Ether phospholipids and glycosylinositolphospholipids are not required for amastigote virulence or for inhibition of macrophage activation by Leishmania major.
Ether phospholipids are major components of the membranes of humans and Leishmania. In protozoan parasites they occur separately or as part of the glycosylphosphatidylinositol (GPI) anchor of molecules implicated in virulence, such as lipophosphoglycan (LPG), smaller glycosylinositolphospholipids (GIPLs), and GPI-anchored proteins. We generated null mutants of the Leishmania major alkyldihydroxyacetonephosphate synthase (ADS), the first committed step of ether lipid synthesis. Enzymatic analysis and comprehensive mass spectrometric analysis showed that ads1- knock-outs lacked all ether phospholipids, including plasmalogens, LPG, and GIPLs. Leishmania ads1- thus represents the first ether lipid-synthesizing eukaryote for which a completely null mutant could be obtained. Remarkably ads1- grew well and maintained lipid rafts (detergent-resistant membranes). In virulence tests it closely resembled LPG-deficient L. major, including sensitivity to complement and an inability to survive the initial phase of macrophage infection. Likewise it retained the ability to inhibit host cell signaling and to form infectious amastigotes from the few parasites surviving the establishment defect. These findings counter current proposals that GIPLs are required for amastigote survival in the mammalian host or that parasite lyso-alkyl or alkylacyl-GPI anchors are solely responsible for inhibition of macrophage activation.


Blockchain Technology Secures Robot Swarms: A Comparison of Consensus Protocols and Their Resilience to Byzantine Robots.
Consensus achievement is a crucial capability for robot swarms, for example, for path selection, spatial aggregation, or collective sensing. However, the presence of malfunctioning and malicious robots (Byzantine robots) can make it impossible to achieve consensus using classical consensus protocols. In this work, we show how a swarm of robots can achieve consensus even in the presence of Byzantine robots by exploiting blockchain technology. Bitcoin and later blockchain frameworks, such as Ethereum, have revolutionized financial transactions. These frameworks are based on decentralized databases (blockchains) that can achieve secure consensus in peer-to-peer networks. We illustrate our approach in a collective sensing scenario where robots in a swarm are controlled via blockchain-based smart contracts (decentralized protocols executed via blockchain technology) that serve as "meta-controllers" and we compare it to state-of-the-art consensus protocols using a robot swarm simulator. Additionally, we show that our blockchain-based approach can prevent attacks where robots forge a large number of identities (Sybil attacks). The developed robot-blockchain interface is released as open-source software in order to facilitate future research in blockchain-controlled robot swarms. Besides increasing security, we expect the presented approach to be important for data analysis, digital forensics, and robot-to-robot financial transactions in robot swarms.


Incidence and classification of new-onset epilepsy and epilepsy syndromes in children in Olmsted County, Minnesota from 1980 to 2004: a population-based study.
To determine the incidence and classification of new-onset epilepsy, as well as the distribution of epilepsy syndromes in a population-based group of children, using the newly proposed Report of the ILAE Commission on Classification and Terminology 2005-2009. We identified all children residing in Olmsted County, MN, 1 month through 17 years with newly diagnosed epilepsy from 1980 to 2004. For each patient, epilepsy was classified into mode of onset, etiology, and syndrome or constellation (if present). Incidence rates were calculated overall and also separately for categories of mode of onset and etiology. The adjusted incidence rate of new-onset epilepsy in children was 44.5 cases per 100,000 persons per year. Incidence rates were highest in the first year of life and diminished with age. Mode of onset was focal in 68%, generalized/bilateral in 23%, spasms in 3% and unknown in 5%. Approximately half of children had an unknown etiology for their epilepsy, and of the remainder, 78 (22%) were genetic and 101 (28%) were structural/metabolic. A specific epilepsy syndrome could be defined at initial diagnosis in 99/359 (28%) children, but only 9/359 (3%) had a defined constellation. Nearly half of childhood epilepsy is of "unknown" etiology. While a small proportion of this group met criteria for a known epilepsy syndrome, 41% of all childhood epilepsy is of "unknown" cause with no clear syndrome identified. Further work is needed to define more specific etiologies for this group.


Theory of carbomorph cycles.
We present a theory of a reversibly deforming sp2-carbon-based system controlled by competing strain, surface, and electrostatic energies, a carbomorph. For example, external forces (such as electrostatic, chemical, interfacial) could convert a bistable carbon nanotube between the collapsed and inflated states. Such a system could operate as a voltage-controlled constant-force spring, a charge-controlled harmonic spring, or an electromechanical engine or generator (with linear stroke up to few microns) driven across a propagating quasi-one-dimensional structural phase transition.


Efficacy of Novel Topical Liposomal Formulation of Cyclosporine in Mild to Moderate Stable Plaque Psoriasis: A Randomized Clinical Trial.
Attempts to use topical cyclosporine in treatment of psoriasis have failed because of unfavorable physicochemical properties and inappropriate formulation design of the conventional dosage forms. To evaluate the efficacy of topical cyclosporine using liposomal nanocarriers (lipogel) in limited chronic plaque psoriasis. A single-center randomized clinical trial was conducted using a 3-arm parallel group, double-blind, vehicle and active comparator design and included 38 patients with chronic plaque psoriasis measuring less than or equal to 100 cm2 performed in a tertiary care hospital. In the first arm, a total of 24 patients were randomized to receive either cyclosporine lipogel, 2.0% weight by weight (w/w), or placebo lipogel. In the second arm, 7 patients were randomized to receive cyclosporine lipogel, 2.0%, or conventional cyclosporine cream, 2.0% w/w. The third arm comprised 7 patients randomized with cyclosporine lipogel, 2.0% or standard clobetasol propionate cream, 0.05% w/w. Patients were examined twice weekly for 14 weeks, or until total lesional clearance was observed, whichever was earlier. The primary outcome measure was the dermatological sum score (DSS) assessing erythema, scaling, and plaque elevation on a 4-point scale (0, absent; 1, minimal; 2, moderate; 3, severe). In 38 patients (23 men and 15 women with a mean [SD] age range from 35 [8] to 40 [13] years), a 19% decrease in DSS score from a mean (SD) of 8.45 (0.67) to 6.82 (0.77) compared with baseline was observed after 2 weeks of treatment with cyclosporine lipogel, 2.0% w/w (P < .001; 95% CI, 13.77-24.51) in 59% of psoriasis lesional sites. At the end of the eighth week, a significant reduction (approximately 83%) in DSS was seen in all sites treated with cyclosporine lipogel, (P < .001; 95% CI, 77.48-88.22). At the end of the study period, complete clearance (ie, DSS = 0) was observed in 16 (41%) psoriasis lesional sites treated with cyclosporine lipogel, 85.7% of sites treated with clobetasol propionate cream, and none of the sites treated with conventional cyclosporine cream or placebo gel. Topical liposomal formulation of cyclosporine, 2.0% w/w, is effective in treatment of limited chronic plaque psoriasis with a satisfactory safety profile. Future clinical trials should assess liposomal cyclosporine in larger study populations. Clinical Trials Registry-India Identifier: CTRI/2011/12/002307.


Phosphorylation Sites of the Gastric Inhibitory Polypeptide Receptor (GIPR) Revealed by Trapped-Ion-Mobility Spectrometry Coupled to Time-of-Flight Mass Spectrometry (TIMS-TOF MS).
The gastric inhibitory polypeptide receptor (GIPR), a G protein-coupled receptor (GPCR) that regulates glucose metabolism and insulin secretion, is a target for the development of therapeutic agents to address type 2 diabetes and obesity. Signal transduction processes mediated by GPCR activation typically result in receptor phosphorylation, but very little is known about GIPR phosphorylation. Mass spectrometry (MS) is a powerful tool for detecting phosphorylation and other post-translational modifications of proteins and for identifying modification sites. However, applying MS methods to GPCRs is challenging because the native expression levels are low and the hydrophobicity of these proteins complicates isolation and enrichment. Here we use a widely available technique, trapped-ion-mobility spectrometry coupled to time-of-flight mass spectrometry (TIMS-TOF MS), to characterize the phosphorylation status of the GIPR. We identified eight serine residues that are phosphorylated, one in an intracellular loop and the remainder in the C-terminal domain. Stimulation with the native agonist GIP enhanced phosphorylation at four of these sites. For comparison, we evaluated tirzepatide (TZP), a dual agonist of the glucagon-like peptide-1 (GLP-1) receptor and the GIPR that has recently been approved for the treatment of type 2 diabetes. Stimulation with TZP enhanced phosphorylation at the same four sites that were enhanced with GIP; however, TZP also enhanced phosphorylation at a fifth site that is unique to this synthetic agonist. This work establishes an important and accessible tool for the characterization of signal transduction via the GIPR and reveals an unanticipated functional difference between GIP and TZP.


Synthesis of nano-sized arsenic-imprinted polymer and its use as As(3+) selective ionophore in a potentiometric membrane electrode: part 1.
In this study, a new strategy was proposed for the preparation of As (III)-imprinted polymer by using arsenic (methacrylate)3 as template. Precipitation polymerization was utilized to synthesize nano-sized As (III)-imprinted polymer. Methacrylic acid and ethylene glycol dimethacrylate were used as the functional monomer and cross-linking agent, respectively. In order to assembly functional monomers around As (III) ion, sodium arsenite and methacrylic acid were heated in the presence of hydroquinone, leading to arsenic (methacrylate)3. The nano-sized As (III) selective polymer was characterized by FT-IR and scanning electron microscopy techniques (SEM). It was demonstrated that arsenic was recognized as As(3+) by the selective cavities of the synthesized IIP. Based on the prepared polymer, the first arsenic cation selective membrane electrode was introduced. Membrane electrode was constructed by dispersion of As (III)-imprinted polymer nanoparticles in poly(vinyl chloride), plasticized with di-nonylphthalate. The IIP-modified electrode exhibited a Nernstian response (20.4±0.5 mV decade(-1)) to arsenic ion over a wide concentration range (7.0×10(-7) to 1.0×10(-1) mol L(-1)) with a lower detection limit of 5.0×10(-7) mol L(-1). Unlike this, the non-imprinted polymer (NIP)-based membrane electrode was not sensitive to arsenic in aqueous solution. The selectivity of the developed sensor to As (III) was shown to be satisfactory. The sensor was used for arsenic determination in some real samples.


