Long-term persistence and effects of fetal microchimerisms on disease onset and status in a cohort of women with rheumatoid arthritis and systemic lupus erythematosus

BACKGROUND: The discovery of a fetal cells transfer to the mother is a phenomenon with multiple implications for autoimmunity and tolerance. The prevalence and meaning of the feto-maternal microchimerism (MC) in rheumatic diseases has not been thoroughly investigated. The aim of this study was to analyze the prevalence of fetal MC in patients with inflammatory rheumatic diseases and to investigate the association of MC with disease onset and current status. METHODS: A total of 142 women who gave birth to at least one male offspring were recruited: 72 women with rheumatoid arthritis (RA), 16 women with systemic lupus erythematosus (SLE), and 54 healthy women. For the detection of fetal microchimerism a nested PCR method was used to amplify a Y chromosome specific sequence (TSPY1). For characterization of disease activity we analyzed autoantibody profiles and X-rays in RA, and in addition complement levels in SLE respectively. RESULTS: A significant higher prevalence of fetal MC was found in RA (18%) and SLE (31%) compared to controls (3.7%) (p = 0.02 and p = 0.006, resp.). The mean age at disease onset was comparable in MC + and MC- RA patients. Disease onset occurred 18.7 (MC +) and 19.8 (MC-) years post partum of the first son, respectively. The presence of anti-CCP and RF did not differ significantly, anti-CCP were found in 75% of MC + and 87% of MC- patients, RF in 75% of both MC + and MC- patients. A slightly higher mean Steinbrocker score in MC + patients was associated with longer disease duration in MC + compared to MC- RA. In SLE patients the mean age at disease onset was 42.6 years in MC + and 49.1 years in MC- patients. Disease onset occurred 24.0 and 26.4 years post partum of the first son for MC + and MC- patients, respectively. The presence of ANA and anti-dsDNA antibodies, C3, C4 and CH50 did not differ significantly. CONCLUSION: Our results indicate a higher frequency of long-term male MC in RA and SLE patients compared with controls without impact on disease onset and status in RA and SLE

Pyrosequencing of the Camptotheca acuminata transcriptome reveals putative genes involved in camptothecin biosynthesis and transport

Background: Camptotheca acuminata is a Nyssaceae plant, often called the "happy tree", which is indigenous in Southern China. C. acuminata produces the terpenoid indole alkaloid, camptothecin (CPT), which exhibits clinical effects in various cancer treatments. Despite its importance, little is known about the transcriptome of C. acuminata and the mechanism of CPT biosynthesis, as only few nucleotide sequences are included in the GenBank database.Results: From a constructed cDNA library of young C. acuminata leaves, a total of 30,358 unigenes, with an average length of 403 bp, were obtained after assembly of 74,858 high quality reads using GS De Novo assembler software. Through functional annotation, a total of 21,213 unigenes were annotated at least once against the NCBI nucleotide (Nt), non-redundant protein (Nr), Uniprot/SwissProt, Kyoto Encyclopedia of Genes and Genomes (KEGG), and Arabidopsis thaliana proteome (TAIR) databases. Further analysis identified 521 ESTs representing 20 enzyme genes that are involved in the backbone of the CPT biosynthetic pathway in the library. Three putative genes in the upstream pathway, including genes for geraniol-10-hydroxylase (CaPG10H), secologanin synthase (CaPSCS), and strictosidine synthase (CaPSTR) were cloned and analyzed. The expression level of the three genes was also detected using qRT-PCR in C. acuminata. With respect to the branch pathway of CPT synthesis, six cytochrome P450s transcripts were selected as candidate transcripts by detection of transcript expression in different tissues using qRT-PCR. In addition, one glucosidase gene was identified that might participate in CPT biosynthesis. For CPT transport, three of 21 transcripts for multidrug resistance protein (MDR) transporters were also screened from the dataset by their annotation result and gene expression analysis.Conclusion: This study produced a large amount of transcriptome data from C. acuminata by 454 pyrosequencing. According to EST annotation, catalytic features prediction, and expression analysis, novel putative transcripts involved in CPT biosynthesis and transport were discovered in C. acuminata. This study will facilitate further identification of key enzymes and transporter genes in C. acuminata

Colorectal and uterine movement and tension of the inferior hypogastric plexus in cadavers

