Search

Peptidylglycine α-amidating monooxygenase (PAM) converts inactive terminal-glycine prohormones into their activated α-amidated forms. PAM is thought to play a role in the development of antiandrogen drug resistance in prostate cancer (CaP) through PAM-activated autocrine growth. On the basis of the previous finding that many lung cancer cell lines excrete PAM into their culture media, this study investigates PAM levels in media collected from human CaP cell line cultures. Androgen-independent DU145 and PC-3 prostate cancer cell lines exhibited readily detectable levels of PAM activity in extracts and media, whereas the androgen-dependent LNCaP cell line showed little or no activity. Because of the much larger volume of media versus cell extracts, more than 90% of the total PAM activity was located in the media for both the PC-3 and DU145 cell lines, providing a readily accessible source of CaP PAM. A simple, scalable method to obtain PAM from the culture media of androgen-independent human prostate cancer cell lines is described in this article. This approach provides a much easier means of collecting CaP-derived PAM than previously described cell fractionation procedures and should facilitate the investigations of the role and targeting of PAM in hormone-independent CaP.

ABSTRACTSerological diagnosis of Lyme disease may be complicated by antigenic differences between infecting organisms and those used as test references. Accordingly, it would be helpful to include antigens whose sequences are well conserved by a broad range of Lyme disease spirochetes. In the present study, line blot analyses were performed using recombinant complement regulator-acquiring surface protein 2 (BbCRASP-2) from Borrelia burgdorferisensu stricto strain B31 and serum samples from human Lyme disease patients from throughout the United States and Germany. The results indicated that a large proportion of the patients had produced antibodies recognizing recombinant BbCRASP-2. In addition, Lyme disease spirochetes isolated from across North America and Europe were found to contain genes encoding proteins with high degrees of similarity to the B. burgdorferitype strain B31 BbCRASP-2, consistent with the high percentage of serologically positive patients. These data indicate that BbCRASP-2 may be valuable for use in a widely effective serological assay.

Serological diagnosis of Lyme disease may be complicated by antigenic differences between infecting organisms and those used as test references. Accordingly, it would be helpful to include antigens whose sequences are well conserved by a broad range of Lyme disease spirochetes. In the present study, line blot analyses were performed using recombinant complement regulator-acquiring surface protein 2 (BbCRASP-2) from Borrelia burgdorferi sensu stricto strain B31 and serum samples from human Lyme disease patients from throughout the United States and Germany. The results indicated that a large proportion of the patients had produced antibodies recognizing recombinant BbCRASP-2. In addition, Lyme disease spirochetes isolated from across North America and Europe were found to contain genes encoding proteins with high degrees of similarity to the B. burgdorferi type strain B31 BbCRASP-2, consistent with the high percentage of serologically positive patients. These data indicate that BbCRASP-2 may be valuable for use in a widely effective serological assay.

The Lyme disease spirochete, Borrelia burgdorferi, is largely resistant to being killed by its hosts’ alternative complement activation pathway. One possible resistance mechanism of these bacteria is to coat their surfaces with host complement regulators, such as factor H. Five different B. burgdorferi outer surface proteins having affinities for factor H have been identified: complement regulator-acquiring surface protein 1 (BbCRASP-1), encoded by cspA; BbCRASP-2, encoded by cspZ; and three closely related proteins, BbCRASP-3, -4, and -5, encoded by erpP, erpC, and erpA, respectively. We now present analyses of the recently identified BbCRASP-2 and cspZ expression patterns throughout the B. burgdorferi infectious cycle, plus novel analyses of BbCRASP-1 and erp-encoded BbCRASPs. Our results, combined with data from earlier studies, indicate that BbCRASP-2 is produced primarily during established mammalian infection, while BbCRASP-1 is produced during tick-to-mammal and mammal-to-tick transmission stages but not during established mammalian infection, and Erp-BbCRASPs are produced from the time of transmission from infected ticks into mammals until they are later acquired by other feeding ticks. Transcription of cspZ and synthesis of BbCRASP-2 were severely repressed during cultivation in laboratory medium relative to mRNA levels observed during mammalian infection, and cspZ expression was influenced by culture temperature and pH, observations which will assist identification of the mechanisms employed by B. burgdorferi to control expression of this borrelial infection-associated protein.

