Complete amino acid sequence of the lentil trypsin-chymotrypsin inhibitor LCI-1.7 and a discussion of atypical binding sites of Bowman-Birk inhibitors

The complete primary structure of the lentil (Lens culinaris) trypsin-chymotrypsin inhibitor LCI-1.7 was determined by conventional methods in order to find relationships between partial sequences and the difference in action against human and bovine chymotrypsin. As other Bowman-Birk type inhibitors, LCI-1.7 contained 68 amino acid residues, seven disulfide bridges, and two reactive sites, Arg16-Ser17 for trypsin and Tyr42-Ser43 for chymotrypsin. Evaluation of sequence homologies showed that it belonged to the group III Bowman-Birk inhibitors. The atypical additional binding site of LCI-1.7 for human chymotrypsin was discussed and compared with such binding sites of two other Bowman-Birk inhibitors, the Bowman-Birk soybean proteinase inhibitor BBI, and the lima bean proteinase inhibitor LBI I, for human and bovine trypsin and chymotrypsin. A concept to reduce the action of these inhibitors against human enzymes by genetic engineering was proposed.     

Comprehensive authentication of (E)-alpha(beta)-ionone from raspberries, using constant flow MDGC-C/P-IRMS and enantio-MDGC-MS

A new coupling system of GC-GC, connected via a Multi Column Switching Device MCS2 for measuring isotope ratios, is introduced. By means of several standard substances the precise and accurate measurement of isotopic values is proved. First applications concerning the authentication of raspberry aroma compounds are established. Consequently, the combination of constant flow multidimensional gas chromatography-combustion/pyrolysis-isotope ratio mass spectrometry (MDGC-C/P-IRMS) is applied to the authenticity assessment of (E)-alpha(beta)-ionone from six different raspberry cultivars. Furthermore, 12 commercially available raspberry products and samples of (E)-alpha(beta)-ionone, some declared to be natural, are investigated. delta(2)Eta(V)(-)(SMOW) and delta(13)C(V)(-)(PDB) values of (E)-alpha(beta)-ionone are determined, and characteristic authenticity ranges were concluded from raspberries by correlation of both delta(2)Eta(V)(-)(SMOW) and delta(13)C( V)(-)(PDB) values. The results are correlated with the determination of enantiomeric purities of (E)-alpha-ionone, using stir bar sorptive extraction enantio-multidimensional gas chromatography mass spectrometry (SBSE-enantio-MDGC-MS).     

Analysis of volatile compounds and triglycerides of seed oils extracted from different poppy varieties (Papaver somniferum L.)

Poppy seed oil (Oleum Papaveris Seminis) is used for culinary and pharmaceutical purposes, as well as for making soaps, paints, and varnishes. Astonishingly, hardly anything was yet known about the volatile compounds of this promising comestible. Likewise, there are no current published data about the triglyceride (TAG) composition of poppy seed oils available. In this investigation solid-phase microextraction (SPME) with DVB/Carboxen/PDMS Stable-Flex fiber was applied to the study of volatile compounds of several seed oil samples from Papaver somniferum L. (Papaveraceae). 1-Pentanol (3.3-4.9%), 1-hexanal (10.9-30.9%), 1-hexanol (5.3-33.7%), 2-pentylfuran (7.2-10.0%), and caproic acid (2.9-11.5%) could be identified as the main volatile compounds in all examined poppy seed oil samples. Furthermore, the TAG composition of these oils was analyzed by MALDI-ReTOF- and ESI-IT-MS/MS. The predominant TAG components were found to be composed of linoleic, oleic, and palmitic acid, comprising approximately 70% of the oils. TAG patterns of the different poppy varieties were found to be very homogeneous, showing also no significant differences in terms of the applied pressing method of the plant seeds.     

