M Gemeiner

Publications (41)76.15 Total impact

• Article: Katecholamin‐ und Cortisolkonzentrationen im Plasma von Rindern um die Geburt

  W.-D. Rausch, A. Hofer, M. Gemeiner, E. Möstl
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  ABSTRACT: Catecholamine and Cortisol Concentrations in Plasma from Cows at Term: Cows with spontaneous parturition (n = 8) and cows with flumethasone (n = 3) or prostaglandin (n = 3) induced parturition were used in this study. Catecholamine concentrations from cows in the periparturient period were measured in order to evaluate the stress of birth on the dam and the calf. The plasma content of both norepinephrine and epinephrine in late pregnancy was not different from control values in the 3rd to 6th month of pregnancy (norepinephrine 3.27 ± 0.99 nmol/l plasma and epinephrine 0.98 ± 0.45 nmol/l plasma), though both parameters appear elevated in the pregnant group. Norepinephrine concentrations rose to a value of 4.50 nmol/l immediately after birth, whereas epinephrine levels did not show an increase post partum. In two neonates norepinephrine and epinephrine values were 2–3 times higher than in the dams and decreased to the values of the dams within 48 hours.Synthetic glucocorticoids can mimic the increase of Cortisol, which leads to birth. Cortisol concentrations in the cows remained unaffected by induction of parturition by prostaglandins. Newborn calves showed values of about 330 nmol/l, which also declined within 48 hours to values similar to those measured in the dam (9–28 nmol/l).As stress and exogenous glucocorticoids may affect antibody production, the IgG-content of the colostrum was examined. Prostaglandin induction of birth did not alter the IgG-content of colostrum, but flumethasone produced a decrease of about 14.5 %. It is therefore concluded that pre-term birth induction does not produce negative effects on the supply of IgG.ZusammenfassungIn dieser Studie wurden Kühe mit physiologischer Geburt (n = 8) sowie Tiere mit Prostaglandin- (n = 3) und Flumethason-induzierter Geburt (n = 3) verwendet. Katecholaminkonzentrationen im Plasma von Kühen um die Geburt wurden gemessen, um anhand dieser Substanzen das Streßgeschehen zu untersuchen, dem Kuh und Kalb während des Geburtsvorganges ausgesetzt sind. Der Plasmagehalt von Noradrenalin und Adrenalin im Endstadium der Trächtigkeit war von Vergleichswerten von Muttertieren im 3. bis 6. Graviditätsmonat nicht signifikant verschieden (Noradrenalin 3,27 ± 0,99 nmol/l und Adrenalin 0,98 ± 0,45 nmol/l Plasma). Die Noradrenlinkonzentrationen des Plasmas stiegen unmittelbar nach der Geburt auf Werte von ca. 4,50 nmol/l an, die Adrenalinwerte blieben unverändert. Bei zwei neugeborenen Kälbern waren die Noradrenalin- und Adrenalinwerte gegenüber den Muttertieren etwa 2–3 fach erhöht. Innerhalb von 48 Stunden nach der Geburt sanken die Hormonkonzentrationen auf die Werte der Muttertiere ab.Synthetische Glukokortikoide können den Cortisolanstieg, der zur Geburt führt, nachahmen. Die Cortisolwerte waren bei Muttertieren mit Prostaglandin-induzierter Geburt, verglichen mit spontan gebärenden Tieren, nicht unterschiedlich. Neugeborene Kälber zeigten Werte um 330 nmol/l. Diese Werte sanken innerhalb von 48 Stunden nach der Geburt gleichfalls in den Bereich der Konzentrationen der Muttertiere (9–28 nmol/l) ab.Da Streß und exogene Glukokortikoide die Antikörperproduktion hemmen können, wurde der IgG-Gehalt des Kolostrums untersucht. Die Prostaglandininduktion hatte keinen Einfluß auf den IgG-Gehalt und auch nach Flumethasonverabreichung war die Antikörperkonzentration des Kolostrums nur um 14,5 % niedriger als in der Kontrollgruppe.Es ist daher anzunehmen, daß sich auch eine frühzeitige Gebunsinduktion nicht nachteilig auf die Immunglobulinversorgung der Kälber auswirkt.
  Journal of Veterinary Medicine Series A 05/2010; 36(1‐10):218 - 224. · 0.93 Impact Factor
• Article: Heparin and EDTA as anticoagulant differentially affect cytokine mRNA level of cultured porcine blood cells.