ABSTRACTThe Lyme disease spirochete, Borrelia burgdorferi, is largely resistant to being killed by its hosts’ alternative complement activation pathway. One possible resistance mechanism of these bacteria is to coat their surfaces with host complement regulators, such as factor H. Five different B. burgdorferiouter surface proteins having affinities for factor H have been identified: complement regulator-acquiring surface protein 1 (BbCRASP-1), encoded by cspA; BbCRASP-2, encoded by cspZ; and three closely related proteins, BbCRASP-3, -4, and -5, encoded by erpP, erpC, and erpA, respectively. We now present analyses of the recently identified BbCRASP-2 and cspZexpression patterns throughout the B. burgdorferiinfectious cycle, plus novel analyses of BbCRASP-1 and erp-encoded BbCRASPs. Our results, combined with data from earlier studies, indicate that BbCRASP-2 is produced primarily during established mammalian infection, while BbCRASP-1 is produced during tick-to-mammal and mammal-to-tick transmission stages but not during established mammalian infection, and Erp-BbCRASPs are produced from the time of transmission from infected ticks into mammals until they are later acquired by other feeding ticks. Transcription of cspZand synthesis of BbCRASP-2 were severely repressed during cultivation in laboratory medium relative to mRNA levels observed during mammalian infection, and cspZexpression was influenced by culture temperature and pH, observations which will assist identification of the mechanisms employed by B. burgdorferito control expression of this borrelial infection-associated protein.

The causative agent of Lyme disease, Borrelia burgdorferi, is naturally resistant to its host's alternative pathway of complement-mediated killing. Several different borrelial outer surface proteins have been identified as being able to bind host factor H, a regulator of the alternative pathway, leading to a hypothesis that such binding is important for borrelial resistance to complement. To test this hypothesis, the development of B. burgdorferi infection was compared between factor H-deficient and wild-type mice. Factor B- and C3-deficient mice were also studied to determine the relative roles of the alternative and classical/lectin pathways in B. burgdorferi survival during mammalian infection. While it was predicted that B. burgdorferi should be impaired in its ability to infect factor H-deficient animals, quantitative analyses of bacterial loads indicated that those mice were infected at levels similar to those of wild-type and factor B- and C3-deficient mice. Ticks fed on infected factor H-deficient or wild-type mice all acquired similar numbers of bacteria. Indirect immunofluorescence analysis of B. burgdorferi acquired by feeding ticks from the blood of infected mice indicated that none of the bacteria had detectable levels of factor H on their outer surfaces, even though such bacteria express high levels of surface proteins capable of binding factor H. These findings demonstrate that the acquisition of host factor H is not essential for mammalian infection by B. burgdorferi and indicate that additional mechanisms are employed by the Lyme disease spirochete to evade complement-mediated killing.

ABSTRACTThe causative agent of Lyme disease, Borrelia burgdorferi, is naturally resistant to its host's alternative pathway of complement-mediated killing. Several different borrelial outer surface proteins have been identified as being able to bind host factor H, a regulator of the alternative pathway, leading to a hypothesis that such binding is important for borrelial resistance to complement. To test this hypothesis, the development of B. burgdorferiinfection was compared between factor H-deficient and wild-type mice. Factor B- and C3-deficient mice were also studied to determine the relative roles of the alternative and classical/lectin pathways in B. burgdorferisurvival during mammalian infection. While it was predicted that B. burgdorferishould be impaired in its ability to infect factor H-deficient animals, quantitative analyses of bacterial loads indicated that those mice were infected at levels similar to those of wild-type and factor B- and C3-deficient mice. Ticks fed on infected factor H-deficient or wild-type mice all acquired similar numbers of bacteria. Indirect immunofluorescence analysis of B. burgdorferiacquired by feeding ticks from the blood of infected mice indicated that none of the bacteria had detectable levels of factor H on their outer surfaces, even though such bacteria express high levels of surface proteins capable of binding factor H. These findings demonstrate that the acquisition of host factor H is not essential for mammalian infection by B. burgdorferiand indicate that additional mechanisms are employed by the Lyme disease spirochete to evade complement-mediated killing.