Collaborative quality improvement in the cardiac intensive care unit: development of the Paediatric Cardiac Critical Care Consortium (PC4).
Despite many advances in recent years for patients with critical paediatric and congenital cardiac disease, significant variation in outcomes remains across hospitals. Collaborative quality improvement has enhanced the quality and value of health care across specialties, partly by determining the reasons for variation and targeting strategies to reduce it. Developing an infrastructure for collaborative quality improvement in paediatric cardiac critical care holds promise for developing benchmarks of quality, to reduce preventable mortality and morbidity, optimise the long-term health of patients with critical congenital cardiovascular disease, and reduce unnecessary resource utilisation in the cardiac intensive care unit environment. The Pediatric Cardiac Critical Care Consortium (PC4) has been modelled after successful collaborative quality improvement initiatives, and is positioned to provide the data platform necessary to realise these objectives. We describe the development of PC4 including the philosophical, organisational, and infrastructural components that will facilitate collaborative quality improvement in paediatric cardiac critical care.


How the ACA's Health Insurance Expansions Have Affected Out-of-Pocket Cost-Sharing and Spending on Premiums.
One important benefit gained by the millions of Americans with health insurance through the Affordable Care Act (ACA) is protection from high out-of-pocket health spending. While Medicaid unambiguously reduces out-of-pocket premium and medical costs for low-income people, it is less certain that marketplace coverage and other types of insurance purchased to comply with the law's individual mandate also protect from high health spending. Goal: To compare out-of-pocket spending in 2014 to spending in 2013; assess how this spending changed in states where many people enrolled in the marketplaces relative to states where few people enrolled; and project the decline in the percentage of people paying high amounts out-of-pocket. Methods: Linear regression models were used to estimate whether people under age 65 spent above certain thresholds. Key findings and conclusions: The probability of incurring high out-of-pocket costs and premium expenses declined as marketplace enrollment increased. The percentage reductions were greatest among those with incomes between 250 percent and 399 percent of poverty, those who were eligible for premium subsidies, and those who previously were uninsured or had very limited nongroup coverage. These effects appear largely attributable to marketplace enrollment rather than to other ACA provisions or to economic trends.


Influence of lipophilicity on the interactions of N-alkyl-4-phenyl-1,2,3,6-tetrahydropyridines and their positively charged N-alkyl-4-phenylpyridinium metabolites with cytochrome P450 2D6.
The relationship between lipophilicity and CYP2D6 affinity of cyclic tertiary (N-alkyl-4-phenyl-1,2,3,6-tetrahydropyridines) and quaternary (N-alkyl-4-phenylpyridinium) amines was examined. The 1,2,3,6-tetrahydropyridine scaffold was chosen due to its common occurrence in the structures of CYP2D6 ligands such as the Parkinsonian neurotoxin 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) and the dehydrated haloperidol metabolite N-[4-(4-fluorophenyl)-4-oxobutyl]-4-(4-chlorophenyl)-1,2,3,6-tetrahydropyridine (HPTP). Likewise, the pyridinium framework is found in and 4-(4-chlorophenyl)-1-[4-(4-fluorophenyl)-4-oxobutyl]pyridinium and N-methyl-4-phenylpyridinium (MPP(+)), the positively charged metabolites of MPTP and haloperidol. The lack of CYP2D6 inhibition by MPTP and its pyridinium metabolite MPP(+) was due to their hydrophilic nature since higher N-alkyl homologs revealed substantial increases in inhibitory potency against recombinant CYP2D6-mediated bufuralol-1'-hydroxylation. The reasonable correlation between lipophilicity and CYP2D6 inhibition by pyridiniums and 1,2,3,6-tetrahydropyridines was only limited to straight chain N-alkyl analogs, since certain N-alkylaryl analogs of lower lipophilicity were better CYP2D6 inhibitors. CYP2D6 substrate properties of straight chain N-alkyltetrahydropyridines were also governed by lipophilicity, and N-heptyl-4-phenyl-1,2,3,6-tetrahydropyridine was the optimal substrate (K(mapp) = 0.63 microM). Metabolism studies indicated that the N-heptyl analog underwent monohydroxylation on the aromatic ring and on the N-heptyl group suggesting that 1,2,3,6-tetrahydropyridines can bind in more than one conformation in the CYP2D6 active site. Increased lipophilicity of haloperidol metabolites did not correlate with inhibitory potency since the more lipophilic HPTP metabolite was less potent as an inhibitor than reduced-haloperidol and reduced-HPTP. Furthermore, HPTP and reduced-HPTP, of comparable lipophilicity to the N-heptyltetrahydropyridine analog were inactive as CYP2D6 substrates. This observation suggests that steric constraints rather than lipophilicity are responsible for the lack of CYP2D6 substrate properties of cyclic tertiary amines tethered to bulky N-substituents. This phenomenon appears to be a common theme among several cyclic tertiary amine-containing anti-depressants and should be taken into consideration when designing central nervous system agents devoid of CYP2D6 substrate properties.


[Alternating signal speech audiometry in the diagnosis of central lesions of the acoustic analyzer].
Diagnostic sensitivity was compared for two binaural methods of "sensitized" speech audiometry--alternating speech audiometry (ASA) and Dichotic. A total of 35 patients were examined having neurosensory hypoacusis resultant from cerebrovascular atherosclerosis and chronic insufficiency of cerebral circulation. ASA indicated defects of the central parts of the acoustic analyzer in 80% of the examinees. The results of the two methods were similar in 85.7% of the patients. The diotic and dichotic alternating speech tests can be used separately and in combination in diagnosis of central acoustic disturbances, of vascular genesis, in particular.


Detection of quantitative trait loci in Danish Holstein cattle affecting clinical mastitis, somatic cell score, udder conformation traits, and assessment of associated effects on milk yield.
The aim of this study was to 1) detect QTL across the cattle genome that influence the incidence of clinical mastitis and somatic cell score (SCS) in Danish Holsteins, and 2) characterize these QTL for pleiotropy versus multiple linked quantitative trait loci (QTL) when chromosomal regions affecting clinical mastitis were also affecting other traits in the Danish udder health index or milk production traits. The chromosomes were scanned using a granddaughter design where markers were typed for 19 to 34 grandsire families and 1,373 to 2,042 sons. A total of 356 microsatellites covering all 29 autosomes were used in the scan. Among the across-family regression analyses, 16 showed chromosome-wide significance for the primary traits incidence of clinical mastitis in first (CM1), second (CM2), and third (CM3) lactations, and SCS. Regions of chromosomes 5, 6, 9, 11, 15, and 26 were found to affect CM and regions of chromosomes 5, 6, 8, 13, 22, 23, 24, and 25 affected SCS. Markers on chromosomes 6, 11, 15, and 26 can be used to perform marker-assisted selection on CM without a direct negative selection on milk yield, because no effects were detected on the milk traits. Comparing multi-trait models assuming either a pleiotropic QTL affecting 2 traits or 2 QTL each affecting 1 trait gave some evidence to distinguish between these models. For Bos taurus autosome 5, the most likely models were a pleiotropic QTL affecting CM2, CM3, and SCS, and a linked QTL affecting fat yield index. For Bos taurus autosome 9, the most likely model is a pleiotropic QTL affecting CM1 and CM2 at approximately 8 cM.


VHL mutations and dysregulation of pVHL- and PTEN-controlled pathways in multilocular cystic renal cell carcinoma.
Multilocular cystic renal cell carcinoma is a rare renal cell carcinoma with an excellent prognosis. To clarify the relationship with typical clear cell renal cell carcinoma, we evaluated 15 cases of multilocular cystic renal cell carcinomas diagnosed according to the 2004 WHO classification. Von Hippel Lindau (VHL) gene mutations were determined by whole genome amplification and direct sequencing. Carbonic anhydrase 9 (CAIX), a hypoxia-inducible factor (HIF) target, paired box gene 2 (PAX2), cyclin-dependent kinase inhibitor p27 and glycogen synthase kinase 3-β (GSK3β) were immunohistochemically evaluated as members of the VHL protein (pVHL)- and phosphatase and tensin homolog (PTEN)-controlled pathways. VHL mutations were identified in 3 of 12 (25%) tumors. Inactivated GSK3β, decreased PTEN expression and PAX2 positivity were observed in the vast majority of the multilocular cystic renal cell carcinomas. Strong nuclear staining of p27 was seen in 14 of 15 cases. Compared with multilocular cystic renal cell carcinomas, expression frequencies of PAX2, p-GSK3β, PTEN and CAIX were similar in a set of low-grade, early-stage clear cell renal cell carcinomas, whereas only 30% had strong p27 positivity. These results are consistent with the hypothesis that multilocular cystic renal cell carcinomas are related at the molecular level with clear cell renal cell carcinomas. Maintenance of a strong subcellular p27 expression in all multilocular cystic renal cell carcinomas analyzed may in part explain the excellent prognosis of these tumor patients.


Detecting common gene expression patterns in multiple cancer outcome entities.
Most oncological microarray studies focus on molecular distinctions in different cancer entities. Recently, researchers started using microarrays for investigating molecular commonalities of multiple cancer types. This poses novel bioinformatics challenges. In this paper we describe a method that detects common molecular mechanisms in different cancer entities. The method extends previously described concepts by introducing Meta-Analysis Pattern Matches. In an analysis of four prognostic cancer studies, involving breast cancer, leukemia, and mesothelioma, we are able to identify 42 genes that show consistent up- or down-regulation in patients with a poor disease outcome. These genes complement the set of previously published candidates for universal prognostic markers in cancer.


