growth, but had the least insulin content. In contrast, MIN6 cells exhibited only a weak furin expression, little furin-type endoprotease activity, and the slowest cell growth, but had the highest insulin content. To test whether furin-expressing cells secrete growth-promoting factors cleaved by furin, we prepared conditioned media from RINm5F and furin cDNA-introduced MIN6 (MIN6-F) cells. The conditioned media from RINm5F and MIN6-F induced increased DNA synthesis and promoted the growth of normal MIN6 cells, compared with the medium from the empty vector-introduced MIN6-0 cells. We then examined the effect of the protease inhibitors alpha1-antitrypsin and its variants by infecting their vaccinia recombinants to the four cell lines. All conditioned media from each cell line expressing the furin-specific alpha1-antitrypsin variant exhibited the least DNA synthetic capacity on normal MIN6 cells. Furthermore, all three sublines of MIN6-F grew faster than MIN6-0 and MIN6. Thus, we suggest that the islet cells with higher furin expression may induce increased production of growth factors, which result in an increase in cell growth, through an autocrine/paracrine mechanism.


Radiosensitive human tumour cell lines show misrepair of DNA termini.
Physical measures of the rejoining of radiation-induced breaks in DNA strands are limited in terms of sensitivity and the fact that they do not assess the fidelity with which the rejoining occurs. In this report, transfection of cleaved plasmid has been used as a probe for repair in three radiosensitive tumour cell lines and shown them to have low repair fidelity compared with resistant cells. Errors in the repair of linear plasmid were found by Southern analysis, in keeping with the measured repair fidelity. Radiosensitive tumour cells showed few errors in the uptake and integration of circular plasmid, in contrast to ataxia-telangiectasia (A-T) cells. In the neuroblastoma HX142, the repair of blunt-ended linear plasmid was associated with deletions of > 1 kb; staggered-ended linear plasmid was repaired with small insertions and circular plasmid integration was intact in > 60% of the copies. The neuroblastoma SKN.SH, processed staggered-ended plasmid by insertions of a variety of sizes, but processed circular plasmid largely error-free. In contrast, A-T cells (AT5BIVA) had the same spectrum of errors irrespective of the form of plasmid transfected. Cell fusion between HX142 and AT5BIVA showed complementation to a resistant phenotype, suggesting that misrepair in the tumour cell did not result from somatic mutation in the ATM gene. In conclusion, radiosensitive tumours show evidence of misrepair of DNA termini, with a mechanism which is functionally and genetically distinct from that in A-T cells.


Coronary Vasospasm-Induced Myocardial Infarction: An Uncommon Presentation of Unrecognized Hyperthyroidism.
Hyperthyroidism can present with cardiac issues, such as tachycardia, atrial fibrillation, and high output congestive heart failure. Rare case reports of coronary vasospasm leading to myocardial infarction (MI) are published. Of these cases, many are known to be hyperthyroid prior to cardiac presentation. We report a female patient with unrecognized thyrotoxicosis who presents with acute MI secondary to coronary vasospasm.


New class of transforming growth factors potentiated by epidermal growth factor: isolation from non-neoplastic tissues.
Proteins potentiated by epidermal growth factor (EGF) to induce a transformed phenotype in non-neoplastic rat kidney fibroblasts in cell culture have been isolated from many non-neoplastic tissues of the adult mouse, including submaxillary gland, kidney, liver, muscle, heart, and brain. They resemble previously described transforming growth factors (TGFs) isolated from neoplastic cells as follows: they are extractable by acid/ethanol and are acid-stable, low molecular weight (6000-10,000) polypeptides requiring disulfide bonds for activity; and they cause anchorage-independent growth of non-neoplastic indicator cells that will not grow in soft agar in their absence. Partial purification of these TGFs from submaxillary glands of male mice shows that they are distinct from EGF. Unlike previously described extracellular TGFs, but like certain cellular TGFs from neoplastic cells, they are potentiated by EGF in their ability to promote anchorage-independent growth. The isoelectric point of the submaxillary gland TGF protein is near neutrality. Chromatography on Bio-Gel P-30 followed by high-pressure liquid chromatography has resulted in a 22,000-fold overall purification. The most purified protein is active in inducing growth in soft agar at 1 ng/ml when assayed in the presence of EGF. The data add further evidence to the concept that neoplasia may result from a quantitative, rather than qualitative, alteration in non-neoplastic biochemical processes.