The possibility that non-sedating antihistamines could elicit sedation in mice due to drug-induced inhibition of brain PgP was evaluated by measuring the ability of desloratadine alone or in combination with verapamil to cause ataxia in mice. Also, the concentrations of desloratadine in plasma and in brain homogenates were measured by liquid chromatography- mass spectrometry. Relative to methylcellulose (control) treatment, verapamil plus desloratadine decreased rotarod performance of mice. Plasma concentrations of desloratadine appeared comparable in the mice treated with either desloratadine or verapamil plus desloratadine, however the rate of decline of desloratadine from brain tissue was slower in mice treated with verapamil plus desloratadine compared to mice treated with desloratadine only. These data suggest that inhibition of brain PgP can convert desloratadine to a sedating antihistamine in mice.

The article presents a synopsis of the play "28 Light Years From Now" directed by Rachel Bykowski that focuses on mothers, daughters, regret and time travel. It mentions about the characters like Josie who sets off to meet Annabelle, leaving her daughter, life and the present behind to revisit the past one more time as she attempts to fix her broken heart from 28 years ago.

We investigated the intra- and interrater reliability of ischemic lesion volumes measurements assessed by different MRI sequences at various times from onset.

HPLC with ICP-MS or ESI-MS detection has been used to investigate metal-containing reaction intermediates in the ligand exchange process leading to formation of the well known Rh2(MEPY)4 catalyst. A variety of intermediates are observed along the pathway to formation of the desired tetrasubstituted product, including isomeric species from di-, tri-, or tetrasubstitution that were previously believed to be absent.

During the natural mammal-tick infection cycle, the Lyme disease spirochete Borrelia burgdorferi comes into contact with components of the alternative complement pathway. B. burgdorferi, like many other human pathogens, has evolved the immune evasion strategy of binding two host-derived fluid-phase regulators of complement, factor H and factor H-like protein 1 (FHL-1). The borrelial complement regulator-acquiring surface protein 1 (CRASP-1) is a surface-exposed lipoprotein that binds both factor H and FHL-1. Analysis of CRASP-1 expression during the mammal-tick infectious cycle indicated that B. burgdorferi expresses this protein during mammalian infection, supporting the hypothesized role for CRASP-1 in immune evasion. However, CRASP-1 synthesis was repressed in bacteria during colonization of vector ticks. Analysis of cultured bacteria indicated that CRASP-1 is differentially expressed in response to changes in pH. Comparisons of CRASP-1 expression patterns with those of other infection-associated B. burgdorferi proteins, including the OspC, OspA, and Erp proteins, indicated that each protein is regulated through a unique mechanism.

ABSTRACTDuring the natural mammal-tick infection cycle, the Lyme disease spirochete Borrelia burgdorfericomes into contact with components of the alternative complement pathway. B. burgdorferi, like many other human pathogens, has evolved the immune evasion strategy of binding two host-derived fluid-phase regulators of complement, factor H and factor H-like protein 1 (FHL-1). The borrelial complement regulator-acquiring surface protein 1 (CRASP-1) is a surface-exposed lipoprotein that binds both factor H and FHL-1. Analysis of CRASP-1 expression during the mammal-tick infectious cycle indicated that B. burgdorferiexpresses this protein during mammalian infection, supporting the hypothesized role for CRASP-1 in immune evasion. However, CRASP-1 synthesis was repressed in bacteria during colonization of vector ticks. Analysis of cultured bacteria indicated that CRASP-1 is differentially expressed in response to changes in pH. Comparisons of CRASP-1 expression patterns with those of other infection-associated B. burgdorferiproteins, including the OspC, OspA, and Erp proteins, indicated that each protein is regulated through a unique mechanism.