Susceptibility of Human Endogenous Retrovirus Type K to Reverse Transcriptase Inhibitors.
Human endogenous retroviruses (HERVs) make up 8% of the human genome. The HERV type K (HERV-K) HML-2 (HK2) family contains proviruses that are the most recent entrants into the human germ line and are transcriptionally active. In HIV-1 infection and cancer, HK2 genes produce retroviral particles that appear to be infectious, yet the replication capacity of these viruses and potential pathogenicity has been difficult to ascertain. In this report, we screened the efficacy of commercially available reverse transcriptase inhibitors (RTIs) at inhibiting the enzymatic activity of HK2 RT and HK2 genomic replication. Interestingly, only one provirus, K103, was found to encode a functional RT among those examined. Several nucleoside analogue RTIs (NRTIs) blocked K103 RT activity and consistently inhibited the replication of HK2 genomes. The NRTIs zidovudine (AZT), stavudine (d4T), didanosine (ddI), and lamivudine (3TC), and the nucleotide RTI inhibitor tenofovir (TDF), show efficacy in blocking K103 RT. HIV-1-specific nonnucleoside RTIs (NNRTIs), protease inhibitors (PIs), and integrase inhibitors (IIs) did not affect HK2, except for the NNRTI etravirine (ETV). The inhibition of HK2 infectivity by NRTIs appears to take place at either the reverse transcription step of the viral genome prior to HK2 viral particle formation and/or in the infected cells. Inhibition of HK2 by these drugs will be useful in suppressing HK2 infectivity if these viruses prove to be pathogenic in cancer, neurological disorders, or other diseases associated with HK2. The present studies also elucidate a key aspect of the life cycle of HK2, specifically addressing how they do, and/or did, replicate.IMPORTANCE Endogenous retroviruses are relics of ancestral virus infections in the human genome. The most recent of these infections was caused by HK2. While HK2 often remains silent in the genome, this group of viruses is activated in HIV-1-infected and cancer cells. Recent evidence suggests that these viruses are infectious, and the potential exists for HK2 to contribute to disease. We show that HK2, and specifically the enzyme that mediates virus replication, can be inhibited by a panel of drugs that are commercially available. We show that several drugs block HK2 with different efficacies. The inhibition of HK2 replication by antiretroviral drugs appears to occur in the virus itself as well as after infection of cells. Therefore, these drugs might prove to be an effective treatment by suppressing HK2 infectivity in diseases where these viruses have been implicated, such as cancer and neurological syndromes.


Household migration, social support, and psychosocial health: the perspective from migrant-sending areas.
An extensive literature demonstrates various negative health consequences of family disruption in Western societies, which is largely due to marital dissolution. In developing settings, family disruption commonly arises in the context of labor out-migration. However, studies on household emigration often focus on the economic benefits from remittances, overlooking emigration as a source of stress and loss of social support. This research examines the psychosocial consequences of internal out-migration using longitudinal survey data collected in Indonesia between 1993 and 2007. Results demonstrate considerable psychosocial costs of out-migration, with adults left behind by migrants more susceptible to stress-related health impairments such as hypertension and to psychological distress such as depressive symptoms. These findings largely hold when specific relations are investigated, including spouses left behind and parents left behind by adult children. This study also finds some support for the stress-buffering role of social support from extended families and the differential psychosocial processes for men and women.


Diagnostic value of rapid test for malaria among febrile neonates in a tertiary hospital in North-East Nigeria: a prospective cross-sectional study.
The WHO recommends testing using microscopy or rapid diagnostic test (RDT) before treatment for malaria. However, the use of RDT to diagnose neonatal malaria has not been widely validated with most studies limited to the first week of life. Thus, we conducted this study to determine the utility of RDT in the diagnosis of congenital and acquired malaria in febrile neonates in Nigeria. This prospective cross-sectional descriptive study consecutively recruited 131 febrile neonates at the Special Care Baby Unit (SCBU) of the Federal Teaching Hospital Gombe, Nigeria. All study participants concurrently had RDT (HRP2, LDH) and malaria microscopy. The performance of both methods was then compared. Seventy-eight of 131 neonates tested for malaria by blood smear microscopy demonstrated malaria parasites; a prevalence of 59.5%. Parasite count ranged from 16 to 520 /μL and the median parasite count was 81.0 /μL with IQR (40.0-134.5). The majority of patients (93.5%) had low-density parasitaemia (≤2+). All species identified were Plasmodium falciparum. None of the 131 neonates tested positive on RDT. The sensitivity and positive predictive value of RDT for neonatal malaria was zero. Congenital malaria was the most common form of neonatal malaria, accounting for 75.6%, while acquired and transfusion-related malaria were estimated at 12.8% and 11.6%, respectively. The RDT used in this study was not sensitive in the diagnosis of congenital or acquired neonatal malaria; therefore, microscopy remains the preferred method of diagnosis of neonatal malaria.


Acrosomal reaction of thyone sperm. I. Changes in the sperm head visualized by high resolution video microscopy.
Structural changes inside the head of Thyone sperm undergoing the acrosomal reaction were followed with a high-resolution, differential interference contrast (DIC) video microscope. The beating sperm, adhering by their midpiece to the cover slip of a wedge perfusion chamber, were activated by a calcium ionophore (20 microM A23187) suspended in sea water containing 50 mM excess CaCl2. Before activation of the sperm, the acrosomal region appears as a 1.1-microM diameter sphere, slightly less dense than the rest of the sperm head. Upon activation, the acrosome pops; the acrosomal region suddenly swells and its refractive index drops. After approximately 1 s, a crescent-shaped periacrosomal cup appears behind the acrosomal vacuole. In the next several seconds, the cup loses more refractive index and expands forward as the acrosomal process extends. The acrosomal vacuole becomes smaller, but without appreciable drop in refractive index. These observations, coupled with the behavior of the extending acrosomal process reported in the companion paper, and in electron microscopy (EM) and early physiological studies, suggest that the acrosomal process is extended by a combination of the explosive polymerization of actin and the osmotic swelling of the periacrosomal cup material. In this paper, we also consider the meaning of the enhanced DIC image seen in the high-resolution video microscope, and discuss the reliability of measurements on small linear dimensions made with the DIC microscope.


A serum factor for macrophage activation after in vitro dodecylglycerol treatment of mouse lymphocytes.
Alkylglycerols, inflammation products of cancerous tissues, are potent macrophage activating agents. A brief in vitro treatment (30 min) of mouse peritoneal cells with a low concentration (50 ng/mL) of dodecylglycerol (DDG) in 10% foetal calf serum supplemented RPMI-1640 medium (FCS medium) activates macrophages for Fc-receptor mediated ingestion activity. A serum factor(s) was shown to be required for the activation of macrophages. When non-adherent cells were treated with rac-sn-1(3)-dodecylglycerol (DDG) in a serum free-0.1% egg albumin supplemented RPMI medium (EA medium) for 30 min and cultured in FCS medium for 2 h, the resultant conditioned medium contained a signal factor able to activate macrophages (macrophage activating factor). A conditioned medium prepared with electrophoresed serum alpha 2-globulin fraction in EA medium markedly enhanced activation of macrophages. Incubation of DDG-treated non-adherent cell ghosts in EA medium containing alpha 2-globulin also produced the macrophage activating signal factor. Therefore, it is concluded that a serum factor in alpha 2-globulin fraction is processed by pre-existing functions or enzymes of DDG-treated non-adherent cell membrane to yield a macrophage activating signal factor.


Isolation, purification and characterization of naturally derived Crocetin beta-d-glucosyl ester from Crocus sativus L. against breast cancer and its binding chemistry with ER-alpha/HDAC2.
Saffron plant (Crocus sativus L.) is being used as a source of saffron spice and medicine to cure or prevent different types of diseases including cancers. We report the isolation, characterization of bioactive small molecule ([crocetin (β-d-glucosyl) ester] from the leaf biowastes of saffron plant of Kashmir, India. MTTC assay and Bio-autography aided approach were used to assess anti-oxidant activity and anti-cancer properties of crocin (s) against DPPH free radical and breast cancer cell line respectively. Crocetin beta-d-glucosyl ester restrained proliferation of human breast adeno-carcinoma cell model (MCF-7) without significantly affecting normal cell line (L-6). Further studies involving molecular mechanics generalized born surface area and molecular docking showed that crocetin beta-d-glucosyl ester exhibits strong affinity for estrogen receptor alpha and histone deacetylase 2 (crucial receptors involved in breast cancer signalling) as evidenced by the negative docking score and binding free energy (BFE) values. Therefore, crocetin beta-d-glucosyl ester from Crocus sativus biowastes showed antiproliferative effect possibly by inhibiting estrogen receptor alpha and HDAC2 mediated signalling cascade.


Effect of indomethacin on edema following single and repetitive cerebral ischemia in the gerbil.
Repetitive periods of cerebral ischemia result in more severe injury than a single period of ischemia of similar total duration. We investigated the possibility of prostaglandin mediation of this increased injury by attempting to modify brain edema formation with indomethacin pretreatment. Under halothane/N2O anesthesia, groups of gerbils underwent bilateral carotid occlusion to induce forebrain ischemia. Group I underwent a single 15-minute period of carotid occlusion. Group II underwent three 5-minute periods of occlusion at hourly intervals. Groups III and IV were similar to groups I and II, respectively, but received 0.2 mg/kg indomethacin before carotid occlusion. Cortical and cerebellar water and sodium contents were determined in control animals (n = 6) at time zero and in experimental animals 24, 48, and 72 hours after ischemia (n = 6-10 gerbils/group at each time point). Cortical water and sodium contents in group II peaked 48 hours after insult (82.15 +/- 0.31% and 420 +/- 14 meq/kg dry wt, respectively) and were significantly higher than control and group I values at both 24 and 48 hours. Cortical water did not change from control in group I animals. Indomethacin pretreatment significantly attenuated increases in water and sodium content seen at 48 hours in gerbils undergoing repetitive ischemia (peak 80.02 +/- 0.45% and 300 +/- 39 meq/kg dry wt), but did not affect mortality. Indomethacin lessens edema after repetitive cerebral ischemia, suggesting that elevations of cyclooxygenase products are responsible, at least in part, for severe brain edema following repetitive ischemia.


Identification of domains of the insulin-like growth factor I receptor that are required for protection from apoptosis.
Using a series of insulin-like growth factor I (IGF-I) receptor mutants, we have attempted to define domains required for transmitting the antiapoptotic signal from the receptor and to compare these domains with those required for mitogenesis or transformation. In FL5.12 cells transfected with wild-type IGF-I receptors, IGF-I affords protection from interleukin 3 withdrawal but is not mitogenic. An IGF-I receptor lacking a functional ATP binding site provided no protection from apoptosis. However, receptors mutated at tyrosine residue 950 or in the tyrosine cluster (1131, 1135, and 1136) within the kinase domain remained capable of suppressing apoptosis, although such mutations are known to inactivate transforming and mitogenic functions. In the C terminus of the IGF-I receptor, two mutations, one at tyrosine 1251 and one which replaced residues histidine 1293 and lysine 1294, abolished the antiapoptotic function, whereas mutation of the four serines at 1280 to 1283 did not. Interestingly, receptors truncated at the C terminus had enhanced antiapoptotic function. In Rat-1/ c-MycER fibroblasts, the Y950F mutant and the tyrosine cluster mutant could still provide protection from c-Myc-induced apoptosis, whereas mutant Y1250/1251F could not. These studies demonstrate that the domains of the IGF-I receptor required for its antiapoptotic function are distinct from those required for its proliferation or transformation functions and suggest that domains of the receptor required for inhibition of apoptosis are necessary but not sufficient for transformation.


