Italian consensus Guidelines for the management of hepatitis B virus infections in patients with rheumatoid arthritis.
Hepatitis B (HBV) infection, which is prevalent worldwide, is also frequently seen in patients with rheumatoid arthritis (RA). The Italian Society of Rheumatology (SIR) and the Italian Society of Infectious and Tropical Diseases (SIMIT) endorsed a national consensus process to review the available evidence on HBV management in RA patients and to produce practical, hospital-wide recommendations. The consensus panel consisted of infectious disease consultants, rheumatologists and epidemiologists and used the criteria of the Oxford Center for Evidence-based Medicine to assess the quality of the evidence and the strength of their recommendations. A core-set of statements has been developed to help clinicians in the management of patients with RA and HBV infection. Vaccination and prophylaxis of RA patients treated with biological drugs have been also discussed. HBV infection is not rare in clinical practice; a screening for HBV in all patients with early arthritis is not universally accepted, while it is considered mandatory before starting any immunosuppressive or hepatotoxic treatment. In fact, a specific risk, associated with the use of biologic treatments, exists for patients with HBV infection, although longitudinal studies of viral reactivation are generally reassuring. RA patients with HBV infection should be referred to the hepatologist and correctly classified into active or inactive carriers. Patients with active hepatitis B should undergo antiviral treatment before starting immunosuppressive treatments. Occult HBV carriers should be monitored or receive prophylaxis on the basis of the risk of reactivation associated with the administered treatment.


DNA gyrase stimulates transcription.
The nuclear DNA of HeLa cells can now be isolated unbroken and supercoiled. Using DNA gyrase and the untwisting enzyme, we have prepared an allomorphic series of templates derived from this nuclear DNA, and also from the circular DNA of the bacterial virus, PM2. We have then transcribed these templates using 2 different RNA polymerases--from wheat germ and Escherichia coli. Relaxed DNA is transcribed slowly by both polymerases. Supertwisting the naturally-supercoiled templates with gyrase slightly inhibits transcription by the bacterial polymerase but stimulates dramatically transcription by RNA polymerase II from wheat germ.


Development of the United Kingdom physician associate profession.
The world of healthcare is changing, and patient needs are changing with it. Traditional doctor-driven models of workforce planning are no longer sustainable in the United Kingdom (UK) healthcare economy, and newer models are needed. In the multiprofessional, multiskilled clinical workforce of the future, the physician associate (PA) has a fundamental role to play as an integrated, frontline, generalist clinician. As of 2016, about 350 PAs were practicing in the UK, with 550 PAs in training and plans to expand rapidly. This report describes the development of the PA profession in the UK from 2002, with projections through 2020, and includes governance, training, and the path to regulation. With rising demands on the healthcare workforce, the PA profession is predicted to positively influence clinical workforce challenges across the UK healthcare economy.


Live attenuated varicella vaccine use in immunocompromised children and adults.
Live attenuated varicella vaccine has been administered to 307 children with leukemia in remission and to 86 healthy adults. The vaccine was well tolerated and immunogenic. The major side effect in leukemic children receiving maintenance chemotherapy was development of a vaccine-associated rash. Vaccinees in whom a rash developed were potentially somewhat infectious to others about 1 month after immunization. Vaccination was not associated with an increase in the incidence of herpes zoster or in relapse of leukemia. Vaccination provided excellent protection against severe varicella. It was associated with a significant decrease in the attack rate of chickenpox following an intimate exposure to varicella-zoster virus, conferring about 80% protection in leukemic children. The cases of breakthrough varicella that occurred were mild. Thus, the vaccine may either prevent or modify varicella in high-risk individuals. It may also have use for prevention of nosocomial varicella.


Studying a Moving Target in Development: The Challenge and Opportunity of Heterotypic Continuity.
Many psychological constructs show heterotypic continuity-their behavioral manifestations change with development but their meaning remains the same (e.g., externalizing problems). However, research has paid little attention to how to account for heterotypic continuity. Conceptual and methodological challenges of heterotypic continuity may prevent researchers from examining lengthy developmental spans. Developmental theory requires that measurement accommodate changes in manifestation of constructs. Simulation and empirical work demonstrate that failure to account for heterotypic continuity when collecting or analyzing longitudinal data results in faulty developmental inferences. Accounting for heterotypic continuity may require using different measures across time with approaches that link measures on a comparable scale. Creating a developmental scale (i.e., developmental scaling) is recommended to link measures across time and account for heterotypic continuity, which is crucial in understanding development across the lifespan. The current synthesized review defines heterotypic continuity, describes how to identify it, and presents solutions to account for it. We note challenges of addressing heterotypic continuity, and propose steps in leveraging opportunities it creates to advance empirical study of development.


HTS discovery of PARP1-HPF1 complex inhibitors in cancer.
PARP1/2 inhibitors (PARPi) are effective clinically used drugs for the treatment of cancers with BRCA deficiencies. PARPi have had limited success and applicability beyond BRCA deficient cancers, and their effect is diminished by resistance mechanisms. The recent discovery of Histone PARylation Factor (HPF1) and the role it plays in the PARylation reaction by forming a shared active site with PARP1 raises the possibility that novel inhibitors that target the PARP1-HPF1 complex can be identified. Herein we describe a simple and cost-effective high-throughput screening (HTS) method aimed at discovering inhibitors of the PARP1-HPF1 complex. Upon HTS validation, we first applied this method to screen a small PARP-focused library of compounds and then scale up our approach using robotic automation to conduct a pilot screen of 10,000 compounds and validating >100 hits. This work demonstrates for the first time the capacity to discover potent inhibitors of the PARP1-HPF1 complex, which may have utility as probes to better understand the DNA damage response and as therapeutics for cancer.


Gastric microbial community profiling reveals a dysbiotic cancer-associated microbiota.
Gastric carcinoma development is triggered by Helicobacter pylori. Chronic H. pylori infection leads to reduced acid secretion, which may allow the growth of a different gastric bacterial community. This change in the microbiome may increase aggression to the gastric mucosa and contribute to malignancy. Our aim was to evaluate the composition of the gastric microbiota in chronic gastritis and in gastric carcinoma. The gastric microbiota was retrospectively investigated in 54 patients with gastric carcinoma and 81 patients with chronic gastritis by 16S rRNA gene profiling, using next-generation sequencing. Differences in microbial composition of the two patient groups were assessed using linear discriminant analysis effect size. Associations between the most relevant taxa and clinical diagnosis were validated by real-time quantitative PCR. Predictive functional profiling of microbial communities was obtained with PICRUSt. The gastric carcinoma microbiota was characterised by reduced microbial diversity, by decreased abundance of Helicobacter and by the enrichment of other bacterial genera, mostly represented by intestinal commensals. The combination of these taxa into a microbial dysbiosis index revealed that dysbiosis has excellent capacity to discriminate between gastritis and gastric carcinoma. Analysis of the functional features of the microbiota was compatible with the presence of a nitrosating microbial community in carcinoma. The major observations were confirmed in validation cohorts from different geographic origins. Detailed analysis of the gastric microbiota revealed for the first time that patients with gastric carcinoma exhibit a dysbiotic microbial community with genotoxic potential, which is distinct from that of patients with chronic gastritis.


Adjunctive canakinumab reduces peripheral inflammation markers and improves positive symptoms in people with schizophrenia and inflammation: A randomized control trial.
Clinical trials of anti-inflammatories in schizophrenia do not show clear and replicable benefits, possibly because patients were not recruited based on elevated inflammation status. Interleukin 1-beta (IL-1β) mRNA and protein levels are increased in serum, plasma, cerebrospinal fluid, and brain of some chronically ill patients with schizophrenia, first episode psychosis, and clinical high-risk individuals. Canakinumab, an approved anti-IL-1β monoclonal antibody, interferes with the bioactivity of IL-1β and interrupts downstream signaling. However, the extent to which canakinumab reduces peripheral inflammation markers, such as, high sensitivity C-reactive protein (hsCRP) and symptom severity in schizophrenia patients with inflammation is unknown. We conducted a randomized, placebo-controlled, double-blind, parallel groups, 8-week trial of canakinumab in chronically ill patients with schizophrenia who had elevated peripheral inflammation. Twenty-seven patients with schizophrenia or schizoaffective disorder and elevated peripheral inflammation markers (IL-1β, IL-6, hsCRP and/or neutrophil to lymphocyte ratio: NLR) were randomized to a one-time, subcutaneous injection of canakinumab (150 mg) or placebo (normal saline) as an adjunctive antipsychotic treatment. Peripheral blood hsCRP, NLR, IL-1β, IL-6, IL-8 levels were measured at baseline (pre injection) and at 1-, 4- and 8-weeks post injection. Symptom severity was assessed at baseline and 4- and 8-weeks post injection. Canakinumab significantly reduced peripheral hsCRP over time, F(3, 75) = 5.16, p = 0.003. Significant hsCRP reductions relative to baseline were detected only in the canakinumab group at weeks 1, 4 and 8 (p's = 0.0003, 0.000002, and 0.004, respectively). There were no significant hsCRP changes in the placebo group. Positive symptom severity scores were significantly reduced at week 8 (p = 0.02) in the canakinumab group and week 4 (p = 0.02) in the placebo group. The change in CRP between week 8 and baseline (b = 1.9, p = 0.0002) and between week 4 and baseline (b = 6.0, p = 0.001) were highly significant predictors of week 8 change in PANSS Positive Symptom severity scores. There were no significant changes in negative symptoms, general psychopathology or cognition in either group. Canakinumab was well tolerated and only 7 % discontinued. Canakinumab quickly reduces peripheral hsCRP serum levels in patients with schizophrenia and inflammation; after 8 weeks of canakinumab treatment, the reductions in hsCRP are related to reduced positive symptom severity. Future studies should consider increased doses or longer-term treatment to confirm the potential benefits of adjunctive canakinumab in schizophrenia. Australian and New Zealand Clinical Trials Registry number: ACTRN12615000635561.


A small phospholipase A2-α from castor catalyzes the removal of hydroxy fatty acids from phosphatidylcholine in transgenic Arabidopsis seeds.
Ricinoleic acid, an industrially useful hydroxy fatty acid (HFA), only accumulates to high levels in the triacylglycerol fraction of castor (Ricinus communis) endosperm, even though it is synthesized on the membrane lipid phosphatidylcholine (PC) from an oleoyl ester. The acyl chains of PC undergo intense remodeling through the process of acyl editing. The identities of the proteins involved in this process, however, are unknown. A phospholipase A2 (PLA2) is thought to be involved in the acyl-editing process. We show here a role for RcsPLA2α in the acyl editing of HFA esterified to PC. RcsPLA2α was identified by its high relative expression in the castor endosperm transcriptome. Coexpression in Arabidopsis (Arabidopsis thaliana) seeds of RcsPLA2α with the castor fatty acid hydroxylase RcFAH12 led to a dramatic decrease in seed HFA content when compared with RcFAH12 expression alone in both PC and the neutral lipid fraction. The low-HFA trait was heritable and gene dosage dependent, with hemizygous lines showing intermediate HFA levels. The low seed HFA levels suggested that RcsPLA2α functions in vivo as a PLA2 with HFA specificity. Activity assays with yeast (Saccharomyces cerevisiae) microsomes showed a high specificity of RcsPLA2α for ricinoleic acid, superior to that of the endogenous Arabidopsis PLA2α. These results point to RcsPLA2α as a phospholipase involved in acyl editing, adapted to specifically removing HFA from membrane lipids in seeds.