No Abstract

Sustained response to antipsychotic therapy is an important outcome measure for patients with psychotic disorders. Placebo control in studies of relapse prevention contributes valuable information yet provokes much debate. This study, using placebo as a control, evaluated olanzapine’s efficacy in preventing a psychotic relapse. Participants were stable minimally symptomatic outpatients with schizophrenia or schizoaffective disorder. The study included 4 phases (1) 4-day to 9-day screening/evaluation (N583), (2) 6-week conversion to open-label olanzapine (N493; 10-20 mg/d), (3) 8-week stabilization on olanzapine (N458; 10-20 mg/d), and (4) 52-week randomized (21), double-blind maintenance with olanzapine (N224; 10–20 mg/d) or placebo (N102). Primary relapse criteria were clinically significant changes in the Brief Psychiatric Rating Scale (BPRS) positive item cluster or rehospitalization due to positive symptoms. Statistical methodology allowed sequential real-time estimation of efficacy across blinded treatment groups and multiple interim analyses, which permitted study termination when efficacy was significantly different between treatments. A significant between-treatment difference emerged 210 days after first patient randomization to double-blind treatment. Thus, 151 (46.3) of the randomized patients were discontinued early and 34 (10.4) of the planned patient enrollment were not required. The olanzapine group had a significantly longer time to relapse (P< 0.0001) than the placebo group. The 6-month cumulative estimated relapse rate (Kaplan-Meier) was 5.5 for olanzapine-treated patients versus 55.2 for placebo-treated patients. The design of this study enabled appropriate statistical testing of the primary hypothesis while minimizing exposure of patients to a less effective treatment than olanzapine. In remitted stabilized patients with schizophrenia or schizoaffective disorder, olanzapine demonstrated a positive benefit-to-risk profile in relapse prevention.

Background:Comprehensive Stroke Centers (CSCs) strive to narrow rt-PA and “Door To Groin” (DTG) neurointerventional (NIR) times. Process improvement workflows have been put in place for rt-PA. While similar processes have been implemented to streamline workflows for hyperacute NIR cases, complex pathways, disparate imaging locations, and fragmented communications all highlight a need for continued improvements.Methods:This quality improvement initiative (IRB #210525) was implemented to assess our transition to the Viz.ai platform for immediate image review and centralized communication and its effect on key performance indicators (KPIs) in an already robust CSC. We compared 6 month periods prior to and following deployment. Sequential stroke NIR patients were included. Both Direct Arriving LVO (DALVO) and telemedicine transfer LVO (BEMI) cases were assessed. We assessed subgroups of DALVO-OnHours, DALVO-OffHours, BEMI-OnHours, and BEMI-OffHours. Mann-Whitney U was utilized.Results:Eighty-two NIR cases were analyzed pre v. post Viz.ai implementation (DALVO-OnHours 7 v. 7, DALVO-OffHours 10 v. 5, BEMI-OnHours 13 v. 6, BEMI-OffHours 17 v. 17). DALVO-OnHours improved 19% (97min, 79min; p=0.201) in median DTG times. DALVO-OffHours had a significant 39% reduction (157min, 95min; p=0.009). DALVO-“All” showed a significant 32% reduction (127min, 86 min; p=0.006). BEMI-OnHours improved 18% (37min, 31min; p=0.337). BEMI-OffHours improved 38% (45min, 28min; p=0.077). BEMI-“All” significantly improved 33% (42min, 28min; p=0.036). Overall, there was a 22% reduction (50min, 39min; p=0.066) after Viz.ai implementation.Conclusions:There was an immediate KPI improvement following Viz.ai implementation for both direct arrival and telemedicine transfer NIR cases (32% and 33% respectively). In the greatest opportunity subset (direct arriving cases requiring team mobilization off hours without benefit of telemedicine transfer lead time) we noted a 39% improvement. With Viz.ai, we noted immediate access to images and streamlined group communications, even in an already well-functioning CSC. These results have implications for future care processes and can be a model for centers striving to optimize workflow and improve NIR timeliness.

When human skin is irradiated with ultraviolet radiation (340–100 nm) there is an immediate pigment response, termed Immediate Pigment Darkening (IPD). This reaction is thought to be due to photooxidation of preexisting melanin, precursors and/or melanin metabolites because it appears during exposure. It has been demonstrated that UVA‐induced skin reactions, including erythema and pigmentation, are oxygen dependent. Therefore, these reactions should also be irradiance (fluence rate) dependent. The purpose of this investigation was to determine the dependence of the IPD threshold on fluence rate. We exposed the forearm of 12 volunteers (skin type II‐V) to monochromatic UVA radiation (362 nm) at a range of fluence rates of 6–115 mW/cm2and determined the fluence at which pigment was perceptible. The threshold fluence for the IPD reaction increased by a factor of 2.7 as the fluence rate increased by a factor of 18. Therefore, we conclude that the IPD reaction following exposure to 362 nm radiation is dependent on fluence rate, and independent of skin type.