Minimally invasive approach for redo mitral valve surgery.
Redo cardiac surgery represents a clinical challenge due to a higher rate of peri-operative morbidity and mortality. Mitral valve re-operations can be particularly demanding in patients with patent coronary artery bypass grafts, previous aortic valve replacement, calcified aorta or complications following a previous operation (abscesses, perivalvular leaks, or thrombosis). Risk of graft injuries, hemorrhage, the presence of dense adhesions and complex valve exposure can make redo valve operations challenging through a median sternotomy. In this review article we provide an overview of minimally invasive approaches for redo mitral valve surgery discussing indications, techniques, outcomes, concerns and controversies. Scientific literature about minimally invasive approach for redo mitral surgery was reviewed with a MEDLINE search strategy combining "mitral valve" with the following terms: 'minimally invasive', 'reoperation', and 'alternative approach'. The search was limited to the last ten years. A total of 168 papers were found using the reported search. From these, ten papers were identified to provide the best evidence on the subject. Mitral valve reoperations can be safely and effectively performed through a smaller right thoracotomy in the fourth intercostal space termed "mini" thoracotomy or "port access". The greatest potential benefit of a right mini-thoracotomy is the avoidance of sternal re-entry and limited dissection of adhesions, avoiding the risk of injury to cardiac structures or patent grafts. Good percentages of valve repair can be achieved. Mortality is low as well as major complications. Minimally invasive procedures with an unclamped aorta have the potential to combine the benefits of minimally invasive access and continuous myocardial perfusion. Less invasive trans-catheter techniques could be considered as the natural future evolution for management of structural heart disease and mitral reoperations. The safety and efficacy of these procedures has never been compared to open reoperations in a randomized trial, although published case series and comparisons to historical cohorts suggest that they are an effective and feasible alternative. Ongoing follow-up on current series will further define these procedures and provide valuable clinical outcome data.


Clinic-Based Food Assistance is Associated with Increased Medication Adherence among HIV-Infected Adults on Long-Term Antiretroviral Therapy in Zambia.
BACKGROUND: There has been limited research to date on the effects of food assistance provided to HIV-infected adults in resource-constrained settings with a high prevalence of malnutrition and chronic food insecurity. We compare antiretroviral therapy (ART) adherence, weight gain, and CD4+ lymphocyte count change among HIV-infected adults enrolled in a clinic-based food assistance program in Lusaka, Zambia versus a control group of non-recipients. METHODS: We conducted a cohort study incorporating interviewer-administered surveys and retrospective clinical data to compare ART patients receiving food assistance with a control group of non-recipients. Medication adherence was assessed using pharmacy dispensation records. We use propensity score matching to assess the effect of food assistance on outcome measures. RESULTS: After 6 months, food assistance recipients (n=145) had higher ART adherence compared to non-recipients (n=147, 98.3% versus 88.8%, respectively; p<0.01), but no significant effects were observed for weight or CD4+ lymphocyte count change. The improvement in adherence rates was greater for participants on ART for less than 230 days, and those with BMI<18.5 kg/m(2), a higher HIV disease stage, or a CD4+ lymphocyte count ≤ 350 cells/μl. CONCLUSIONS: Promoting optimal medication adherence among persons on ART is relevant to public health and the success of HIV control efforts. The provision of food assistance to HIV-infected adults on ART may have an incentivizing effect which can improve medication adherence, particularly among patients recently initiated on treatment and those with poor nutrition or advanced disease. The effects on body weight and immune reconstitution appear minimal.


Burden of infections on older patients presenting to general practice: a registry-based study.
Estimates on the incidence rates of infections are needed to assess the burden of disease in the community. To assess incidence rates of potentially serious infections in patients aged 65 years and over presenting to Flemish general practice from 2000 to 2015, and to describe patient characteristics. We performed a retrospective study, based on data provided by the Intego morbidity registry of the KU Leuven, which includes the electronic medical records of 111 general practitioners. Incidence rates were calculated taking person-time at risk into account, and longitudinal trends from 2000 to 2015 were analysed using autoregressive time-series analyses. On average, a person aged 65 years or older has an 8.0% risk of getting a potentially serious infection each year. Acute cystitis was the most often occurring potentially serious infection [39.8/1000 person-years; 95% confidence interval (CI): 39.4-40.2], followed by influenza like illness (ILI, 24.3/1000 person-years; 95% CI: 24.0-24.6) and pneumonia (9.7/1000 person-years; 95% CI: 9.5-9.9). The incidence rates of pneumonia were higher in older age groups and in men, whereas they were markedly lower for ILI at older ages, in both genders. From 2000 to 2015, overall incidence rates decreased significantly for ILI, while they increased in women for pneumonia, acute cystitis and pyelonephritis. Common chronic comorbidities were non-insulin dependent diabetes, chronic obstructive pulmonary disease, asthma, heart failure and chronic renal insufficiency. Potentially serious infections are quite common in an older patient population presenting to primary care. They are accompanied by several chronic comorbidities, which may differ by infection type.


Medicaid managed care: the last bastion of the HMO?
States rely on health maintenance organizations (HMOs) for their Medicaid beneficiaries because they offer guaranteed access to comprehensive benefits at a predictable cost. This is true despite movement away from HMOs, or at least the more restrictive variants, in the private sector. Plans that focus on Medicaid are becoming more central to states' programs as commercial plans exit. Publicly traded, Medicaid-focused plans are also emerging. Medicaid participating plans are aggressively managing costs and care, contrasting sharply with commercial insurance where the trend is toward less intrusive managed care. In this context, state Medicaid managed care programs are facing important policy challenges related to plan participation, mainstreaming, and product design.


Analysis of ceRNA network identifies prognostic circRNA biomarkers in bladder cancer.
Bladder cancer remains a very challenging disease to treat with the high rates of recurrence and progression associated with current therapies. Although the association between bladder cancer pathology and circRNAs remains undetermined, circRNAs signatures may be useful as prognostic and predictive factors and clinical tools for assessing disease state and outcome. This study investigates if these circRNAs can be used as biomarkers for bladder cancer diagnosis. Using bioinformatics method to analysis GEO databases (GSE37815, GSE39093, GSE97239, and GSE92675) for differentially expressed RNAs in bladder cancer and normal bladder tissues were screened from. The related volcanic maps and the interaction network maps of differentially expressed RNAs were drawn, and the mRNA-miRNA and miRNA-circRNA interaction were predicted to establish mRNA-miRNA-circRNA competitive endogenous RNA (ceRNA) network. The differential circRNAs related to prognosis of bladder cancer patients were screened based on the influence of miRNA interacting with the circRNA above on survival rate. The expression of miRNA (hsa-mir-214), circRNA (hsa_circ_0076704, hsa_circ_0081963, hsa_circ_0001361) in bladder cancer tissues, adjacent tissues, bladder cancer cells and normal bladder epithelial cells were validated by qRT-PCR. Kaplan Meier curve analysis confirmed the relationship between circRNA (hsa_circ_0076704) and overall survival and prognosis of bladder cancer patients. Through database screening and analysis, we found 19231 differentially expressed genes, 847 differentially expressed miRNAs, 7282 differentially expressed circRNAs. The establishment of ceRNA network consisted of 28 DERNAs (differentially- expressed RNAs), 12 Demi-RNAs and 12 DEcircRNAs. Further prognostic analysis showed that circRNA interacted miRNA hsa-miR-106b, hsa-miR-145 and hsa-miR-214 were associated with overall survival in patients with bladder cancer (P < 0.05). Among them, hsa_circ_0076704, hsa_circ_0081963 and hsa_circ_0001361 are potential circRNA related to OS in bladder cancer and expressed in bladder cancer. The expression of hsa-mir-214 was contrary. Further Kaplan Meier survival analysis showed that hsa_circ_0076704 had significant prognostic value (P < 0.05). In conclusion, hsa_circ_0076704 is independent prognostic factor for bladder cancer.


Caffeic acid phenethyl ester against cadmium induced toxicity mediated by CircRNA modulates autophagy in HepG2 cells.
Cadmium pollution and poisoning are serious environmental and pharmacological concerns, and effective drugs can alleviate or offset cadmium-induced toxicity are badly needed. In this study, Caffeic acid phenethyl ester (CAPE), a major active component of propolis, showed protective effect against CdCl2-induced toxicology by suppressing autophagy in HepG2 cells. CircRNAs are increasingly perceived as vital regulators in the process of autophagy. However, it remain unclear whether circRNAs are involved in CAPE's protection against CdCl2-induced autophagy. Under this context, the roles of CircRNA (hsa_circ_0040768) in CAPE's protection against CdCl2-induced damage were investigated by PCR and Western blot. Results showed that CAPE significantly (P < 0.05) increased cell viability via inhibiting CdCl2-induced autophagy, and this process was regulated by hsa_circ_0040768/MAP1LC3B axis. Overexpressing hsa_circ_0040768 led to reduced cell viability and increased autophagy in CAPE-treated HepG2 cells exposed to CdCl2. In contrast, silencing hsa_circ_0040768 showed similar protective effect to CAPE. These results show for the first time the involvement of the hsa_circ_0040768/MAP1LC3B axis in the CAPE's protection against CdCl2-induced autophagy, and provide novel insights into the pathogenesis and potential prevention/treatment of cadmium-associated diseases.


Osmotically stable L forms of Haemophilus influenzae and their significance in testing sensitivity to penicillins.
The sensitivity of Haemophilus influenzae to penicillins in vitro, determined either by serial antibiotic dilution in broth or by the disc method on agar, is apparently profoundly influenced by inoculum size if the results are read by macroscopic inspection. Microscopic inspection of the growth, however, reveals that the turbidity in heavily inoculated broth containing concentrations higher than the minimal inhibitory concentration is the product of L forms which have failed to succumb to osmotic lysis. Similarly, minute colonies appearing in the ;inhibition zone' of disc tests are composed of L forms. In both broth and agar tests reduction of the osmolality of the medium from 340 to 144 mOsm per kg failed to bring about lysis of organisms exposed either to ampicillin or amoxycillin. The significance of this remarkable osmotic stability of haemophilus L forms is discussed in relation both to testing of sensitivity of this organism to penicillins and to persistence of chronic haemophilus infections of the lower respiratory tract.