Immune responses of lung transplant recipients against SARS-CoV-2 and common respiratory coronaviruses: Evidence for pre-existing cross-reactive immunity.
Humoral and cellular immune responses to SARS-CoV-2 and other coronaviruses in lung transplant recipients are unknown. We measured antibodies and T cell responses against the SARS-CoV-2 spike S2 and nucleocapsid antigens and spike antigens from common respiratory coronaviruses (229E, NL63, OC43, and HKU1) after vaccination or infection of LTxRs. 148 LTxRs from single center were included in this study: 98 after vaccination and 50 following SARS-CoV-2 infection. Antibodies were quantified by enzyme-linked immunosorbent assay. The frequency of T cells secreting IL2, IL4, IL10, IL17, TNFα, and IFNγ were enumerated by enzyme-linked immunospot assay. Our results have shown the development of antibodies to SARS-CoV-2 spike protein in infected LTxRs (39/50) and vaccinated LTxRs (52/98). Vaccinated LTxRs had higher number of T cells producing TNFα but less cells producing IFNγ than infected LTxRs in response to the nucleocapsid antigen and other coronavirus spike antigens. We didn't find correlation between the development of antibodies and cellular immune responses against the SARS-CoV-2 spike protein after vaccination. Instead, LTxRs have pre-existing cellular immunity to common respiratory coronaviruses, leading to cross-reactive immunity against SARS-CoV-2 which likely will provide protection against SARS-Cov-2 infection.


Evoked Potentials in Multiple Sclerosis Diagnosis and Management.
We high lightened the changes in Visual Evoked Potentials (VEP) and Brainstem Auditory Evoked Potentials (BAEP) in Multiple Sclerosis (MS). The recordings of evoked potentials were repeated at an interval of 2 months for a total of 12 months. Prolonged latencies of the VEP waves (mostly P100) gas confirmed the multifocal character of MS lesions. The positive effect of corticostheroids in MS flare and the changing course of the disease following administration of interferon. The BAEP recordings have not shown reproducible modifications. Succesive VEP monitoring brings on valuable information by highlighting infraclinical lesions and monitoring the evolution of the disease.


Preparation and induction of immune responses by a DNA AIDS vaccine.
In an effort to evaluate the feasibility of developing a safe DNA vaccine for acquired immunodeficiency syndrome (AIDS), we have prepared a plasmid-based immunogen modeled after a naturally occurring noninfectious mutant of the simian immunodeficiency virus (SIV). The mutant SIV genome produces defective virus particles that are noninfectious in vitro and nonpathogenic in vivo in rhesus macaques. Analysis of the mutant genome revealed a 1.6 kb deletion that is in frame and spans integrase, vif, vpx, and most of vpr and results in a pol/vpr gene fusion. This deletion was introduced into the parental pathogenic molecular clone and the U3 region of the 5' LTR was replaced with a cytomegalovirus promoter to produce a candidate DNA vaccine, pIV. After transfection with this plasmid, SIV gag and envelope proteins are expressed and properly processed in vitro. When injected into rabbits, pIV elicited an antibody response to SIV gp130 envelope glycoprotein with titers reaching 1:2048, and a strong lymphoproliferative response to SIV gp130 and whole SIV. The potential to produce defective virus particles in vivo without integrating into the host genome should result in both a strong humoral and cellular immune response in rhesus macaques. In addition, this approach offers a safe alternative to live attenuated vaccines and DNA vaccines that are capable of integration.


[Lacrimal duct reconstruction in tumor surgery and following trauma].
Traumatic injuries of the lid often affect the lower lid and the lower canaliculus. Lid destruction by tumours also frequently takes place in the medial part of the lower lid, involving the canaliculus inferior. In order to prevent chronic epiphora the lower canaliculus has to be reconstructed. The most elegant and effective method is the bicanalicular silicon ring intubation (Murube de Castillo). This technique can be utilized in traumatic cases and in tumour surgery. The intubation technique is described and the results are reported.


Relating solubility data of parabens in liquid PEG 400 to the behaviour of PEG 4000-parabens solid dispersions.
The solid state behaviour of polyethylene glycol 4000 (PEG 4000) and dispersions of a homologous series of parabens (methyl- (MP), ethyl- (EP), propyl- (PP) and butyl- (BP)) were examined and compared to the paraben solubility in liquid PEG 400. Dispersions were prepared by co-melting different amounts of paraben (5-80% (w/w)) and PEG 4000 and were studied using a combination of differential scanning calorimetry (DSC) and small and wide angle X-ray diffraction (SAXD/WAXD). Depending on the concentration of parabens in the dispersions, DSC showed melting peaks from folded and unfolded forms of PEG, a eutectic melting and melting of pure parabens. The fraction of folded PEG increased and the melting temperatures of both PEG forms decreased with increasing paraben content. In an apparent phase diagram of PP-PEG dispersions a eutectic mixture appeared above 5% PP. In addition, a melting peak corresponding to the paraben appeared for dispersion containing more than 60% PP. Similar phase diagrams were shown for the other parabens. The SAXD data and a 1D correlation function analysis revealed that MP and BP were incorporated into the amorphous domains of the lamellae of solid PEG to a higher degree than EP and PP. In addition, the lamellae thickness of PEG and the fraction of amorphous domains increased more for MP and BP compared to EP and PP. BP showed the highest solubility of the parabens followed by MP, EP and PP in both liquid and solid PEG. Furthermore, the thickness of the amorphous domains of the PEG in the different parabens-PEG dispersions could be correlated to the solubility in liquid PEG 400.


Two routes of iron accumulation in astrocytes: ascorbate-dependent ferrous iron uptake via the divalent metal transporter (DMT1) plus an independent route for ferric iron.
Astrocytes are central to iron and ascorbate homoeostasis within the brain. Although NTBI (non-transferrin-bound iron) may be a major form of iron imported by astrocytes in vivo, the mechanisms responsible remain unclear. The present study examines NTBI uptake by cultured astrocytes and the involvement of ascorbate and DMT1 (divalent metal transporter 1). We demonstrate that iron accumulation by ascorbate-deficient astrocytes is insensitive to both membrane-impermeant Fe(II) chelators and to the addition of the ferroxidase caeruloplasmin. However, when astrocytes are ascorbate-replete, as occurs in vivo, their rate of iron accumulation is doubled. The acquisition of this additional iron depends on effluxed ascorbate and can be blocked by the DMT1 inhibitor ferristatin/NSC306711. Furthermore, the calcein-accessible component of intracellular labile iron, which appears during iron uptake, appears to consist of only Fe(III) in ascorbate-deficient astrocytes, whereas that of ascorbate-replete astrocytes comprises both valencies. Our data suggest that an Fe(III)-uptake pathway predominates when astrocytes are ascorbate-deficient, but that in ascorbate-replete astrocytes, at least half of the accumulated iron is initially reduced by effluxed ascorbate and then imported by DMT1. These results suggest that ascorbate is intimately involved in iron accumulation by astrocytes, and is thus an important contributor to iron homoeostasis in the mammalian brain.


Economic fabrication of a novel hybrid planar Grating/Fresnel lens for miniature spectrometers.
We propose a new technique to fabricate a highly specialized optical element, a hybrid planar Grating/Fresnel lens (G-Fresnel), which is particularly useful to improve or enable more-affordable miniature/portable spectrometers. Both the Fresnel and the grating surface are fabricated simultaneously by sandwiching soft PDMS between a hard grating and a pre-replicated negative Fresnel surface. Several adhesion reduction techniques are also investigated that help improve both fabrication and cost efficiency (by reducing the solidification time) as well as the lifetime of the mold. Alignment errors are systematically analyzed, and their effects on the G-Fresnel lens evaluated. A compact fabrication platform was built, which is smaller than a volume of 160☓140☓106 mm3 to fit into a conventional vacuum drying oven, for the fabrication of a G-Fresnel lens with a diameter of 25.4 mm, an equivalent focal length of 25 mm, and a blazed grating pattern with 600 lines/mm spacing. The solidification time was reduced to 2 hours thanks to the improved adhesion reduction technique that permits a PDMS drying-temperature as high as 65 °C. The fabricated G-Fresnel lens was evaluated with regard to both geometrical fabrication precision and optical performance. The measured results, using a step gauge and atomic force microscopy, confirm that this replication technique produces high-quality replicates of the master surface-profile. Furthermore, a prototype spectrometer that uses a G-Fresnel lens was built and evaluated. The spectrometer fits within a volume of about 100 mm☓50 mm☓30 mm, and it operates across a wide wavelength spectrum (450 nm to 650 nm). Both the calculation based on the optical software ZEMAX and the experimental measurements are consistent and confirm that the spectrometer with the G-Fresnel lens can provide a spectral resolution of better than 1.2nm.


Chemical Composition, Antibacterial and Phytotoxic Activities of Peganum harmala Seed Essential Oils from Five Different Localities in Northern Africa.
Peganum harmala L., also known as Syrian rue or Pègano, is a herbaceous plant belonging to the Zygohpyllaceae family, and is widely used in traditional medicine. The chemical composition of essential oils of P. harmala seeds from five different regions of Northern Africa (Algeria, Egypt, Libya, Morocco and Tunisia) was studied by GC and GC-MS analyses. A total of 105 compounds were identified, the main components being oxygenated monoterpenes and oxygenated sesquiterpenes. Eugenol is the main component in all oils. The antimicrobial activity of the essential oils was assayed against some bacterial strains: Staphylococcus aureus (DSM 25693), Bacillus cereus (DSM 4313), Bacillus cereus (DSM4384), Escherichia coli (DMS 857) and Pseudomonas aeruginosa (ATCC 50071). All the oils showed different inhibitory activity. In the twentieth century this is an important result; we need possible new botanical drugs because the problem of resistance to antimicrobial drugs has become apparent. Moreover, the essential oils were evaluated for their possible in vitro phytotoxic activity against germination and initial radicle growth of Raphanus sativus L., Lepidium sativum L., and Ruta graveolens L. The results showed that both germination and radical elongation were sensitive to the oils.