The growth properties of an Escherichia coli strain carrying a chromosomal deletion of the ssuEADCB genes (formerly designated ycbPONME) indicated that the products of this gene cluster are required for the utilization of sulfur from aliphatic sulfonates. Sequence similarity searches indicated that the proteins encoded by ssuA, ssuB, and ssuC are likely to constitute an ABC type transport system, whereas ssuD and ssuE encode an FMNH(2)-dependent monooxygenase and an NAD(P)H-dependent FMN reductase, respectively (Eichhorn, E., van der Ploeg, J. R., and Leisinger, T. (1999) J. Biol. Chem. 274, 26639-26646). Synthesis of beta-galactosidase from a transcriptional chromosomal ssuE'-lacZ fusion was repressed by sulfate or cystine and depended on the presence of a functional cbl gene, which encodes a LysR-type transcriptional regulator. Electrophoretic mobility shift assays with the ssu promoter region and measurements of beta-galactosidase from plasmid-encoded ssuE'-'lacZ fusions showed that full expression of the ssu operon required the presence of a Cbl-binding site upstream of the -35 region. CysB, the LysR transcriptional regulator for the cys genes, was not required for expression of a chromosomal ssuE'-lacZ fusion although the ssu promoter region contained three CysB-binding sites. Integration host factor could also occupy three binding sites in the ssu promoter region but had no influence on expression of a chromosomal ssuE'-lacZ fusion.

The growth properties of an Escherichia colistrain carrying a chromosomal deletion of thessuEADCBgenes (formerly designated ycbPONME) indicated that the products of this gene cluster are required for the utilization of sulfur from aliphatic sulfonates. Sequence similarity searches indicated that the proteins encoded by ssuA,ssuB,and ssuCare likely to constitute an ABC type transport system, whereas ssuDand ssuEencode an FMNH2-dependent monooxygenase and an NAD(P)H-dependent FMN reductase, respectively (Eichhorn, E., van der Ploeg, J. R., and Leisinger, T. (1999) J. Biol. Chem.274, 26639–26646). Synthesis of β-galactosidase from a transcriptional chromosomal ssuE′-lacZfusion was repressed by sulfate or cystine and depended on the presence of a functional cblgene, which encodes a LysR-type transcriptional regulator. Electrophoretic mobility shift assays with the ssupromoter region and measurements of β-galactosidase from plasmid-encoded ssuE′-′lacZfusions showed that full expression of the ssuoperon required the presence of a Cbl-binding site upstream of the −35 region. CysB, the LysR transcriptional regulator for the cysgenes, was not required for expression of a chromosomal ssuE′-lacZfusion although the ssupromoter region contained three CysB-binding sites. Integration host factor could also occupy three binding sites in the ssupromoter region but had no influence on expression of a chromosomal ssuE′-lacZfusion.

Erich arch bars facilitate intermaxillary fixation, and traditional teaching questions and often argues against the use of arch bars in patients during primary and mixed dentition. Dental anatomy, risk of tooth avulsion, and potential disruption of developing permanent dentition are reasons cited. This study provides the only outcome data on the use of arch bars in children during primary and mixed dentition.

The Lewis acid–base adducts MoF5(NC5H5)nand MoOF4(NC5H5)n(n= 1, 2) were synthesized from the reactions of MoF5and MoOF4with C5H5N and structurally characterized by X-ray crystallography. Whereas the crystal structures of MoF5(NC5H5)2and MoOF4(NC5H5)2are isomorphous containing pentagonal-bipyramidal molecules, the fluorido-bridged, heptacoordinate [MoF5(NC5H5)]2dimer differs starkly from monomeric, hexacoordinate MoOF4(NC5H5). For the weaker Lewis base CH3CN, only the 1:1 adduct, MoF5(NCCH3), could be isolated. All adducts were characterized by Raman spectroscopy in conjunction with vibrational frequency calculations. Multinuclear NMR spectroscopy revealed an unprecedented isomerism of MoOF4(NC5H5)2in solution, with the pyridyl ligands occupying adjacent or nonadjacent positions in the equatorial plane of the pentagonal bipyramid. Paramagnetic MoF5(NC5H5)2was characterized by electron paramagnetic resonance (EPR) spectroscopy as a dispersion in solid adamantane as well as in a diamagnetic host lattice of MoOF4(NC5H5)2; EPR parameters were computed using ZORA with the BPW91 functional using relativistic all-electron wave functions for Mo and simulated using EasySpin. Density functional theory calculations (B3LYP) and natural bond orbital analyses were conducted to elucidate the distinctive bonding and structural properties of all adducts reported herein and explore fundamental differences observed in the Lewis acid behavior of MoF5and MoOF4.