Burden of obstructive lung disease study in Tehran: Prevalence and risk factors of chronic obstructive pulmonary disease.
Chronic obstructive pulmonary disease (COPD) was ranked the sixth-most common cause of death worldwide in 1990, but now it is the third-most common cause. The goal of the present study was to assess the prevalence and determine the causes and risk factors of COPD in Tehran. This study followed a stratified cluster sampling strategy with proportional allocation within strata. The target population was all non-institutionalized inhabitants, aged 18 to 40 in one group and over 40 in another who resided in Tehran in 2013. The core questionnaire was developed from pre-existing validated questionnaires that had already been used in multi-national studies. The single most important outcome measure obtained as part of this protocol was spirometry before and after the administration of 200 mg (two puffs) of salbutamol. The most commonly reported respiratory symptoms were: sputum production in 291 patients (16.2%) [95% confidence interval (CI): 14.5-17.9%], chronic cough in 171 (9.5%) (95% CI: 8.2-10.9%), wheezing in 377 (21.0%) (95%CI: 19.1-22.9%) and dyspnea in 388 patients (21.6%) (95% CI: 19.7-23.5%). The overall COPD prevalence defined by the post-bronchodilator spirometric functional criteria was 9.2%. This value in men (10.1%) was higher than in women (8.5%); the prevalence was significantly higher in subjects aged over 55 years (P ≤ 0.002). The prevalence of COPD was strongly dependent on smoking status, especially in ex-smokers, and increased considerably with age. 69% of patients with COPD were non-smoker. The high prevalence of verified COPD, a great deal of which was undiagnosed before by a physician, highlights the necessity of raising awareness of this disease among health professionals, and use of spirometry in the primary care setting. A future cross-sectional and prospective cohort study should be performed to explore all risk factors and their impact on decline in lung function and worsening of respiratory symptoms especially in non-smokers.


Neighborhood selection and the social reproduction of concentrated racial inequality.
In this paper, we consider neighborhood selection as a social process central to the reproduction of racial inequality in neighborhood attainment. We formulate a multilevel model that decomposes multiple sources of stability and change in longitudinal trajectories of achieved neighborhood income among nearly 4000 Chicago families followed for up to seven years wherever they moved in the United States. Even after we adjust for a comprehensive set of fixed and time-varying covariates, racial inequality in neighborhood attainment is replicated by movers and stayers alike. We also study the emergent consequences of mobility pathways for neighborhood-level structure. The temporal sorting by individuals of different racial and ethnic groups combines to yield a structural pattern offlows between neighborhoods that generates virtually nonoverlapping income distributions and little exchange between minority and white areas. Selection and racially shaped hierarchies are thus mutually constituted and account for an apparent equilibrium of neighborhood inequality.


The diagnosis of brain tumours in children: a guideline to assist healthcare professionals in the assessment of children who may have a brain tumour.
Brain tumours are the commonest solid tumour in children. Children with brain tumours are frequently unwell for months prior to diagnosis. A prolonged period between symptom onset and diagnosis is associated with increased morbidity. To develop an evidence-based clinical guideline to support healthcare professionals in the identification, assessment and investigation of children presenting with symptoms and signs that could be due to a brain tumour. A systematic literature review with a meta-analysis and cohort study provided the guideline evidence base. A multi-disciplinary workshop and Delphi consensus voting were used to translate the evidence into a clinical guideline. The results of the literature review and cohort study have been previously published. 20 healthcare professionals and parents participated in the workshop. 77 statements were generated detailing the presenting features of childhood brain tumours, factors that could be used to discriminate brain tumours from other less serious conditions and possible referral pathways for children with brain tumours. 156 healthcare professionals agreed to participate in the Delphi process; 112 completed the first round and 88 completed all three rounds (attrition rate 21%). 64 statements reached consensus. The final guideline comprises 76 recommendations advising on the symptomatology of childhood brain tumours, assessment of children who may have a brain tumour and recommendations for selection for and timing of central nervous system imaging. Implementation of this guideline may support clinicians in the identification and timely imaging of children with brain tumours. This may reduce the morbidity currently experienced by many children with brain tumours.