An abnormal response of retinoblastoma cells (Y-79) to neurotrophins.
To clarify the expression of neurotrophins and their receptors in retinoblastoma (Rb) cells, to elucidate their potential role in the proliferation of neuroectodermal tumor cells, and to establish conditions for Rb cell differentiation. The Rb-derived cell line Y-79 was grown in serum-free suspension or monolayer culture. Proliferating and differentiated cells were isolated and submitted to semiquantitative reverse transcription-polymerase chain reaction (RT-PCR) analysis, immunostaining, and flow cytometry. The proliferation rate of the cells was estimated by 5-bromo-2'-deoxyuridine (BrdU) incorporation, and the effects of neurotrophins and laminin on BrdU-incorporation, process outgrowth, or immunostaining were determined. In contrast to previously studied normal retinal precursor cells, Y-79 cells not only express nerve growth factor (NGF), brain-derived neurotrophic factor (BDNF), neurotrophin-3 (NT-3), and p75, but also the corresponding high affinity receptors TrkA, TrkB, and TrkC. Proliferation was stimulated by exogenous and endogenous neurotrophin receptor ligands. Inhibition of protein kinase phosphorylation with K252a blocked proliferation and promoted differentiation. The effect of K252a on differentiation was enhanced by the addition of soluble laminin. After 9 days of combined treatment, the fraction of differentiated cells amounted to 30%, differentiation being characterized by improved attachment, neurite outgrowth, expression of NF-68, and a loss of glial fibrillary acidic protein (GFAP) and parvalbumin immunoreactivity. These changes were accompanied by a downregulation of TrkB and TrkC, but not TrkA or p75. Differentiated cells were isolated and further grown in the absence of K252a. However, despite the high level of TrkA expression in differentiated cells, the addition of NGF had no effect on their survival. A mitogenic action of neurotrophins could contribute to retinal tumor growth.


Species abundance and temporal variation of arbovirus vectors in Brownsville, Texas.
The recent outbreaks of the dengue fever and West Nile viruses and the looming threats of the Zika and chikungunya viruses highlight the importance of establishing effective, proactive arboviral surveillance in communities at high risk of transmission, such as those on the Texas-Mexico border. Currently, there are no approved human vaccines available for these mosquito-borne diseases, so entomological control and case management are the only known methods for decreasing disease incidence. The principal vectors, which include Culex quinquefasciatus, Aedes aegypti, and Ae. Albopictus, all have an established presence in South Texas. The public health response to most arbovirus outbreaks in the region has been reactionary rather than proactive. However, after the 2005 dengue outbreak and subsequent fatality, the City of Brownsville Public Health Department began collecting data on mosquito vector abundance and incidence. The objective of this study was to describe the various species of mosquitoes found in vector surveillance in Brownsville, Texas, during 2009-2013; quantify their prevalence; and identify any associations with temporal or weather-related variations. The results confirm a significant mosquito population in Brownsville in late winter months, indicating a high risk of arbovirus transmission in South Texas year-round, and not just until November, previously considered the end date of arbovirus season by state health services. The data from Brownsville's surveillance program can help characterize local vector ecology and facilitate more proactive mitigation of future arboviral threats in South Texas.


Impact of contact lens group on antifungal efficacy of multipurpose disinfecting contact lens solutions.
To evaluate and compare the in vitro effectiveness of multipurpose contact lens solutions (MPDS) to inhibit fungal colonization of Food and Drug Administration (FDA) contact lens groups. Contact lenses from FDA groups I (low water content, nonionic), II (high water content, nonionic), III (low water content, ionic), and IV (high water content, ionic) were placed in sterile contact lens cases with 2 mL of ReNu MultiPlus, ReNu with MoistureLoc, or OPTI-FREE RepleniSH. Each contact lens-multipurpose solution combination was challenged with 0.1 mL of 10(6) colony-forming units/mL of Fusarium oxysporum and incubated at room temperature. Contact lenses and aliquots from phosphate-buffered saline rinse solutions, multipurpose solutions in cases, and lens cases were sampled at 24, 48, and 72 hours, plated on Sabouraud's agar, incubated at room temperature, and observed for growth (i.e., adhesion) for 1 to 7 days. Used multipurpose contact lens solutions met or exceeded the 1-log reduction requirement of the FDA stand-alone test. Viable fungal colonies were recovered from group II lenses after 24 hours of storage in all three multipurpose solutions. Colonization was also detected with at least one contact lens-multipurpose solution pairing from groups I, III, and IV. Fungi were not recovered from lens-solution combinations after 24 hours or from phosphate-buffered saline solutions or lens cases at any period. Antifungal activity of contaminated multipurpose solutions may be insufficient to prevent fungal colonization of contact lens materials despite meeting or exceeding the FDA stand-alone test criteria. Colonized lens may increase the risk of fungal keratitis.


Epitope mapping by chemical modification of free and antibody-bound protein antigen.
A monoclonal antibody bound to a protein antigen slows the rate of chemical modification of amino acid residues located at the epitope. By comparing the degree of acetylation of 18 lysine and 7 threonine residues in free and antibody-bound horse cytochrome c, a discontiguous, conformational epitope was characterized on this protein antigen. The new approach is particularly suitable to probe discontiguous and conformational epitopes, which are difficult to analyze by other procedures.


Effect of Hfq on RprA-rpoS mRNA pairing: Hfq-RNA binding and the influence of the 5' rpoS mRNA leader region.
The rpoS mRNA encodes a stress response transcription factor in Escherichia coli. It is one of a growing number of mRNAs found to be regulated by small RNAs (sRNA). Translation initiation of rpoS mRNA is enhanced by two sRNAs, DsrA and RprA, that pair to the same site near the rpoS start codon in the presence of the Hfq protein. In this work, we examine the interaction of E. coli Hfq with RprA and two portions of the rpoS mRNA leader region. One rpoS RNA, rpoS-L, contained the entire 565-nucleotide leader region, while the other, rpoS-S, contained the 199-nucleotide sequence surrounding the start codon. An RNase H assay indicated both rpoS RNAs have similar secondary structures in the translation initiation region. Hfq formed two complexes with RprA in a gel mobility assay with binding parameters similar to values previously determined for DsrA. Unlike DsrA, Hfq binding to RprA was inhibited by poly(A) and influenced by Hfq mutations on both the distal and proximal surfaces. Hfq increased the level of RprA binding to both rpoS RNAs but showed a much larger enhancement when rpoS-L, the entire leader region, was examined. The lower affinity of RprA for rpoS-L versus rpoS-S in the absence of Hfq suggests that Hfq overcomes an inhibitory structure within rpoS-L in stimulating RprA binding. Similar results were obtained with DsrA. The results indicate that the full upstream leader sequence of rpoS mRNA influences Hfq-facilitated annealing of RprA and DsrA and is likely to be involved in its regulation.


Pancreatic insulinomas. A 15-year experience.
To describe our experience in the management of patients with pancreatic insulinomas, emphasizing the need for preoperative localization and the outcome of surgical treatment. A case series. A university hospital in Hong Kong. From 1981 to 1995, 27 patients with pancreatic insulinomas were surgically treated; the mean follow-up was 25.4 months. Postoperative morbidity and euglycemia during the follow-up period. The accuracy of tumor localization by ultrasonography, computed tomography, and angiography was 33%, 44%, and 52%, respectively. Venous sampling for an insulin assay regionalized 90% of the tumors. In 24 patients with solitary tumors, most lesions detected or missed by preoperative localization could be either seen (n = 14) or palpated (n = 22). Intraoperative ultrasonography (n = 17) has been routinely performed since 1987; nonpalpable tumors were imaged in 2 of 15 patients with solitary tumors. Eight solitary occult tumors were detected by palpation alone or a combination of palpation and intraoperative ultrasonography. Operative mortality occurred in 1 (3.7%) of the 27 patients, while major morbidity developed in 9 (33%) of the patients. Euglycemia was achieved in 25 patients. Surgery cured all patients with benign insulinomas, whereas the cure rate for patients with malignant neoplasms was only 33%. The type of surgical treatment or correct preoperative localization did not affect the outcome of surgery. Pancreatic insulinomas can be readily localized intraoperatively despite failed preoperative localization studies. Surgical treatment cured benign adenomas but was associated with notable morbidity.


Identification of novel plant cysteine oxidase inhibitors from a yeast chemical genetic screen.
Hypoxic responses in plants involve Plant Cysteine Oxidases (PCOs). They catalyze the N-terminal cysteine oxidation of Ethylene Response Factors VII (ERF-VII) in an oxygen-dependent manner, leading to their degradation via the cysteine N-degron pathway (Cys-NDP) in normoxia. In hypoxia, PCO activity drops, leading to the stabilization of ERF-VIIs and subsequent hypoxic gene upregulation. Thus far, no chemicals have been described to specifically inhibit PCO enzymes. In this work, we devised an in vivo pipeline to discover Cys-NDP effector molecules. Budding yeast expressing AtPCO4 and plant-based ERF-VII reporters was deployed to screen a library of natural-like chemical scaffolds and was further combined with an Arabidopsis Cys-NDP reporter line. This strategy allowed us to identify three PCO inhibitors, two of which were shown to affect PCO activity in vitro. Application of these molecules to Arabidopsis seedlings led to an increase in ERF-VII stability, induction of anaerobic gene expression, and improvement of tolerance to anoxia. By combining a high-throughput heterologous platform and the plant model Arabidopsis, our synthetic pipeline provides a versatile system to study how the Cys-NDP is modulated. Its first application here led to the discovery of at least two hypoxia-mimicking molecules with the potential to impact plant tolerance to low oxygen stress.


Integration of a high finesse cryogenic build-up cavity with an ion trap.
We report on the realization of a hemispherical optical cavity with a finesse of F = 13 000 and sustaining inter-cavity powers of 10 kW, which we operate in a closed-cycle cryostat vacuum system close to 4 K. This was designed and built with an integrated radio-frequency Paul trap in order to combine optical and radio-frequency trapping. The cavity provides a power build-up factor of 2300. We describe a number of aspects of the system's design and operation, including low-vibration mounting and locking and thermal effects at high powers. Thermal self-locking in the high intracavity power regime was observed to enhance passive stability below 1 kHz. Observations made over repeated cool-downs over the course of a year show a repeatable shift between the ion trap center and the cavity mode.


Rare Huge Tubular Papillary Adenoma on the Buttock Demonstrated by Magnetic Resonance Imaging (MRI).
BACKGROUND Tubular papillary adenoma is a rare eccrine-derivate dermal adnexal tumor, located in the scalp mostly. Earlier, the sebaceous cyst size was larger than 5 cm in diameter and may be confused with subcutaneous tumor. Cases of the tumor on the buttock with rapid growth are rare, therefore magnetic resonance imaging (MRI) may be helpful to determine their precise size, nature, and invasion of nearby organs for further confirmation. CASE REPORT We report on a case of a 35-year-old male with a rapid-growth tumor on the buttock. Initially, he had received drainage by syringe and the amount of drainage was 50 mL. MRI favored diagnosis of sebaceous cyst. As for recurrent tumor, the pathology revealed tubular papillary adenoma. CONCLUSIONS MRI might play an important role on imaging of soft tissue to exclude some uncertain malignant tumors. This case report indicated a rare case of a large rapid-growing tubular papillary adenoma on the buttock that required further management.