  J C Duvigneau, W Sipos, R T Hartl, M Bayer, R Moldzio, L Stevenson, B Adair, M Gemeiner
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  ABSTRACT: Cytokine mRNA expression profiles serve to characterize immune cell activation in different test systems. Both, diluted whole blood and isolated PBMC are widely applied for these studies. Comprehensive data regarding the suitability of different anticoagulants for profiling cytokine expression are not available for the pig. Therefore the aim of this study was to compare the effect of two commonly used anticoagulants (heparin and EDTA) on the cytokine expression pattern of porcine blood cells. IL-1alpha, IL-2, IL-4, IL-6, IL-10 and IFN-gamma mRNA levels were detected ex-vivo and upon in-vitro stimulation in diluted porcine whole blood and isolated PBMC by real-time PCR. The cells were stimulated with ConA or LPS, known to act on different target cells and implying different signalling pathways. Additionally the integrity of the isolated RNA was investigated. Ex-vivo cytokine expression pattern of fresh whole blood were not affected by the investigated anticoagulants. In contrast, stimulation of cultured diluted whole blood or PBMC resulted in significant differences depending on the applied anticoagulant. Using EDTA we found a significantly decreased capacity of whole blood to express cytokines. However, isolated PBMC from EDTA anticoagulated blood showed a higher cytokine expression capacity than PBMC from heparinized blood. Comparing diluted whole blood and PBMC we found that cultured porcine whole blood responded better to bacterial products than isolated PBMC, probably because sufficient auxiliary plasma derived factors such as LPS-binding protein, are present. However, isolated PBMC showed a higher T-cell response than diluted whole blood. In conclusion, our findings underline that each application demands a specific assay system.
  Journal of Immunological Methods 08/2007; 324(1-2):38-47. · 2.20 Impact Factor
• Source

  Available from: René van den Hoven

  Article: Clenbuterol affects the expression of messenger RNA for interleukin 10 in peripheral leukocytes from horses challenged intrabronchially with lipopolysaccharides.

  R van den Hoven, J C Duvigneau, R T Hartl, M Gemeiner
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  ABSTRACT: On four occasions, four horses with heaves and four horses with small airway inflammatory diseases inhaled 0.9% saline based aerosol mixtures with or without lipopolysaccharides (LPS). Prior to the first saline and LPS inhalation, horses were untreated, while three and a half days prior to the third and forth inhalation horses had received 0.8 microg/kg clenbuterol intravenously twice daily. The messenger RNA (mRNA) expression of tumour necrosis factor-alpha (TNF-alpha), interleukin (IL)-1beta, IL-4, IL-6, IL-8, IL-10 and interferon- gamma (IFN- gamma) was investigated by RT-PCR, all of which were expressed in the white blood cells of samples collected. Inhalation of LPS only changed the cytokine expression profile of IL-10, IL-4 and TNF-alpha mRNA which were higher after challenge with LPS. However in those horses that were treated with clenbuterol the LPS-induced IL-10 mRNA expression was shown to be suppressed. Further changes in IL-4 and TNF-alpha were not significant. Thus the results of this study indicated that clenbuterol can modulate the expression of IL-10 mRNA in peripheral white blood cells in those horses with small airway diseases that have been exposed to LPS.
  Veterinary Research Communications 12/2006; 30(8):921-8. · 0.82 Impact Factor
• Article: Quantitative simultaneous multiplex real-time PCR for the detection of porcine cytokines.

