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1. Mycobacterium tuberculosis evolution from monoresistance to pre-extensive drug resistance during a prolonged household outbreak

Author

G. Greif, C. Coithino, M.N. Bentancor, and C. Robello

Subjects
Tuberculosis, Epidemiology, Drug resistance, Diseases of the respiratory system, RC705-779, Infectious and parasitic diseases, RC109-216
Abstract

Whole genome sequencing (WGS) is sensitive tool for the analysis of tuberculosis transmission and drug-resistance. We used WGS to analyze the Mycobacterium tuberculosis evolution from isoniazid monoresistance to MDR/preXDR during a prolonged household outbreak. The outbreak started with a isoniazid resistant strain (katG S315T mutation) and evolve in two cases to pre-XDR phenotype (with mutations in katG, rpoB, embB, pncA and gyrA genes). Based on WGS data and epidemiological interview we proposed a possible chain of transmission an evolution of the strains.Similar intra-patient and inter-patient acquisition of variability was observed, making difficult to distinguish reinfection or reactivation. Analysis of WGS data together with epidemiological clinical history are discussed in order to distinguish between prolonged infections or transition from latency to reactivation. Classical interview and clinical history taking should be consider to fully understanding WGS data. With a still low incidence of TB cases, Uruguay could use universal WGS of all isolates to reduce time of diagnosis, detect outbreaks and perform public actions to reduce TB incidence.

Published
2024
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2. First case of Mycobacterium heckeshornense cavitary lung disease in the Latin America and Caribbean region

Author

C. Coitinho, G. Greif, J. van Ingen, P. Laserra, C. Robello, and C. Rivas

Subjects
Epidemiology, infection, mycobacterium, Mycobacterium heckeshornense, pulmonary, Infectious and parasitic diseases, RC109-216
Abstract

A case of cavitary pulmonary disease caused by Mycobacterium heckeshornense in Uruguay is described. This is the first case reported in the Latin America and Caribbean region, showing that this species is a worldwide opportunistic human pathogen.

Published
2016
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3. Štúidium rastu a produkcie biogénnych amínov niektorými mikroorganizmami za modelových podmienok

Author

G. Greif, M. Greifova, J. Dvoran, J. Karovicova, and V. Buchtova

Subjects
cadaverine, putrescine, histamine, escherichia coli, enterobacter aerogenes, klebsiella pneumoniae, Agriculture
Abstract

The study was aimed at the growth of selected strains from the family Enterobacteriaceae (Escherichia coli, Enterobacter aerogenes and Klebsiella pneumoniae) in meat-peptone (MPB) broth and cabbage juice at different cultivation temperatures, and at the production of biogenic amines (cadaverine, putrescine, histamine). Bacterial growth was evaluated on the basis of specific growth rate (IJm) and lag phase (A.) calculated from growth curves. Cadaverine was produced as the first amine in MPB and cabbage juice by all studied st rains at the cultivation temperatures and at Jiving cell densities 10 6 KTJ/cm3. Putrescine was produced by E. coli only in both substrates at the cultivation temperatures. Histamine was produced by E. coli at 18 °C in cabbage juice and by Enterobacter aerogenes in both substrates at the cultivation temperatures.

Published
1999
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4. Differences in human and minipig platelet number, volume and activation induced by borosilicate glass beads in a modified chandler loop-system

Author

F. Jung, G. Greif, M. Wendt, Bernhard Hiebl, H. Meyer-Sievers, and C. Mrowietz

Subjects
Blood Platelets, Platelet Count, Swine, Physiology, Borosilicate glass, Chemistry, Activation markers, Cardiovascular research, Hematology, Blood flow, Flow Cytometry, Platelet Activation, Physiology (medical), Animals, Humans, Swine, Miniature, Platelet, Platelet activation, Mean platelet volume, Cardiology and Cardiovascular Medicine, Large animal, Biomedical engineering
Abstract

The pig is the most widely used large animal model in Europe, with cardiovascular research being one of the main areas of application. Adequate refinement of interventional studies in this field, meeting the requirements of Russel and Burchs’ 3 R concept, can only be performed if blood-contacting medical devices are hemocompatible. Because most medical devices for cardiovascular interventional procedures are developed for humans they are tested mostly for compatibility with human blood. The aim of this study was therefore to determine whether there are differences in behavior of porcine and human platelets when they come into contact with glass, which was used as an exemplary thrombogenic material. For this purpose changes of platelet count, platelet volume and platelet expression of the activation markers CD61, CD62P and CD63 were measured using a modified chandler loop-system simulating the fluidic effects of the blood flow. Minipig and human platelets showed significant differences in number and volume, but not in activation after 4–8 min exposure to glass.

Published
2021
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5. Urinary ethyl glucuronide (uEtG) as a marker for alcohol consumption in liver transplant candidates: a real-world cohort

Author

Friedrich Foerster, Hauke Lang, JM Schattenberg, M. A. Woerns, G. Greif-Higer, F. Darstein, Daniel Grimm, Tim Zimmermann, Johanna Vollmar, Peter R. Galle, Karl J. Lackner, Philipp Mildenberger, Franziska Stern, Jens Mittler, and Jens U. Marquardt

Subjects
Male, medicine.medical_specialty, Alcohol Drinking, Waiting Lists, Urinary system, medicine.medical_treatment, Enzyme-Linked Immunosorbent Assay, Glucuronates, Liver transplantation, Sensitivity and Specificity, Gastroenterology, Cohort Studies, 03 medical and health sciences, chemistry.chemical_compound, 0302 clinical medicine, Ethyl glucuronide, Liver Cirrhosis, Alcoholic, Predictive Value of Tests, Tandem Mass Spectrometry, Internal medicine, Humans, Mass Screening, Medicine, Prospective Studies, 030212 general & internal medicine, Prospective cohort study, Mean corpuscular volume, Aged, Creatinine, Ethanol, medicine.diagnostic_test, business.industry, Middle Aged, Liver Transplantation, chemistry, Predictive value of tests, Female, 030211 gastroenterology & hepatology, business, Biomarkers, Chromatography, Liquid, Alcohol Abstinence
Abstract

In order to reduce alcohol relapse after liver transplantation (LT), the German national guidelines for waiting-list maintenance and organ allocation demand a minimum 6-month period of alcohol abstinence pre-LT, confirmed by measuring urinary ethyl glucuronide (uEtG). Between January 2015 and June 2016, uEtG was measured at least once in 339 cirrhotic patients with an indication for LT at the University Medical Center Mainz. uEtG was measured with an enzyme-linked immunosorbent assay (ELISA) screening test (cutoff value: 500 µg/L). For uEtG values ≥ 500 µg/L, liquid chromatography-mass spectrometry (LC-MS/MS) was performed as a confirmatory assay. Data were collected prospectively in a transplant database. Of the 339 potential liver transplant candidates, uEtG was negative in 86.4 %. Most patients were male (64.3 %), with an average age of 56.42 ± 10.1 years. In the multivariate analysis, mean corpuscular volume (p = 0.001), urinary creatinine (p = 0.001), gamma-glutamyl transferase (p = 0.001), and hemoglobin (p = 0.003) were significantly associated with a positive uEtG test result. The sensitivity of the ELISA screening test was 100 % for uEtG values 2000 µg/L, as confirmed by LC-MS/MS. uEtG is an effective parameter to reveal alcohol consumption by patients on the waiting list for LT. The sensitivity of the ELISA is excellent for uEtG values 2000 µg/L, for which LC-MS/MS confirmation could be omitted. In der deutschen Richtlinie für die Wartelistenführung und die Organvermittlung gem. § 16 Abs. 1 S. 1 Nrn. 2 u. 5 TPG wird eine durch Bestimmung von Ethylglucuronid im Urin (uEtG) nachweisliche, absolute Alkoholabstinenzzeit von mindestens 6 Monaten als Voraussetzung für eine Lebertransplantation (LT) bei einer ethyltoxischen Leberzirrhose gefordert. Zwischen Januar 2015 und Juni 2016 wurde an der Universitätsmedizin Mainz uEtG mindestens einmal bei 339 Patienten mit Leberzirrhose und Indikation für eine LT mittels ELISA Screening Test (cut off: 500 µg/l) gemessen. Bei Werten ≥ 500 µg/l erfolgte ein massenspektrometrischer Bestätigungstest. Die Daten wurden prospektiv in einer LT-Datenbank erhoben. Von den 339 potenziellen LT Kandidaten waren 86,4 % uEtG-negativ. Die meisten Patienten waren männlich (64,3 %) und durchschnittlich 56,42 ± 10,1 Jahre alt. In der multivariaten Analyse waren MCV (p = 0,001), Urin-Kreatinin (p = 0,001), Gamma-Glutamyltransferase (GGT) (p = 0,001) und Hämoglobin (Hb) (p = 0,003) signifikant mit einem positiven uEtG assoziiert. Die Sensitivität des ELISA Screeningtest war bei uEtG-Werten 2000 µg/l 100 %. In unserer Real-World-Kohorte konnte uEtG als valider und zuverlässiger Marker zum Nachweis eines Alkoholkonsums bei Patienten auf der Warteliste zur Lebertransplantation bestätigt werden. Zudem konnten wir zeigen, dass der ELISA-Screenintest bei uEtG-Werten 2000 µg/l eine exzellente Sensitivität hat und somit auf den massenspektrometrischen Bestätigungstest verzichtet werden kann.