BIOCHEMICAL CHANGES DURING THE GROWTH OF FUNGI. II. ERGOSTEROL AND FATTY ACIDS IN PENICILLIUM ATROVENETUM.
Van Etten, James L. (University of Illinois, Urbana), and David Gottlieb. Biochemical changes during the growth of fungi. II. Ergosterol and fatty acids in Penicillium atrovenetum. J. Bacteriol. 89:409-414. 1965.-Changes in the lipid constituents of Penicillium atrovenetum were studied during the growth and development of this fungus. The stages in development, as measured by the dry weight, were divided into four phases: lag, log, stationary, and death. The total fatty acids on a dry-weight basis increased from a minimum in the spores to a maximum near the end of the log phase of growth. The major fatty acids were palmitic, stearic, oleic, and linoleic. Myristic, pentadecanoic, palmitoleic, heptadecanoic, linolenic, arachidic, and heptadecenoic acids, together with two unidentified components, were also present in the fatty acid fraction. Compared to ungerminated spores, young mycelium contained a much lower percentage (on the basis of total fatty acids) of linoleic acid. There was a corresponding increase of oleic acid. Except for palmitic acid, which remained constant, the remaining fatty acids increased slightly. During subsequent growth of the fungus, linleic acid decreased, whereas the percentage of palmitic and stearic acids increased steadily from the lag phase to the end of the log phase. Ergosterol was the only sterol detected. The percentage of ergosterol, on a dry-weight basis, increased to a maximum at the start of the log phase and then steadily decreased. The following changes in lipids appear to be associated with the development and aging of fungi: (i) the presence of a relatively high content of nonsaponifiable lipid and ergosterol in the young mycelium, and their later decrease with age, and (ii) a shift from more unsaturated to less unsaturated fatty acids with age.


Design, fabrication and perivascular implantation of bioactive scaffolds engineered with human adventitial progenitor cells for stimulation of arteriogenesis in peripheral ischemia.
Cell therapy represents a promising option for revascularization of ischemic tissues. However, injection of dispersed cells is not optimal to ensure precise homing into the recipient's vasculature. Implantation of cell-engineered scaffolds around the occluded artery may obviate these limitations. Here, we employed the synthetic polymer polycaprolactone for fabrication of 3D woodpile- or channel-shaped scaffolds by a computer-assisted writing system (pressure assisted micro-syringe square), followed by deposition of gelatin (GL) nanofibers by electro-spinning. Scaffolds were then cross-linked with natural (genipin, GP) or synthetic (3-glycidyloxy-propyl-trimethoxy-silane, GPTMS) agents to improve mechanical properties and durability in vivo. The composite scaffolds were next fixed by crown inserts in each well of a multi-well plate and seeded with adventitial progenitor cells (APCs, 3 cell lines in duplicate), which were isolated/expanded from human saphenous vein surgical leftovers. Cell density, alignment, proliferation and viability were assessed 1 week later. Data from in vitro assays showed channel-shaped/GPTMS-crosslinked scaffolds confer APCs with best alignment and survival/growth characteristics. Based on these results, channel-shaped/GPTMS-crosslinked scaffolds with or without APCs were implanted around the femoral artery of mice with unilateral limb ischemia. Perivascular implantation of scaffolds accelerated limb blood flow recovery, as assessed by laser Doppler or fluorescent microspheres, and increased arterial collaterals around the femoral artery and in limb muscles compared with non-implanted controls. Blood flow recovery and perivascular arteriogenesis were additionally incremented by APC-engineered scaffolds. In conclusion, perivascular application of human APC-engineered scaffolds may represent a novel option for targeted delivery of therapeutic cells in patients with critical limb ischemia.


Communication and healthcare complexity in people with little or no English: the Communication Complexity Score.
To investigate the level of healthcare complexity of patients with limited English and the probability of use of professional interpreters, at a tertiary hospital in a multicultural area of Sydney, Australia. A multilingual telephone survey was matched with medical record reviews to assess modes of communication in a sample of 258 patients with limited English from nine language groups. The Model of Interpreter Usage and Complexity was used to explain the association between clinical complexity and interpreter usage. To measure clinical complexity, a summative rating scale, the Communication Complexity Score (CCS) was constructed comprising eight weighted scores related to admission status, urgency of admission, triage category, medical emergencies/events, HDU/ICU admission, numbers of medications, invasiveness of procedures, and diagnosis. Robust Multiple Logistic Regression was used for modelling the likelihood of interpreter usage with different levels of complexity. Of the 205 patients (79%) who spoke little or no English, 31% reported usage of professional interpreters. There was a positive association between CCS and usage of interpreters. Patients with high and moderate CCSs were significantly more likely to have used interpreters (OR=32.9, 95% CI 12.4-87.1 and OR=10.8, 95% CI 3.2-36.5, respectively). The CCS was found to have good psychometric properties and to be internally consistent (Cronbach's Alpha 0.84). The model was found to have good predictive power (Area under the Receiver Operating Characteristic curve 0.90). While most patients (69%) had no interpreter, patients with high or moderate CCSs were more likely to have an interpreter, demonstrating a robust relationship between these two variables. Poor usage of interpreters in the Emergency Department was observed. Family and friends had a significant role in communication. These findings have important safety, policy, and practice implications for patients with limited English proficiency.


Structure of the insulin receptor of rat adipocytes. The three interconvertible redox forms.
Isolated intact rat adipocytes were photoaffinity labeled with radioactive and photoreactive N alpha B1-monoazidobenzoyl insulin (B1-MABI) or N epsilon B29-monoazidobenzoyl insulin (B29-MABI). Polyacrylamide gel electrophoresis of the labeled plasma membranes solubilized in sodium dodecyl sulfate revealed the specific labeling of three receptor species of 380 kDa, 300 kDa, and 230 kDa. Reduction of each species individually produced the subunits of 130 kDa, 90 kDa, and 40 kDa. Exposure of the adipocytes or plasma membranes after photolabeling to sulfhydryl alkylating agents such as N-ethylmaleimide or p-chloromercuriphenylsulfonate resulted in the appearance of the receptor quantitatively in the 380-kDa form. The effect of the sulfhydryl reagent was concentration dependent and in the case of p-chloromercuriphenylsulfonate the three receptor species reappeared when high concentrations of the reagent were used. Incubation of the adipocytes with low concentrations of dithiothreitol before photolabeling reduced these receptors to discrete lower-molecular-weight forms. In addition, an 85-kDa subunit was now photolabeled by B1-MABI. This subunit was demonstrated to be different from the 90-kDa subunit normally labeled by B29-MABI. We conclude that on the cell surface of the adipocyte, there is one molecular-weight form of insulin receptor of 380 kDa composed of one 130-kDa, one 90-kDa, one 85-kDa, and two 40-kDa subunits. The 300 kDa and 230 kDa are partially reduced forms of the 380-kDa species. We further postulate that a membrane factor or factors sensitive to sulfhydryl alkylating reagents may be involved in the partial reduction and oxidation of these three redox receptor species. The distribution of these redox receptor species may be related to the cellular or tissue sensitivity to insulin.


Matching oceanography and genetics at the basin scale. Seascape connectivity of the Mediterranean shore crab in the Adriatic Sea.
Investigating the interactions between the physical environment and early life history is crucial to understand the mechanisms that shape the genetic structure of marine populations. Here, we assessed the genetic differentiation in a species with larval dispersal, the Mediterranean shore crab (Carcinus aestuarii) in the Adriatic Sea (central Mediterranean), and we investigated the role of oceanic circulation in shaping population structure. To this end, we screened 11 polymorphic microsatellite loci from 431 individuals collected at eight different sites. We found a weak, yet significant, genetic structure into three major clusters: a northern Adriatic group, a central Adriatic group and one group including samples from southern Adriatic and Ionian seas. Genetic analyses were compared, under a seascape genetics approach, with estimates of potential larval connectivity obtained with a coupled physical-biological model that integrates a water circulation model and a description of biological traits affecting dispersal. The cross-validation of the results of the two approaches supported the view that genetic differentiation reflects an oceanographic subdivision of the Adriatic Sea into three subbasins, with circulation patterns allowing the exchange of larvae through permanent connections linking north Adriatic sites and ephemeral connections like those linking the central Adriatic with northern and southern locations.


A story of chelatase evolution: identification and characterization of a small 13-15-kDa "ancestral" cobaltochelatase (CbiXS) in the archaea.
The cobaltochelatase required for the synthesis of vitamin B12 (cobalamin) in the archaeal kingdom has been identified as CbiX through similarity searching with the CbiX from Bacillus megaterium. However, the CbiX proteins in the archaea are much shorter than the CbiX proteins found in eubacteria, typically containing less than half the number of amino acids in their primary structure. For this reason the shorter CbiX proteins have been termed CbiXS and the longer versions CbiXL. The CbiXS proteins from Methanosarcina barkeri and Methanobacter thermoautotrophicum were overproduced in Escherichia coli as recombinant proteins and characterized. Through complementation studies of a defined chelatase-deficient strain of E. coli and by direct in vitro assays the function of CbiXS as a sirohydrochlorin cobaltochelatase has been demonstrated. On the basis of sequence alignments and conserved active site residues we suggest that CbiXS may represent a primordial chelatase, giving rise to larger chelatases such as CbiXL, SirB, CbiK, and HemH through gene duplication and subsequent variation and selection. A classification scheme for chelatases is proposed.


[Role of platelets in the pathogenesis of Coxiella infection].
To determine the relationship of hemostatic disorders to the direct impact of Coxiella burnetii on platelets as one of the key mechanisms of the pathogenesis of Q fever. SUBJECTS AND METHODS; Platelet functional activity, plasma hemostatic parameters, von Willibrand factor (vWF) were investigated; and polymerase chain reaction assay was used to determine C. burnetii DNA in the leukocyte and platelet sediments of 41 patients aged 39.9 +/- 0.8 years diagnosed with Q fever at the Astrakhan Regional Clinical Hospital in 2009 to 2010. The examinees were recorded to have hemorrhagic phenomena (34.7%) as a hematoma (27.2%), gingival (2.4%) and nasal (9.2%) hemorrhages, vomiting blood streaks (3.4%), melena (4.5%), roseolous-papular (22.1%) and hemorrhagic (9.3%) rashes on the skin. Examination of hemostasis revealed thrombocytopenia and platelet hypoaggregation, increased plasma fibrinogen homeostasis, and significantly elevated vWF during convalescence. C. burnetii genomic DNA was isolated from platelets in all the examinees, from leukocytes and platelets in 78% of cases and only from platelets in 22%. A fluorescence signal indicating the pathogen genome was more early recorded in 54.8% of cases in the platelets than in the leukocytes. At week 1 of the disease, the absence of significant plasma hemostatic changes and the retention of the control level vWF with the lower count of platelets and their aggregatory activity suggest that the platelets are able to interact with this pathogen, which is confirmed by the results of genodiagnosis of this rickettsiosis with the pathogens being isolated from the platelet sediment. The determination of platelet aggregatory activity is a primary diagnostic test to detect disorders in the hemostatic system. The higher detection rate of C. burnetii genomic DNA from the platelets than from the leukocyte sediment can recommend that platelets be used as biological material in the diagnosis of Q fever.