  J C Duvigneau, R T Hartl, S Groiss, M Gemeiner
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  ABSTRACT: We have established an easy real-time PCR assay, which allows the precise quantification of changes in the expression level of 6 relevant porcine cytokines, and 3 housekeeping genes. This assay simultaneously detects 9 sequences by measuring 3 x 3 targets in a triplex-format. The mRNA of the lymphokines IL-2, IL-4, IL-10, and IFN-gamma, of the proinflammatory cytokines IL-1alpha and IL-6, and of the housekeeping genes are quantified using TaqMan-probes by means of standard dilution series on the iCycler iQ. The standard consists of equal aliquots of the experimental cDNAs under investigation. Simultaneously the most suitable combination of 3 out of the four housekeeping genes beta-actin, HPRT, GAPDH, and cyclophilin can be selected, and their averaged expression values constitute a normalisation factor. The raw data of all targets of interest is then calculated relative to this normalisation factor, making eventual changes of the relative expression level of the single housekeeping genes controllable and quantifiable. We have applied this assay to quantify changes in the cytokine mRNA levels of porcine stimulated with various concentrations of LPS and ConA, known to induce different cytokine expression patterns. We have shown, that even small differences in the expression level (less than 2-fold) can be precisely quantified, and reveal statistically significant changes, when using the normalisation factor. This assay will be useful for studying changes in the expression of relevant porcine cytokines and will help to further improve the investigation of immune responses in the pig.
  Journal of Immunological Methods 12/2005; 306(1-2):16-27. · 2.20 Impact Factor
• Article: Variations in the concentration of zinc in the blood of Icelandic horses.

  G Kolm, A Helsberg, M Gemeiner
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  ABSTRACT: The effect of factors including the horses' farm environment, their sex and age and whether they suffered from summer seasonal recurrent dermatitis (sweet itch) on the concentrations of zinc in the plasma, whole blood and blood cells of 104 Icelandic horses was investigated. Its concentration in plasma varied significantly between farms (P<0.01), but its concentration in blood and blood cells was not influenced by any of the variables. The concentration of zinc in the blood cells was 10.5 times greater than in plasma, but its concentration in plasma was not correlated with its concentration in whole blood or blood cells owing to the variability in the proportion of whole blood zinc present in plasma (relative plasma zinc), which ranged between 9 and 24 per cent. This variability was significantly influenced by a three-way interaction between farm, sex and sweet itch (P<0.05). Relative plasma zinc was positively correlated with absolute plasma zinc (r=0.78, P<0.001) and negatively correlated with whole blood and blood cellular zinc (r=-0.58, r=-0.71, P<0.001).
  The Veterinary record 11/2005; 157(18):549-51. · 1.25 Impact Factor
• Article: Redox options in two-dimensional electrophoresis.

  R Wait, S Begum, D Brambilla, A M Carabelli, F Conserva, A Rocco Guerini, I Eberini, R Ballerio, M Gemeiner, I Miller, E Gianazza
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  ABSTRACT: Two-dimensional electrophoresis is usually run on fully reduced samples. Under these conditions even covalently bound oligomers are dissociated and individual polypeptide chains may be fully unfolded by both, urea and SDS, which maximizes the number of resolved components and allows their pI and M(r) to be most accurately evaluated. However, various electrophoretic protocols for protein structure investigation require a combination of steps under varying redox conditions. We review here some of the applications of these procedures. We also present some original data about a few related samples -- serum from four species: Homo sapiens, Mus musculus, Rattus norvegicus, Bos taurus -- which we run under fully unreduced and fully reduced conditions as well as with reduction between first and second dimension. We demonstrate that in many cases the unreduced proteins migrate with a better resolution than reduced proteins, mostly in the crowded 'alpha-globulin' area of pI 4.5-6 and M(r) 50-70 kDa.
  Amino Acids 06/2005; 28(3):239-72. · 3.25 Impact Factor
• Article: Ligula intestinalis infection as a potential source of bias in the bioindication of endocrine disruption in the European chub Leuciscus cephalus.