Addressing social determinants of health inequities through settings: a rapid review.
Changing settings to be more supportive of health and healthy choices is an optimum way to improve population health and health equity. This article uses the World Health Organisation's (1998) (WHO Health Promotion Glossary. WHO Collaborating Centre for Health Promotion, Department of Public Health and Community Medicine, University of Sydney, NSW) definition of settings approaches to health promotion as those focused on modifying settings' structure and nature. A rapid literature review was undertaken in the period June-August 2014, combining a systematically conducted search of two major databases with targeted searches. The review focused on identifying what works in settings approaches to address the social determinants of health inequities, using Fair Foundations: the VicHealth framework for health equity. This depicts the social determinants of health inequities as three layers of influence, and entry points for action to promote health equity. The evidence review identified work in 12 settings (cities; communities and neighbourhoods; educational; healthcare; online; faith-based; sports; workplaces; prisons; and nightlife, green and temporary settings), and work at the socioeconomic, political and cultural context layer of the Fair Foundations framework (governance, legislation, regulation and policy). It located a relatively small amount of evidence that settings themselves are being changed in ways which address the social determinants of health inequities. Rather, many initiatives focus on individual behaviour change within settings. There is considerable potential for health promotion professionals to focus settings work more upstream and so replace or integrate individual approaches with those addressing daily living conditions and higher level structures, and a significant need for programmes to be evaluated for differential equity impacts and published to provide a more solid evidence base.


The healthcare burden imposed by liver disease in aging Baby Boomers.
The Baby Boomer generation is composed of 78 million Americans who are just beginning to reach their retirement years. Most Boomers have at least one chronic health problem, and these significantly increase the expense of providing medical care. Liver disease is the 12th most common cause of death in the United States, representing a relatively small portion of overall healthcare costs compared with cardiovascular disease and malignancy. Nonetheless, hepatitis C and fatty liver disease are more common in the Boomers and may play a more dominant role as they age. As a consequence, primary liver cancer is likely to become more prevalent. As with most chronic illnesses, prevention rather than disease management is likely to have the greatest impact. For those already afflicted by chronic liver disease, recognition and treatment can reduce the incidence of late complications, as was clearly demonstrated with chronic hepatitis B and C. Perhaps obesity is the greatest threat to our future health, and fatty liver disease, although likely preventable, will probably become the disease that fills the waiting rooms of future hepatologists.


Bone-Forming Capabilities of a Newly Developed NanoHA Composite Alloplast Infused with Collagen: A Pilot Study in the Sheep Mandible.
Lateral or vertical bone augmentation has always been a challenge, since the site is exposed to constant pressure from the soft tissue, and blood supply only exists from the donor site. Although, for such clinical cases, onlay grafting with autogenous bone is commonly selected, the invasiveness of the secondary surgical site and the relatively fast resorption rate have been reported as a drawback, which motivated the investigation of alternative approaches. This study evaluated the bone-forming capability of a novel nanoHA alloplast infused with collagen graft material made from biodegradable polylactic acid/polyglycolic acid versus a control graft material with the same synthesized alloplast without the nanoHA component and collagen infiltration. The status of newly formed bone and the resorption of the graft material were evaluated at 6 weeks in vivo histologically and three dimensionally by means of 3D microcomputed tomography. The histologic observation showed that newly formed bone ingrowth and internal resorption of the block were observed for the experimental blocks, whereas for the control blocks less bone ingrowth occurred along with lower resorption rate of the block material. The three-dimensional observation indicated that the experimental block maintained the external geometry, but at the same time successfully altered the graft material into bone. It is suggested that the combination of numerous factors contributed to the bone ingrowth and the novel development could be an alternative bone grafting choice.