Background: Hypotheses on somatovisceral dysfunction often assume interference by stretch or compression of the nerve supply to visceral structures. The purpose of this study is to examine the potential of pelvic visceral movement to create tension of the loose connective tissue that contains the fine branches of the inferior hypogastric nerve plexus. Methods: Twenty eight embalmed human cadavers were examined. Pelvic visceral structures were displaced by very gentle 5 N unidirectional tension and the associated movement of the endopelvic fascia containing the inferior hypogastric plexus that this caused was measured. Results: Most movement of the fascia containing the inferior hypogastric plexus was obtained by pulling the rectosigmoid junction or broad ligament of the uterus. The plexus did not cross any vertebral joints and the fascia containing it did not move on pulling the hypogastric nerve. Conclusions: Uterine and rectosigmoid displacement produce most movement of the fascia containing the hypogastric nerve plexus, potentially resulting in nerve tension. In the living this might occur as a consequence of menstruation, pregnancy or constipation. This may be relevant to somatovisceral reflex theories of the effects of manual therapy on visceral conditions.Ian P Johnso

Vampires in the village Žrnovo on the island of Korčula: following an archival document from the 18th century

Središnja tema rada usmjerena je na raščlambu spisa pohranjenog u Državnom arhivu u Mlecima (fond: Capi del Consiglio de’ Dieci: Lettere di Rettori e di altre cariche) koji se odnosi na događaj iz 1748. godine u korčulanskom selu Žrnovo, kada su mještani – vjerujući da su se pojavili vampiri – oskvrnuli nekoliko mjesnih grobova. U radu se podrobno iznose osnovni podaci iz spisa te rečeni događaj analizira u širem društvenom kontekstu i prate se lokalna vjerovanja.The main interest of this essay is the analysis of the document from the State Archive in Venice (file: Capi del Consiglio de’ Dieci: Lettere di Rettori e di altre cariche) which is connected with the episode from 1748 when the inhabitants of the village Žrnove on the island of Korčula in Croatia opened tombs on the local cemetery in the fear of the vampires treating. This essay try to show some social circumstances connected with this event as well as a local vernacular tradition concerning superstitions

Orwell, Jones and the New York Times:: Conversations Shaping the Fundamental Rights of Privacy

The conversations among Orwell’s Nineteen Eight-Four, the US Supreme Court decision in United States v. Jones, and the New York Times compellingly illustrate the ways “law feeds and is fed by the world around it,” as Judge Guido Calabresi once put it. A suspected drug dealer, Antoine Jones was convicted and sentenced to life in prison at trial, but the case was overturned on appeal. The U.S. Supreme Court had to decide whether using a GPS tracking unit on the suspect’s vehicle violated Jones’ constitutional rights. Both lawyers and judges spoke often about whether this surveillance technique brought the U.S. closer to a state akin to the one in Nineteen Eight-Four. The New York Times covered this story, adding media sensationalism to the events. This article explores the legal development of privacy and surveillance at a specific historical moment, as it relates to both Orwell’s text, and the media at the nexus of privacy. One of the main issues was whether a U.S. citizen might reasonably expect a GPS tracking unit to trace their every movement, and exactly where that reasonable expectation of privacy ends. We see that technology has far outstripped even what Orwell envisioned in his dystopian masterpiece, and that only conscious resistance and legal protection can reverse what has often and ironically become a “voluntary” surrendering of the right to privacy

Kristallisation der Arbutin-Synthase und der Strictosidin Glukosidase - zwei Enzyme aus dem sekundären Glykosidstoffwechsel von Rauvolfia serpentina