  M Schabuss, M Gemeiner, A Gleiss, J W Lewis, I Miller, E Möstl, U Schober, W Tschulenk, I Walter, B Grillitsch
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  ABSTRACT: European chub Leuciscus cephalus collected from five localities in the lowland and subalpine regions of Austria were analysed for oestrogenic effects of endocrine-disrupting chemicals and the presence of the plerocercoid of the tapeworm Ligula intestinalis. Of 1494 chub analysed, only seven (six males, one female) were found to be infected with single, but large plerocercoids up to 15 cm in length. Ligula-infected fish showed comparatively immature gonads, as demonstrated by the gonadosomatic index and gamete developmental stages. Plasma levels of the egg precursor protein vitellogenin also showed concentrations ranging below the detection limit. The present results indicate that chub infected with L. intestinalis and exposed to exogenous oestrogenic compounds can result in reduced gonadal maturation and produce false oestrogen-positive diagnoses in male fish. For plasma vitellogenin levels, L. intestinalis infections can result in false oestrogen-negative diagnoses in male and female fish.
  Journal of Helminthology 04/2005; 79(1):91-4. · 1.38 Impact Factor
• Article: Effect of clenbuterol on the clearance of particles of charcoal (4 to 90 microm) from the uteri of mares.

  G Kolm, M Gemeiner, K Deichsel, S Budik, J Aurich, C Aurich
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  ABSTRACT: The failure of clearance mechanisms in the mare's uterus results in persistent inflammation and is considered a major cause of subfertility. Eighteen mares, of which three were susceptible to endometritis and four had been ovariectomised, underwent charcoal clearance testing to evaluate their clearance mechanisms. This consisted of installing 500 mg of charcoal (particle size 4 to 90 microm) added to 50 ml of phosphate-buffered saline (PBS) into the uterus. Forty-eight hours later the uterus was flushed out with 0.0012 per cent methylene blue in 50 ml of PBS for determination of the diluting factor by photospectrometry. Flush volume, pH, specific gravity and pellet size were all analysed. To investigate the effect of a beta2-adrenergic agonist on the ability of genitally healthy oestrous mares to eliminate a suspension of charcoal from the uterus, four genitally healthy mares with appropriate charcoal clearance were also subjected to clearance testing following intravenous administration of 0.8 microg/kg of clenbuterol every 12 hours and 1 microg/kg of clenbuterol every eight hours. All parameters were within their normal range following clenbuterol treatment. However, minimal but significant differences in pre- and post-treatment values regarding fluid volume and extinction rate were recorded.
  The Veterinary record 03/2005; 156(9):279-81. · 1.25 Impact Factor
• Article: Hypothyroid goitre in a ram: chemical analysis gives indirect evidence for a structurally altered type of ovine thyroglobulin.

  W Sipos, I Miller, M Fountoulakis, F Schmoll, M Patzl, I Schwendenwein, E Rapp, A Taxacher, M Gemeiner
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  ABSTRACT: The thyroglobulin of a ram of the East Friesian milk sheep breed suffering from goitre was investigated by physico- and immunochemical methods. The respective ram was the only animal amongst the other sheep of the flock, that exhibited severe goitre, additionally showing depressed behaviour. Results of the thyroid-stimulating hormone response test were indicative of hypothyroidism. The dysfunction of the thyroid gland could be treated by additional iodine supplementation quite successfully, although all sheep had been given iodinated cattle salt throughout the course of the history. Without reducing conditions sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) of isolated thyroglobulin molecules of the ram and control sheep did not reveal different band patterns, but under reducing conditions different band patterns were evident for the respective animals: the ram's thyroglobulin displayed two main bands, those of healthy reference sheep only one. Both bands reacted equally with anti-thyroglobulin antibodies, even with those produced by immunizing rabbits with single bands. The reduced single thyroglobulin band of healthy sheep corresponded to a truncated form of that molecule, whereas the additional main band of the ram was a more resistant, intact thyroglobulin subunit, as was shown by mass spectrometry. In conclusion, results of physico- and immunochemical investigations gave evidence of a modification of thyroglobulin with suspected different iodine binding properties in the ram. The latter finding may have clinical relevance in similar cases in other species, as it is an example of the impact that a minor change in a protein molecule may have on a complete metabolic pathway. Additionally, it could be shown, that in the ovine species the generally found single main band of thyroglobulin after reduction is a truncated form and not an intact subunit. This truncation seems to be induced in vitro by the reductive sample pretreatment prior to SDS-PAGE.
  Journal of Veterinary Medicine Series A 04/2004; 51(2):90-6. · 0.93 Impact Factor
• Article: Endocrine disrupters in the aquatic environment: the Austrian approach--ARCEM.