Published
2020
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6. Differences in human and sheep platelet adherence, aggregation and activation induced by glass beads in a modified chandler loop-system

Author

M. Ganter, H. Meyer-Sievers, G. Greif, C. Mrowietz, Bernhard Hiebl, and F. Jung

Subjects
Change over time, Blood Platelets, Sheep, CD63, Platelet Aggregation, Physiology, Chemistry, Platelet Count, Platelet-Rich Plasma, Swine, Cardiovascular research, Hematology, Platelet Activation, Andrology, Physiology (medical), Models, Animal, Animals, Humans, Platelet, Mean platelet volume, Cardiology and Cardiovascular Medicine, CD61, Silicone tube, Large animal
Abstract

In human cardiovascular research, sheep in particular are used as a large animal model in addition to pigs. In these animals, medical products, developed and tested for human medical purposes, are almost exclusively used in interventional studies. Therefore, the extent to which platelets from human and ovine blood differ in terms of adherence, aggregation and activation after a 4- or 8-minutes exposure to glass was investigated. Testing was performed with platelet-rich plasma (PRP) and a modified chandler loop-system, with 4- and 8-minute blood-material exposure times corresponding to 20 and 40 test cycles, respectively, through the entire silicone tube loop of the test system. In sheep and human PRP, contact with the silicone tubing resulted in a decrease in platelet count after 4 minutes and 20 test cycles, respectively. Four more minutes (20 additional test cycles) caused a further decrease of the platelet count only in sheep PRP. When the silicon tube was partly filled with glass beads, these effects were more pronounced and stronger in sheep then in human PRP. The mean platelet volume, which was used as parameter for platelet aggregation, did not change over time in human PRP without glass exposure. With glass exposure in human and sheep PRP the mean platelet volume increased within 40 test cycles, but this increase was stronger in sheep than in human PRP. Regarding activation behavior, the activation markers CD62P and CD63 were detectable only in

Published
2021

7. A Comparative, Multicenter, Observational Study of Medication Adherence in Liver Cirrhosis Patients and Dialysis Patients Using Electronic Event Measurement

Author

Irene Krämer, Tim Zimmermann, Jens Lutz, G. Greif-Higer, Rita Marina Heeb, Manfred Berres, Patrick Harloff, and Thomas Mettang

Subjects
Not evaluated, medicine.medical_specialty, Cirrhosis, business.industry, medicine.medical_treatment, 05 social sciences, medicine.disease, 0506 political science, End stage renal disease, Transplantation, 03 medical and health sciences, Liver disease, 0302 clinical medicine, Internal medicine, 050602 political science & public administration, medicine, 030212 general & internal medicine, Dosing, Intensive care medicine, business, Dialysis, Patient education
Abstract

Introduction: Medication adherence was only studied in a limited number of potential organ recipients. So far medication adherence was not evaluated by utilizing electronic monitoring systems in dialysis and liver cirrhosis patients. The main objective of the present study was to measure the medication adherence of patients suffering from end stage kidney or liver disease by an objectified method. Methods: Adult liver cirrhosis patients taking propranolol and dialysis patients taking phosphate binders, each medication 3 times daily, were eligible to be enrolled in the study protocol. Medication adherence was measured electronically with MEMS™-containers over a period of 6 months in each patient. Results: 34 patients suffering from liver cirrhosis and 36 dialysis patients participated in the study and were analysed per protocol. The Dosing Adherence (DA) rate differed significantly (p

Published
2017
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8. Psychological Conflicts Between Relatives During the Long-Term Course After Successful Living Organ Donation

Author

E. Wandel, M. E. Beutel, G. Greif-Higer, Peter R. Galle, and Gerd Otto

Subjects
Adult, Male, medicine.medical_specialty, Time Factors, medicine.medical_treatment, Family conflict, Stalemate, Liver transplantation, Nephrectomy, Conflict, Psychological, Interviews as Topic, Interpersonal relationship, Living organ donation, Living Donors, medicine, Hepatectomy, Humans, Family, Interpersonal Relations, Organ donation, Spouses, Transplantation, business.industry, Psychosomatic medicine, Middle Aged, Surgery, Family medicine, Female, business, Follow-Up Studies
Abstract

The German transplantation law prefers living organ donation between close relatives and spouses, which is assumed to guarantee unequivocal altruistic motivation. Since 2001, 68 recipient-donor-pairs, who aspired to have a renal or liver transplantation, underwent a systematic psychosomatic evaluation. Meanwhile, 43 transplantations were performed including 34 renal and 9 liver cases. Seventeen recipient-donor-pairs were readministered evaluations by the department of psychosomatic medicine after 1 to 6 years after transplantation for long-term follow-up. In 10 cases of medically successful transplantation, we identified severe conflicts between donor, recipient, and next-of-kin. Major conflicts are presented by case vignettes regarding deterioration of a previously conflicted marriage, noncompliance of the recipient due to a marital stalemate, and family conflict revolving around refusal to donate. Based on these findings, concise assessments of donor-recipient-pairs are recommended regardless of family relationships. Particular attention must be paid to signs of conflict both before and after transplantation.

Published
2008
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10. Einstellungen zur postmortalen Organspende - Ergebnisse einer Repräsentativerhebung der deutschen Bevölkerung

Author

PR Galle, A. Haselbacher, Gerd Otto, M. E. Beutel, and G. Greif-Higer

Subjects
medicine.medical_specialty, Next of kin, business.industry, Gastroenterology, Possession (law), Social class, Public opinion, Surgery, Transplantation, German population, Family medicine, medicine, Consolation, Organ donation, business
Abstract

The increasing deficit of organs causes a drastic decline in the quality of life and survival of numerous patients in need of a transplantation. The purpose of this representative community study was to survey attitudes toward transplantation in the German population and to identify underlying determinants. Unlike previous surveys, fears and concerns were elicited based on a concrete case vignette. Among the 1,002 participants, 90 % were in favour of organ donation in general; 21 % reported having signed an organ donor card. Consent to organ donation was associated with younger age and higher social class; the same was true for the possession of an organ donor card. In the virtual decision situation, the majority (77 %) voted in favour of an organ donation based on saving lives, consolation for bereaved and the absence of disadvantages for the donor. Common (up to 50 %), however, were also fears and concerns regarding determination of the time of death, displacement of medical concern from the donor to the recipient of the organ, utilisation of organs for other purposes, or explantation before death. The knowledge of the determinants identified, of existing fears and concerns are helpful not only for informing the public, but also for the dialogue with the next of kin of potential donors. Here, it may be helpful not only to address arguments pro organ donation, but also to address potential fears and concerns.

Published
2006
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11. Acid tolerance response and survival by oral bacteria

Author

Dennis G. Cvitkovitch, E. C. G. Greif, U.-B. Larsson, G. Svensater, and I. R. Hamilton

Subjects
Microbiology (medical), Lactobacillus casei, Immunology, Adaptation, Biological, Biology, Prevotella intermedia, Microbiology, Enterococcus faecalis, Prevotella, Actinomyces, Humans, General Dentistry, Mouth, Streptococcus, Actinomycetaceae, Drug Tolerance, Hydrogen-Ion Concentration, biology.organism_classification, Streptococcaceae, Streptococcus mutans, Culture Media, Lactobacillus, Enterococcus, Acids, Neisseria, Bacteria
Abstract

Using 21 species of oral bacteria, representing six acidogenic genera, we undertook to determine whether the pH-limiting exponential growth is related to the ability of the organisms to generate an acid-tolerance response that results in enhanced survival at low pH. The lower pH limit of exponential growth varied by more than two units with that of Neisseria A182 at pH 6.34; growth of Lactobacillus casei RB1014 stopped at pH 3.81, with species of Actinomyces, Enterococcus, Prevotella and Streptococcus falling between these limits. The working hypothesis was that the organisms with the higher pH limits for growth are unable to respond to acidic environments in order to survive, whereas the more aciduric organisms would possess or acquire acid tolerance. Adaptation to acid tolerance was tested by determining whether the prior exposure of exponential-phase cells to a low, sub-lethal pH would trigger the induction of a mechanism that would enhance survival at a pH killing pH 7.5 control cells. The killing pH varied from pH 4.5 for Prevotella intermedia ATCC 25611 to pH 2.3 for the three Lactobacillus casei strains in the study, with the three Streptococcus mutans strains killed at pH 3.0 for 3 h. The adaptation experiments revealed three groups of organisms: non-acid-responders, generally representing strains with the highest terminal pH values; weak acid-responders in the middle of the pH list, generating low numbers of survivors at one or two pH values, and the aciduric, strong responders generating a high number of survivors at pH values in the range 6.0 to 3.5, but not at pH 7.5. Predominant among the latter group were the S. mutans and Lactobacilli casei strains, with the most significant adaptive response exhibited by S. mutans LT11 and S. mutans Ingbritt, involving a process that required protein synthesis. Time course experiments with the latter organisms indicated that 90-120 min was required after exposure to the triggering pH before the acid response was fully functional. These results indicate that the sudden exposure of strains of oral streptococci and lactobacilli, as well as Enterococcus faecalis, to pH values between 6.0 and 3.5 results in the induction of an acid tolerance response that enhances the survival of these strains at or below pH 3.5.

Published
1997
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13. Molecular analysis of isoniazid-resistant Mycobacterium tuberculosis isolates in Uruguay

Author

G, Greif, C, Coitinho, C, Rivas, J, van Ingen, and C, Robello

Subjects
DNA, Bacterial, Genotyping Techniques, Mutation, Tuberculosis, Multidrug-Resistant, Antitubercular Agents, Isoniazid, Humans, Uruguay, Mycobacterium tuberculosis, Sequence Analysis, DNA
Abstract

The incidence of tuberculosis (TB) is increasing in high-risk populations in Uruguay, possibly owing to emerging resistance. Mycobacterial interspersed repetitive units (MIRU) genotyping and katG sequence analysis of isoniazid (INH) resistance-associated mutations were performed in 45 INH-resistant Mycobacterium tuberculosis isolates in Uruguayan patients. The genotype distribution among INH-resistant isolates shares features of that of neighbouring countries, with a predominance of Latin American and Mediterranean, T and Haarlem genotypes, although the S genotype was particularly frequent among our isolates. Forty-four per cent of INH-resistant strains harboured the S315T mutation in katG; we found novel katG mutations (W321X, G269T, P232R and G221Wfs1) that could explain INH resistance.

Published
2012

15. Psychologische und beziehungsdynamische Langzeitfolgen für Organempfänger und -spender vor und nach medizinisch gelungener Transplantation mittels Lebendorganspende–eine kritische Untersuchung

Author

G. Greif-Higer, Wolfgang Söllner, G. Otto, M. E. Beutel, Peter R. Galle, P. Dobe-Tauchert, and U. Eckart

Subjects
Psychiatry and Mental health, Clinical Psychology, Applied Psychology
Abstract

In der Transplantationsmedizin stellt die Lebendorganspende (LOS) eine wichtige Methode dar, schwer erkrankte Patienten schnell und mit gutem Erfolg mit einem Spenderorgan zu versorgen. In den Transplantationszentren Mainz und Erlangen-Nurnberg wurden in den letzten 5 Jahren 251 Empfanger(E)-Spender(S)-Paare vor der geplanten Transplantation psychosomatisch evaluiert, bei 123 Fallen wurde eine Transplantation realisiert. Bereits in der diagnostischen Phase vor Transplantation ergaben sich in 38 E- S- Paaren (15%) Belastungen, die sich 1. durch Beziehungskonflikte, 2. Rucktritt eines Beteiligten von der Spende u.a. entwickelten und sich auf die seelische Befindlichkeit und die Beziehung der Beteiligten auswirkte. Im Langzeitverlauf ergaben sich bei 31 der E- S- Paaren trotz gelungener Transplantation 1. gravierende seelische Belastungen von Empfanger und Spender, 2. beziehungsdynamische Konflikte zwischen Empfanger und Spender und der erweiterten Familie bis hin zu aggressiven Ubergriffen. 3. In einigen Fallen, bei denen E und S nach der Transplantation dauerhaft zusammenlebten, nahm das gespendete Organ im Laufe der Zeit eine bedeutende Rolle im Beziehungsgefuge ein mit erheblichen Folgeproblemen. Zusammengefasst ist bei mindestens 25% der Empfanger-Spenderpaaren mit psychologischen Konfliktlagen im Langzeitverlauf zu rechnen, die einer weiterfuhrenden Behandlung bedurfen. Die dominierenden Befunde werden dargestellt und anhand von Fall-Vignetten exemplarisch analysiert.