Kristallisation der Arbutin-Synthase und der Strictosidin Glukosidase - zwei Enzyme aus dem sekundären Glykosidstoffwechsel von Rauvolfia serpentina Die vorliegende Arbeit befasst sich mit der Kristallisation und der strukturellen Auswertung der Arbutin-Synthase (AS) und der Strictosidin Glukosidase (SG). Beide Enzyme stammen aus der Medizinalpflanze Rauvolfia serpentina. Für die Kristallisation der Arbutin-Synthase wurden ca. 2500 verschiedene Beding-ungen experimentell untersucht. Für einige dieser Experimente wurde das Enzym molekularbiologisch und chemisch verändert. Trotzdem konnten keine Kristalle erhalten werden. Die bei diesen Veränderungen erhaltenen Ergebnisse wurden anhand von Vergleichen mit Strukturen anderer Glykosyltransferasen der gleichen Familie analysiert. Bei der Reinigung der AS konnte mit verschiedenen Trennsystemen nie eine homogene Lösung produziert werden. Der wahrscheinliche Grund für diese schlechte Isolierbarkeit, und damit der wahrscheinliche Grund für die schwierige Kris-tallisation, liegt in der überdurchschnittlich hohen Anzahl an Cysteinen in der Proteinsequenz. Mit den Aminosäuren Cys171, Cys253 und Cys461 wurden drei Cysteine gefunden, die einem Strukturvergleich nach an der Proteinoberfläche liegen und möglicherweise durch Quervernetzungen mit anderen Proteinmolekülen ein heterogenes Gemisch bilden, das nicht geordnet kristallisieren kann. Durch gezielte Mutationen dieser drei Aminosäuren könnte die Kristallisation zukünftig ermöglicht werden. Für die SG waren bereits Bedingungen bekannt bei denen nicht vermessbare Enzymkristalle (Nadeln) wuchsen. In weit gefächerten Versuchen konnten diese Kristalle jedoch nicht zu 3D-Wachstum angeregt werden. Es wurden mit einem HTS-Screening neue Bedingungen zur Kristallisation gefunden. Anschließend konnten die native Struktur und der Strictosidin/Enzym-Komplex vermessen und aufgeklärt werden. Die SG gehört zur Familie 1 der Glukosidasen (GH-1) und besitzt die in dieser Familie konservierte (beta/alpha)8-Barrel-Faltung. Im Vergleich mit 16 bekannten Glykosidasen der Familie GH-1 wurde die Substratbindung untersucht. Dabei wurde die in der Familie konservierte Zuckerbindung vorgefunden, jedoch große Unterschiede in der Aglykonbindung entdeckt. Es wurden Bedingungen für die Konformationsänderung des Trp388 erkannt. Diese Konformationsänderung dirigiert den Aglykonteil des Substrates auf verschiedene Seiten der Substratbindungstasche und teilt so die Familie GH-1 in zwei Gruppen.Crystallization of the Arbutin-Synthase and the Strictosidine Glucosidase - two enzymes in the secondary glycoside metabolism of Rauvolfia serpentina In this dissertation the crystallization and structural interpretation of the enzymes Arbutin-Synthase (AS) and Strictosidine Glucosidase (SG) is described. Both enzymes originate from the medicinal plant Rauvolfia serpentina. With about 2500 different conditions the crystallization of AS was tested. Therefore the enzyme was also chemically and molecularbiologically modified. Nevertheless no crystals were obtained. The acquired results were evaluated in comparison to known structures of glycosyltransferases of the same family. Although a wide variety of purification systems was used during the purification of the heterologously expressed AS, a homogeneous solution could never be produced. The most likely cause for the bad separability of AS, and thus the most likely cause for the missing crystallization, is the higher than average number of cysteins in the sequence of AS. Three cysteins (Cys171, Cys253 and Cys461) were identified as positioned on the surface of the protein. Thereby they provide the possibility for crosslink’s between several protein molecules, forming a heterologous solution and inhibiting crystal growth. After point mutations of these three amino acids the crystallization of AS could be possible. In the case of SG there were conditions known that produces needle-shaped enzyme crystals. But these were not suitable for X-ray measurements. In wide variations it was tried to optimize the conditions towards 3D-growth. Only a new High-Throughput-Screening was successful. With the acquired crystals the native structure of SG was solved and after soaking the complex of SG-Glu207Gln/strictosidine was elucidated. SG belongs to family 1 of glycosidases (GH-1) and shows the conserved (beta/alpha)8-barrel fold. The substrate binding of SG was compared with all known structures of that family. Thereby was conserved sugar binding site found, but great differences in the aglycone binding were observed. Conformational changes at Trp388 were studied through point mutations and causes for the changes recorded. By the conformational changes of the large indole part of Trp388 the substrate is directed into two different pockets and thereby is the family divided into two groups of different substrate binding