  W Bursch, M Fuerhacker, M Gemeiner, B Grillitsch, A Jungbauer, N Kreuzinger, E Moestl, S Scharf, E Schmid, S Skutan, I Walter
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  ABSTRACT: A consortium of Austrian scientists (ARCEM) carried out a multidisciplinary environmental study on Austrian surface and ground waters including chemical monitoring, bioindication, risk assessment and risk management for selected endocrine disrupters: 17beta-estradiol, estriol, estrone, 17alpha-ethinylestradiol, 4-nonylphenol, 4-nonylphenol ethoxylates (4-NP1EO, 4-NP2EO) and their degradation products, ocytlphenol, ocytlphenol ethoxylates (OP1EO, OP2EO) as well as bisphenol A. To obtain data representative for Austria, a material flow analysis served to select relevant compounds and water samples were collected monthly over one year at those sites routinely used in Austrian water quality control. The following results were obtained and conclusions drawn: 1. Chemical monitoring: As compared to other countries, relatively low levels of pollution with endocrine disrupters were detected. 2. Bioindication: In the surface waters under study, male fish showed significant signs of feminization and demasculinization (increased production of the egg-yolk protein and histological changes of the gonads. 3. Risk assessment: For humans, exposure via either drinking water abstraction (ground water) or fish consumption was considered. The exposure levels of the compounds under study were below those considered to result in human health risks. Likewise, for bisphenol A and octylphenols, there was no indication for risk posed upon the aquatic environment (fish). However, nonylphenol or 17alpha-ethinylestradiol exposure along with results of bioindication (2) suggest a borderline estrogenic activity in a considerable number of surface waters. Consequently the emissions of these substances into the surface waters affected have to be reduced. 4. Risk management: Waste water treatment experiments revealed a positive correlation between the removal rate of endocrine disrupters from the waste water and the sludge retention time in the treatment plants. These substances are removed to a higher extend at low loaded plants designed for nutrient removal than at plants that remove carbon and/or employ nitrification only. As to drinking water treatment, chlorine dioxide and ozone were found to eliminate all investigated substances, except nonylphenol ethoxylates. (For the complete study see: www.arcem.at)
  Water Science & Technology 02/2004; 50(5):293-300. · 1.12 Impact Factor
• Article: Delay in processing porcine whole blood affects cytokine expression.

  J C Duvigneau, R T Hartl, M Teinfalt, M Gemeiner
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  ABSTRACT: Quantification of cytokine messenger ribonucleic acid (mRNA) in blood samples has become an important tool in the investigation of immune cell activation in a variety of clinical settings. It has been shown that the method of sample collection and processing influences the levels of several cytokine mRNAs. Therefore, it is generally accepted that blood samples for analysis of cytokine expression be processed as soon as possible and under standardised conditions. Since immediate sample processing is not always possible, we investigated the effect of different storage conditions (room temperature (Rt) and 4 degrees C) and storage times (1, 2, 4, 6 and 24 h) on the mRNA level of different cytokines (IL-1alpha, IL-2, IL-6, IL-8, IL-10, IFN-gamma), as well as the IL-2 receptor (IL-2R) in porcine whole blood samples (n=8). Quantification of cytokine expression was performed using simultaneous reverse transcription PCR (RT-PCR) combined with the expression of the housekeeping gene glyceraldehyde-3-phosphate dehydrogenase (GAPDH) as a reference. Our data demonstrate that delays in sample processing longer than 1 h result in significant changes of the mRNA levels of individual cytokines. Expression of the monokines IL-1alpha, IL-6 and IL-10 were increased by storage at both room temperature and 4 degrees C. Expression of IL-8 was increased only in the samples stored at room temperature, and expression of IFN-gamma was raised exclusively in the samples stored at 4 degrees C. We conclude that porcine blood samples should be processed within 2 h to prevent undesired stimulatory effects on the cytokine expression pattern. However, if only selected cytokines are investigated, the undesired effects of prolonged storage can be selectively suppressed by choosing the appropriate temperature of sample storage.
  Journal of Immunological Methods 02/2003; 272(1-2):11-21. · 2.20 Impact Factor
• Article: Zinc and copper plasma levels in Icelandic horses with Culicoides hypersensitivity.