Published
2008
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17. [Attitudes towards cadaveric organ donation--results from a representative survey of the German population]

Author

M E, Beutel, G, Greif-Higer, A, Haselbacher, P R, Galle, and G, Otto

Subjects
Health Knowledge, Attitudes, Practice, Tissue and Organ Procurement, Germany, Public Opinion, Surveys and Questionnaires, Cadaver, Attitude to Health, Tissue Donors
Abstract

The increasing deficit of organs causes a drastic decline in the quality of life and survival of numerous patients in need of a transplantation. The purpose of this representative community study was to survey attitudes toward transplantation in the German population and to identify underlying determinants. Unlike previous surveys, fears and concerns were elicited based on a concrete case vignette. Among the 1,002 participants, 90 % were in favour of organ donation in general; 21 % reported having signed an organ donor card. Consent to organ donation was associated with younger age and higher social class; the same was true for the possession of an organ donor card. In the virtual decision situation, the majority (77 %) voted in favour of an organ donation based on saving lives, consolation for bereaved and the absence of disadvantages for the donor. Common (up to 50 %), however, were also fears and concerns regarding determination of the time of death, displacement of medical concern from the donor to the recipient of the organ, utilisation of organs for other purposes, or explantation before death. The knowledge of the determinants identified, of existing fears and concerns are helpful not only for informing the public, but also for the dialogue with the next of kin of potential donors. Here, it may be helpful not only to address arguments pro organ donation, but also to address potential fears and concerns.

Published
2006

23. D

Author

Franz von Bruchhausen, Eberhard Hackenthal, Siegfried Ebel, Ulrike Holzgrabe, August Wilhelm Frahm, M. Albinus, G. Amschler, E. von Angerer, null Arras-Reiter, P. Barth, W. Barthel, K. Bauer, P. Bauer, I. Baumann, J. Beckmann, W. Beil, J. Reitz, K. Binder, F. Bossle, F. Bracher, H. Bräunlich, E. Bretschneider, R. Brigelius-Flohé, K. Brinkmann, F. von Bruchhausen, A. Rüge, W. Christ, M. Cimbollek, R. Daniels, G. Dannhardt, H. Duchstein, S. Ebel, K. Eger, P. Eichhorn, U. Eiben, T. Erker, P. Felfe, A. Frahm, M. Frahm, V. Franke, K. Freundt, D. Geffken, U. Geis, E. Glusa, B. Göber, P. Gobina, W. Golder, M. Goppelt­Strübe, K. Götte, E. Gottstein, G. Greif, A. Grisk, M. Grosam, H. Gustmann, M. Gütschow, E. Hackenthal, A. Häfner, B. Haluszczynski, A. Harder, H. Häusler, D. Heber, M. Heidenreich, G. Heinemeyer, E. Heller, D. von Herrath, J. Hilfenhaus, H. Hoffmann, U. Hoffmann-Schollmayer, B. Hofmann, C. Holpert, U. Holzgrabe, U. Hübner-Steiner, M. Hug, E. Inkmann, A. Jördens, J. Jürgens, B. Kaiser, D. Kalbhen, H. Kemmler, P. Kisser, D. Kleinsorge, C. Klett, S. Klett, M. Klingmüller, H. Klöcking, A. Kramer, B. Krammer, M. Kreher, M. Krüger, M. Kuhn, D. Landsiedel-Maier, P. Lauven, J. Lehmann, M. Lehner, D. Leopoldt, A. Maurer, W. Meindl, K. Menges, P. Mes­singer, F. Meyer, W. Meyerhof, R. Morgenstern, U. Mühlhans, A. Müller, C. Müller, K. Müller, A. Mülsch, C. Nachtsheim, M. Neugebauer, W. Neupert, P. Nickel, P. Nuhn, B. Nürnberg, H. Oelschläger, J. Oertel, M. Oettel, R. Ott, T. Ott, T. Otzen, P. Pachaly, H. Pelzer, K. Petersen, R. Pick, M. Pickert, A. Pies, H. Priewer, O. Queckenberg, G. Radau, E. Reimann, J. Remien, M. Reuß, W. Reuß, J. Richter, P. Richter, K. Riecke, H. Rommelspacher, U. Rose, G. Roth, D. Rothley, G. Rücker, J. Schäfer, J. Schantl, H. Schlager, H. Schleinitz, W. Schlichter, M. Schmauß, H. Schmidhammer, G. Schmidt, T. Schmidt, H. Schmitt, J. Sehräder, T. Schulz, H. Schwilden, M. Serke, G. Skopp, G. Skorka, K. Smolinka, U. Speck, M. Spohn, R. Stahlmann, J. Stasch, C. Steffen, H. Stein, J. Steinmeyer, K. Stiefvater, G. Strippel, K. Surborg, U. Stürig, H. Szelényi, I. Szelényi, A. Täufel, R. Thieroff-Ekerdt, R. Troschütz, H. Ungeheuer, B. Unterhalt, E. Verspohl, S. Vogel, F. Volk, T. Vorwerk, J. Wallmann, H. Weber, M. Wenzel, M. Weyandt-Spangenberg, S. Wich, R. Wintersteiger, B. Wüst, and D. Youssef

Published
1999
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24. A

Author

Franz von Bruchhausen, Eberhard Hackenthal, Siegfried Ebel, Ulrike Holzgrabe, August Wilhelm Frahm, M. Albinus, G. Amschler, E. von Angerer, null Arras-Reiter, P. Barth, W. Barthel, K. Bauer, P. Bauer, I. Baumann, J. Beckmann, W. Beil, J. Reitz, K. Binder, F. Bossle, F. Bracher, H. Bräunlich, E. Bretschneider, R. Brigelius-Flohé, K. Brinkmann, F. von Bruchhausen, A. Rüge, W. Christ, M. Cimbollek, R. Daniels, G. Dannhardt, H. Duchstein, S. Ebel, K. Eger, P. Eichhorn, U. Eiben, T. Erker, P. Felfe, A. Frahm, M. Frahm, V. Franke, K. Freundt, D. Geffken, U. Geis, E. Glusa, B. Göber, P. Gobina, W. Golder, M. Goppelt­Strübe, K. Götte, E. Gottstein, G. Greif, A. Grisk, M. Grosam, H. Gustmann, M. Gütschow, E. Hackenthal, A. Häfner, B. Haluszczynski, A. Harder, H. Häusler, D. Heber, M. Heidenreich, G. Heinemeyer, E. Heller, D. von Herrath, J. Hilfenhaus, H. Hoffmann, U. Hoffmann-Schollmayer, B. Hofmann, C. Holpert, U. Holzgrabe, U. Hübner-Steiner, M. Hug, E. Inkmann, A. Jördens, J. Jürgens, B. Kaiser, D. Kalbhen, H. Kemmler, P. Kisser, D. Kleinsorge, C. Klett, S. Klett, M. Klingmüller, H. Klöcking, A. Kramer, B. Krammer, M. Kreher, M. Krüger, M. Kuhn, D. Landsiedel-Maier, P. Lauven, J. Lehmann, M. Lehner, D. Leopoldt, A. Maurer, W. Meindl, K. Menges, P. Mes­singer, F. Meyer, W. Meyerhof, R. Morgenstern, U. Mühlhans, A. Müller, C. Müller, K. Müller, A. Mülsch, C. Nachtsheim, M. Neugebauer, W. Neupert, P. Nickel, P. Nuhn, B. Nürnberg, H. Oelschläger, J. Oertel, M. Oettel, R. Ott, T. Ott, T. Otzen, P. Pachaly, H. Pelzer, K. Petersen, R. Pick, M. Pickert, A. Pies, H. Priewer, O. Queckenberg, G. Radau, E. Reimann, J. Remien, M. Reuß, W. Reuß, J. Richter, P. Richter, K. Riecke, H. Rommelspacher, U. Rose, G. Roth, D. Rothley, G. Rücker, J. Schäfer, J. Schantl, H. Schlager, H. Schleinitz, W. Schlichter, M. Schmauß, H. Schmidhammer, G. Schmidt, T. Schmidt, H. Schmitt, J. Sehräder, T. Schulz, H. Schwilden, M. Serke, G. Skopp, G. Skorka, K. Smolinka, U. Speck, M. Spohn, R. Stahlmann, J. Stasch, C. Steffen, H. Stein, J. Steinmeyer, K. Stiefvater, G. Strippel, K. Surborg, U. Stürig, H. Szelényi, I. Szelényi, A. Täufel, R. Thieroff-Ekerdt, R. Troschütz, H. Ungeheuer, B. Unterhalt, E. Verspohl, S. Vogel, F. Volk, T. Vorwerk, J. Wallmann, H. Weber, M. Wenzel, M. Weyandt-Spangenberg, S. Wich, R. Wintersteiger, B. Wüst, and D. Youssef

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1999
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25. H