Superovulationsinduktion bei Merinolandschafen nach einmaliger oder multipler FSH-Applikation

Inhalt: Zyklussynchronisierte Merinolandschafe (n=79) wurden mit FSH-P in einer Gesamtdosis von 25 mg oder 17 mg superovuliert. Während die Versuchsgruppe 1 das Gonadotropin in Form einer mehrfachen Applikation in 12 stündigen Abständen erhielt (Gruppe 1/25 mg und 1/17 mg), wurde die Gesamtdosis bei Versuchsgruppe 2 36 Stunden vor Gestagenentzug (Gruppe 2/25 mg und 2/17 mg) und bei Versuchsgruppe 3 bei Gestagenentzug (Gruppe 3/25 mg und 3/17 mg) verabreicht. Die Embryonengewinnung wurde nach intrauteriner Eksamung der Tiere am 7.-11. Zyklustag durchgefülut.
Die Anzahl der Corpora lutea betrug in den einzelnen Stimulationsgruppen jeweils 11,8 ± 4,6; 9,2 ± 55; 8,4 ± 6,4; 7,2 ± 7,2; 8,8 ± 6,5 und 4,2 ± 2,9 C.l., wobei die Reaktion der Gruppe 3/17 mg signifikant niedriger als die der Gruppe 1/25 mg (p < 0,001), 1/17 mg (p < 0,01), 2/25 mg (p< 0,05) und 3/25 mg (p < 0,05) war. Die anderen Gruppe unterscheiden sich nicht signifikant.
Sowohl die Anzahl der erzielten Ovulationen als auch die Zahl der gewonnenen Eizellen/Embryonen lassen darauf schließen, daß die einmalige FSH-Applikation in höherer Dosierung, insbesondere bei Synchronisationsende, eine sinnvolle Alternative zu den gebräuchlichen Superovulationsmethoden beim Schaf darstellt.     

Concentrations of Oestrone and Oestradiol-l 7β in the Peripheral Plasma of the Nanny Goat during the Oestrous Cycle, Gestation and Puerperium

The changes in the plasma levels of oestrone sulphate and oestradiol-17β during the oestrous cycle, gestation and puerperium in the goat are described. The oestrone sulphate concentrations remained fairly constant (250–350 pg/ml) throughout the oestrous cycle until day 20 when a sharp increase of the oestrone sulphate plasma levels occurred in pregnant goats which became significantly different at day 38 of gestation from nonpregnant values. Oestradiol-17β plasma levels were significantly lower at days 17–20 in pregnant than in nonpregnant does. Oestrone sulphate and oestradiol-17β concentrations rose until the 12th week of gestation and then declined to about 50% of the former ranges of concentrations before rising again to high values at weeks 17–20 of gestation. Increasing plasma levels of oestrone sulphate and oestradiol-17β were determined during the last ten days preceding parturition. The concentrations of oestrone sulphate returned to basal levels by the 2nd—4th day post partum whereas oestradiol-17β values reached base values 24 hours after parturition. Both oestrogen concentrations remained constant during the puerperium until day 51 post partum .     

Die Reifungs- und Befruchtungspotenz atretischer Schweineeizellen in vivo und in vitro1

Inhalt An 475 atretischen Schweineoozyten wurde das Kernreifungsvermögen und die Befruchtungsfähigkeit in vitro bzw. in vivo geprüft. Etwa die Hälfte der Eizellen befanden sich bereits intrafollikulär in fortgeschrittenen Stadien der Reifeteilungen (Prometaphase 1 – Metaphase 2). Nach der in vitro bzw. in vivo Kultivierung stieg der Anteil sekundärer Oozyten auf 25,3 bzw. 42,9 % an. Parthenogenetische Vorkernausbildungen und Fragmentationen traten bei etwa 10 % der Oozyten auf. Nach der Übertragung atretischer Eizellen in zuvor gedeckte Empfänger konnten zahlreiche pathologische Befruchtungsformen beobachtet werden, deren Entstehung im Hinblick auf die Kern- und Zytoplasmareifung in degenerierenden und extrakorporal gereiften Oozyten diskutiert wird. Contents Maturation and fertilization capability of atretic pig oocytes in vitro and in vivo 475 atretic porcine oocytes were tested in vitro and in vivo to determine the stage of nuclear maturation and the capacity for fertilization. In more than half of the degenerated follicular oocytes, advanced stages of maturation division (Prometaphase I – Metaphase II) were found. Following culture the number of secondary oocytes in vitro and in vivo rose to 25,3 % or 42,9 % respectively. In about 10 % of the oocytes parthenogenetic pronuclear development and fragmentation occurred. Following the transfer of the atretic oocytes to previously mated recipient sows, many pathological abnormalities of fertilization were found. The development of these abnormalities with respect to nuclear and cytoplasmatic maturation of degenerate and in vitro matured oocytes was discussed.     