In this article, the author shares her experience of managing her work as a playwright during COVID-19 pandemic; and pandemic has also made her think about the delivery of her plays.

The article presents several responses to the question that asks how can a mother, who is obliged to stay at home to nurture her child, still maintain friendships or social activities with friends. A mother explains that she found new friends, who are also mothers, when she was attending meetings in La Leche League.

The World Wide Web has provided the public with easy and affordable access to a vast range of information. However, claims may be unsubstantiated and misleading. The purpose of this study was to use cutaneous laser surgery as a model to assess the availability and reliability of Web sites and to evaluate this resource for the quality of patient and provider education. Three commercial methods of searching the Internet were used, identifying nearly 500,000 possible sites. The first 100 sites listed by each search engine (a total of 300 sites) were compared. Of these, 126 were listed repeatedly within a given retrieval method, whereas only 3 sites were identified by all 3 search engines. After elimination of duplicates, 40 sites were evaluated for content and currency of information. The most common features included postoperative care suggestions, options for pain management or anesthesia, a description of the way in which lasers work, and the types of lasers used for different procedures. Potential contraindications to laser procedures were described on fewer than 30% of the sites reviewed. None of the sites contained substantiation of claims or referrals to peer-reviewed publications or research. Because of duplication and the prioritization systems of search engines, the ease of finding sites did not correlate with the quality of the site’s content. Our findings show that advertisements for services exceed useful information. (J Am Acad Dermatol 2000;42:784-6.)

The article offers information on a visit through the city of Chicago, Illinois. Topics of discussion include the Art institute of Chicago, the DuSable Harbor named after the city's founder, Jean Baptiste Point du Sable, the Navy Pier, and the Loop. A brief overview of the Fulton River District is also included.

This article describes common laboratory procedures that can reduce the risk of culture contamination (sepsis), collectively referred as “aseptic technique.” Two major strategies for aseptic work are described: using a Bunsen burner and using a laminar flow hood. Both methods are presented in the form of general protocols applicable to a variety of laboratory tasks such as pipetting and dispensing aliquots, preparing growth media, and inoculating, passaging, and spreading microorganisms on petri dishes. © 2020 by John Wiley & Sons, Inc.

The article reviews the ""Schalcken. Gemalte Verführung" exhibition of the works of Dutch painter Godfried Schalcken at the Wallraf-Richartz-Museum art museum in Cologne, Germany until January 24, 2016.

The article reviews the exhibition of the works of post-impressionist Dutch painter Vincent van Gogh "Van Gogh & Co." in the Kröller Müller Museum art museum in Otterlo, the Netherlands until September 27, 2015.

Introduction:

The article previews the Expo Chicago art fair in Chicago, Illinois from September 13 to 17, 2017 featuring art galleries and dealers including Dittrich & Schlechtriem and Dennis Kimmerich of the Galerie Kimmerich art gallery.

The article presents information on the watercolor and tempera portrait painting "Mädchen mit Skimütze" by Alfons Walde for sale at the Kunsthandel Freller art gallery in Linz, Austria. The painting's depiction of a girl wearing a knitted skiing cap is analyzed and positioned within Walde's oeuvre and compared to his typical paintings of landscapes.

The article discusses the art trade show Art Nocturne to be held in Knocke, Belgium from August 10-18, 2013.

The article presents a preview of the 2015 Art Bodensee art fair in the Lake Constance region near Dornbirn, Austria from June 10 to 12, 2015 featuring exhibitons of works by artists including Christoph Luger and Michael Kutschbach.

The article offers a profile of German art dealer Thomas Derda. Derda's specialization in German art of the 1920s and 1930s is discussed, the design and decoration of Derda's derdaberlin art gallery in Berlin, Germany is reviewed, and works sold by Derda by artists including Marcel Breuer, Heinrich Hoerle, and László Moholy-Nagy are described.