Discovery of 3-oxo-1,2,3,4-tetrahydropyrido[1,2-a]pyrazin derivatives as SARS-CoV-2 main protease inhibitors through virtual screening and biological evaluation.
The COVID-19 caused by SARS-CoV-2 has led to a global pandemic that continues to impact societies and economies worldwide. The main protease (Mpro) plays a crucial role in SARS-CoV-2 replication and is an attractive target for anti-SARS-CoV-2 drug discovery. Herein, we report a series of 3-oxo-1,2,3,4-tetrahydropyrido[1,2-a]pyrazin derivatives as non-peptidomimetic inhibitors targeting SARS-CoV-2 Mpro through structure-based virtual screening and biological evaluation. Further similarity search and structure-activity relationship study led to the identification of compound M56-S2 with the enzymatic IC50 value of 4.0 μM. Moreover, the molecular simulation and predicted ADMET properties, indicated that non-peptidomimetic inhibitor M56-S2 might serve as a useful starting point for the further discovery of highly potent inhibitors targeting SARS-CoV-2 Mpro.


Sexual dimorphism in adrenergic regulation of hepatic glycogenolysis.
The total phosphorylase a plus b of hepatocytes isolated from females and incubated in the absence or presence of estradiol and progesterone at concentrations found in vivo does not vary during the estrous cycle. However, there is a slight but significant influence of the estrous cycle on basal and epinephrine-stimulated phosphorylase a activity, with a nadir being seen on diestrus. The relative contributions of the alpha- and beta-mediated pathways to phosphorylase a activation do not vary with the estrous cycle but are constant at 75 and 56%, respectively, of the response to 5 X 10(-8) M epinephrine. When the epinephrine-stimulated glucose release from glycogen stores in cells from females and males is compared, the release from the female is greater than that from the male, while the alpha-receptor-mediated stimulation in the female is comparable with that in the male. The beta-mediated pathway causes glucose release which averages 45% of that stimulated by epinephrine alone in cells from females. Basal cytosolic free calcium is similar in cells from males and females (142 vs. 149 nM, respectively). The epinephrine-stimulated increase in cytostolic free calcium (Cai) is greater in the male than the female at 10(-6) M (489 vs. 380 nM) but greater in the female than the male at 5 X 10(-9) M (54 vs. 27 nM). The changes in Cai are equivalent at intermediate epinephrine concentrations. When considered with our prior analysis of 45Ca efflux after adrenergic stimulation, this suggests there may be a sexual dimorphism in hepatocyte calcium transport systems. The glucose release for a given increase in Cai is greater in the female than the male probably due to the concomitant action of the beta-mediated increase in cAMP and the alpha-mediated increase in Cai. This supports the conclusion that the beta-mediated component does make a significant contribution to the catecholamine regulation of glycogenolysis in hepatocytes from adult female rats.


Social Engagement and Antipsychotic Use in Addressing the Behavioral and Psychological Symptoms of Dementia in Long-Term Care Facilities.
Objectives The use of antipsychotics, mainly to address the behavioral and psychological symptoms of dementia (BPSD), remains a common and frequent practice in long-term care facilities (LTCFs) despite their associated risks. The objective of this study was to explore the association between social engagement (SE) and the use of antipsychotics in addressing the BPSD in newly admitted residents to LTCFs. Methods A cross-sectional study was undertaken using administrative data, primarily the Resident Assessment Instrument Minimum Data Set (Version 2.0) that collected between 2008 and 2011 (Fraser Health region, British Columbia, Canada). The data analysis conducted on a sample of 2,639 newly admitted residents aged 65 or older with a diagnosis of Alzheimer's disease or other dementias as of their first full or first quarterly assessment. Multivariate logistic regression analyses were undertaken to predict antipsychotic use based on SE. Results SE was found to be a statistically significant predictor of antipsychotic use when controlling for sociodemographic variables (odds ratio ( OR) = .86, p < .0001, confidence interval (CI) [0.82, 0.90]). However, the association disappeared when controlling for health variables ( OR = .97, p = .21, CI [0.97, 1.0]). Conclusion The prediction of antipsychotic use in newly admitted residents to LTCFs by SE is complex. Further research is warranted for further examination of the association of antipsychotic use in newly admitted residents to LTCFs.


Glycerol kinase stimulates uncoupling protein 1 expression by regulating fatty acid metabolism in beige adipocytes.
Browning of adipose tissue is induced by specific stimuli such as cold exposure and consists of up-regulation of thermogenesis in white adipose tissue. Recently, it has emerged as an attractive target for managing obesity in humans. Here, we performed a comprehensive analysis to identify genes associated with browning in murine adipose tissue. We focused on glycerol kinase (GYK) because its mRNA expression pattern is highly correlated with that of uncoupling protein 1 (UCP1), which regulates the thermogenic capacity of adipocytes. Cold exposure-induced Ucp1 up-regulation in inguinal white adipose tissue (iWAT) was partially abolished by Gyk knockdown (KD) in vivo Consistently, the Gyk KD inhibited Ucp1 expression induced by treatment with the β-adrenergic receptors (βAR) agonist isoproterenol (Iso) in vitro and resulted in impaired uncoupled respiration. Gyk KD also suppressed Iso- and adenylate cyclase activator-induced transcriptional activation and phosphorylation of the cAMP response element-binding protein (CREB). However, we did not observe these effects with a cAMP analog. Therefore Gyk KD related to Iso-induced cAMP products. In Iso-treated Gyk KD adipocytes, stearoyl-CoA desaturase 1 (SCD1) was up-regulated, and monounsaturated fatty acids such as palmitoleic acid (POA) accumulated. Moreover, a SCD1 inhibitor treatment recovered the Gyk KD-induced Ucp1 down-regulation and POA treatment down-regulated Iso-activated Ucp1 Our findings suggest that Gyk stimulates Ucp1 expression via a mechanism that partially depends on the βAR-cAMP-CREB pathway and Gyk-mediated regulation of fatty acid metabolism.


Influence of platen temperatures and relative humidity during storage on the survival of freeze-dried Salmonella typhimurium.
Salmonella typhimurium survived freeze-drying at a platen temperature of 120 F (48.9 C) and also, though to a much lesser degree, at 160 F (82.6 C). The extent of the survival at these temperatures was dependent on the composition of the model system employed. The incidence of damage immediately after freeze-drying was greater for cells dried at the higher platen temperature and was influenced by the composition of the menstruum in which the cells were dried. In model systems having protein-dominant isotherms, survival during subsequent storage depended greatly on relative humidity, with recovery highest at relative humidities below those corresponding to moisture contents at which a monomolecular layer is formed. In menstrua having a higher sugar content, survival was best at low relative humidities corresponding to a very low equilibrium moisture content in the model system used. Damage during storage tended to be a function of the composition of the gels in which the organisms were freeze-dried, and also depended greatly on the presence of air and on the relative humidity. The maximal percentage of damage usually occurred at the low relative humidities as storage time increased.


Functional differences between the 40 kDa and 50 to 70 kDa IgG Fc receptors on human neutrophils revealed by elastase treatment and antireceptor antibodies.
Most previous studies of IgG FcR on neutrophils (PMN) have focused on a single FcR of Mr = 50 to 70 kDa, which is recognized by mAb 3G8 and anti-Leu-11a. In the course of studying the effects of extracellular proteases on PMN receptor expression and function, we found that treatment with human leukocyte elastase reduced the expression of this FcR on the PMN surface by as much as 85% in flow cytometric studies, but did not inhibit ingestion of IgG-coated particles or O2- production induced by multivalent IgG complexes, and caused only a 35% decrease in IC binding to PMN. Since a second FcR with Mr = 40 kDa recognized by mAb IV-3, recently has been identified on PMN, we sought to determine if this FcR was resistant to elastase and thus accounted for the elastase stability of IgG-mediated PMN functions. Elastase treatment that reduced 3G8 binding by 85% caused no decrease in binding of mAb IV-3. For non-elastase-treated PMN, mAb IV-3 against the 40 kDa FcR caused as much as 79 +/- 7% inhibition of IgG-induced O2- production, whereas mAb 3G8 against the 50 to 70 kDa FcR caused only 32 +/- 5% inhibition. In contrast, for IC-binding, mAb IV-3 caused only 15 +/- 6% inhibition, whereas mAb 3G8 caused as much as 80 +/- 9% inhibition, a reversal of their relative effects on O2- production. In parallel studies with elastase-treated PMN, mAb IV-3 actually blocked more IC binding than did mAb 3G8, 55 +/- 4% vs 40 +/- 6%, respectively, presumably because most of the 50 to 70 kDa FcR molecules had been cleaved. The effect of the two mAb together in blocking IC binding was additive, whereas for blocking of O2- production, mAb 3G8 added little or nothing to the effect of mAb IV-3 alone. Direct 125I-labeled Ab binding studies with intact PMN revealed seven times as many 50 to 70 kDa as 40kDa FcR, 110,200 +/- 9600 and 15,100 +/- 700 sites/cell, respectively. Our findings suggest that the elastase-resistant 40 kDa FcR is primarily responsible for IgG-mediated activation of human PMN, whereas the elastase-sensitive 50 to 70 kDa FcR predominates in IC binding, by virtue of its numerical superiority, but does not directly activate the cell. The latter may serve to hold IgG-coated microorganisms or other multivalent IC in place at the PMN surface, enhancing contact with the 40 kDa FcR and thus facilitating cell activation in a cooperative manner.


Novel system for the simultaneous analysis of geminivirus DNA replication and plant interactions in Nicotiana benthamiana.
The origin of replication of African cassava mosaic virus (ACMV) and a gene expression vector based on Potato virus X were exploited to devise an in planta system for functional analysis of the geminivirus replication-associated protein (Rep) in transgenic Nicotiana benthamiana line pOri-2. This line contains an integrated copy of a tandem repeat of the ACMV origin of replication flanking nonviral sequences that can be mobilized and replicated by Rep as an episomal replicon. A Rep-GFP fusion protein can also mobilize and amplify the replicon, facilitating Rep detection in planta. The activity of Rep and its mutants, Rep-mediated host response, and the correlation between Rep intracellular localization and biological functions could be effectively assessed by using this in planta system. Our results indicate that modification of amino acid residues R(2), R(5), R(7) and K(11) or H(56), L(57) and H(58) prevent Rep function in replication. This defect correlates with possible loss of Rep nuclear localization and inability to trigger the host defense mechanism resembling a hypersensitive response.