Hybrid stochastic simulations of intracellular reaction-diffusion systems.
With the observation that stochasticity is important in biological systems, chemical kinetics have begun to receive wider interest. While the use of Monte Carlo discrete event simulations most accurately capture the variability of molecular species, they become computationally costly for complex reaction-diffusion systems with large populations of molecules. On the other hand, continuous time models are computationally efficient but they fail to capture any variability in the molecular species. In this study a hybrid stochastic approach is introduced for simulating reaction-diffusion systems. We developed an adaptive partitioning strategy in which processes with high frequency are simulated with deterministic rate-based equations, and those with low frequency using the exact stochastic algorithm of Gillespie. Therefore the stochastic behavior of cellular pathways is preserved while being able to apply it to large populations of molecules. We describe our method and demonstrate its accuracy and efficiency compared with the Gillespie algorithm for two different systems. First, a model of intracellular viral kinetics with two steady states and second, a compartmental model of the postsynaptic spine head for studying the dynamics of Ca+2 and NMDA receptors.


Downstaging chemotherapy and alteration in the classic computed tomography/magnetic resonance imaging signs of vascular involvement in patients with pancreaticobiliary malignant tumors: influence on patient selection for surgery.
To determine whether computed tomography (CT)/magnetic resonance imaging (MRI) signs of vascular involvement are accurate after downstaging chemotherapy (DCTx) and to highlight factors associated with survival in patients who have undergone resection. Retrospective cohort study; prospective database. University pancreatic disease center. Patients with unresectable pancreaticobiliary cancer who underwent curative intent surgery after completing DCTx. Use of CT/MRI scan, pancreatic resection, and palliative bypass. Resectability after DCTx and disease-specific survival. We operated on 41 patients (1992-2009) with locally advanced periampullary malignant tumors after a median of 8.5 months of DCTx. Before DCTx, most patients (38 [93%]) were unresectable because of evidence of vascular contact on CT/MRI scan or operative exploration. Criteria for exploration after DCTx were CT/MRI evidence of tumor shrinkage and/or change in signs of vascular involvement, cancer antigen 19-9 decrease, and good functional status. None had progressive disease. At operation, we resected tumors in 34 of 41 patients (83%), and 6 had persistent vascular involvement. Surprisingly, CT/MRI scan was only 71% sensitive and 58% specific to detect vascular involvement after DCTx. "Involvement" on imaging was often from tumor fibrosis rather than viable cancer. Radiographic decrease in tumor size also did not predict resectability (P = .10). Patients with tumors that were resected had a median 87% decrease in cancer antigen 19-9 (P = .04) during DCTx. The median follow-up (all survivors) was 31 months, and disease-specific survival was 52 months for patients with resected tumors. In patients with initially unresectable periampullary malignant tumors, original CT/MRI signs of vascular involvement may persist after successful DCTx. Patients should be chosen for surgery on the basis of lack of disease progression, good functional status, and decrease in cancer antigen 19-9.


Key Elements of a Family Intervention for Schizophrenia: A Qualitative Analysis of an RCT.
Schizophrenia is a complex biopsychosocial condition in which expressed emotion in family members is a robust predictor of relapse. Not surprisingly, family interventions are remarkably effective and thus recommended in current treatment guidelines. Their key elements seem to be common therapeutic factors, followed by education and coping skills training. However, few studies have explored these key elements and the process of the intervention itself. We conducted a qualitative and quantitative analysis of the records from a pioneering family intervention trial addressing expressed emotion, published by Leff and colleagues four decades ago. Records were analyzed into categories and data explored using descriptive statistics. This was complemented by a narrative evaluation using an inductive approach based on emotional markers and markers of change. The most used strategies in the intervention were addressing needs, followed by coping skills enhancement, advice, and emotional support. Dealing with overinvolvement and reframing were the next most frequent. Single-family home sessions seemed to augment the therapeutic work conducted in family groups. Overall the intervention seemed to promote cognitive and emotional change in the participants, and therapists were sensitive to the emotional trajectory of each subject. On the basis of our findings, we developed a longitudinal framework for better understanding the process of this treatment approach.