Author

Franz von Bruchhausen, Eberhard Hackenthal, Siegfried Ebel, Ulrike Holzgrabe, August Wilhelm Frahm, M. Albinus, G. Amschler, E. von Angerer, null Arras-Reiter, P. Barth, W. Barthel, K. Bauer, P. Bauer, I. Baumann, J. Beckmann, W. Beil, J. Reitz, K. Binder, F. Bossle, F. Bracher, H. Bräunlich, E. Bretschneider, R. Brigelius-Flohé, K. Brinkmann, F. von Bruchhausen, A. Rüge, W. Christ, M. Cimbollek, R. Daniels, G. Dannhardt, H. Duchstein, S. Ebel, K. Eger, P. Eichhorn, U. Eiben, T. Erker, P. Felfe, A. Frahm, M. Frahm, V. Franke, K. Freundt, D. Geffken, U. Geis, E. Glusa, B. Göber, P. Gobina, W. Golder, M. Goppelt­Strübe, K. Götte, E. Gottstein, G. Greif, A. Grisk, M. Grosam, H. Gustmann, M. Gütschow, E. Hackenthal, A. Häfner, B. Haluszczynski, A. Harder, H. Häusler, D. Heber, M. Heidenreich, G. Heinemeyer, E. Heller, D. von Herrath, J. Hilfenhaus, H. Hoffmann, U. Hoffmann-Schollmayer, B. Hofmann, C. Holpert, U. Holzgrabe, U. Hübner-Steiner, M. Hug, E. Inkmann, A. Jördens, J. Jürgens, B. Kaiser, D. Kalbhen, H. Kemmler, P. Kisser, D. Kleinsorge, C. Klett, S. Klett, M. Klingmüller, H. Klöcking, A. Kramer, B. Krammer, M. Kreher, M. Krüger, M. Kuhn, D. Landsiedel-Maier, P. Lauven, J. Lehmann, M. Lehner, D. Leopoldt, A. Maurer, W. Meindl, K. Menges, P. Mes­singer, F. Meyer, W. Meyerhof, R. Morgenstern, U. Mühlhans, A. Müller, C. Müller, K. Müller, A. Mülsch, C. Nachtsheim, M. Neugebauer, W. Neupert, P. Nickel, P. Nuhn, B. Nürnberg, H. Oelschläger, J. Oertel, M. Oettel, R. Ott, T. Ott, T. Otzen, P. Pachaly, H. Pelzer, K. Petersen, R. Pick, M. Pickert, A. Pies, H. Priewer, O. Queckenberg, G. Radau, E. Reimann, J. Remien, M. Reuß, W. Reuß, J. Richter, P. Richter, K. Riecke, H. Rommelspacher, U. Rose, G. Roth, D. Rothley, G. Rücker, J. Schäfer, J. Schantl, H. Schlager, H. Schleinitz, W. Schlichter, M. Schmauß, H. Schmidhammer, G. Schmidt, T. Schmidt, H. Schmitt, J. Sehräder, T. Schulz, H. Schwilden, M. Serke, G. Skopp, G. Skorka, K. Smolinka, U. Speck, M. Spohn, R. Stahlmann, J. Stasch, C. Steffen, H. Stein, J. Steinmeyer, K. Stiefvater, G. Strippel, K. Surborg, U. Stürig, H. Szelényi, I. Szelényi, A. Täufel, R. Thieroff-Ekerdt, R. Troschütz, H. Ungeheuer, B. Unterhalt, E. Verspohl, S. Vogel, F. Volk, T. Vorwerk, J. Wallmann, H. Weber, M. Wenzel, M. Weyandt-Spangenberg, S. Wich, R. Wintersteiger, B. Wüst, and D. Youssef

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1999
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26. I

Author

Franz von Bruchhausen, Eberhard Hackenthal, Siegfried Ebel, Ulrike Holzgrabe, August Wilhelm Frahm, M. Albinus, G. Amschler, E. von Angerer, null Arras-Reiter, P. Barth, W. Barthel, K. Bauer, P. Bauer, I. Baumann, J. Beckmann, W. Beil, J. Reitz, K. Binder, F. Bossle, F. Bracher, H. Bräunlich, E. Bretschneider, R. Brigelius-Flohé, K. Brinkmann, F. von Bruchhausen, A. Rüge, W. Christ, M. Cimbollek, R. Daniels, G. Dannhardt, H. Duchstein, S. Ebel, K. Eger, P. Eichhorn, U. Eiben, T. Erker, P. Felfe, A. Frahm, M. Frahm, V. Franke, K. Freundt, D. Geffken, U. Geis, E. Glusa, B. Göber, P. Gobina, W. Golder, M. Goppelt­Strübe, K. Götte, E. Gottstein, G. Greif, A. Grisk, M. Grosam, H. Gustmann, M. Gütschow, E. Hackenthal, A. Häfner, B. Haluszczynski, A. Harder, H. Häusler, D. Heber, M. Heidenreich, G. Heinemeyer, E. Heller, D. von Herrath, J. Hilfenhaus, H. Hoffmann, U. Hoffmann-Schollmayer, B. Hofmann, C. Holpert, U. Holzgrabe, U. Hübner-Steiner, M. Hug, E. Inkmann, A. Jördens, J. Jürgens, B. Kaiser, D. Kalbhen, H. Kemmler, P. Kisser, D. Kleinsorge, C. Klett, S. Klett, M. Klingmüller, H. Klöcking, A. Kramer, B. Krammer, M. Kreher, M. Krüger, M. Kuhn, D. Landsiedel-Maier, P. Lauven, J. Lehmann, M. Lehner, D. Leopoldt, A. Maurer, W. Meindl, K. Menges, P. Mes­singer, F. Meyer, W. Meyerhof, R. Morgenstern, U. Mühlhans, A. Müller, C. Müller, K. Müller, A. Mülsch, C. Nachtsheim, M. Neugebauer, W. Neupert, P. Nickel, P. Nuhn, B. Nürnberg, H. Oelschläger, J. Oertel, M. Oettel, R. Ott, T. Ott, T. Otzen, P. Pachaly, H. Pelzer, K. Petersen, R. Pick, M. Pickert, A. Pies, H. Priewer, O. Queckenberg, G. Radau, E. Reimann, J. Remien, M. Reuß, W. Reuß, J. Richter, P. Richter, K. Riecke, H. Rommelspacher, U. Rose, G. Roth, D. Rothley, G. Rücker, J. Schäfer, J. Schantl, H. Schlager, H. Schleinitz, W. Schlichter, M. Schmauß, H. Schmidhammer, G. Schmidt, T. Schmidt, H. Schmitt, J. Sehräder, T. Schulz, H. Schwilden, M. Serke, G. Skopp, G. Skorka, K. Smolinka, U. Speck, M. Spohn, R. Stahlmann, J. Stasch, C. Steffen, H. Stein, J. Steinmeyer, K. Stiefvater, G. Strippel, K. Surborg, U. Stürig, H. Szelényi, I. Szelényi, A. Täufel, R. Thieroff-Ekerdt, R. Troschütz, H. Ungeheuer, B. Unterhalt, E. Verspohl, S. Vogel, F. Volk, T. Vorwerk, J. Wallmann, H. Weber, M. Wenzel, M. Weyandt-Spangenberg, S. Wich, R. Wintersteiger, B. Wüst, and D. Youssef

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1999
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27. C

Author

Franz von Bruchhausen, Eberhard Hackenthal, Siegfried Ebel, Ulrike Holzgrabe, August Wilhelm Frahm, M. Albinus, G. Amschler, E. von Angerer, null Arras-Reiter, P. Barth, W. Barthel, K. Bauer, P. Bauer, I. Baumann, J. Beckmann, W. Beil, J. Reitz, K. Binder, F. Bossle, F. Bracher, H. Bräunlich, E. Bretschneider, R. Brigelius-Flohé, K. Brinkmann, F. von Bruchhausen, A. Rüge, W. Christ, M. Cimbollek, R. Daniels, G. Dannhardt, H. Duchstein, S. Ebel, K. Eger, P. Eichhorn, U. Eiben, T. Erker, P. Felfe, A. Frahm, M. Frahm, V. Franke, K. Freundt, D. Geffken, U. Geis, E. Glusa, B. Göber, P. Gobina, W. Golder, M. Goppelt­Strübe, K. Götte, E. Gottstein, G. Greif, A. Grisk, M. Grosam, H. Gustmann, M. Gütschow, E. Hackenthal, A. Häfner, B. Haluszczynski, A. Harder, H. Häusler, D. Heber, M. Heidenreich, G. Heinemeyer, E. Heller, D. von Herrath, J. Hilfenhaus, H. Hoffmann, U. Hoffmann-Schollmayer, B. Hofmann, C. Holpert, U. Holzgrabe, U. Hübner-Steiner, M. Hug, E. Inkmann, A. Jördens, J. Jürgens, B. Kaiser, D. Kalbhen, H. Kemmler, P. Kisser, D. Kleinsorge, C. Klett, S. Klett, M. Klingmüller, H. Klöcking, A. Kramer, B. Krammer, M. Kreher, M. Krüger, M. Kuhn, D. Landsiedel-Maier, P. Lauven, J. Lehmann, M. Lehner, D. Leopoldt, A. Maurer, W. Meindl, K. Menges, P. Mes­singer, F. Meyer, W. Meyerhof, R. Morgenstern, U. Mühlhans, A. Müller, C. Müller, K. Müller, A. Mülsch, C. Nachtsheim, M. Neugebauer, W. Neupert, P. Nickel, P. Nuhn, B. Nürnberg, H. Oelschläger, J. Oertel, M. Oettel, R. Ott, T. Ott, T. Otzen, P. Pachaly, H. Pelzer, K. Petersen, R. Pick, M. Pickert, A. Pies, H. Priewer, O. Queckenberg, G. Radau, E. Reimann, J. Remien, M. Reuß, W. Reuß, J. Richter, P. Richter, K. Riecke, H. Rommelspacher, U. Rose, G. Roth, D. Rothley, G. Rücker, J. Schäfer, J. Schantl, H. Schlager, H. Schleinitz, W. Schlichter, M. Schmauß, H. Schmidhammer, G. Schmidt, T. Schmidt, H. Schmitt, J. Sehräder, T. Schulz, H. Schwilden, M. Serke, G. Skopp, G. Skorka, K. Smolinka, U. Speck, M. Spohn, R. Stahlmann, J. Stasch, C. Steffen, H. Stein, J. Steinmeyer, K. Stiefvater, G. Strippel, K. Surborg, U. Stürig, H. Szelényi, I. Szelényi, A. Täufel, R. Thieroff-Ekerdt, R. Troschütz, H. Ungeheuer, B. Unterhalt, E. Verspohl, S. Vogel, F. Volk, T. Vorwerk, J. Wallmann, H. Weber, M. Wenzel, M. Weyandt-Spangenberg, S. Wich, R. Wintersteiger, B. Wüst, and D. Youssef

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1999
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28. G