Functional annotation of chemical libraries across diverse biological processes.
Chemical-genetic approaches offer the potential for unbiased functional annotation of chemical libraries. Mutations can alter the response of cells in the presence of a compound, revealing chemical-genetic interactions that can elucidate a compound's mode of action. We developed a highly parallel, unbiased yeast chemical-genetic screening system involving three key components. First, in a drug-sensitive genetic background, we constructed an optimized diagnostic mutant collection that is predictive for all major yeast biological processes. Second, we implemented a multiplexed (768-plex) barcode-sequencing protocol, enabling the assembly of thousands of chemical-genetic profiles. Finally, based on comparison of the chemical-genetic profiles with a compendium of genome-wide genetic interaction profiles, we predicted compound functionality. Applying this high-throughput approach, we screened seven different compound libraries and annotated their functional diversity. We further validated biological process predictions, prioritized a diverse set of compounds, and identified compounds that appear to have dual modes of action.


Importance of Metal-Ion Exchange for the Biological Activity of Coordination Complexes of the Biomimetic Ligand N4Py.
Metal coordination complexes can display interesting biological activity, as illustrated by the bleomycins (BLMs), a family of natural antibiotics that when coordinated to a redox-active metal ion, show antitumor activity. Yet, which metal ion is required for the activity in cells is still subject to debate. In this study, we described how different metal ions affect the intracellular behavior and activity of the synthetic BLM-mimic N, N-bis(2-pyridylmethyl)- N-bis(2-pyridyl)methylamine (N4Py). Our study shows that a mixture of iron(II), copper(II), and zinc(II) complexes can be generated when N4Py is added to cell cultures but that the metal ion can also be exchanged by other metal ions present in cells. Moreover, the combination of chemical data, together with the performed biological experiments, shows that the active complex causing oxidative damage to cells is the FeII-N4Py complex and not per se the metal complex that was initially added to the cell culture medium. Finally, it is proposed that the high activity observed upon the addition of the free N4Py ligand is the result of a combination of scavenging of biologically relevant metals and oxidative damage caused by the iron(II) complex.


ETV6 (TEL1) regulates embryonic hematopoiesis in zebrafish.
Chromosomal translocations involving fusions of the human ETV6 (TEL1) gene occur frequently in hematologic malignancies. However, a detailed understanding of the normal function of ETV6 remains incomplete. This study has employed zebrafish as a relevant model to investigate the role of ETV6 during embryonic hematopoiesis. Zebrafish possessed a single conserved etv6 ortholog that was expressed from 12 hpf in the lateral plate mesoderm, and later in hematopoietic, vascular and other tissues. Morpholino-mediated gene knockdown of etv6 revealed the complex contribution of this gene toward embryonic hematopoiesis. During primitive hematopoiesis, etv6 knockdown resulted in reduced levels of progenitor cells, erythrocyte and macrophage populations, but increased numbers of incompletely differentiated heterophils. Definitive hematopoiesis was also perturbed, with etv6 knockdown leading to decreased erythrocytes and myeloid cells, but enhanced lymphopoiesis. This study suggests that ETV6 plays a broader and more complex role in early hematopoiesis than previously thought, impacting on the development of multiple lineages.


Prognostic indicators in pediatric clinically isolated syndrome.
To assess prognostic factors for a second clinical attack and a first disability-worsening event in pediatric clinically isolated syndrome (pCIS) suggestive of multiple sclerosis (MS) patients. A cohort of 770 pCIS patients was followed up for at least 10 years. Cox proportional hazard models and Recursive Partitioning and Amalgamation (RECPAM) tree-regression were used to analyze data. In pCIS, female sex and a multifocal onset were risk factors for a second clinical attack (hazard ratio [HR], 95% confidence interval [CI] = 1.28, 1.06-1.55; 1.42, 1.10-1.84, respectively), whereas disease-modifying drug (DMD) exposure reduced this risk (HR, 95% CI = 0.75, 0.60-0.95). After pediatric onset MS (POMS) diagnosis, age at onset younger than 15 years and DMD exposure decreased the risk of a first Expanded Disability Status Scale (EDSS)-worsening event (HR, 95% CI = 0.59, 0.42-0.83; 0.75, 0.71-0.80, respectively), whereas the occurrence of relapse increased this risk (HR, 95% CI = 5.08, 3.46-7.46). An exploratory RECPAM analysis highlighted a significantly higher incidence of a first EDSS-worsening event in patients with multifocal or isolated spinal cord or optic neuritis involvement at onset in comparison to those with an isolated supratentorial or brainstem syndrome. A Cox regression model including RECPAM classes confirmed DMD exposure as the most protective factor against EDSS-worsening events and relapses as the most important risk factor for attaining EDSS worsening. This work represents a step forward in identifying predictors of unfavorable course in pCIS and POMS and supports a protective effect of early DMD treatment in preventing MS development and disability accumulation in this population. Ann Neurol 2017;81:729-739.


Methylnitrosourea-induced tumorigenesis in MGMT gene knockout mice.
Gene targeting was used to obtain mice defective in the MGMT gene, encoding O6-methylguanine-DNA methyltransferase [Tsuzuki et al., Carcinogenesis (Lond.), 17: 1215-1220, 1996]. These MGMT-/- mice were most sensitive to the alkylating carcinogen, methylnitrosourea; when varied doses of methylnitrosourea were administered to 6-week-old mice and survivals at the 30th day were determined, LD50s of MGMT-/- and MGMT+/+ mice were 20 and 240 mg/kg of body weight, respectively. MGMT+/- mice were as resistant as MGMT+/+ mice, but some difference in survival time was noted when the two genotypes of mice were exposed to a relatively high dose of methylnitrosourea. A large number of thymic lymphomas, as well as lung adenomas, occurred in MGMT-/- mice exposed to methylnitrosourea at a dose of 2.5 mg/kg of body weight. In case of exposure to the same dose of drug, no or few tumors occurred in the MGMT+/+ and MGMT+/- mice. It appears that the DNA repair methyltransferase protein protected these mice from methylnitrosourea-induced tumorigenesis.


Metal-based nanoparticles and their toxicity assessment.
Nanoparticles (NPs) can potentially cause adverse effects on organ, tissue, cellular, subcellular, and protein levels due to their unusual physicochemical properties (e.g., small size, high surface area to volume ratio, chemical composition, crystallinity, electronic properties, surface structure reactivity and functional groups, inorganic or organic coatings, solubility, shape, and aggregation behavior). Metal NPs, in particular, have received increasing interest due to their widespread medical, consumer, industrial, and military applications. However, as particle size decreases, some metal-based NPs are showing increased toxicity, even if the same material is relatively inert in its bulk form (e.g., Ag, Au, and Cu). NPs also interact with proteins and enzymes within mammalian cells and they can interfere with the antioxidant defense mechanism leading to reactive oxygen species generation, the initiation of an inflammatory response and perturbation and destruction of the mitochondria causing apoptosis or necrosis. As a result, there are many challenges to overcome before we can determine if the benefits outweigh the risks associated with NPs.


The application of Oncotype DX in early-stage lymph-node-positive disease.
The recurrence score derived from the 21-gene Oncotype DX assay is both prognostic and predictive of adjuvant chemotherapy benefit in node-negative, estrogen-receptor-positive breast cancer patients treated with tamoxifen. This has led to a remarkable shift in the treatment paradigm, with a sizeable number of patients being able to avoid adjuvant chemotherapy. The recurrence score was then analyzed in a large retrospective study with node-positive, estrogen-receptor-positive patients, in which it demonstrated both prognostic and predictive abilities. This review introduces the clinical trials that validated the Oncotype DX assay in the node-negative population, highlights the studies evaluating the utility of the assay in node-positive patients, examines the impact of the assay results on treatment decisions, and discusses the health outcomes and health care expenditures associated with this assay.


Coarse-grained model for phospholipid/cholesterol bilayer employing inverse Monte Carlo with thermodynamic constraints.
The authors introduce a coarse-grained (CG) model for a lipid membrane comprised of phospholipids and cholesterol at different molar concentrations, which allows them to study systems that are approximately 100 nm in linear size. The systems are studied in the fluid phase above the main transition temperature. The effective interactions for the CG model are extracted from atomic-scale molecular dynamics simulations using the inverse Monte Carlo (IMC) technique, an approach similar to the one the authors used earlier to construct another CG bilayer model [T. Murtola et al., J. Chem. Phys. 121, 9156 (2004)]. Here, the authors improve their original CG model by employing a more accurate description of the molecular structure for the phospholipid molecules. Further, they include a thermodynamic constraint in the IMC procedure to yield area compressibilities in line with experimental data. The more realistic description of the molecular structure of phospholipids and a more accurate representation of the interaction between cholesterols and phospholipid tails are shown to improve the behavior of the model significantly. In particular, the new model predicts the formation of denser transient regions in a pure phospholipid system, a finding that the authors have verified through large scale atomistic simulations. They also find that the model predicts the formation of cholesterol-rich and cholesterol-poor domains at intermediate cholesterol concentrations, in agreement with the original model and the experimental phase diagram. However, the domains observed here are much more distinct compared to the previous model. Finally, the authors also explore the limitations of the model, discussing its advantages and disadvantages.


[Psychological types and learning styles of students entering medical school at the Pontificia Universidad Católica de Chile].
Psychological type and learning style influence the way students perceive and process information. However, research in medical education in Chile still does not put enough emphasis in the study of these variables. To characterize the psychological types and learning styles of the students admitted to a Medical School. The Myers Briggs Type Indicator (MBTI) and the Kolb's Learning Styles Inventory (IEA) were administered to the 270 students admitted from 2000 to 2002 to the medical school of the Pontificia Universidad Católica de Chile. Fifty five percent of our students are concentrated in 4 of the 16 psychological types described. These students are characterized by the ability to base their decisions upon logical and objective reasoning (Thinking [T]) and to face life in a structured and decided way (Judging [J]). Only 10% of the students have preferences opposite to T and J. These students base their decisions on the preservation of harmony and teamwork (Feeling [F]) and have a flexible attitude towards life (Perceiving [P]). The remaining 35% have types with pairs of preferences TP and FJ. With regard to learning styles, more than two thirds of our students are Assimilators or Convergers. These learners tend to assimilate large amounts of information and abstract the main concepts, rather than to pay attention to concrete details. In general, our students are more reflective than active; they evaluate thoroughly all alternatives before making a decision. The psychological types and learning styles of medical students cluster around specific patterns whose features may either favor or hamper a specific learning. Knowledge of the differences in psychological types and learning styles of students may provide teachers with a new and valuable tool for improving learning and contributing to the academic success of students.