Real-World Study of the Incidence, Risk Factors, and Prognostic Factors Associated with Bone Metastases in Women with Uterine Cervical Cancer Using Surveillance, Epidemiology, and End Results (SEER) Data Analysis.
BACKGROUND The aims of this study were to investigate the incidence and risk factors for the development of bone metastases and prognosis in women with cancer of the uterine cervix using database analysis. MATERIAL AND METHODS The National Cancer Institute (NCI) Surveillance, Epidemiology, and End Results (SEER) database was analyzed for the incidence and survival rates of women diagnosed with uterine cervical cancer in the United States between 2010-2015. Multivariate logistic regression analysis identified risk factors for bone metastases. Kaplan-Meier analysis estimated the overall survival. Proportional hazard regression analysis estimated prognostic factors associated with bone metastases. RESULTS There were 19,363 women with uterine cervical cancer, and 469 women were diagnosed with bone metastases on initial diagnosis (2.42%). Increased T-stage, N-stage, non-squamous and non-adenocarcinoma histology, high-grade tumors, and the presence of lung, liver, and brain metastases were all significantly associated with early bone metastases. There were 364 patients with cervical cancer and bone metastases on initial diagnosis who were followed-up for at least one year. Multivariate Cox regression analysis showed that unmarried status and lung, liver, and brain metastases were significantly associated with reduced overall survival. No other significant risk or prognostic associations were found. CONCLUSIONS SEER data analysis showed that women with uterine cervical cancer had some standard risk factors associated with bone metastases, and with prognosis, but a heterogeneous group of risk factors was also present. The findings of this study may have clinical application in screening for bone metastases in women with cervical cancer.


Rewarding yet demanding: client perspectives on enabling occupations during early stages of recovery from schizophrenia.
The purpose of this study, by exploring client perspectives, was to achieve a better understanding of how people with schizophrenia experience an occupational therapy intervention designed to enable them to carry out meaningful occupations in the early phases of recovery. A qualitative design comprising an eight-week client-centred occupational therapy intervention with semi-structured interviews with five of the six clients out of 10 who completed the intervention. Braun and Clark's thematic analysis was applied to the transcripts. Adherence rate and dropouts were recorded in a logbook. The Canadian Occupational Performance Measure was used to initiate and guide the intervention and the Canadian Model of Client-Centred Enablement for the client-therapist relationship. Participants described the intervention, which presupposed a certain level of patient readiness, as demanding. Participants valued engaging in real-life occupations while anchoring new strategies but also the occupational therapist's role in dealing with failure. Participants felt the intervention assisted in their recovery process and enabled them to engage in meaningful occupations. The study provided unique insight into how participants experienced a client-centred partnership with an occupational therapist in the early phases of recovery. The intervention was feasible and supported the participants' recovery process.