Author

Franz von Bruchhausen, Eberhard Hackenthal, Siegfried Ebel, Ulrike Holzgrabe, August Wilhelm Frahm, M. Albinus, G. Amschler, E. von Angerer, null Arras-Reiter, P. Barth, W. Barthel, K. Bauer, P. Bauer, I. Baumann, J. Beckmann, W. Beil, J. Reitz, K. Binder, F. Bossle, F. Bracher, H. Bräunlich, E. Bretschneider, R. Brigelius-Flohé, K. Brinkmann, F. von Bruchhausen, A. Rüge, W. Christ, M. Cimbollek, R. Daniels, G. Dannhardt, H. Duchstein, S. Ebel, K. Eger, P. Eichhorn, U. Eiben, T. Erker, P. Felfe, A. Frahm, M. Frahm, V. Franke, K. Freundt, D. Geffken, U. Geis, E. Glusa, B. Göber, P. Gobina, W. Golder, M. Goppelt­Strübe, K. Götte, E. Gottstein, G. Greif, A. Grisk, M. Grosam, H. Gustmann, M. Gütschow, E. Hackenthal, A. Häfner, B. Haluszczynski, A. Harder, H. Häusler, D. Heber, M. Heidenreich, G. Heinemeyer, E. Heller, D. von Herrath, J. Hilfenhaus, H. Hoffmann, U. Hoffmann-Schollmayer, B. Hofmann, C. Holpert, U. Holzgrabe, U. Hübner-Steiner, M. Hug, E. Inkmann, A. Jördens, J. Jürgens, B. Kaiser, D. Kalbhen, H. Kemmler, P. Kisser, D. Kleinsorge, C. Klett, S. Klett, M. Klingmüller, H. Klöcking, A. Kramer, B. Krammer, M. Kreher, M. Krüger, M. Kuhn, D. Landsiedel-Maier, P. Lauven, J. Lehmann, M. Lehner, D. Leopoldt, A. Maurer, W. Meindl, K. Menges, P. Mes­singer, F. Meyer, W. Meyerhof, R. Morgenstern, U. Mühlhans, A. Müller, C. Müller, K. Müller, A. Mülsch, C. Nachtsheim, M. Neugebauer, W. Neupert, P. Nickel, P. Nuhn, B. Nürnberg, H. Oelschläger, J. Oertel, M. Oettel, R. Ott, T. Ott, T. Otzen, P. Pachaly, H. Pelzer, K. Petersen, R. Pick, M. Pickert, A. Pies, H. Priewer, O. Queckenberg, G. Radau, E. Reimann, J. Remien, M. Reuß, W. Reuß, J. Richter, P. Richter, K. Riecke, H. Rommelspacher, U. Rose, G. Roth, D. Rothley, G. Rücker, J. Schäfer, J. Schantl, H. Schlager, H. Schleinitz, W. Schlichter, M. Schmauß, H. Schmidhammer, G. Schmidt, T. Schmidt, H. Schmitt, J. Sehräder, T. Schulz, H. Schwilden, M. Serke, G. Skopp, G. Skorka, K. Smolinka, U. Speck, M. Spohn, R. Stahlmann, J. Stasch, C. Steffen, H. Stein, J. Steinmeyer, K. Stiefvater, G. Strippel, K. Surborg, U. Stürig, H. Szelényi, I. Szelényi, A. Täufel, R. Thieroff-Ekerdt, R. Troschütz, H. Ungeheuer, B. Unterhalt, E. Verspohl, S. Vogel, F. Volk, T. Vorwerk, J. Wallmann, H. Weber, M. Wenzel, M. Weyandt-Spangenberg, S. Wich, R. Wintersteiger, B. Wüst, and D. Youssef

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1999
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29. F

Author

Franz von Bruchhausen, Eberhard Hackenthal, Siegfried Ebel, Ulrike Holzgrabe, August Wilhelm Frahm, M. Albinus, G. Amschler, E. von Angerer, null Arras-Reiter, P. Barth, W. Barthel, K. Bauer, P. Bauer, I. Baumann, J. Beckmann, W. Beil, J. Reitz, K. Binder, F. Bossle, F. Bracher, H. Bräunlich, E. Bretschneider, R. Brigelius-Flohé, K. Brinkmann, F. von Bruchhausen, A. Rüge, W. Christ, M. Cimbollek, R. Daniels, G. Dannhardt, H. Duchstein, S. Ebel, K. Eger, P. Eichhorn, U. Eiben, T. Erker, P. Felfe, A. Frahm, M. Frahm, V. Franke, K. Freundt, D. Geffken, U. Geis, E. Glusa, B. Göber, P. Gobina, W. Golder, M. Goppelt­Strübe, K. Götte, E. Gottstein, G. Greif, A. Grisk, M. Grosam, H. Gustmann, M. Gütschow, E. Hackenthal, A. Häfner, B. Haluszczynski, A. Harder, H. Häusler, D. Heber, M. Heidenreich, G. Heinemeyer, E. Heller, D. von Herrath, J. Hilfenhaus, H. Hoffmann, U. Hoffmann-Schollmayer, B. Hofmann, C. Holpert, U. Holzgrabe, U. Hübner-Steiner, M. Hug, E. Inkmann, A. Jördens, J. Jürgens, B. Kaiser, D. Kalbhen, H. Kemmler, P. Kisser, D. Kleinsorge, C. Klett, S. Klett, M. Klingmüller, H. Klöcking, A. Kramer, B. Krammer, M. Kreher, M. Krüger, M. Kuhn, D. Landsiedel-Maier, P. Lauven, J. Lehmann, M. Lehner, D. Leopoldt, A. Maurer, W. Meindl, K. Menges, P. Mes­singer, F. Meyer, W. Meyerhof, R. Morgenstern, U. Mühlhans, A. Müller, C. Müller, K. Müller, A. Mülsch, C. Nachtsheim, M. Neugebauer, W. Neupert, P. Nickel, P. Nuhn, B. Nürnberg, H. Oelschläger, J. Oertel, M. Oettel, R. Ott, T. Ott, T. Otzen, P. Pachaly, H. Pelzer, K. Petersen, R. Pick, M. Pickert, A. Pies, H. Priewer, O. Queckenberg, G. Radau, E. Reimann, J. Remien, M. Reuß, W. Reuß, J. Richter, P. Richter, K. Riecke, H. Rommelspacher, U. Rose, G. Roth, D. Rothley, G. Rücker, J. Schäfer, J. Schantl, H. Schlager, H. Schleinitz, W. Schlichter, M. Schmauß, H. Schmidhammer, G. Schmidt, T. Schmidt, H. Schmitt, J. Sehräder, T. Schulz, H. Schwilden, M. Serke, G. Skopp, G. Skorka, K. Smolinka, U. Speck, M. Spohn, R. Stahlmann, J. Stasch, C. Steffen, H. Stein, J. Steinmeyer, K. Stiefvater, G. Strippel, K. Surborg, U. Stürig, H. Szelényi, I. Szelényi, A. Täufel, R. Thieroff-Ekerdt, R. Troschütz, H. Ungeheuer, B. Unterhalt, E. Verspohl, S. Vogel, F. Volk, T. Vorwerk, J. Wallmann, H. Weber, M. Wenzel, M. Weyandt-Spangenberg, S. Wich, R. Wintersteiger, B. Wüst, and D. Youssef

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1999
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30. B

Author

Franz von Bruchhausen, Eberhard Hackenthal, Siegfried Ebel, Ulrike Holzgrabe, August Wilhelm Frahm, M. Albinus, G. Amschler, E. von Angerer, null Arras-Reiter, P. Barth, W. Barthel, K. Bauer, P. Bauer, I. Baumann, J. Beckmann, W. Beil, J. Reitz, K. Binder, F. Bossle, F. Bracher, H. Bräunlich, E. Bretschneider, R. Brigelius-Flohé, K. Brinkmann, F. von Bruchhausen, A. Rüge, W. Christ, M. Cimbollek, R. Daniels, G. Dannhardt, H. Duchstein, S. Ebel, K. Eger, P. Eichhorn, U. Eiben, T. Erker, P. Felfe, A. Frahm, M. Frahm, V. Franke, K. Freundt, D. Geffken, U. Geis, E. Glusa, B. Göber, P. Gobina, W. Golder, M. Goppelt­Strübe, K. Götte, E. Gottstein, G. Greif, A. Grisk, M. Grosam, H. Gustmann, M. Gütschow, E. Hackenthal, A. Häfner, B. Haluszczynski, A. Harder, H. Häusler, D. Heber, M. Heidenreich, G. Heinemeyer, E. Heller, D. von Herrath, J. Hilfenhaus, H. Hoffmann, U. Hoffmann-Schollmayer, B. Hofmann, C. Holpert, U. Holzgrabe, U. Hübner-Steiner, M. Hug, E. Inkmann, A. Jördens, J. Jürgens, B. Kaiser, D. Kalbhen, H. Kemmler, P. Kisser, D. Kleinsorge, C. Klett, S. Klett, M. Klingmüller, H. Klöcking, A. Kramer, B. Krammer, M. Kreher, M. Krüger, M. Kuhn, D. Landsiedel-Maier, P. Lauven, J. Lehmann, M. Lehner, D. Leopoldt, A. Maurer, W. Meindl, K. Menges, P. Mes­singer, F. Meyer, W. Meyerhof, R. Morgenstern, U. Mühlhans, A. Müller, C. Müller, K. Müller, A. Mülsch, C. Nachtsheim, M. Neugebauer, W. Neupert, P. Nickel, P. Nuhn, B. Nürnberg, H. Oelschläger, J. Oertel, M. Oettel, R. Ott, T. Ott, T. Otzen, P. Pachaly, H. Pelzer, K. Petersen, R. Pick, M. Pickert, A. Pies, H. Priewer, O. Queckenberg, G. Radau, E. Reimann, J. Remien, M. Reuß, W. Reuß, J. Richter, P. Richter, K. Riecke, H. Rommelspacher, U. Rose, G. Roth, D. Rothley, G. Rücker, J. Schäfer, J. Schantl, H. Schlager, H. Schleinitz, W. Schlichter, M. Schmauß, H. Schmidhammer, G. Schmidt, T. Schmidt, H. Schmitt, J. Sehräder, T. Schulz, H. Schwilden, M. Serke, G. Skopp, G. Skorka, K. Smolinka, U. Speck, M. Spohn, R. Stahlmann, J. Stasch, C. Steffen, H. Stein, J. Steinmeyer, K. Stiefvater, G. Strippel, K. Surborg, U. Stürig, H. Szelényi, I. Szelényi, A. Täufel, R. Thieroff-Ekerdt, R. Troschütz, H. Ungeheuer, B. Unterhalt, E. Verspohl, S. Vogel, F. Volk, T. Vorwerk, J. Wallmann, H. Weber, M. Wenzel, M. Weyandt-Spangenberg, S. Wich, R. Wintersteiger, B. Wüst, and D. Youssef

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1999
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31. Animal Models of Coccidia Infection

Author

A. Haberkorn and G. Greif

Subjects
Veterinary medicine, biology, Laboratory mouse, biology.organism_classification, medicine.disease, Small intestine, Breed, Caecum, Isospora, Coccidiosis, medicine.anatomical_structure, Coccidia, medicine, Parasite hosting
Abstract

Publisher Summary This chapter focuses on animal models of coccidia infection. Coccidiosis, an infection of the small intestine, is relatively rarely diagnosed in humans, where it is caused by Isospora belli. Two models have proved particularly successful in work to identify and evaluate new anticoccidials, which include coccidiosis of poultry and mammalian coccidiosis. In case of the former, any defined breed of domestic fowl (Gallus gallus) is suitable in principle. A guaranteed supply of birds of the same breed from a well-run breeding operation is essential. When the groups used in the studies are small, the chicks should be of the same sex. The animals should be kept separate from others before the start of the study in order to prevent spontaneous infection. Surgical interventions before and during the study are not necessary. Infections with E. tenella, E. acervulina, and E. maxima have proved particularly satisfactory as these three species, which parasitize different parts of the intestine, are particularly important from an economic point of view and, with a little practice, their oocysts are readily distinguishable from one another under a microscope. In case of mammalian model, a mouse model is used, where E.falciformis is used as the parasite. E. falciformis develops in the caecum and especially in the upper colon of the laboratory mouse. Oral infection via oesophageal tube is best carried out on Tuesday morning, with medication with the feed on Monday. The rate of mortality from the infection should be around 50% and the suspension used for the infection must be adjusted so that no more than 0.5 ml per mouse has to be administered.