Near-infrared diffuse optical tomography.
Diffuse optical tomography (DOT) is emerging as a viable new biomedical imaging modality. Using near-infrared (NIR) light, this technique probes absorption as well as scattering properties of biological tissues. First commercial instruments are now available that allow users to obtain cross-sectional and volumetric views of various body parts. Currently, the main applications are brain, breast, limb, joint, and fluorescence/bioluminescence imaging. Although the spatial resolution is limited when compared with other imaging modalities, such as magnetic resonance imaging (MRI) or X-ray computerized tomography (CT), DOT provides access to a variety of physiological parameters that otherwise are not accessible, including sub-second imaging of hemodynamics and other fast-changing processes. Furthermore, DOT can be realized in compact, portable instrumentation that allows for bedside monitoring at relatively low cost. In this paper, we present an overview of current state-of-the -art technology, including hardware and image-reconstruction algorithms, and focus on applications in brain and joint imaging. In addition, we present recent results of work on optical tomographic imaging in small animals.


Renal 25-hydroxyvitamin D-1 alpha-hydroxylase activity and mitochondrial phosphate transport in Hyp mice.
The Hyp mouse is a homologue of the X chromosome-linked human disease, familial hypophosphatemic rickets (FHR). In FHR, reduced renal tubular brush-border membrane transport of phosphate results in hypophosphatemia and rickets. Both humans with FHR and Hyp mice have abnormal regulation of 25-hydroxyvitamin D-1 alpha-hydroxylase (1 alpha-hydroxylase), a mitochondrial enzyme found in proximal renal tubular cell epithelia, the apparent site of defective brush-border membrane phosphate transport. No common pathophysiology for these defects has been demonstrated. We hypothesized that phosphate transport may be present in renal mitochondria from Hyp mice and that its regulation may be deranged in parallel with the mitochondrial 1 alpha-hydroxylase. Using inhibitor-stop techniques described for measurement of phosphate transport in liver mitochondria, we examined mitochondria in normal and Hyp mouse kidney and found them to be comparable. We performed manipulations known to alter 1 alpha-hydroxylase differentially in normal and Hyp mice, i.e., phosphorus deprivation and phosphorus loading, and found no effect on mitochondrial phosphate transport. We also subjected Hyp and normal mice to calcium and vitamin D deprivation; this maneuver resulted in no significant changes in mitochondrial phosphate transport in Hyp or normal mice but confirmed the earlier observation that 1 alpha-hydroxylase activity is stimulated to a greater degree in normal mice than Hyp mice after this diet. Furthermore, administration of 1,25-hydroxyvitamin D3 depresses 1 alpha-hydroxylase activity in mitochondria from both normal and Hyp mice but has no effect on mitochondrial phosphate transport. We conclude that the mechanism of abnormal vitamin D metabolism in Hyp mice is not related to a primary defect in renal mitochondrial phosphate transport.


[Economic evaluations of health technologies: a global perspective for their implementation in Latin America].
Phenomena as the progressive increase of health expenditure and the population aging have lead many countries to consider economic methodologies in order to obtain bigger sanitary benefits in contexts of limited resources. This article describes the basic components to consider in a health technology assessment , it analyses the process of decision making with cost-effectiveness analysis and reports how this methodology has been widely implemented in Latin America and the rest of the world.


The postoperative analgesic efficacy of intraperitoneal tramadol compared to normal saline or intravenous tramadol in laparoscopic cholecystectomy.
The aim of this study was to compare the postoperative analgesic efficacy of intraperitoneal tramadol with intravenous tramadol or normal saline in patients undergoing laparoscopic cholecystectomy. Sixty-one patients undergoing laparoscopic cholecystectomy were randomized to one of three groups in a double-blind manner via coded syringes. All patients received an intravenous and an intraperitoneal injection after installation of the pneumoperitoneum and again before removal of the trocars. In the control group, all injections were with normal saline. In the intravenous tramadol group, patients received intravenous tramadol 100 mg and intraperitoneal saline. In the intraperitoneal tramadol group, patients received intravenous saline and intraperitoneal tramadol 100 mg. All patients had a standard anaesthetic. Postoperative analgesia was with morphine. Postoperatively, numeric pain scores for parietal and visceral pain, 1 h and 24 h morphine consumption, and adverse effects were recorded. Parietal and visceral pain scores were lowest in the intravenous tramadol group during the first postoperative hour (P < 0.016 compared with control). The delay until the first analgesic administration was longest in the intravenous tramadol group (median 23 min, range 1-45), when compared with the intraperitoneal tramadol group (10, 1-120 min, P = 0.263) or with the control group (1, 1-30 min, P = 0.015). One-hour morphine consumption was significantly lower in the intravenous tramadol group (mean +/- SD; 3.4 mg +/- 2.5) and in the intraperitoneal tramadol group (4.4 +/- 4.3 mg) compared with the control group (6 +/- 2 mg) (P = 0.044). There was no difference between the three groups regarding pain scores, morphine consumption and incidence of shoulder pain or adverse effects at 24 h. Intravenous tramadol provides superior postoperative analgesia in the early postoperative period after laparoscopic cholecystectomy compared with an equivalent dose of tramadol administered intraperitoneally and with normal saline in patients undergoing laparoscopic cholecystectomy.


Adolescent Family Factors Promoting Healthy Adult Functioning: A Longitudinal Community Study.
BACKGROUND: Although long-held wisdom and current research suggests that accepting and supportive family relationships may positively influence adult psychosocial functioning, few studies have prospectively investigated these associations. This study examined whether positive family factors during adolescence are associated with healthy adult functioning. METHOD: The 353 participants were part of a single-age cohort whose psychosocial development has been prospectively traced. Two aspects of family functioning - feeling highly valued as a family member and having a family confidant - were measured at age 15. Developmentally-relevant areas of functioning were assessed at age 30. RESULTS: Both positive family factors were predictive of adaptive adult functioning across several domains, including mental health and social/interpersonal functioning. CONCLUSIONS: Findings provide evidence about the salient relationships between positive family relationships and later healthy functioning.


Merochlorins A-D, cyclic meroterpenoid antibiotics biosynthesized in divergent pathways with vanadium-dependent chloroperoxidases.
Meroterpenoids are mixed polyketide-terpenoid natural products with a broad range of biological activities. Herein, we present the structures of four new meroterpenoid antibiotics, merochlorins A-D, produced by the marine bacterium Streptomyces sp. strain CNH-189, which possess novel chemical skeletons unrelated to known bacterial agents. Draft genome sequencing, mutagenesis, and heterologous biosynthesis in the genome-minimized model actinomycete Streptomyces coelicolor provided the 57.6 kb merochlorin gene cluster that contains two genes encoding rare bacterial vanadium-dependent haloperoxidase (VHPO) genes. Pathway expression of two different fosmid clones that differ largely by the presence or absence of the VHPO gene mcl40 resulted in the differential biosynthesis of merochlorin C, suggesting that Mcl40 catalyzes an unprecedented 15-membered chloronium-induced macrocyclization reaction converting merochlorin D to merochlorin C.


The protein kinase A signaling pathway mediates the effect of electroacupuncture on excessive contraction of the bladder detrusor in a rat model of neurogenic bladder.
Neurogenic bladder (NB) is a form of neurological bladder dysfunction characterized by excessive contraction of the bladder detrusor. Protein kinase A (PKA) signaling is involved in the contraction of the detrusor muscle. To investigate whether PKA signaling mediates the effect of electroacupuncture (EA) on the excessive contraction of the bladder detrusor in NB. Sixty rats were randomly divided into control, sham, NB, NB + EA, and NB + EA + H89 (a PKA receptor antagonist) groups. The modified Hassan Shaker spinal cord transection method was used to generate a NB model. After EA intervention for one week, urodynamic tests were used to evaluate bladder function, hematoxylin and eosin staining was conducted to assess morphological changes, enzyme-linked immunosorbent assay (ELISA) was performed to measure the concentration of PKA, and Western blotting was conducted to measure the protein levels of phosphorylated myosin light chain kinase (p-MLCK)/p-MLC. The results showed that NB resulted in morphological disruption, impairment of urodynamics, and decreases in the concentration of PKA and the protein levels of p-MLCK/p-MLC. EA reversed the changes induced by NB dysfunction. However, the improvement in urodynamics and the increases in the concentration of PKA and the protein levels of p-MLCK/p-MLC were inhibited by H89. Our findings indicate that the PKA signaling pathway mediates the beneficial effect of EA on excessive contraction of the bladder detrusor in a rat model of NB.


Observations on urinary frequency in exclusively breastfed neonates.
This study was conducted at the University Teaching Hospital, Benin City, Nigeria to investigate urinary frequency in exclusively breastfed neonates. Amongst exclusively breastfed neonates, 59.0% passed urine eight times or more per day while 14.5% passed urine less than five times a day. The comparative figures in partially breastfed neonates were 55.8% and 12.8% respectively. There was no significant association between the type of breastfeeding and urinary frequency. Sex was also not significantly associated with urinary frequency amongst exclusively breastfed infants. However, birthweight showed significant association with urinary frequency in the analysed sample. It is suggested that birthweight, which correlates with gestational maturity may influence urinary frequency because of its effect on renal functional maturation and possibly differential threshold for intravesical pressures required for initiating the micturition reflex. The observed association is, however, not an indication for supplementary water in low-birthweight infants since all the infants had normal hydration status. It is, therefore, recommended that all neonates should not receive supplementary water irrespective of their birthweights.


Identification of the N-acetylneuraminyllactose-specific laminin-binding protein of Helicobacter pylori.
The interaction of the gastroduodenal pathogen Helicobacter pylori with the glycoprotein laminin was investigated. Binding of 125I-radiolabelled laminin in a liquid-phase assay by both hemagglutinating and poorly hemagglutinating strains was rapid, saturable, specific, partially reversible, of high affinity, and insensitive to pH. Inhibition of laminin binding by fetuin, but not asialofetuin, and reduced bacterial binding to periodate- or sialidase-treated laminin indicated that glycosylation, particularly sialylation, was important for laminin binding by H. pylori. Inhibition experiments with monosaccharides, disaccharides, and trisaccharides showed that the strains bound to a region spanning a trisaccharide. In particular, inhibition and displacement studies showed that binding to the trisaccharide N-acetylneuraminyl-alpha(2-3)-lactose [NeuAc(2-3)Lac] was preferential to that to the NeuAc(2-6)Lac isomer. Complete inhibition of laminin binding by both hemagglutinating and poorly hemagglutinating strains was achieved only when isolated lipopolysaccharide (LPS) was used as an inhibitor in combination with heat or protease treatment of H. pylori cells, thereby confirming the involvement of both LPS and a protein adhesin in laminin binding. Further inhibition experiments indicated that the protein receptor, rather than LPS, on H. pylori bound NeuAc(2-3)Lac. By using a Western blotting procedure, a 25-kDa outer membrane protein was identified as mediating laminin binding by both hemagglutinating and poorly hemagglutinating H. pylori strains. The specificity of binding was confirmed by complete inhibition of laminin binding by the 25-kDa protein with NeuAc(2-3)Lac. The data collectively suggest that a 25-kDa outer membrane protein acts in a lectin-like manner with LPS to mediate attachment of H. pylori to laminin.