Impact of multimodal interventions on medication nonadherence among elderly hypertensives: a randomized controlled study.
Purpose: The study was conducted to investigate the effects of multimodal interventions on medication nonadherence, quality of life (QoL), hypertension (HTN), self-efficacy, and clinical outcome in terms of blood pressure (BP) among elderly people with HTN. Methods: An experimental design using a randomized controlled trial was adopted (N=80+80). The experimental group received multimodal interventions and the control group received routine care. Both groups were followed up at baseline and at 3 and 6 months. The data collection tools of demographic and clinical proforma, structured knowledge questionnaire on HTN (r=0.84), Revised Medication Adherence Self-Efficacy Scale (r=0.94), Morisky Medication Adherence Scale (r=0.83), World Health Organization Quality of Life-BREF scale (r=0.87), and digital BP apparatus were used. Necessary administrative permission was obtained for the study. Results: The study results proved that nurse-led multimodal interventions led to an improvement in medication adherence [F(1.75,214.30)=774.18, p<0.001], knowledge on HTN [F(2,244)=43.83, p<0.001], and self-efficacy [F(1,122)=3.99, p=0.04] of elderly people on antihypertensives over a period of 6 months. Overall QoL did not exhibit any statistically significant improvement, and no statistically significant reductions in the systolic BP (SBP) and diastolic BP (DBP) scores were obtained (p>0.05) in the experimental group over a period of 6 months. However, the clinical significance of multimodal interventions for improvements in medication adherence, QoL, knowledge on HTN, and self-efficacy was more favorable compared with the reduction in SBP and DBP scores. Conclusion: Nurses play a crucial role in improving medication adherence among elderly people with HTN. Trial details: Ethical clearance was obtained (IEC no. KH IEC 253/2012) from the Institutional Ethical Committee of Manipal University, Manipal, and the study was conducted in accordance with the Declaration of Helsinki. The study is registered under Clinical Trials Registry of India (CTRI/2017/04/008405). Informed consent was obtained from participants, and the confidentiality of information was assured.


Dietary fish oil reduces survival and impairs bacterial clearance in C3H/Hen mice challenged with Listeria monocytogenes.
1. To investigate the effect of dietary fat source on host resistance to intracellular pathogens, weanling female C3H/Hen mice were fed one of three experimental diets containing, 20% by weight, lard, soybean oil or 17% menhaden fish oil plus 3% corn oil. After 4 weeks, survival of mice (n = 12/treatment group) injected intraperitoneally with 2 x 10(6) colony forming units of live Listeria monocytogenes was determined. In a second study, bacterial clearance from the liver and spleen at 2, 4 and 7 days post-challenge was determined (n = 8/treatment group). 2. We found that the survival of mice fed the diets with soybean oil or menhaden fish oil was significantly lower than those fed lard (P < 0.05). Survival rates were 58% (7/12), 33% (4/12) and 100% (12/12), respectively, for mice fed soybean oil, menhaden fish oil and lard. In the second study, mice fed menhaden fish oil had approximately 1 log10 greater bacteria in their spleens at day 4 than mice fed lard or soybean oil (P < 0.001). There were no significant treatment differences in the number of bacteria recovered from liver samples. 3. In summary, dietary fat source significantly affects murine resistance to Listeria, with diets rich in n-3 polyunsaturated fatty acids, such as from fish oil, having the most detrimental effect.


Lung cancer in relation to exposure to silica dust, silicosis and uranium production in South African gold miners.
A nested case-control study for lung cancer was performed on a cohort of 2260 South African gold miners in whom an association between exposure to silica dust and risk of lung cancer was previously reported. The objective was to investigate an expanded set of risk factors and also cancer cell type. The 78 cases of lung cancer found during the follow up period from 1970 to 1986 were matched with 386 controls. Risk of lung cancer was related to smoking, exposure to silica dust, incidence of silicosis, and uranium production and the uranium content of the mine ore. The risk of lung cancer was associated with tobacco smoking, cumulative dust exposure, duration of underground mining, and with silicosis. The best predictive model included pack years of cigarette consumption (adjusted relative risk (RR) = 1.0 for < 6.5 pack years, 3.5 (95% confidence interval (CI) 0.7 to 16.8) for 6.5-20 pack years, 5.7 (95% CI 1.3 to 25.8) for 21-30 pack years, and 13.2 (95% CI 3.1 to 56.2) for more than 30 pack years) and silicosis (RR = 2.45 (95% CI 1.2 to 5.2)). No association was found with uranium production. The lung tumour cell type distribution was 40.3% small cell carcinoma, 38.8% squamous cell, 16.4% adenocarcinoma, and 4.5% large cell carcinoma. Small and large cell cancer combined were associated with exposure to dust. The results cannot be interpreted definitively in terms of causal association. Possible interpretations are: (1) subjects with high dust exposure who develop silicosis are at increased risk of lung cancer; (2) high levels of exposure to silica dust on its own is important in the pathogenesis of lung cancer and silicosis is coincidental; and (3) high levels of silica dust exposure may be a surrogate for the exposure to radon daughters.