Published
1999
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32. K

Author

Franz von Bruchhausen, Eberhard Hackenthal, Siegfried Ebel, Ulrike Holzgrabe, August Wilhelm Frahm, M. Albinus, G. Amschler, E. von Angerer, null Arras-Reiter, P. Barth, W. Barthel, K. Bauer, P. Bauer, I. Baumann, J. Beckmann, W. Beil, J. Reitz, K. Binder, F. Bossle, F. Bracher, H. Bräunlich, E. Bretschneider, R. Brigelius-Flohé, K. Brinkmann, F. von Bruchhausen, A. Rüge, W. Christ, M. Cimbollek, R. Daniels, G. Dannhardt, H. Duchstein, S. Ebel, K. Eger, P. Eichhorn, U. Eiben, T. Erker, P. Felfe, A. Frahm, M. Frahm, V. Franke, K. Freundt, D. Geffken, U. Geis, E. Glusa, B. Göber, P. Gobina, W. Golder, M. Goppelt­Strübe, K. Götte, E. Gottstein, G. Greif, A. Grisk, M. Grosam, H. Gustmann, M. Gütschow, E. Hackenthal, A. Häfner, B. Haluszczynski, A. Harder, H. Häusler, D. Heber, M. Heidenreich, G. Heinemeyer, E. Heller, D. von Herrath, J. Hilfenhaus, H. Hoffmann, U. Hoffmann-Schollmayer, B. Hofmann, C. Holpert, U. Holzgrabe, U. Hübner-Steiner, M. Hug, E. Inkmann, A. Jördens, J. Jürgens, B. Kaiser, D. Kalbhen, H. Kemmler, P. Kisser, D. Kleinsorge, C. Klett, S. Klett, M. Klingmüller, H. Klöcking, A. Kramer, B. Krammer, M. Kreher, M. Krüger, M. Kuhn, D. Landsiedel-Maier, P. Lauven, J. Lehmann, M. Lehner, D. Leopoldt, A. Maurer, W. Meindl, K. Menges, P. Mes­singer, F. Meyer, W. Meyerhof, R. Morgenstern, U. Mühlhans, A. Müller, C. Müller, K. Müller, A. Mülsch, C. Nachtsheim, M. Neugebauer, W. Neupert, P. Nickel, P. Nuhn, B. Nürnberg, H. Oelschläger, J. Oertel, M. Oettel, R. Ott, T. Ott, T. Otzen, P. Pachaly, H. Pelzer, K. Petersen, R. Pick, M. Pickert, A. Pies, H. Priewer, O. Queckenberg, G. Radau, E. Reimann, J. Remien, M. Reuß, W. Reuß, J. Richter, P. Richter, K. Riecke, H. Rommelspacher, U. Rose, G. Roth, D. Rothley, G. Rücker, J. Schäfer, J. Schantl, H. Schlager, H. Schleinitz, W. Schlichter, M. Schmauß, H. Schmidhammer, G. Schmidt, T. Schmidt, H. Schmitt, J. Sehräder, T. Schulz, H. Schwilden, M. Serke, G. Skopp, G. Skorka, K. Smolinka, U. Speck, M. Spohn, R. Stahlmann, J. Stasch, C. Steffen, H. Stein, J. Steinmeyer, K. Stiefvater, G. Strippel, K. Surborg, U. Stürig, H. Szelényi, I. Szelényi, A. Täufel, R. Thieroff-Ekerdt, R. Troschütz, H. Ungeheuer, B. Unterhalt, E. Verspohl, S. Vogel, F. Volk, T. Vorwerk, J. Wallmann, H. Weber, M. Wenzel, M. Weyandt-Spangenberg, S. Wich, R. Wintersteiger, B. Wüst, and D. Youssef

Published
1999
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33. Eimeria tenella: localisation of rhoptry antigens during parasite-host cell interactions by a rhoptry-specific monoclonal antibody in PCKC culture

Author

G, Greif and R, Entzeroth

Subjects
Organelles, Mice, Inbred BALB C, Immunoblotting, Antibodies, Protozoan, Antigens, Protozoan, Host-Parasite Interactions, Mice, Tumor Cells, Cultured, Animals, Electrophoresis, Polyacrylamide Gel, Microscopy, Immunoelectron, Chickens, Cells, Cultured, Eimeria tenella
Abstract

A rhoptry-specific monoclonal antibody (mab) A4C6 was obtained by fusion of X63-Ag 8.653 plasmacytoma cells with splenocytes of BALB/c mice. Mab A4C6 reacted with three protein bands at about 94, 66 and 45 kDa under non-reducing conditions. Gel electrophoresis under reducing conditions separated these protein complexes into several protein fragments ranging between 20 and 200 kDa. Antigens recognised by mab A4C6 were concentrated in the rhoptry sacs located in the apical region of sporozoites as shown by immunoelectron microscopy. During parasite-host cell interaction and development in PCKC culture, mab A4C6 located rhoptry antigens in the parasitophorous vacuole space between intracellular sporozoites and the vacuole membrane.

Published
1996

35. Der Crithidia luciliae-Immunfluoreszenztest zum Nachweis von Antikörpern gegen doppelsträngige DNA

Author

G. Greif, P. Herzer, and E. M. Lemmel

Subjects
business.industry, Drug Discovery, Molecular Medicine, Medicine, General Medicine, business, Molecular biology, Genetics (clinical)
Abstract

Der Crithidia luciliae-Immunfluoreszenztest (Cl-IFT) zum Nachweis von Antikorpern gegen doppelstrangige DNA (ds DNA) wurde in der Diagnostik und Verlaufsbeurteilung des systemischen Lupus erythematodes (SLE) in seiner Wertigkeit mit der radioimmunologischen Bestimmung im Farr-Test verglichen. Es zeigte sich, das der Cl-IFT lediglich bei Patienten ohne Zeichen von Krankheitsaktivitat von geringerer Sensitivitat ist, in Phasen von klinischer Krankheitssymptomatik dem Farr-Test jedoch gleichwertig ist. Eine Ubereinstimmung beider Testverfahren bestand in 76,9% der untersuchten Patienten-Seren; fehlende Ubereinstimmung war nicht nur eine Frage der Sensitivitat, in einigen Seren konnten nur mit dem Cl-IFT Antikorper nachgewiesen werden.

Published
1982
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36. Rheumatologie

Author

G. Greif, K. Bandilla, D. Berg, G. Schimsheimer, H. Schmidt, K.-H. Schicketanz, S. Zarges, R. Dreher, K. Federlin, G. Holz, B. Rotzler, R. Ziegler, U. Sprandel, H. Kremer, K. Wallnöfer, F. Weber, and M. Schattenkirchner

Published
1984
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37. [The Crithidia luciliae immunofluorescence test for detection of antibodies to double-stranded DNA (author's transl)]

Author

P, Herzer, G, Greif, and E M, Lemmel

Subjects
Lupus Erythematosus, Discoid, Sjogren's Syndrome, Antibodies, Antinuclear, Crithidia, Fluorescent Antibody Technique, Humans, Immunoglobulins, Lupus Erythematosus, Systemic, Complement System Proteins, DNA
Abstract

For measurement of antibodies of double-stranded DNA (ds DNA) the Crithidia luciliae immunofluorescence test (C1-IFT) was compared with the Farr precipitation technique in diagnosis and management of systemic lupus erythematosus (SLE). The C1-IFT proved to be less sensitive in SLE patients without clinical disease activity, but the sensitivity of the test in those with active disease was comparable to that of the Farr test. In 76.9% of the patients' sera identical test results were obtained by both assays. In some sera antibodies were detected only by the C1-IFT. Positive test results in patients with diseases other than SLE were found more often with the Farr test. Thus, The C1-IFT seemed to be more specific for diagnosis of SLE. Measurement of titers using the C1-IFT provides a simple method to survey SL E disease activity. It allows to analyze the immunoglobulin classes of the reacting antibodies and their complement fixation ability in vitro. All three major immunoglobulin classes (IgG, IgM, IgA) were found to be present in the sera with high antibody titers, complement fixation in the C1-IFT could predominantly be detected is sera with high antibody titers.

Published
1982

39. Efficacy of an escalating dose regimen of pegylated interferon ?-2a plus ribavirin in the early phase of HCV reinfection after liver transplantation

Author

Stefan Biesterfeld, Peter R. Galle, Ansgar W. Lohse, Stefan Kanzler, Gerd Otto, Christian Mönch, G. Greif-Higer, Ana Paula Barreiros, W. O. Böcher, Marcus Schuchmann, Martin F. Sprinzl, Tim Zimmermann, Anca Zimmermann, and Marcus A. Wörns

Subjects
Liver Cirrhosis, Male, medicine.medical_specialty, Cirrhosis, Hepatitis C virus, medicine.medical_treatment, Hepacivirus, Liver transplantation, Interferon alpha-2, medicine.disease_cause, Gastroenterology, Antiviral Agents, Polyethylene Glycols, Cohort Studies, chemistry.chemical_compound, Postoperative Complications, Pegylated interferon, Recurrence, Internal medicine, Ribavirin, medicine, Humans, Adverse effect, Aged, Transplantation, business.industry, Interferon-alpha, Alanine Transaminase, Hepatitis C, Middle Aged, medicine.disease, Recombinant Proteins, Liver Transplantation, Regimen, Treatment Outcome, chemistry, Immunology, RNA, Viral, Female, business, medicine.drug
Abstract

We evaluated the safety and efficacy of an escalating dose regimen of pegylated interferon alpha-2a (PEG-IFN(alpha-2a)) and ribavirin in the early phase of recurrent hepatitis C after orthotopic liver transplantation (OLT). In this prospective study, 26 patients transplanted for hepatitis C virus cirrhosis with recurrent hepatitis C were treated 3.4 +/- 3.6 months after OLT and compared with an untreated historical control. PEG-IFN(alpha-2a) was initiated as monotherapy, following stepwise dose escalation up to 180 mug/week and the addition of ribavirin up to 1200 mg/day or maximally tolerated doses for 48 weeks. In the intent-to-treat analysis, 38% showed an early virological response (EVR), 35% an end of treatment response (ETR) and 19% a sustained virological response (SVR). SVR was associated with EVR (P = 0.0001) and cumulative PEG-IFN(alpha-2a) dose (P = 0.04). There was no significant histological improvement compared with untreated patients. There were no treatment-related serious adverse events. Adverse events included leucopenia (77%) and thrombocytopenia (46%). Three patients discontinued therapy due to side effects, fourteen were nonresponders and four relapsers. Treatment with PEG-IFN(alpha-2a) and ribavirin in the acute phase of post-transplant recurrent hepatitis C yielded an EVR of 38% and an SVR of 19%. The combination was safe and well tolerated.