Diversity in the chiropractic profession: preparing for 2050.
As the diversity of the United States (US) population continues to change, concerns about minority health and health disparities grow. Health professions must evolve to meet the needs of the population. The purpose of this editorial is to review current trends in the diversity of chiropractic students, faculty, and practitioners in the United States. This editorial was informed by a search of the literature, to include PubMed, using the terms chiropractic and diversity, minority, and cultural competency. Demographic information for the chiropractic profession was obtained from the National Board of Chiropractic Examiners and The Chronicle of Higher Education. These data were compared to diversity data for medical doctors and the national and state populations from the American Association of Medical Colleges and the US Census, respectively. Surprisingly little has been published in the peer-reviewed literature on the topic of diversity in the chiropractic profession. For the variables available (sex and race), the data show that proportions in the US chiropractic profession do not match the population. State comparisons to associated chiropractic colleges show similar relationships. No reliable data were found on other diversity characteristics, such as gender identity, religion, and socioeconomic status. The chiropractic profession in the United States currently does not represent the national population with regard to sex and race. Leaders in the profession should develop a strategy to better meet the changing demographics of the US population. More attention to recruiting and retaining students, such as underrepresented minorities and women, and establishing improved cultural competency is needed.


Computerized adaptive test for patients with foot or ankle impairments produced valid and responsive measures of function.
We tested the item response theory (IRT) model assumptions of the original item bank, and evaluated the practical and psychometric adequacy, of a computerized adaptive test (CAT) for patients with foot or ankle impairments seeking rehabilitation in outpatient therapy clinics. Data from 10,287 patients with foot or ankle impairments receiving outpatient physical therapy were analyzed. We first examined the unidimensionality, fit, and invariance IRT assumptions of the CAT item bank. Then we evaluated the efficiency of the CAT administration and construct validity and sensitivity of change of the foot/ankle CAT measure of lower-extremity functional status (FS). Results supported unidimensionality, model fit, and invariance of item parameters and patient ability estimates. On average, the CAT used seven items to produce precise estimates of FS that adequately covered the content range with negligible floor and ceiling effects. Patients who were older, had more chronic symptoms, had more surgeries, had more comorbidities, and did not exercise prior to receiving rehabilitation reported worse discharge FS. Seventy-one percent of patients obtained statistically significant change at follow-up. Change of 8 FS units (scale 0-100) represented minimal clinically important improvement. We concluded that the foot/ankle item bank met IRT assumptions and that the CAT FS measure was precise, valid, and responsive, supporting its use in routine clinical application.


Beta-lactamase expression in Yersinia enterocolitica biovars 1A, 1B, and 3.
Characteristic patterns of beta-lactam susceptibility are associated with different biovars of Yersinia enterocolitica. In a previous study differences in beta-lactam susceptibility among biovar 2, 4 and 5 strains were largely attributed to differences in expression of beta-lactamase A (BlaA) and beta-lactamase B (BlaB). The basis for differences in beta-lactam susceptibility of strains of biovars 1A, 1B and 3 is now considered. All the strains examined had blaB; nine of 31 biovar 3 strains and two of 13 biovar 1B strains had blaA, but PCR did not amplify blaA from biovar 1A strains. Nevertheless, inhibition data indicated that the majority of uninduced biovar 1A strains expressed BlaA and BlaB in similar amounts. Strong inducibility was seen in all these strains. Biovar 1B strains (which were less inducible than strains of biovar 1A) predominantly produced BlaA without induction; ticarcillin-sensitive strains of biovar 3 produced only BlaB but were not inducible; without induction biovar 3 strains resistant to ticarcillin and amoxycillin/clavulanate produced either predominantly BlaA, predominantly BlaB or exclusively BlaB and induction was demonstrated except for strains producing BlaB alone; biovar 3 strains resistant to ticarcillin but sensitive to amoxycillin/clavulanate predominantly produced BlaA without induction and were inducible for beta-lactamase activity. After induction, nearly all strains predominantly or exclusively produced BlaB. Although PCR amplification fragments with primers specific for blaA were obtained only from some strains, the induction and inhibition data suggest that all Y. enterocolitica strains possess enzymes related to BlaA- as well as BlaB. Nevertheless, expression of the beta-lactamase is regulated differently in different biovars and varies within most biovars. Failure to predict beta-lactamase expression profiles from MIC data indicates the presence of additional mechanisms contributing to differences in susceptibility.


Effects of feeding frequency and feeding level on nutrient utilization in heavy preruminant calves.
The objective of this study was to determine the effects of feeding frequency (FF) and feeding level (FL) on protein and energy metabolism in adapted, heavy preruminant calves. It was hypothesized that an increased FF would increase protein utilization by an improved synchrony between the supply of and requirements for protein during the day when a quickly hydrolyzable protein source was used. Eighteen Holstein Friesian calves of 136 +/- 3 kg of body weight were assigned to FF (1, 2, or 4 meals daily) at 2 FL (1.5 or 2.5 times the metabolizable energy requirements for maintenance), except for calves fed once daily (only at a low FL). Calves were individually housed in respiration chambers during 2 experimental periods of 10 d. Whey protein was the only protein source in the diet. Neither FL nor FF affected apparent fecal nutrient digestibility. Increasing FF increased the efficiency with which digestible protein was utilized in calves. The increase was greater at a high FL (+11% from 2 to 4 meals/d) than at a low FL (+5% from 2 to 4 meals/d), but no significant interaction occurred between FL and FF. An increased FF and a higher FL enhanced fat deposition. Heat production was not affected by FF, but its circadian rhythm differed considerably between FF. Activity-related heat production was not affected by FF or FL. Thus, increasing FF improved the efficiency with which protein and energy were utilized in heavy preruminant calves when a quickly hydrolyzable protein source was used.


Skin Delivery of Hydrophilic Biomacromolecules Using Marine Sponge Spicules.
We report the development of sponge Haliclona sp. spicules, referred to as SHS, and its topical application in skin delivery of hydrophilic biomacromolecules, a series of fluorescein isothiocyanate-dextrans (FDs). SHS are silicious oxeas which are sharp-edged and rod-shaped (∼120 μm in length and ∼7 μm in diameter). SHS can physically disrupt skin in a dose-dependent manner and retain within the skin over at least 72 h, which allows sustained skin penetration of hydrophilic biomacromolecules. The magnitude of enhancement of FD delivery into skin induced by SHS treatment was dependent on its molecular weight. Specifically, SHS topical application enhanced FD-10 (MW: 10 kDa) penetration into porcine skin in vitro by 33.09 ± 7.16-fold compared to control group (p < 0.01). SHS dramatically increased the accumulation of FD-10 into and across the dermis by 62.32 ± 13.48-fold compared to the control group (p < 0.01). In vivo experiments performed using BALB/c mice also confirmed the effectiveness of SHS topical application; the skin absorption of FD-10 with SHS topical application was 72.14 ± 48.75-fold (p < 0.05) and 15.39 ± 9.91-fold (p < 0.05) higher than those from the PBS and Dermaroller microneedling, respectively. Further, skin irritation study and transepidermal water loss (TEWL) measurement using guinea pig skin in vivo indicated that skin disruption induced by SHS treatment is self-limited and can be recovered with time and efficiently. SHS can offer a safe, effective, and sustained skin delivery of hydrophilic biomacromolecules and presents a promising platform technology for a wide range of cosmetic and medical applications.


Reconstructive indications of simultaneous double free flaps in the head and neck: a case series and literature review.
Extensive and complex defects of the head and neck involving multiple anatomical and functional subunits are a reconstructive challenge. The purpose of this study is to elucidate the reconstructive indications of the use of simultaneous double free flaps in head and neck oncological surgery. This is a retrospective review of 21 consecutive cases of head and neck malignancies treated surgically with resection and reconstruction with simultaneous use of double free flaps. Nineteen of 21 patients had T4 primary tumor stage. Eleven patients had prior history of radiotherapy or chemo-radiotherapy. Forty-two free flaps were used in these patients. The predominant combination was that of free fibula osteo-cutaneous flap with free anterolateral thigh (ALT) fascio-cutaneous flap. The indications of the simultaneous use of double free flaps can be broadly classified as: (a) large oro-mandibular bone and soft tissue defects (n = 13), (b) large oro-mandibular soft tissue defects (n = 4), (c) complex skull-base defects (n = 2), and (d) dynamic total tongue reconstruction (n = 2). Flap survival rate was 95%. Median follow-up period was 11 months. Twelve patients were alive and free of disease at the end of the follow-up. Eighteen of 19 patients with oro-mandibular and glossectomy defects were able to resume an oral diet within two months while one patient remained gastrostomy dependant till his death due to disease not related to cancer. This patient had a combination of free fibula flap with free ALT flap, for an extensive oro-mandibular defect. The associated large defect involving the tongue accounted for the swallowing difficulty. Simultaneous use of double free flap aided the reconstruction in certain large complex defects after head and neck oncologic resections. Such combination permits better complex multiaxial subunit reconstruction. An algorithm for choice of flap combination for the appropriate indications is proposed.


Impacts of retinal polyene (de)methylation on the photoisomerization mechanism and photon energy storage of rhodopsin.
Ab initio multiconfigurational quantum chemical methodology combined with molecular mechanics (CASPT2//CASSCF/AMBER) was applied to probe impacts of retinal protonated Schiff base (RPSB) polyene methylation and/or demethylation on the mechanism of photochemical isomerization in bovine rhodopsin. We have examined structural and spectroscopic properties of wild-type rhodopsin (with 11-cis-9,13-dimethyl-RPSB) and artificial rhodopsins, hosting four 11-cis-RPSB derivatives, 13-demethyl-, 9-demethyl-, 10-methyl-13-demethyl-, and 10-methyl-RPSB, evolving along the photoisomerization coordinate. It is found that the addition of 10-methyl or/and deletion of 9-/13-methyl groups do not appear to interfere structurally with the photoisomerization pathway in the S1 excited state. Remarkably, the two-mode space-saving mechanism initiated by bond order inversion and followed by asynchronous bicycle-pedal distortion in the RPSB backbone drives the photoreaction in all rhodopsin analogues studied here. However, methylation and/or demethylation is responsible for perturbation of excess energy deposited in the conical intersection structures. The analysis of photon energy stored by bathorhodopsin in synthetic pigments reveals that it is affected by steric crowding of methyl substituents in the RPSB backbone.