  G Stark, B Schneider, M Gemeiner
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  ABSTRACT: Zinc concentration has been shown to have a potent immunomodulatory capacity, particularly influencing T helper cell organisation and cytokine secretion. Culicoides hypersensitivity (CHS) in horses resembles the early and late phase of type I hypersensitive reactions in man, characterised by a shift from T helper cell subtype 1 to T helper cell subtype 2 cytokine profile. In this pilot study, zinc and copper levels were measured in the plasma of 48 CHS-affected and 56 healthy Icelandic horses age 4-25 years (mean approximately 11 years) kept on 7 farms. Affected horses were divided into 3 groups according to the severity of disease. Time of blood collection and feeding management was constant. No differences in zinc or copper plasma levels and plasma copper/zinc ratio were determined among CHS horses and controls by univariate analysis of variance. Therefore, the most significant influences on zinc and copper plasma levels were affected by the location of housing. However, Spearman correlation showed a negative coefficient between the plasma zinc concentration and the severity of CHS (r = -0.31). Due to a probability value of P = 0.002 the null hypothesis r = 0 is rejected, although only 9% of the total variation of plasma zinc is presently explained by its relationship to CHS. In contrast, the Spearman correlation coefficient between plasma copper levels and severity of CHS was not significant (r = -0.14; P = 0.16). The minor deviations in plasma zinc concentrations in association with the severity of CHS may be real or due to neurohumoral or cytokine-mediated mechanisms, but appear too minimal to be relevant.
  Equine Veterinary Journal 10/2001; 33(5):506-9. · 1.46 Impact Factor
• Source

  Available from: unimi.it

  Article: Proteins of rat serum, urine, and cerebrospinal fluid: VI. Further protein identifications and interstrain comparison.

  R Wait, E Gianazza, I Eberini, L Sironi, M J Dunn, M Gemeiner, I Miller
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  ABSTRACT: We have investigated the biological fluids--serum, cerebrospinal fluid, and urine--of three strains of rats; the present data extend our database (also available on-line) and may be of interest for pharmacological and toxicological investigation. Specifically, we have defined reference maps of the major protein components in cerebrospinal fluid and urine. Compartment-specific isoforms were recognized for transferrin and transthyretin. Mass spectrometric data established the cleavage site of the signal peptide and identified the N-terminal blocking group of prostaglandin D synthase from rat cerebrospinal fluid. A previously undescribed member of the family of low molecular mass rat urinary proteins was characterized as containing a sequence similar, but not identical, to the N-terminal region of rat urinary protein-2 (RUP-2), and divergent from RUP-1.
  Electrophoresis 09/2001; 22(14):3043-52. · 3.30 Impact Factor
• Source

  Available from: Uliano Guerrini

  Article: Acute-phase proteins before cerebral ischemia in stroke-prone rats: identification by proteomics.