Evaluation of the Rahel Hirsch habilitation program

Hintergrund: Habilitationen durch Frauen sind erst seit ca. 100 Jahren in Deutschland möglich. Bei einem Frauenanteil an Habilitationen von unter 30% ist die Wissenschaft in Deutschland von einer Parität weit entfernt. Im Bemühen um internationale Anschlussfähigkeit wird innerhalb von Reformen das Ziel der Gleichstellung von Frauen in der Wissenschaft über den Einsatz von Förderinstrumenten verfolgt. Nur in medizinischen Fächern hat die Habilitation als Voraussetzung für eine Professur kaum Bedeutungsverlust erfahren. Zur Erhöhung des Frauenanteils an Professuren an der Charité - Universitätsmedizin Berlin wird seit 1999 eine Förderung durch das Rahel-Hirsch-Habilitationsprogramm an herausragende Wissenschaftlerinnen vergeben. Ziel dieser Untersuchung ist es erstmals den Erfolg, fördernde und hemmende Einflussfaktoren auf Erfolg und die Einschätzung der Geförderten zum Programm empirisch zu erheben. Methode: Die Evaluation ist angelegt als retrospektive Querschnittserhebung und nutzt Sekundärdatenanalysen sowie eine Online-Befragung ehemaliger Programmgeförderter, um über den Erwerb einer PD bzw. Professur Erfolg zu ermitteln. Erfolgswahrscheinlichkeiten werden bestimmt und fördernde und hemmende Einflussfaktoren auf Erfolg untersucht. Die Einschätzungen der Geförderten zum Programm werden über deskriptive Analysen dargestellt. Ergebnisse: Zwischen 2018 - 2019 beteiligen sich 77 von 101 ehemaligen Geförderten an einer Befragung. Mit einer Zielerreichung von 75% wird der Erfolg des Rahel-Hirsch-Programmes belegt (Sekundärdaten). Resultierende Wahrscheinlichkeiten für Erfolg liegen in Abhängigkeit des Förderbeginns zwischen 76% und 100%. Engagement in Gremien und Kommissionen, in der Lehre und bei der Betreuung wissenschaftlicher Arbeiten zeigt fördernden Einfluss auf Erfolg (p ≤ 0,05). Alter und Mutterschaft zeigen keinen statistisch signifikanten Zusammenhang mit Erfolg. Dagegen wirkt die Übernahme von Pflegeverantwortung, eine hohe Belastung der Vereinbarkeit und eine starke Einbindung in die Patient:innenversorgung, die den Fortgang des Habilitationsprojektes ungünstig beeinflussen, hindernd auf Erfolg. Die Rahmenbedingungen des Programmes werden von 92% der Befragten positiv bewertet. Auf Basis festgestellter Einflussfaktoren und in Verbindung mit den Einschätzungen der Geförderten können wichtige Erkenntnisse zur Weiterentwicklung des Programmes abgeleitet werden.Background: Habilitations by women have only been possible in Germany for around 100 years. With a percentage of women doing post-doctoral qualifications of less than 30%, science in Germany is far from parity. In an effort to achieve international connectivity, reforms pursue the goal of equality for women in science through the use of funding instruments. Only in medical subjects has the habilitation as a prerequisite for a professorship experienced hardly any loss of importance. To increase the proportion of women in professorships at Charité - Universitätsmedizin Berlin, the Rahel Hirsch habilitation program has been awarded to outstanding female scientists since 1999. The aim of this study is, for the first time, to empirically ascertain the success, supporting and inhibiting factors influencing success and the assessment of those receiving the program. Method: The evaluation is designed as a retrospective cross-sectional survey and uses secondary data analyzes as well as an online survey of former program participants in order to determine success through the acquisition of a private lecturer (PD) or professorship. The probabilities of success are determined and factors that promote and inhibit success are examined. The participants’ assessments of the program are presented using descriptive analyzes. Results: Between 2018 and 2019, 77 out of 101 former participants took part in a survey. The success of the Rahel Hirsch program is proven with a target achievement of 75% (secondary data). The resulting probabilities of success lie between 76% and 100% depending on the start of funding. Commitment in commissions and committees, in teaching and in the supervision of scientific work shows a positive influence on success (p ≤ 0.05). Age and motherhood show no statistically significant association with success. On the other hand, the assumption of care responsibility, a high burden of work-life balance and a strong involvement in patient care, which has an unfavorable influence on the progress of the habilitation project, prevent success. The framework conditions of the program are rated positively by 92% of those surveyed. On the basis of determined influencing factors and in connection with the assessments of the participants, important findings for the further development of the program can be derived