Dietary intervention of flaxseed: effect on serum levels of IGF-1, IGF-BP3, and C-peptide.
Higher levels of insulin and insulin-like growth factor 1 (IGF-1) increase cancer risk by stimulating cell proliferation and increasing survival of DNA-damaged cells through antiapoptotic mechanisms. Laboratory studies suggest that flaxseed added to the diet may lower circulating levels of insulin and IGF-1, but there is limited information on the effects of dietary flaxseed on these biomarkers of cancer risk in humans. The aim of this study was to determine the effect of flaxseed supplementation in postmenopausal women on serum levels of insulin-like growth factor 1 (IGF-1), insulin-like growth factor binding protein 3 (IGF-BP3), and C-peptide, a marker of insulin production. Forty-eight postmenopausal women participated in this 12-wk baseline to postintervention study. Participants were asked to consume 7.5 g per day of ground flaxseed for 6 wk and 15 g per day for an additional 6 wk. No significant changes were observed in blood levels of IGF-1, IGF-BP3, or C-peptide over the study intervention. Flaxseed supplementation did not impact circulating levels of IGF-1, IGF-BP3, or C-peptide. Longer duration of intake may be necessary to observe changes in these biomarkers of cancer risk.


Air travel and venous thromboembolism.
There has recently been increased publicity on the risk of venous thrombosis after long-haul flights. This paper reviews the evidence base related to the association between air travel and venous thromboembolism. The evidence consists only of case reports, clinical case-control studies and observational studies involving the use of intermediate end-points, or expert opinion. Some studies have suggested that there is no clear association, whereas others have indicated a strong relationship. On the whole it appears that there is probably a link between air travel and venous thrombosis. However, the link is likely to be weak, mainly affecting passengers with additional risk factors for venous thromboembolism. The available evidence is not adequate to allow quantification of the risk. There are insufficient scientific data on which to base specific recommendations for prevention, other than that leg exercise should be taken during travel. Further studies are urgently needed in order to identify prospectively the incidence of the condition and those at risk.


Skin graft survival--the bacterial answer.
An in vitro wound model was created to determine the mechanism by which bacteria cause skin graft failure. A wound surface was simulated by a human fibrin clot. Staphylococcus aureus or group A streptococcus was incubated over the clot. Either saline, human plasminogen, aprotinin, or epsilon-aminocaproic acid (EACA), or a combination of these, was added to the tubes. After 30 hours, the tubes were examined for the presence of the clot. The supernatant was then examined for the presence of fibrin degradation products (FDP). S. aureus was incapable of destroying the fibrin clot without the presence of plasminogen in the culture media. Group A streptococcus was capable of some clot degradation, but this was markedly improved in the presence of plasminogen. High FDP levels correlated with the destruction of the clot. Both aprotinin and EACA were capable of preserving the fibrin clots. In this study we carefully controlled the chemical and bacterial milieu on a simulated wound.


Association between Irregular Meal Timing and the Mental Health of Japanese Workers.
Breakfast skipping and nighttime snacking have been identified as risk factors for obesity, diabetes, and cardiovascular diseases. However, the effects of irregularity of meal timing on health and daily quality of life are still unclear. In this study, a web-based self-administered questionnaire survey was conducted involving 4490 workers (73.3% males; average age = 47.4 ± 0.1 years) in Japan to investigate the association between meal habits, health, and social relationships. This study identified that irregular meal timing was correlated with higher neuroticism (one of the Big Five personality traits), lower physical activity levels, and higher productivity loss. Irregular meal timing was also associated with a higher incidence of sleep problems and lower subjective health conditions. Among health outcomes, a high correlation of irregular meal timing with mental health factors was observed. This study showed that irregularity of meal timing can be explained by unbalanced diets, frequent breakfast skipping, increased snacking frequency, and insufficient latency from the last meal to sleep onset. Finally, logistic regression analysis was conducted, and a significant contribution of meal timing irregularity to subjective mental health was found under adjustment for other confounding factors. These results suggest that irregular meal timing is a good marker of subjective mental health issues.


Genetic analysis of chromosome region 63 of Drosophila melanogaster.
The salivary chromosome region including cytological division 63 of Drosophila melanogaster was genetically analyzed in order to (1) characterize this previously unstudied region and (2) attempt to isolate mutations in the hsp82 gene. Seven deletions which span this region were isolated, including four which remove the hsp82 gene. A Minute mutation was mapped to this region and this Minute was used to isolate duplications in the 63 region. These duplications map the Minute to 63B8-C1. F2 screens were initiated using deletions which remove the hsp82 gene. Over 15,000 chromosomes were screened, yielding 40 lethal mutations which comprise 14 complementation groups. Several of these mutations map outside the 63 region and appear to give second site interaction with the Minute locus. Four loci, including the Minute gene, are candidates for hsp82 mutations by cytogenetic mapping. These loci were tested for complementation with a P element carrying the hsp82 gene. However, none of the mutations was rescued.


Predicting muscle forces of individuals with hemiparesis following stroke.
Functional electrical stimulation (FES) has been used to improve function in individuals with hemiparesis following stroke. An ideal functional electrical stimulation (FES) system needs an accurate mathematical model capable of designing subject and task-specific stimulation patterns. Such a model was previously developed in our laboratory and shown to predict the isometric forces produced by the quadriceps femoris muscles of able-bodied individuals and individuals with spinal cord injury in response to a wide range of clinically relevant stimulation frequencies and patterns. The aim of this study was to test our isometric muscle force model on the quadriceps femoris, ankle dorsiflexor, and ankle plantar-flexor muscles of individuals with post-stroke hemiparesis. Subjects were seated on a force dynamometer and isometric forces were measured in response to a range of stimulation frequencies (10 to 80-Hz) and 3 different patterns. Subject-specific model parameter values were obtained by fitting the measured force responses from 2 stimulation trains. The model parameters thus obtained were then used to obtain predicted forces for a range of frequencies and patterns. Predicted and measured forces were compared using intra-class correlation coefficients, r2 values, and model error relative to the physiological error (variability of measured forces). Results showed excellent agreement between measured and predicted force-time responses (r2 >0.80), peak forces (ICCs>0.84), and force-time integrals (ICCs>0.82) for the quadriceps, dorsiflexor, and plantar-fexor muscles. The model error was within or below the +95% confidence interval of the physiological error for >88% comparisons between measured and predicted forces. Our results show that the model has potential to be incorporated as a feed-forward controller for predicting subject-specific stimulation patterns during FES.


The malonate decarboxylase enzyme system of Malonomonas rubra: evidence for the cytoplasmic location of the biotin-containing component.
Malonate decarboxylase of Malonomonas rubra is a complex enzyme system involving cytoplasmic and membrane-bound components. One of these is a biotin-containing protein of Mr 120'000, the location of which in the cytoplasm was deduced from the following criteria: (i) If the cytoplasm was incubated with avidin and the malonate decarboxylase subsequently completed with the membrane fraction the decarboxylase activity was abolished. The corresponding incubation of the membrane with avidin, however, was without effect. (ii) Western blot analysis identified the single biotin-containing polypeptide of Mr 120'000 within the cytoplasm. (iii) Transmission electron micrographs of immuno-gold labeled M. rubra cells clearly showed the location of the biotinyl protein within the cytoplasm, whereas the same procedure with Propionigenium modestum cells indicated the location of the biotin enzyme methylmalonyl-CoA decarboxylase in the cell membrane. The biotin-containing protein of the M. rubra malonate decarboxylase enzyme system was not retained by monomeric avidin-Sepharose columns but could be isolated with this column in a catalytically inactive form in the presence of detergents. If the high binding affinity of tetrameric avidin towards biotin was reduced by destructing part of the tryptophan residues by irradiation or oxidation with periodate, the inhibition of malonate decarboxylase by the modified avidin was partially reversed with an excess of biotin. Attempts to purify the biotin protein in its catalytically active state using modified avidin columns were without success.


Model of birdsong learning based on gradient estimation by dynamic perturbation of neural conductances.
We propose a model of songbird learning that focuses on avian brain areas HVC and RA, involved in song production, and area LMAN, important for generating song variability. Plasticity at HVC --> RA synapses is driven by hypothetical "rules" depending on three signals: activation of HVC --> RA synapses, activation of LMAN --> RA synapses, and reinforcement from an internal critic that compares the bird's own song with a memorized template of an adult tutor's song. Fluctuating glutamatergic input to RA from LMAN generates behavioral variability for trial-and-error learning. The plasticity rules perform gradient-based reinforcement learning in a spiking neural network model of song production. Although the reinforcement signal is delayed, temporally imprecise, and binarized, the model learns in a reasonable amount of time in numerical simulations. Varying the number of neurons in HVC and RA has little effect on learning time. The model makes specific predictions for the induction of bidirectional long-term plasticity at HVC --> RA synapses.


A Scoring Method for Immunohistochemical Staining on Ki67.
An accurate interpretation of immunohistochemistry (IHC) staining results is crucial for precise disease diagnosis. In this study, we present a novel scoring method for interpreting and reporting of IHC staining assay results for the nuclear-type molecule. On the basis of the histologic characteristics, the samples were subdivided into 3 basic structural units and tissue subtypes including covered, mosaic, and mesenchymal subtypes. A cut-off of moderate-positive (2+) cells and 10% as the differential expression were applied to stratify the results into 11 grade scoring system (0 to X level). The observer can directly identify and count the number and percentage of positive cells from IHC staining data. Furthermore, Ki67 staining results in 88 carcinoma specimens were re-evaluated to determine the ease, reliability, reproducibility, and variance among different observers. The results indicated the consistency ratio of 68.0% for the mosaic subtype and 80% for the mesenchymal subtype, and 68.2% for the covered subtype by 5 experienced pathologists independently. Using 10% as the cut-off threshold, the consistency ratio of 92.5%, 96.8%, and 92.9% was noted for mosaic, mesenchymal, and covered subtypes, respectively. Besides, the correlation of counts revealed excellent agreement among the 5 independent pathologists. Overall, the proposed IHC scoring method is a novel, simple, reliable, and reproducible grading system for accurate interpretation of IHC staining data. Furthermore, the presented practical grading approach has the potential to improve the clinical evaluation of the IHC staining data for personalized therapy.