Stem cells.
Stem cells are self-renewing cells capable of differentiating into multiple cell lines and are classified according to their origin and their ability to differentiate. Enormous potential exists in use of stem cells for regenerative medicine. To produce effective stem cell-based treatments for a range of diseases, an improved understanding of stem cell biology and better control over stem cell fate are necessary. In addition, the barriers to clinical translation, such as potential oncologic properties of stem cells, need to be addressed. With renewed government support and continued refinement of current stem cell methodologies, the future of stem cell research is exciting and promises to provide novel reconstructive options for patients and surgeons limited by traditional paradigms.


Cognitive sequelae of unilateral posteroventral pallidotomy.
To examine the cognitive sequelae of unilateral posteroventral pallidotomy. Single-group pretest and posttest methodologic assessment with baseline evaluation performed 1 to 2 days prior to surgery and follow-up conducted 3 months after pallidotomy. Movement disorder clinic at a university medical center. Fourteen patients (age range, 43-82 years) with Parkinson disease (average disease duration, 7.4 years). Unilateral posteroventral pallidotomy procedures were performed on the right (n=8) and left (n=6) side of the brain. The protocol consisted of a range of neuropsychological instruments sensitive to subcortical dysfunction, including measures of bi-manual coordination, simple-complex reaction time, visual attention, naming, verbal fluency, learning, recognition memory, and problem solving. No significant deterioration in specific cognitive abilities was observed as a function of the procedure. Patients showed a significant improvement in motor coordination speed for both contralateral and ipsilateral upper extremities. Stereotactic unilateral posteroventral pallidotomy is associated with minimal risk of adverse neuropsychological effects or cognitive decline. Additional research is warranted, with an increased sample size and extended follow-up, to assess any potential lateralized effects of the procedure.


Maslinic acid suppresses osteoclastogenesis and prevents ovariectomy-induced bone loss by regulating RANKL-mediated NF-κB and MAPK signaling pathways.
Activation of NF-κB and MAPK/activator protein 1 (AP-1) signaling pathways by receptor activator NF-κB ligand (RANKL) is essential for osteoclast activity. Targeting NF-κB and MAPK/AP-1 signaling to modulate osteoclast activity has been a promising strategy for osteoclast-related diseases. In this study we examined the effects of maslinic acid (MA), a pentacyclic triterpene acid that is widely present in dietary plants, on RANKL-induced osteoclastogenesis, osteoclast function, and signaling pathways by in vitro and in vivo assay systems. In mouse bone marrow monocytes (BMMs) and RAW264.7 cells, MA inhibited RANKL-induced osteoclastogenesis in a dose-dependent manner within nongrowth inhibitory concentration, and MA decreased osteoclastogenesis-related marker gene expression, including TRACP, MMP9, c-Src, CTR, and cathepsin K. Specifically, MA suppressed osteoclastogenesis and actin ring formation at early stage. In ovariectomized mice, administration of MA prevented ovariectomy-induced bone loss by inhibiting osteoclast activity. At molecular levels, MA abrogated the phosphorylation of MAPKs and AP-1 activity, inhibited the IκBα phosphorylation and degradation, blocked NF-κB/p65 phosphorylation, nuclear translocation, and DNA-binding activity by downregulating RANK expression and blocking RANK interaction with TRAF6. Together our data demonstrate that MA suppresses RANKL-induced osteoclastogenesis through NF-κB and MAPK/AP-1 signaling pathways and that MA is a promising agent in the treatment of osteoclast-related diseases such as osteoporosis.


Intravenous paracetamol improves the quality of postoperative analgesia but does not decrease narcotic requirements.
Paracetamol, a centrally acting inhibitor of cyclooxygenase, has less gastrointestinal and platelet-inhibiting side effects and is clinically better tolerated than nonsteroidal anti-inflammatory drugs. Therefore, it will be ideally suited for postoperative pain relief. In this prospective, double-blind, randomized, placebo-controlled study, we evaluated the analgesic efficacy, opioid-sparing effect and effects on opioid-related adverse effects of intravenous (IV) paracetamol in combination with IV morphine after lumbar laminectomy and discectomy. Forty patients were divided into 2 groups (n=20 each) to receive either paracetamol 1 g (group 1) or 0.9% NaCl 100 ml (group 2) at the end of the operation and at 6-hour intervals over 24 hours. IV patient-controlled analgesia with morphine was used as a rescue analgesic in both groups. Pain was evaluated at rest and on movement at the 1st, 2nd, 4th, 6th, 12th, 18th, and 24th hours using a visual analog scale. Hemodynamic parameters, morphine usage, patient satisfaction, and probable side effects were also evaluated. Pain scores at rest and on movement at the 12th, 18th, and 24th hours were significantly lower in group 1 (P<0.001). Morphine consumption was not statistically significantly different between the groups (P>0.05). Vomiting in group 2 was significantly higher (P=0.027). Significantly more patients in the paracetamol group rated their pain management as excellent (45% vs. 5%). Although repeated IV paracetamol usage after lumbar laminectomy and discectomy did not demonstrate a significant opioid-sparing effect, it did decrease visual analog scale scores at certain evaluation times and incidence of vomiting and increase patient satisfaction.


A comparison of serine and threonine O-glycosylation by UDP-GalNAc:polypeptide N-acetylgalactosaminyltransferase.
O-glycosylated proteins are ubiquitous in eukaryotes and are responsible for a variety of biological functions. O-glycosylation is initiated by the addition of N-acetylgalactosamine to serine or threonine residues, though it is not clear how specific residues are selected for modification. We have compared serine and threonine glycosylation using peptide substrates based on sequences from erythropoietin (EPO) and von Willebrand factor (HVF) that are glycosylated in vivo. UDP-GalNAc:polypeptide N-acetylgalactosaminyltransferase was derived from rat parotid, submandibular, and sublingual glands, liver and kidney as well as from human colostrum. The threonine-containing substrates were glycosylated to a much greater extent than those containing serine for all the enzyme sources. Changes in reaction pH, donor concentration, or divalent cation were unable to increase glycosylation of serine. When the incubation time was extended, serine in the EPO-based peptide was found to incorporate GalNAc at a low level, in contrast to the serine-containing HVF peptide, which did not glycosylate at all. By circular dichroism, the non-glycosylating peptide was the only one of the series that did not exhibit random coil structure. Our data suggest that although the structural and sequence requirements for O-glycosylation of serine and threonine residues are similar, serine sites are glycosylated less effectively than are threonine sites in vitro.


[THE IMPACT OF HEALTH ON THE FORMATION OF HUMAN CAPITAL].
The paper examines the importance of health as a part of human capital, the role of which is that only a healthy person can work efficiently and with full potential; a healthy person can work more efficiently; a healthy person can work productively and thus contribute more to the development of the country's economy; an unhealthy person becomes a burden for both the organization and in general for the country.


Prognostic value of TARC and quantitative PET parameters in relapsed or refractory Hodgkin lymphoma patients treated with brentuximab vedotin and DHAP.
Risk-stratified treatment strategies have the potential to increase survival and lower toxicity in relapsed/refractory classical Hodgkin lymphoma (R/R cHL) patients. This study investigated the prognostic value of serum (s)TARC, vitamin D and lactate dehydrogenase (LDH), TARC immunohistochemistry and quantitative PET parameters in 65 R/R cHL patients who were treated with brentuximab vedotin (BV) and DHAP followed by autologous stem-cell transplantation (ASCT) within the Transplant BRaVE study (NCT02280993). At a median follow-up of 40 months, the 3-year progression free survival (PFS) was 77% (95% CI: 67-88%) and the overall survival was 95% (90-100%). Significant adverse prognostic markers for progression were weak/negative TARC staining of Hodgkin Reed-Sternberg cells in the baseline biopsy, and a high standard uptake value (SUV)mean or SUVpeak on the baseline PET scan. After one cycle of BV-DHAP, sTARC levels were strongly associated with the risk of progression using a cutoff of 500 pg/ml. On the pre-ASCT PET scan, SUVpeak was highly prognostic for progression post-ASCT. Vitamin D, LDH and metabolic tumor volume had low prognostic value. In conclusion, we established the prognostic impact of sTARC, TARC staining, and quantitative PET parameters for R/R cHL, allowing the use of these parameters in prospective risk-stratified clinical trials. Trial registration: NCT02280993.


Serum immunoglobulin free light chain levels in systemic autoimmune rheumatic diseases.
Several reports have highlighted the abnormal increments of serum immunoglobulin free light chains (FLCs) in the course of systemic autoimmune rheumatic diseases (SARD), but a comparative analysis among different conditions is still lacking. A strong association between elevated FLC and hepatitis C virus (HCV)-related mixed cryoglobulinaemia (HCVMC) has been well established. Here, we aimed to analyse serum FLC levels in patients with four different SARD in comparison with HCVMC. Using a turbidimetric assay, free κ and λ chains were quantified in sera from 198 SARD patients (37 rheumatoid arthritis, RA; 47 systemic lupus erythematosus, SLE; 52 anti-phospholipid syndrome, APS; 62 primary Sjogren's syndrome, pSS), 62 HCVMC and 50 healthy blood donors (HD). All patient groups showed increased κ levels when compared to HD: 33·5 ± 2·6 mg/l in HCVMC, 26·7 ± 2·3 mg/l in RA, 29·7 ± 1·9 mg/l in SLE, 23·8 ± 1·1 mg/l in APS, 24·2 ± 1·1 mg/l in pSS; 10·1 ± 0·6 mg/l in HD. Free λ levels displayed a significant increase only for HCVMC (20·4 ± 1·4 mg/l) and SLE (18·4 ± 1·0 mg/l) compared to HD (13·6 ± 0·9 mg/l). The increase of κ compared to λ takes into account a κ /λ ratio of 1·6 for all groups. Our results substantially analyse and strengthen the association between FLC and SARD focusing the questions regarding their role in the pathogenesis and diagnosis of human diseases. Unfortunately, the biochemical differences distinguishing normal from pathological FLC have not been identified. Production of different isotypes is probably connected to still-unknown pathways.