  L Sironi, E Tremoli, I Miller, U Guerrini, A M Calvio, I Eberini, M Gemeiner, M Asdente, R Paoletti, E Gianazza
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  ABSTRACT: A high degree of proteinuria has been reported in stroke-prone spontaneously hypertensive rats (SHRSP). We studied the effect of salt loading on the detailed protein pattern of serum and urine in 3 rat strains: Wistar-Kyoto, spontaneously hypertensive rats, and SHRSP, an inbred animal model for a complex form of cerebrovascular disorder resembling the human disease. Rats were given a permissive diet and received 1% NaCl in drinking water. The protein pattern in body fluids was assessed over time by 2-dimensional electrophoretic analysis. Brain alterations were monitored by MRI and histology. Several proteins were excreted in urine after weeks of treatment and in advance of stroke: transferrin, hemopexin, albumin, alpha(2)-HS-glycoprotein, kallikrein-binding protein, alpha(1)-antitrypsin, Gc-globulin, and transthyretin. Markers of an inflammatory response, including very high levels of thiostatin, were detected in the serum of SHRSP at least 4 weeks before a stroke occurred. In SHRSP subjected to salt loading, an atypical inflammatory condition and widespread alterations of vascular permeability developed before the appearance of anomalous features in the brain detected by MRI. Urinary concentrations of each of the excreted serum proteins correlated positively with time before stroke occurred.
  Stroke 04/2001; 32(3):753-60. · 5.73 Impact Factor
• Article: Proteins of rat serum V: adjuvant arthritis and its modulation by nonsteroidal anti-inflammatory drugs.

  I Ebrini, D Agnello, I Miller, P Villa, M Fratelli, P Ghezzi, M Gemeiner, J Chan, R Aebersold, E Gianazza
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  ABSTRACT: The effect of adjuvant arthritis (AA) on the pattern of rat serum proteins includes the upregulation of haptoglobin, orosomucoid, alpha2-macroglobulin, serine protease inhibitor-3, thiostatin, alpha1-antitrypsin, C-reactive protein, and the downregulation of kallikrein-binding protein, alpha1-inhibitor III, apolipoprotein A-I, alpha2-HS-glycoprotein, albumin, apolipoprotein A-IV, transthyretin and transferrin. Minor changes (+/- 20%) are observed for Gc-globulin, ceruloplasmin, and alpha1-macroglobulin. AA thus grossly resembles the acute inflammatory response elicited by the injection of turpentine, although the changes in the levels of negative acute-phase proteins (APP) are smaller in acute inflammation. Indomethacine and ibuprofen inhibit the effects of arthritis on the synthesis of rat serum proteins in different ways: The former is, on average, three times as effective as the latter. Each drug interferes differently with different proteins. In animals without AA, both nonsteroidal anti-inflammatory drugs (NSAID) mimic the inflammatory pattern to a certain extent, with more effect on the negative than on the positive APPs. Overall, the shifts in serum protein levels parallel changes in inflammatory parameters such as joint swelling and serum interleukin-6 (IL-6) activity. Protein quantitation after two-dimensional electrophoresis (2-DE) reveals some effects of the drugs per se which escape detection by other routine tests.
  Electrophoresis 07/2000; 21(11):2170-9. · 3.30 Impact Factor
• Article: A web site for the rat serum protein study group.

  I Eberini, I Miller, M Gemeiner, P Haynes, R Aebersold, L Puglisi, C R Sirtori, E Gianazza
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  ABSTRACT: We describe a site http://users.unimi.it/-ratserum/homeframed.ht ml with clickable maps of serum proteins of control and inflamed rats as well as quantitative data on the expression of such serum proteins under varying physiological and experimental conditions. This information enhances the value of minimally invasive techniques, thus reducing the number of animals to be treated, and eventually sacrificed, in pharmacological/toxicological research projects.
  Electrophoresis 12/1999; 20(18):3599-602. · 3.30 Impact Factor
• Article: An electrophoretic study on interactions of albumins of different species with immobilized Cibacron Blue F3G A.