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40. Identification of Mycobacterium tuberculosis complex by polymerase chain reaction of Exact Tandem Repeat-D fragment from mycobacterial cultures

Author

C Coitinho, G Greif, C Robello, J van Ingen, and C Rivas

Subjects
ETR-D, M. tuberculosis complex, Identification, Diagnosis, Microbiology, QR1-502
Abstract

This study evaluated an in-house polymerase chain reaction (PCR) for rapid identification of the Mycobacterium tuberculosis complex (MTBC) using the MTBC-specific Exact Tandem Repeat D (ETR-D) as the amplification target. In a prospective study, 801 clinical isolates identified as MTBC and 15 nontuberculous mycobacteria were analyzed. Mycobacterial DNA was extracted from automated broth cultures or from egg-based media. The amplification of the ETR-D showed to a sensitivity of 99.6% and a specificity of 100% for the correct identification of MTBC; improved extractions protocols led to 100% sensitivity. The main utility of this technique is the simplicity, rapidity, low cost and accuracy.

Published
2012
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41. Closing the gap: prognostic and predictive biomarker validation for personalized care in a Latin American hormone-dependent breast cancer cohort.

Author

Alves da Quinta D, Rocha D, Retamales J, Giunta D, Artagaveytia N, Velazquez C, Daneri-Navarro A, Müller B, Abdelhay E, Bravo AI, Castro M, Rosales C, Alcoba E, Acosta Haab G, Carrizo F, Sorin I, Di Sibio A, Marques-Silveira M, Binato R, Caserta B, Greif G, Del Toro-Arreola A, Quintero-Ramos A, Gómez J, Podhajcer OL, Fernández EA, and Llera AS

Subjects
Humans, Female, Prognosis, Middle Aged, Cohort Studies, Adult, Latin America epidemiology, Neoplasm Recurrence, Local pathology, Aged, Receptor, ErbB-2 metabolism, Neoplasms, Hormone-Dependent drug therapy, Neoplasms, Hormone-Dependent pathology, Breast Neoplasms drug therapy, Breast Neoplasms pathology, Biomarkers, Tumor metabolism, Biomarkers, Tumor genetics, Precision Medicine methods
Abstract

Background: Several guidelines recommend the use of different classifiers to determine the risk of recurrence (ROR) and treatment decisions in patients with HR+HER2- breast cancer. However, data are still lacking for their usefulness in Latin American (LA) patients. Our aim was to evaluate the comparative prognostic and predictive performance of different ROR classifiers in a real-world LA cohort., Methods: The Molecular Profile of Breast Cancer Study (MPBCS) is an LA case-cohort study with 5-year follow-up. Stages I and II, clinically node-negative HR+HER2- patients (n = 340) who received adjuvant hormone therapy and/or chemotherapy, were analyzed. Time-dependent receiver-operator characteristic-area under the curve, univariate and multivariate Cox proportional hazards regression (CPHR) models were used to compare the prognostic performance of several risk biomarkers. Multivariate CPHR with interaction models tested the predictive ability of selected risk classifiers., Results: Within this cohort, transcriptomic-based classifiers such as the recurrence score (RS), EndoPredict (EP risk and EPClin), and PAM50-risk of recurrence scores (ROR-S and ROR-PC) presented better prognostic performances for node-negative patients (univariate C-index 0.61-0.68, adjusted C-index 0.77-0.80, adjusted hazard ratios [HR] between high and low risk: 4.06-9.97) than the traditional classifiers Ki67 and Nottingham Prognostic Index (univariate C-index 0.53-0.59, adjusted C-index 0.72-0.75, and adjusted HR 1.85-2.54). RS (and to some extent, EndoPredict) also showed predictive capacity for chemotherapy benefit in node-negative patients (interaction P = .0200 and .0510, respectively)., Conclusion: In summary, we could prove the clinical validity of most transcriptomic-based risk classifiers and their superiority over clinical and immunohistochemical-based methods in the heterogenous, real-world node-negative HR+HER2- MPBCS cohort., (© The Author(s) 2024. Published by Oxford University Press.)

Published
2024
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42. 6-(6-Methyl-1,2,4,5-Tetrazine-3-yl)-2,2'-Bipyridine: A N-Donor Ligand for the Separation of Lanthanides(III) and Actinides(III).

Author

Greif G, Sauerwein FS, Weßling P, Duckworth TM, Patzschke M, Gericke R, Sittel T, März J, Wilden A, Modolo G, Panak PJ, and Roesky PW

Abstract

Here, we report the synthesis of the 6-(6-methyl-1,2,4,5-tetrazine-3-yl)-2,2'-bipyridine (MTB) ligand that has been developed for lanthanide/actinide separation. A multimethod study of the complexation of MTB with trivalent actinide and lanthanide ions is presented. Single-crystal X-ray diffraction measurements reveal the formation of [Ce(MTB) 2 (NO 3 ) 3 ], [Pr(MTB)(NO 3 ) 3 H 2 O], and [Ln(MTB)(NO 3 ) 3 MeCN] (Ln = Nd, Sm, Eu, Gd). In addition, the complexation of Cm(III) with MTB in solution was studied by time-resolved laser fluorescence spectroscopy. The results show the formation of [Cm(MTB) 1-3 ] 3+ complexes, which occur in two different isomers. Quantum chemical calculations reveal an energy difference between these isomers of 12 kJ mol -1 , clarifying the initial observations made by time-resolved laser fluorescence spectroscopy (TRLFS). Furthermore, quantum theory of atoms in molecules (QTAIM) analysis of the Cm(III) and Ln(III) complexes was performed, indicating a stronger covalent contribution in the Cm-N interaction compared to the respective Ln-N interaction. These findings align well with extraction data showing a preferred extraction of Am and Cm over lanthanides (e.g., max. SF Am/Eu = 8.3) at nitric acid concentrations <0.1 mol L -1 HNO 3 .

Published
2024
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43. Improved serodiagnosis of Trypanosoma vivax infections in cattle reveals high infection rates in the livestock regions of Argentina.

Author

Bontempi IA, Arias DG, Castro GV, Peverengo LM, Díaz GF, Allassia M, Greif G, and Marcipar I

Subjects
Animals, Cattle, Argentina epidemiology, Antibodies, Protozoan blood, Sensitivity and Specificity, Trypanosomiasis, African diagnosis, Trypanosomiasis, African veterinary, Trypanosomiasis, African epidemiology, Livestock parasitology, Trypanosoma vivax immunology, Trypanosoma vivax genetics, Trypanosoma vivax isolation & purification, Serologic Tests methods, Serologic Tests veterinary, Cattle Diseases diagnosis, Cattle Diseases epidemiology, Cattle Diseases parasitology, Antigens, Protozoan immunology, Antigens, Protozoan genetics, Enzyme-Linked Immunosorbent Assay methods, Enzyme-Linked Immunosorbent Assay veterinary
Abstract

Bovine trypanosomosis, caused by Trypanosoma vivax, currently affects cattle and has a significant economic impact in sub-Saharan Africa and South America. The development of new diagnostic antigens is essential to improve and refine existing methods. Our study evaluated the efficacy of two recombinant antigens in detecting specific antibodies in cattle. These antigens are derivatives of an invariant surface glycoprotein (ISG) from T. vivax. A fraction of a previously described antigen (TvY486&#95;0045500), designated TvISGAf, from an African strain was evaluated, and a new ISG antigen from an American isolate, TvISGAm, was identified. The two antigens were expressed as fusion proteins in Escherichia coli: TvISGAf was fused to the MBP-His-tag, and TvISGAm was obtained as a His-tag fused protein. An ELISA evaluation was conducted using these antigens on 149 positive and 63 negative bovine samples. The diagnostic performance was enhanced by the use of a combination of both antigens (referred to as TvISG-based ELISA), achieving a sensitivity of 89.6% and specificity of 93.8%. Following the validation of the TvISG-based ELISA, the seroprevalence of T. vivax infection in 892 field samples from cattle in the central region of Argentina was determined. The mean seroprevalence of T. vivax was 53%, with variation ranging from 21% to 69% among the six departments studied. These results support the use of the TvISG ELISA as a valuable serological tool for the detection and monitoring of T. vivax infection in cattle. Furthermore, we report for the first time the seroprevalence of T. vivax in Argentina, which highlights the widespread endemic nature of the disease in the region. In order to effectively manage the increasing spread of T. vivax in the vast livestock production areas of South America, it is essential to implement consistent surveillance programs and to adopt preventive strategies., Competing Interests: I have read the journal’s policy and the authors of this manuscript have the following competing interests:We have filed a patent on the diagnostic antigens described in this paper., (Copyright: © 2024 Bontempi et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Published
2024
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44. Specific Mycobacterium tuberculosis Strain Circulating in Prison Revealed by Cost-Effective Amplicon Sequencing.

Author

Hurtado J, Bentancor MN, Laserra P, Coitinho C, and Greif G

Abstract

This scientific study focuses on tuberculosis (TB) within prison settings, where persons deprived of liberty (PDL) face significantly higher rates of the disease compared to the general population. The research employs the low-cost amplicon sequencing of Mycobacterium tuberculosis strains, aiming first to identify specific lineages and also to detect mutations associated with drug resistance. The method involves multiplex amplification, DNA extraction, and sequencing, providing valuable insights into TB dynamics and resistance-mutation profiles within the prison system at an affordable cost. The study identifies a characteristic lineage (X) circulating among PDLs in the penitentiary system in Uruguay, absent in the general population, and notes its prevalence at prison entry. No high-confidence mutations associated with drug resistance were found. The findings underscore the importance of molecular epidemiology in TB control, emphasizing the potential for intra-prison transmissions and the need for broader studies to understand strain dynamics.

Published
2024
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45. A comparative NMR-based metabolomics study of lung parenchyma of severe COVID-19 patients.

Author

Hurtado JI, López-Radcenco A, Izquierdo-García JL, Rodríguez F, Moyna G, Greif G, and Nin N

Abstract

COVID-19 was the most significant infectious-agent-related cause of death in the 2020-2021 period. On average, over 60% of those admitted to ICU facilities with this disease died across the globe. In severe cases, COVID-19 leads to respiratory and systemic compromise, including pneumonia-like symptoms, acute respiratory distress syndrome, and multiorgan failure. While the upper respiratory tract and lungs are the principal sites of infection and injury, most studies on the metabolic signatures in COVID-19 patients have been carried out on serum and plasma samples. In this report we attempt to characterize the metabolome of lung parenchyma extracts from fatal COVID-19 cases and compare them with that from other respiratory diseases. Our findings indicate that the metabolomic profiles from fatal COVID-19 and non-COVID-19 cases are markedly different, with the former being the result of increased lactate and amino acid metabolism, altered energy pathways, oxidative stress, and inflammatory response. Overall, these findings provide additional insights into the pathophysiology of COVID-19 that could lead to the development of targeted therapies for the treatment of severe cases of the disease, and further highlight the potential of metabolomic approaches in COVID-19 research., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2023 Hurtado, López-Radcenco, Izquierdo-García, Rodríguez, Moyna, Greif and Nin.)