Protective effects of grape seed extract fractions with different degrees of polymerisation on blood glucose, lipids and hepatic oxidative stress in diabetic rats.
The purpose of this study was to evaluate the effect of grape seed procyanidin (GSPE) fractions with different degrees of polymerisation (DPs) on blood glucose, lipids and hepatic oxidative stress in diabetic rats. Diabetic rats received a daily oral supplement of GSPE with different DPs for 6 weeks. During this period, blood glucose, body weight and food intake were assessed weekly. At the end of the experiment, serum lipid and hepatic oxidative stress were assessed compared with those of rats that did not receive GSPE. GSPE significantly decreased blood glucose, serum lipids and hepatic oxidative stress. Moreover, these effects were significantly better in the groups administered the oligomeric rather than the polymeric forms. These results demonstrate that GSPE has a positive effect on diabetes in rats, and the oligomeric form of GSPE may be more protective than other forms.


Cytology of inbreds and f 1 hybrids of pearl millet.
The meiotic behaviour of chromosomes was studied in four inbred lines and their F 1 hybrids of P. typhoides. The inbred lines showed a decrease in mean chiasma frequency when compared with the population plants, but differed from one another in their mean chiasma frequencies. Together with the decrease in mean chiasma frequency the inbreds showed variation in mean chiasma frequencies. The inbred lines showed a number of meiotic abnormalities such as extra chromosomes, extra fragments, desynapsis, persistent nucleoli and differential condensation of chromosomes. The F 1 hybrids of these inbreds exhibited heterosis for chiasma frequency. All the F 1's had mean chiasma frequencies higher than the means of the respective participating parents. The F 1's, however, differed in the degree of heterosis exhibited. In the F 1 hybrids, the variation in mean chiasma frequency, both between plants and between PMC's within plants, was significantly lower than that of the inbred lines. The effect of environment was studied in the inbred lines and their F 1 hybrids. The mean chiasma frequencies of the inbred lines were significantly lower, and the variation in mean chiasma frequencies was greater, in the stress season. The mean chiasma frequencies of F 1's did not show any significant differences between the two seasons. The F 1's exhibited less variation in mean chiasma frequency than the inbred lines, showing that F 1's were developmentally more stable. The F 1's did not show any meiotic abnormalities in either season.


Genome-wide linkage disequilibrium and past effective population size in three Korean cattle breeds.
The routine collection and use of genomic data are useful for effectively managing breeding programs for endangered populations. Linkage disequilibrium (LD) using high-density DNA markers has been widely used to determine population structures and predict the genomic regions that are associated with economic traits in beef cattle. The extent of LD also provides information about historical events, including past effective population size (Ne ), and it allows inferences on the genetic diversity of breeds. The objective of this study was to estimate the LD and Ne in three Korean cattle breeds that are genetically similar but have different coat colors (Brown, Brindle and Jeju Black Hanwoo). Brindle and Jeju Black are endangered breeds with small populations, whereas Brown Hanwoo is the main breeding population in Korea. DNA samples from these cattle breeds were genotyped using the Illumina BovineSNP50 Bead Chip. We examined 13 cattle breeds, including European taurines, African taurines and indicines, and hybrids to compare their LD values. Brown Hanwoo consistently had the lowest mean LD compared to Jeju Black, Brindle and the other 13 cattle breeds (0.13, 0.19, 0.21 and 0.15-0.22 respectively). The high LD values of Brindle and Jeju Black contributed to small Ne values (53 and 60 respectively), which were distinct from that of Brown Hanwoo (531) for 11 generations ago. The differences in LD and Ne for each breed reflect the breeding strategy applied. The Ne for these endangered cattle breeds remain low; thus, effort is needed to bring them back to a sustainable tract.


Risk factors for chronic mastitis in morocco and egypt.
Chronic mastitis is a prolonged inflammatory breast disease, and little is known about its etiology. We identified 85 cases and 112 controls from 5 hospitals in Morocco and Egypt. Cases were women with chronic mastitis (including periductal, lobular, granulomatous, lymphocytic, and duct ectasia with mastitis). Controls had benign breast disease, including fibroadenoma, benign phyllodes, and adenosis. Both groups were identified from histopathologically diagnosed patients from 2008 to 2011, frequency-matched on age. Patient interviews elicited demographic, reproductive, breastfeeding, and clinical histories. Cases had higher parity than controls (OR = 1.75, 1.62-1.90) and more reported history of contraception use (OR = 2.73, 2.07-3.61). Cases were less likely to report wearing a bra (OR = 0.56, 0.47-0.67) and less often used both breasts for breastfeeding (OR = 4.40, 3.39-5.72). Chronic mastitis cases were significantly less likely to be employed outside home (OR = 0.71, 0.60-0.84) and more likely to report mice in their households (OR = 1.63, 1.36-1.97). This is the largest case-control study reported to date on risk factors for chronic mastitis. Our study highlights distinct reproductive risk factors for the disease. Future studies should further explore these factors and the possible immunological and susceptibility predisposing conditions.


Acyl-based anandamide uptake inhibitors cause rapid toxicity to C6 glioma cells at pharmacologically relevant concentrations.
Compounds blocking the uptake of the endogenous cannabinoid anandamide (AEA) have been used to explore the functions of the endogenous cannabinoid system in the CNS both in vivo and in vitro. In this study, the effects of four commonly used acyl-based uptake inhibitors [N-(4-hydroxyphenyl)arachidonylamide (AM404), N-(4-hydroxy-2-methylphenyl) arachidonoyl amide (VDM11), (5Z,8Z,11Z,14Z)-N-(3-furanylmethyl)-5,8,11,14-eicosatetraenamide (UCM707) and (9Z)-N-[1-((R)-4-hydroxybenzyl)-2-hydroxyethyl]-9-octadecen-amide (OMDM2)] and the related compound arvanil on C6 glioma cell viability were investigated. All five compounds reduced the ability of the cells to accumulate calcein, reduced the total nucleic acid content and increased the activity of lactate dehydrogenase recovered in the cell medium. AM404 (10 microm) and VDM11 (10 microm) acted rapidly, reducing cell viability after 3 h of exposure when cell densities of 5,000 per well were used. In contrast, UCM707 (30 microm), OMDM2 (10 microm) and the related compound arvanil (10 microm) produced a more slowly developing effect on cell viability, although robust effects were seen after 6-9 h of exposure. At higher cell densities, the toxicities of AM404 and UCM707 were reduced. Comparison of the compounds with arachidonic acid, arachidonic acid methyl ester, AEA, arachidonoyl glycine and oleic acid suggested that the toxicity of the arachidonoyl-based compounds was related primarily to the acyl side-chain rather than the head group. A variety of pre-treatments blocking possible metabolic pathways and receptor targets were tested, but the only consistent protective treatment against the effects of these compounds was the antioxidant N-acetyl-L-cysteine. It is concluded that AM404, VDM11, UCM707 and OMDM2 produce a rapid loss of C6 glioma cell viability over the same concentration range as is required for the inhibition of AEA uptake in vitro, albeit with a longer latency. Such effects should be kept in mind when acyl-derived compounds are used to probe the function of the endocannabinoid system in the CNS, particularly in chronic administration protocols.


Over-, under- and misuse of pain treatment in Germany.
The HTA-report (Health Technology Assessment) deals with over- and undertreatment of pain therapy. Especially in Germany chronic pain is a common reason for the loss of working hours and early retirement. In addition to a reduction in quality of life for the affected persons, chronic pain is therefore also an enormous economic burden for society. Which diseases are in particular relevant regarding pain therapy?What is the social-medical care situation regarding pain facilities in Germany?What is the social-medical care situation in pain therapy when comparing on international level?Which effects, costs or cost-effects can be seen on the micro-, meso- and macro level with regard to pain therapy?Among which social-medical services in pain therapy is there is an over- or undertreatment with regard to the micro-, meso- and macro level?Which medical and organisational aspects that have an effect on the costs and/or cost-effectiveness have to be particularly taken into account with regard to pain treatment/chronic pain?What is the influence of the individual patient's needs (micro level) in different situations of pain (e. g. palliative situation) on the meso- and macro level?Which social-medical and ethical aspects for an adequate treatment of chronic pain on each level have to be specially taken into account?Is the consideration of these aspects appropriate to avoid over- or undertreatment?Are juridical questions included in every day care of chronic pain patients, mainly in palliative care?On which level can appropriate interventions prevent over- or undertreatment? A systematic literature research is done in 35 databases. In the HTA, reviews, epidemiological and clinical studies and economic evaluations are included which report about pain therapy and in particular palliative care in the years 2005 till 2010. 47 studies meet the inclusion criteria. An undertreatment of acupuncture, over- and misuse with regard to opiate prescription and an overuse regarding unspecific chest pain and chronic low back pain (LBP) can be observed. The results show the benefit and the cost-effectiveness of interdisciplinary as well as multi-professional approaches, multimodal pain therapy and cross-sectoral integrated medical care. Only rough values can be determined about the care situation regarding the supply of pain therapeutic and palliative medical facilities as the data are completely insufficient. Due to the broad research question the HTA-report contains inevitably different outcomes and study designs which partially differ qualitatively very strong from each other. In the field of palliative care hospices for in-patients and palliative wards as well as hospices for out-patients are becoming more and more important. Palliative care is a basic right of all terminally ill persons. Despite the relatively high number of studies in Germany the HTA-report shows a massive lack in health care research. Based on the studies a further expansion of out-patient pain and palliative care is recommended. Further training for all involved professional groups must be improved. An independent empirical analysis is necessary to determine over or undertreatment in pain care.


Comparison of Obturation Quality between Calcium Silicate-Based Sealers and Resin-Based Sealers for Endodontic Re-treatment.
The objective of this micro-computed tomography (micro-CT)-based study was to compare the filling quality of endodontic treatment and endodontic Re-treatment between two sealers with matched obturation techniques: calcium silicate-based sealer (Endoseal TCS) with a single-cone technique (SCT) and resin-based sealer (AH Plus) with a continuous wave technique (CWT). Forty maxillary premolars were selected and assigned into four groups, according to the obturation methods of the first endodontic treatment and Re-treatment (n = 10). The AP/AP group was first treated with AH Plus/CWT, then re-treated with AH Plus/CWT. The AP/ET group was first treated with AH Plus/CWT, then re-treated with Endoseal TCS/SCT. The ET/AP group was first treated with Endoseal TCS/SCT, then re-treated with AH Plus/CWT, and the ET/ET group was first treated with Endoseal TCS/SCT, then re-treated with Endoseal TCS/SCT. The specimens were scanned using micro-CT at three time points: after the first endodontic treatment, after gutta-percha (GP) cone removal, and after Re-treatment. The void volume of root canal obturation and the volume of the remaining filling materials were calculated. Data were analyzed using Student's t-tests and ANOVA. The Endoseal TCS groups (ET/AP and ET/ET) showed a lower percentage of voids than the AH plus groups (AP/AP and AP/ET) on the whole canal and the apical third, after first obturation (p < 0.05). The AH plus group showed significantly fewer remnants than the Endoseal TCS group after GP removal (p < 0.05). Re-treated canals and initially treated canals had similar void volumes (p > 0.05). There was no significant difference in void volume after Re-treatment, regardless of whether the same or different sealers were used for the first treatment and Re-treatment (p < 0.05). Endoseal TCS sealer and AH Plus sealer had a similar Re-treatment efficacy, regardless of which sealer was used in the previous treatment.