  I Miller, M Gemeiner
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  ABSTRACT: Albumins of different species, varying in electrophoretic mobility, were compared in their interaction with the dye Cibacron Blue F3G A. Immobilized by coupling to a high molecular weight dextran ("blue dextran"), the dye was used as a ligand in affinity electrophoresis in different setups. One-dimensional electrophoresis with blue dextran entrapped in an intermediate gel and two-dimensional applications with transverse gradients (affinity titration curves, zonal electrophoresis in linear ligand gradients) were performed. Compared to the human homologue, animal albumins albumins display more complex patterns and interaction profiles, depending on pH and ionic strength of the buffers. Results may differ considerably from those obtained by affinity chromatography, illustrating the additional screening potential of the electrophoretic methods. Comparison of different samples under the influence of ligand competition, reducing conditions, or denaturing agents gives supplementary information on conformational behavior of the proteins.
  Electrophoresis 11/1998; 19(14):2506-14. · 3.30 Impact Factor
• Article: Proteins of rat serum: I. Establishing a reference two-dimensional electrophoresis map by immunodetection and microbore high performance liquid chromatography-electrospray mass spectrometry.

  P Haynes, I Miller, R Aebersold, M Gemeiner, I Eberini, M R Lovati, C Manzoni, M Vignati, E Gianazza
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  ABSTRACT: In the present investigation, we have identified 56 major spots, or spot rows, corresponding to 22 proteins, in the 2-DE pattern of adult male rats. This was done mainly by applying two complementary techniques, namely immunoblotting and high performance liquid chromatography-mass spectrometry (HPLC-MS) peptide mapping. Glycoproteins were characterized by affinity blotting with six lectins. We have also detailed how rat serum differs from human serum in two main respects: (i) relative abundance of individual proteins, which amounts in some cases to a complete absence in either sample, and (ii) varying molecular parameters for homologous proteins. It was thus possible to establish a first-generation reference map of rat serum proteins, which can be accessed through http://weber.u.washington.edu/ruedilab/aebersold++ +.html. We hope the present database will be a useful reference for the evaluation of changes in serum protein distribution in the course of pharmacological and toxicological studies. The recognition of species-specific proteins appears of special relevance in this respect.
  Electrophoresis 07/1998; 19(8-9):1484-92. · 3.30 Impact Factor
• Article: Proteins of rat serum: II. Influence of some biological parameters of the two-dimensional electrophoresis pattern.

  I Miller, P Haynes, M Gemeiner, R Aebersold, C Manzoni, M R Lovati, M Vignati, I Eberini, E Gianazza
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  ABSTRACT: This report complements the database already detailed for serum proteins of healthy adult male rats (P. Haynes et al., Electrophoresis 1998, 19, 1484-1492). The influence on the two-dimensional electrophoresis (2-DE) pattern of certain physiological conditions (sex, age) was studied as well as of changes in thyroid metabolism. We have extended the information about the major components of rat serum by identifying the proteins typical for the response to acute inflammation. Analyzing 27 spots, six proteins not found in control sera could be recognized; migration at overlapping or close positions with five already characterized proteins was observed for some. A compilation of all our rat data can be accessed through: http://weber.u.washington.edu/ruedilab/ aebersold.html.
  Electrophoresis 07/1998; 19(8-9):1493-500. · 3.30 Impact Factor
• Article: Two-dimensional electrophoresis for the study of blood/serum proteins of the otter, an endangered species.

  I Miller, A C Gutleb, M Gemeiner
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  ABSTRACT: Studies on serum/blood samples from European otters (Lutra lutra), a species under protection in most countries, were undertaken. Building of a two-dimensional electrophoresis (2-DE) protein identification map was started, identifying serum proteins by pattern comparison with cat or human serum protein maps and by immunoblotting using cross-reacting antibodies against the respective human serum proteins. The capture of endangered species is very restricted, thus making fresh samples almost unavailable; most investigations therefore had to be performed on samples collected from animals found dead. Patterns obtained from specimens of this source were compared to the established pattern of serum proteins and of blood or liver cells. Some of the major differences observed can be explained by contamination with cellular proteins and/or the onset of lytic processes generating breakdown products, as shown by preliminary in vitro experiments. Better knowledge of "normal" serum protein patterns and interfering products will facilitate interpretation of data collected from samples with a less defined history.
  Electrophoresis 08/1995; 16(7):1193-8. · 3.30 Impact Factor