Evidence for baseline retinal pigment epithelium pathology in the Trp1-Cre mouse.
The increasing popularity of the Cre/loxP recombination system has led to the generation of numerous transgenic mouse lines in which Cre recombinase is expressed under the control of organ- or cell-specific promoters. Alterations in retinal pigment epithelium (RPE), a multifunctional cell monolayer that separates the retinal photoreceptors from the choroid, are prevalent in the pathogenesis of a number of ocular disorders, including age-related macular degeneration. To date, six transgenic mouse lines have been developed that target Cre to the RPE under the control of various gene promoters. However, multiple lines of evidence indicate that high levels of Cre expression can be toxic to mammalian cells. In this study, we report that in the Trp1-Cre mouse, a commonly used transgenic Cre strain for RPE gene function studies, Cre recombinase expression alone leads to RPE dysfunction and concomitant disorganization of RPE layer morphology, large areas of RPE atrophy, retinal photoreceptor dysfunction, and microglial cell activation in the affected areas. The phenotype described herein is similar to previously published reports of conditional gene knockouts that used the Trp1-Cre mouse, suggesting that Cre toxicity alone could account for some of the reported phenotypes and highlighting the importance of the inclusion of Cre-expressing mice as controls in conditional gene targeting studies.


CUDASW++: optimizing Smith-Waterman sequence database searches for CUDA-enabled graphics processing units.
The Smith-Waterman algorithm is one of the most widely used tools for searching biological sequence databases due to its high sensitivity. Unfortunately, the Smith-Waterman algorithm is computationally demanding, which is further compounded by the exponential growth of sequence databases. The recent emergence of many-core architectures, and their associated programming interfaces, provides an opportunity to accelerate sequence database searches using commonly available and inexpensive hardware. Our CUDASW++ implementation (benchmarked on a single-GPU NVIDIA GeForce GTX 280 graphics card and a dual-GPU GeForce GTX 295 graphics card) provides a significant performance improvement compared to other publicly available implementations, such as SWPS3, CBESW, SW-CUDA, and NCBI-BLAST. CUDASW++ supports query sequences of length up to 59K and for query sequences ranging in length from 144 to 5,478 in Swiss-Prot release 56.6, the single-GPU version achieves an average performance of 9.509 GCUPS with a lowest performance of 9.039 GCUPS and a highest performance of 9.660 GCUPS, and the dual-GPU version achieves an average performance of 14.484 GCUPS with a lowest performance of 10.660 GCUPS and a highest performance of 16.087 GCUPS. CUDASW++ is publicly available open-source software. It provides a significant performance improvement for Smith-Waterman-based protein sequence database searches by fully exploiting the compute capability of commonly used CUDA-enabled low-cost GPUs.


Codes in the codons: construction of a codon/amino acid periodic table and a study of the nature of specific nucleic acid-protein interactions.
The theory of "codon-amino acid coevolution" was first proposed by Woese in 1967. It suggests that there is a stereochemical matching - that is, affinity - between amino acids and certain of the base triplet sequences that code for those amino acids. We have constructed a common periodic table of codons and amino acids, where the nucleic acid table showed perfect axial symmetry for codons and the corresponding amino acid table also displayed periodicity regarding the biochemical properties (charge and hydrophobicity) of the 20 amino acids and the position of the stop signals. The table indicates that the middle (2/sup nd/) amino acid in the codon has a prominent role in determining some of the structural features of the amino acids. The possibility that physical contact between codons and amino acids might exist was tested on restriction enzymes. Many recognition site-like sequences were found in the coding sequences of these enzymes and as many as 73 examples of codon-amino acid co-location were observed in the 7 known 3D structures (December 2003) of endonuclease-nucleic acid complexes. These results indicate that the smallest possible units of specific nucleic acid-protein interaction are indeed the stereochemically compatible codons and amino acids.