Fatigue-resistant adhesion of hydrogels.
The adhesion of soft connective tissues (tendons, ligaments, and cartilages) on bones in many animals can maintain high toughness (∽800 J m-2) over millions of cycles of mechanical loads. Such fatigue-resistant adhesion has not been achieved between synthetic hydrogels and engineering materials, but is highly desirable for diverse applications such as artificial cartilages and tendons, robust antifouling coatings, and hydrogel robots. Inspired by the nanostructured interfaces between tendons/ligaments/cartilages and bones, we report that bonding ordered nanocrystalline domains of synthetic hydrogels on engineering materials can give a fatigue-resistant adhesion with an interfacial fatigue threshold of 800 J m-2, because the fatigue-crack propagation at the interface requires a higher energy to fracture the ordered nanostructures than amorphous polymer chains. Our method enables fatigue-resistant hydrogel coatings on diverse engineering materials with complex geometries. We further demonstrate that the fatigue-resistant hydrogel coatings exhibit low friction and low wear against natural cartilages.


[Circadian rhythms in hepatic biotransformation of drugs].
The temporal variations in hepatic drug biotransformation are reviewed. Important diurnal variations in the activity of different cytochrome P-450 isoenzymes and in enzyme systems involved in conjugation to glucuronide, sulfate and acetate are described. A circadian rhythm in the hepatic concentration of glutathione is also described. These temporal variations explain the chronopharmacokinetic and chronohepatotoxicity of some drugs or toxic agents.


Escitalopram in the treatment of major depressive disorder: a meta-analysis.
To assess the relative antidepressant efficacy of escitalopram and comparator antidepressants. A meta-analysis was performed using studies in major depressive disorder (MDD) comparing escitalopram with active controls, including selective serotonin reuptake inhibitors [SSRIs] (citalopram, fluoxetine, paroxetine, sertraline) and serotonin/noradrenaline reuptake inhibitors [SNRIs] (venlafaxine, duloxetine). Adult patients had to meet DSM-IV criteria for MDD. The primary outcome measure was the treatment difference in Montgomery-Asberg Depression Rating Scale (MADRS) total score at week 8. Secondary outcome measures were response and remission (MADRS total score < or = 12) rates. Individual patient data (N = 4549) from 16 randomized controlled trials were included in the analyses (escitalopram n = 2272, SSRIs n = 1750, SNRIs n = 527). Escitalopram was significantly more effective than comparators in overall treatment effect, with an estimated mean treatment difference of 1.1 points on the MADRS (p < 0.0001), and in responder (63.7 vs. 58.3%, p < 0.0001) and remitter (53.1 vs. 49.4%, p < 0.0059) analyses. Escitalopram was significantly superior to SSRIs, with an estimated difference in response of 62.1 vs. 58.4% and remission of 51.6 vs. 49.0%. In comparison to SNRIs, the estimated difference in response was 68.3 vs. 59.0% (p = 0.0007) and for remission the difference was 57.8 vs. 50.5% (p = 0.0088). These results were similar for severely depressed patients (baseline MADRS > or = 30). Sensitivity analyses were performed with data from articles reporting Hamilton Rating Scale for Depression (HAMD) scores. The 8-week withdrawal rate due to adverse events was 5.4% for escitalopram and 7.9% for the comparators (p < 0.01). This difference was accounted for by statistically significant higher attrition rates in the SNRI comparisons. This work may be limited by the clinical methodology underlying meta-analytic studies, in particular, the exclusion of trials that fail to meet predetermined criteria for inclusion. In this meta-analysis, superior efficacy of escitalopram compared to SSRIs and SNRIs was confirmed, although the superiority over SSRIs was largely explained by differences between escitalopram and citalopram.


Bioactive peptides from milk: animal determinants and their implications in human health.
Milk is an important protein source in human diets, providing around 32 g protein/l (for bovine milk, which constitutes some 85% of global consumption). The most abundant milk proteins are α-lactalbumin, β-lactoglobulin, αs-casein, β-casein, and κ-casein. Besides their nutritional value, milk proteins play a crucial role in the processing properties of milk, such as solubility, water bonding, heat stability, renneting and foaming, among others. In addition, and most importantly for this review, these proteins are the main source of bioactive components in milk. Due to the wide range of proposed beneficial effects on human health, milk proteins are considered as potential ingredients for the production of health-promoting functional foods. However, most of the evidence for bioactive effects comes from in vitro studies, and there is a need for further research to fully evaluate the true potential of milk-derived bioactive factors. Animal genetics and animal nutrition play an important role in the relative proportions of milk proteins and could be used to manipulate the concentration of specific bioactive peptides in milk from ruminants. Unfortunately, only a few studies in the literature have focused on changes in milk bioactive peptides associated to animal genetics and animal nutrition. The knowledge described in the present review may set the basis for further research and for the development of new dairy products with healthy and beneficial properties for humans.


Integration of metabolomics and expression of glycerol-3-phosphate acyltransferase (GPAM) in breast cancer-link to patient survival, hormone receptor status, and metabolic profiling.
Changes in lipid metabolism are an important but not well-characterized hallmark of cancer. On the basis of our recent findings of lipidomic changes in breast cancer, we investigated glycerol-3-phosphate acyltransferase (GPAM), a key enzyme in the lipid biosynthesis of triacylglycerols and phospholipids. GPAM protein expression was evaluated and linked to metabolomic and lipidomic profiles in a cohort of human breast carcinomas. In addition, GPAM mRNA expression was analyzed using the GeneSapiens in silico transcriptiomics database. High cytoplasmic GPAM expression was associated with hormone receptor negative status (p = 0.013). On the protein (p = 0.048) and mRNA (p = 0.001) levels, increased GPAM expression was associated with a better overall survival. Metabolomic analysis by GC-MS showed that sn-glycerol-3-phosphate, the substrate of GPAM, was elevated in breast cancer compared to normal breast tissue. LC-MS based lipidomic analysis identified significantly higher levels of phospholipids, especially phosphatidylcholines in GPAM protein positive tumors. In conclusion, our results suggest that GPAM is expressed in human breast cancer with associated changes in the cellular metabolism, in particular an increased synthesis of phospholipids, the major structural component of cellular membranes.


Orchestrating osteogenic differentiation of mesenchymal stem cells--identification of placental growth factor as a mechanosensitive gene with a pro-osteogenic role.
Skeletogenesis is initiated during fetal development and persists through adult life as either a remodeling process in response to homeostatic regulation or as a regenerative process in response to physical injury. Mesenchymal stem cells (MSCs) play a crucial role providing progenitor cells from which osteoblasts, bone matrix forming cells are differentiated. The mechanical environment plays an important role in regulating stem cell differentiation into osteoblasts, however, the mechanisms by which MSCs respond to mechanical stimuli are yet to be fully elucidated. To increase understanding of MSC mechanotransuction and osteogenic differentiation, this study aimed to identify novel, mechanically augmented genes and pathways with pro-osteogenic functionality. Using collagen glycoaminoglycan scaffolds as mimics of native extracellular matrix, to create a 3D environment more representative of that found in bone, MSC-seeded constructs were mechanically stimulated in a flow-perfusion bioreactor. Global gene expression profiling techniques were used to identify potential candidates warranting further investigation. Of these, placental growth factor (PGF) was selected and expression levels were shown to strongly correlate to both the magnitude and duration of mechanical stimulation. We demonstrated that PGF gene expression was modulated through an actin polymerization-mediated mechanism. The functional role of PGF in modulating MSC osteogenic differentiation was interrogated, and we showed a concentration-dependent response whereby low concentrations exhibited the strongest pro-osteogenic effect. Furthermore, pre-osteoclast migration and differentiation, as well as endothelial cell tubule formation also maintained concentration-dependent responses to PGF, suggesting a potential role for PGF in bone resorption and angiogenesis, processes key to bone remodeling and fracture repair.


T-box-mediated control of the anabolic proline biosynthetic genes of Bacillus subtilis.
Bacillus subtilis possesses interlinked routes for the synthesis of proline. The ProJ-ProA-ProH route is responsible for the production of proline as an osmoprotectant, and the ProB-ProA-ProI route provides proline for protein synthesis. We show here that the transcription of the anabolic proBA and proI genes is controlled in response to proline limitation via a T-box-mediated termination/antitermination regulatory mechanism, a tRNA-responsive riboswitch. Primer extension analysis revealed mRNA leader transcripts of 270 and 269 nt for the proBA and proI genes, respectively, both of which are synthesized from SigA-type promoters. These leader transcripts are predicted to fold into two mutually exclusive secondary mRNA structures, forming either a terminator or an antiterminator configuration. Northern blot analysis allowed the detection of both the leader and the full-length proBA and proI transcripts. Assessment of the level of the proBA transcripts revealed that the amount of the full-length mRNA species strongly increased in proline-starved cultures. Genetic studies with a proB-treA operon fusion reporter strain demonstrated that proBA transcription is sensitively tied to proline availability and is derepressed as soon as cellular starvation for proline sets in. Both the proBA and the proI leader sequences contain a CCU proline-specific specifier codon prone to interact with the corresponding uncharged proline-specific tRNA. By replacing the CCU proline specifier codon in the proBA T-box leader with UUC, a codon recognized by a Phe-specific tRNA, we were able to synthetically re-engineer the proline-specific control of proBA transcription to a control that was responsive to starvation for phenylalanine.


A unified approach to universal prediction: generalized upper and lower bounds.
We study sequential prediction of real-valued, arbitrary, and unknown sequences under the squared error loss as well as the best parametric predictor out of a large, continuous class of predictors. Inspired by recent results from computational learning theory, we refrain from any statistical assumptions and define the performance with respect to the class of general parametric predictors. In particular, we present generic lower and upper bounds on this relative performance by transforming the prediction task into a parameter learning problem. We first introduce the lower bounds on this relative performance in the mixture of experts framework, where we show that for any sequential algorithm, there always exists a sequence for which the performance of the sequential algorithm is lower bounded by zero. We then introduce a sequential learning algorithm to predict such arbitrary and unknown sequences, and calculate upper bounds on its total squared prediction error for every bounded sequence. We further show that in some scenarios, we achieve matching lower and upper bounds, demonstrating that our algorithms are optimal in a strong minimax sense such that their performances cannot be improved further. As an interesting result, we also prove that for the worst case scenario, the performance of randomized output algorithms can be achieved by sequential algorithms so that randomized output algorithms do not improve the performance.