Placebo-controlled blinded study of postoperative pain therapy with carprofen and levomethadone in dogs with fractures

It was the aim of this placebo-controlled study to evaluate the analgesic efficacy of the NASAID carprofen and the pure m-agonist levomthadone over a five-day postoperative evaluation period in dogs with fractures of the humerus or the femur (n = 30). Pain and sedation evaluation was carried out with a visual analogues system (VAS) and with the aid of a numerical estimation scale(NRS). The degree of lameness, the pain treshhold, the glucose and cortisol concentration curves as well as the respiration and heart rate and the systolic blood pressure were used as further pain indicators and to identify drug side effects. The levomethadon group displayed the lowest degree of pain on postoperative examination on the first day. On days 2 to 5, the carprofen group showed the lowest degree of pain in comparison to the placebo group. The levomethadon- and the carprofen group showed no statistically proven differences from day 2 on. Due to great variations in the pain scores and comparatively high median pain score especially on the first day of this study, the efficacy of all analgesics evaluated here must be regarded as insufficient in many cases. Only the parameter nociceptive pain treshhold showed a little, the degree of lameness, the glucose and cortisol levels showed no close correlation to the VAS and NRS pain scores and were therefore of little usefulness as postoperative pain indicators. No relevant clinical side effects caused by the used analgesics were detected in the kidney, the liver, the gastrointestinal tract and the circulatory system in this study. Rather, traumatically induced elevation of enzyme levels improves or normalised until the 5th day of the study. In addition, no negative effect on wound healing was noted, especially for carprofen. Therefore, the evaluated analgesics seems to be adequate for postoperative pain therapy also in fracture patients (trauma patients). However, the efficacy of all analgesics evaluated here must be regarded as insufficient in many cases.     

The use of sarmazenil in the treatment of a moxidectin intoxication in a foal

A13-day-old Arabian Thoroughbred filly weighing 40 kg (88 1b) was presented to the University of Zurich Equine Clinic with a history of depression after deworming with moxidectin at a dose of 2 mg/kg (recommended dose 0.4 mg/kg body weight)^a the day before admission. The foal was found recumbent 12 hours after drug administration and was in an unconscious state 6 hours later     

Characterisation and quantification of equine interferon gamma

Interferon-gamma (IFN-gamma) is a key cytokine in cell-mediated immunity. To measure IFN-gamma production of equine lymphocytes (eqIFN-gamma), we developed a quantitative ELISA. Monoclonal antibodies (mAb) were produced against bacterially derived eqIFN-gamma. The mAbs recognised recombinant and lymphocyte-derived eqIFN-gamma in ELISA, Western blotting, as well as flow cytometric and microscopic analysis. In contrast to bacterially derived material, mammalian and insect cell-derived eqIFN-gamma was biologically active but could be neutralised by one of the monoclonal antibodies. Unexpectedly, glycosylation seemed to be required for antiviral activity of eqIFN-gamma.     

Evaluation of a canine cranial cruciate ligament repair system for use in equine laryngoplasty

OBJECTIVE: To report the use of a nylon suture system (Canine Cranial Cruciate Ligament Repair System; Securos Inc Veterinary Orthopedics) as a prosthesis for equine laryngoplasty. STUDY DESIGN: Experimental and prospective clinical study. ANIMALS: Cadaver specimens (n = 5) and 7 horses with left laryngeal hemiplegia. METHODS: A commercially available monofilament nylon suture system was implanted as a laryngeal prosthesis. Arytenoid cartilage abduction was achieved with a tensioning device applied to the suture prosthesis during transnasal endoscopic observation. Suture fixation was achieved with crimping clamps and a crimping device. RESULTS: The nylon suture system was suitable as a laryngeal prosthesis for arytenoid cartilage abduction. The ratchet mechanism of the tensioning device facilitated abduction of the arytenoid cartilage and suture fixation was achieved by the crimped clamp without any loss of tension. Postoperatively, there was a slight loss of tension in 4 horses and complete loss of tension in 1 horse because of cartilage failure. After convalescence, none of the horses had abnormal respiratory noise, exercise intolerance or cough. CONCLUSIONS: A nylon suture system designed for canine cranial cruciate ligament repair was used successfully as a laryngeal prosthesis and facilitated control of the degree of arytenoid cartilage abduction during laryngoplasty. CLINICAL RELEVANCE: For improved control of the degree of arytenoid cartilage abduction during laryngoplasty, use of a nylon suture system with metal crimps should be considered.