Shugoshin promotes sister kinetochore biorientation in Saccharomyces cerevisiae.
Chromosome segregation must be executed accurately during both mitotic and meiotic cell divisions. Sgo1 plays a key role in ensuring faithful chromosome segregation in at least two ways. During meiosis this protein regulates the removal of cohesins, the proteins that hold sister chromatids together, from chromosomes. During mitosis, Sgo1 is required for sensing the absence of tension caused by sister kinetochores not being attached to microtubules emanating from opposite poles. Here we describe a differential requirement for Sgo1 in the segregation of homologous chromosomes and sister chromatids. Sgo1 plays only a minor role in segregating homologous chromosomes at meiosis I. In contrast, Sgo1 is important to bias sister kinetochores toward biorientation. We suggest that Sgo1 acts at sister kinetochores to promote their biorientation.


The notch ligand delta-like 4 regulates multiple stages of early hemato-vascular development.
In mouse embryos, homozygous or heterozygous deletions of the gene encoding the Notch ligand Dll4 result in early embryonic death due to major defects in endothelial remodeling in the yolk sac and embryo. Considering the close developmental relationship between endothelial and hematopoietic cell lineages, which share a common mesoderm-derived precursor, the hemangioblast, and many key regulatory molecules, we investigated whether Dll4 is also involved in the regulation of early embryonic hematopoiesis. Using Embryoid Bodies (EBs) derived from embryonic stem cells harboring hetero- or homozygous Dll4 deletions, we observed that EBs from both genotypes exhibit an abnormal endothelial remodeling in the vascular sprouts that arise late during EB differentiation, indicating that this in vitro system recapitulates the angiogenic phenotype of Dll4 mutant embryos. However, analysis of EB development at early time points revealed that the absence of Dll4 delays the emergence of mesoderm and severely reduces the number of blast-colony forming cells (BL-CFCs), the in vitro counterpart of the hemangioblast, and of endothelial cells. Analysis of colony forming units (CFU) in EBs and yolk sacs from Dll4(+/-) and Dll4(-/-) embryos, showed that primitive erythropoiesis is specifically affected by Dll4 insufficiency. In Dll4 mutant EBs, smooth muscle cells (SMCs) were seemingly unaffected and cardiomyocyte differentiation was increased, indicating that SMC specification is Dll4-independent while a normal dose of this Notch ligand is essential for the quantitative regulation of cardiomyogenesis. This study highlights a previously unnoticed role for Dll4 in the quantitative regulation of early hemato-vascular precursors, further indicating that it is also involved on the timely emergence of mesoderm in early embryogenesis.


DNA methylome reveals cellular origin of cell-free DNA in spent medium of human preimplantation embryos.
The discovery of embryonic cell-free DNA (cfDNA) in spent embryo culture media (SECM) has brought hope for noninvasive preimplantation genetic testing. However, the cellular origins of SECM cfDNA are not sufficiently understood, and methods for determining maternal DNA contamination are limited. Here, we performed whole-genome DNA methylation sequencing for SECM cfDNA. Our results demonstrated that SECM cfDNA was derived from blastocysts, cumulus cells, and polar bodies. We identified the cumulus-specific differentially methylated regions (DMRs) and oocyte/polar body-specific DMRs, and established an algorithm for deducing the cumulus, polar body, and net maternal DNA contamination ratios in SECM. We showed that DNA methylation sequencing accurately detected chromosome aneuploidy in SECM and distinguished SECM samples with low and high false negative rates and gender discordance rates, after integrating the origin analysis. Our work provides insights into the characterization of embryonic DNA in SECM and provides a perspective for noninvasive preimplantation genetic testing in reproductive medicine.


Why pain hurts.
We have made great strides in understanding how the human brain constructs the multidimensional experience of pain - both acute and chronic - over the past few decades. Pain wears many guises, but at its core, it hurts. How is this core component of pain represented in the brain, and how can we target it for relief?