Published
2023
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46. Canine leproid granuloma caused by a member of the Mycobacterium tuberculosis complex.

Author

Giannitti F, Dorsch MA, Fernández-Ciganda S, Rabaza A, Vázquez S, César D, Hurtado J, Greif G, Rabeneck DB, Bhatnagar J, and Ritter JM

Subjects
Humans, Dogs, Animals, RNA, Ribosomal, 16S genetics, Granuloma veterinary, Granuloma microbiology, Mycobacterium Infections microbiology, Mycobacterium Infections veterinary, Mycobacterium tuberculosis genetics, Tuberculosis veterinary, Tuberculosis diagnosis, Dermatitis veterinary
Abstract

Canine leproid granuloma (CLG) is a chronic form of dermatitis that has been associated with nontuberculous mycobacterial infections in Africa, Oceania, the Americas, and Europe. We report here a case of CLG associated with a member of the Mycobacterium tuberculosis complex (MTBC), which could be of public health concern. An 8-y-old pet dog developed 0.5-1-cm diameter, raised, firm, nonpruritic, alopecic, painless skin nodules on the external aspects of both pinnae. Histologic examination revealed severe pyogranulomatous dermatitis with intracellular Ziehl-Neelsen-positive bacilli that were immunoreactive by immunohistochemistry using a polyclonal primary antibody that recognizes tuberculous and nontuberculous Mycobacterium species. DNA extracted from formalin-fixed, paraffin-embedded skin sections was tested by a Mycobacterium genus-specific nested PCR assay targeting the 16S rRNA gene. BLAST sequence analysis of 214-bp and 178-bp amplicons showed 99.5% identity with members of the MTBC; however, the agent could not be identified at the species level. Although CLG has been associated traditionally with nontuberculous mycobacterial infections, the role of Mycobacterium spp. within the MTBC as a cause of this condition, and the role of dogs with CLG as possible sources of MTBC to other animals and humans, should not be disregarded given its zoonotic potential.

Published
2023
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47. Trypanosoma cruzi Isolates Naturally Adapted to Congenital Transmission Display a Unique Strategy of Transplacental Passage.

Author

Faral-Tello P, Greif G, Romero S, Cabrera A, Oviedo C, González T, Libisch G, Arévalo AP, Varela B, Verdes JM, Crispo M, Basmadjián Y, and Robello C

Abstract

Chagas disease is mainly transmitted by vertical transmission (VT) in nonendemic areas and in endemic areas where vector control programs have been successful. For the present study, we isolated natural Trypanosoma cruzi strains vertically transmitted through three generations and proceeded to study their molecular mechanism of VT using mice. No parasitemia was detected in immunocompetent mice, but the parasites were able to induce an immune response and colonize different organs. VT experiments revealed that infection with different strains did not affect mating, pregnancy, or resorption, but despite low parasitemia, VT strains reached the placenta and resulted in higher vertical transmission rates than strains of either moderate or high virulence. While the virulent strain modulated more than 2,500 placental genes, VT strains modulated 150, and only 29 genes are shared between them. VT strains downregulated genes associated with cell division and replication and upregulated immunomodulatory genes, leading to anti-inflammatory responses and tolerance. The virulent strain stimulated a strong proinflammatory immune response, and this molecular footprint correlated with histopathological analyses. We describe a unique placental response regarding the passage of T. cruzi VT isolates across the maternal-fetal interphase, challenging the current knowledge derived mainly from studies of laboratory-adapted or highly virulent strains. IMPORTANCE The main findings of this study are that we determined that there are Trypanosoma cruzi strains adapted to transplacental transmission and completely different from the commonly used laboratory reference strains. This implies a specific strategy for the vertical transmission of Chagas disease. It is impressive that the strains specialized for vertical transmission modify the gene expression of the placenta in a totally different way than the reference strains. In addition, we describe isolates of T. cruzi that cannot be transmitted transplacentally. Taken together, these results open up new insights into the molecular mechanisms of this insect vector-independent transmission form.

Published
2023
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48. Contrast Neurotoxicity and its Association with Symptomatic Intracranial Hemorrhage After Mechanical Thrombectomy.

Author

Lopez-Navarro ER, Delfs C, Jarre A, Sanio V, Greif G, Gutierrez J, Ringelstein EB, Meuth SG, Haensch CA, Ringelstein A, and Ringelstein M

Subjects
Humans, Retrospective Studies, Treatment Outcome, Intracranial Hemorrhages diagnostic imaging, Intracranial Hemorrhages etiology, Thrombectomy adverse effects, Thrombectomy methods, Glucose, Brain Ischemia etiology, Stroke etiology, Ischemic Stroke complications, Arterial Occlusive Diseases complications
Abstract

Purpose: Despite improved techniques and sophisticated postinterventional care, symptomatic intracranial hemorrhage (sICH) remains the most feared complication of mechanical thrombectomy (MT). Based on peri-interventional parameters, we aimed to discover which patients have a higher risk of sICH., Methods: From March 2017 until March 2020 consecutive patients with acute ischemic stroke (AIS) and confirmed large-vessel occlusion who underwent MT were analyzed retrospectively. Demographic, clinical, and radiological variables and parameters specific to thrombectomy were reviewed. A univariate analysis was performed and statistically significant variables were included in a logistic regression model to identify independent factors predictive of sICH., Results: A total of 236 patients with confirmed large-vessel occlusion were included and 22 (9.3%) had sICH. Univariate predictors of sICH included diabetes mellitus, glucose > 11.1 mmol/L, creatinine clearance (CrCl) ≤ 30 ml/min/1.73, ASPECTS indicating pretreatment infarct size, acute internal carotid artery (ICA) occlusion, stent implantation, tirofiban use, time from symptom onset to groin puncture > 4.5 h and high contrast medium consumption. In the adjusted analysis, ASPECTS < 6 (OR 3.673, p = 0.041), and amount of contrast injected ≥ 140 ml (OR 5.412, p = 0.003) were independent predictors of sICH, but not any more baseline glucose > 11.1 mmol/L (OR 1.467, p = 0.584), CrCl ≤ 30 ml/min/1.73 (OR 4.177, p = 0.069), acute ICA occlusion (OR 2.079, p = 0.181), stent implantation (OR 0.465, p = 0.512), tirofiban use (OR 5.164, p = 0.167), and time from onset-to-groin puncture (OR 1.453, p = 0.514)., Conclusion: The amount of contrast medium used is a modifiable factor associated with sICH. This association is novel and may be related to the neurotoxicity of the contrast medium disrupting the blood-brain barrier., (© 2022. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany.)

Published
2022
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49. Origin of New Lineages by Recombination and Mutation in Avian Infectious Bronchitis Virus from South America.

Author

Marandino A, Vagnozzi A, Tomás G, Techera C, Gerez R, Hernández M, Williman J, Realpe M, Greif G, Panzera Y, and Pérez R

Subjects
Animals, Chickens, Phylogeny, Mutation, Recombination, Genetic, Brazil, Infectious bronchitis virus genetics, Poultry Diseases, Coronavirus Infections
Abstract

The gammacoronavirus avian infectious bronchitis virus (IBV) is a highly contagious respiratory pathogen of primary economic importance to the global poultry industry. Two IBV lineages (GI-11 and GI-16) have been widely circulating for decades in South America. GI-11 is endemic to South America, and the GI-16 is globally distributed. We obtained full-length IBV genomes from Argentine and Uruguayan farms using Illumina sequencing. Genomes of the GI-11 and GI-16 lineages from Argentina and Uruguay differ in part of the spike coding region. The remaining genome regions are similar to the Chinese and Italian strains of the GI-16 lineage that emerged in Asia or Europe in the 1970s. Our findings support that the indigenous GI-11 strains recombine extensively with the invasive GI-16 strains. During the recombination process, GI-11 acquired most of the sequences of the GI-16, retaining the original S1 sequence. GI-11 strains with recombinant genomes are circulating forms that underwent further local evolution. The current IBV scenario in South America includes the GI-16 lineage, recombinant GI-11 strains sharing high similarity with GI-16 outside S1, and Brazilian GI-11 strains with a divergent genomic background. There is also sporadic recombinant in the GI-11 and GI-16 lineages among vaccine and field strains. Our findings exemplified the ability of IBV to generate emergent lineage by using the S gene in different genomic backgrounds. This unique example of recombinational microevolution underscores the genomic plasticity of IBV in South America.

Published
2022
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50. Genomic and proteomic analysis of Tausonia pullulans reveals a key role for a GH15 glucoamylase in starch hydrolysis.

Author

Trochine A, Bellora N, Nizovoy P, Duran R, Greif G, de García V, Batthyany C, Robello C, and Libkind D

Subjects
Basidiomycota, Chromatography, Liquid, Glucose, Hydrolysis, Starch, Tandem Mass Spectrometry, Glucan 1,4-alpha-Glucosidase genetics, Glucan 1,4-alpha-Glucosidase metabolism, Proteomics
Abstract

Basidiomycetous yeasts remain an almost unexplored source of enzymes with great potential in several industries. Tausonia pullulans (Tremellomycetes) is a psychrotolerant yeast with several extracellular enzymatic activities reported, although the responsible genes are not known. We performed the genomic sequencing, assembly and annotation of T. pullulans strain CRUB 1754 (Perito Moreno glacier, Argentina), a gene survey of carbohydrate-active enzymes (CAZymes), and analyzed its secretome by liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS) after growth in glucose (GLU) or starch (STA) as main carbon sources. T. pullulans has 7210 predicted genes, 3.6% being CAZymes. When compared to other Tremellomycetes, it contains a high number of CAZy domains, and in particular higher quantities of glucoamylases (GH15), pectinolytic enzymes (GH28) and lignocellulose decay enzymes (GH7). When the secretome of T. pullulans was analyzed experimentally after growth in starch or glucose, 98 proteins were identified. The 60% of total spectral counts belonged to GHs, oxidoreductases and to other CAZymes. A 65 kDa glucoamylase of family GH15 (TpGA1) showed the highest fold change (tenfold increase in starch). This enzyme contains a conserved active site and showed extensive N-glycosylation. This study increases the knowledge on the extracellular hydrolytic enzymes of basidiomycetous yeasts and, in particular, establishes T. pullulans as a potential source of carbohydrate-active enzymes. KEY POINTS: • Tausonia pullulans genome harbors a high number of genes coding for CAZymes. • Among CAZy domains/families, the glycoside hydrolases are the most abundant. • Secretome analysis in glucose or starch as main C sources identified 98 proteins. • A 65 kDa GH15 glucoamylase showed the highest fold increase upon culture in starch., (© 2022. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Published
2022
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