Quantitative analysis and detection of chaptalization and watering down of wine using isotope ratio mass spectrometry

In early 1990's European Union has established new isotopic approach for detection of wine authenticity. In this article we setup the possibility of using new approach using new EIM - Module - IRMS (Ethanol Isotope Measurement - Module - Isotope Ratio Mass Spectrometry) instrumental technique and new analytical parameter delta D-n(delta H-2(n)) wine ethanol value, which represents delta H-2 value of non-exchangeable hydrogen stable isotope ratio in ethanol, with other isotopic species (delta O-18 values in wine water) to improve detection of illegal wine production practices such as addition of sugar and/or dilution with water. Total of 42 wine samples were analyzed. 10 wine samples (out of 42) were prepared from grapes by alcoholic fermentation and analyzed for delta D-n values of ethanol. 19 wine samples (out of 42) were collected from wine producers in Serbia plus 1 wine samples designated from United States was taken from Serbian market and analyzed for delta D-n values of ethanol, delta O-18 values in wine water and also delta C-13 values in wine ethanol. Furthermore 9 wine samples (out of 42) were taken from Hungarian market and analyzed for delta D-n values of ethanol, and also 3 wine samples (out of 42) were taken from Austrian market and analyzed for delta D-n values of ethanol. All experiments were done in 4 isotope laboratories located in US, Austria, Hungary and People's Republic of China. delta D-n values of ethanol were measured by using EIM Module connected to FlashHT 2000 pyrolizer (one laboratory - Imprint Analytics GmbH, Austria), while in other 3 laboratories (US - COIL - Cornell University, Stable Isotope Laboratory, Hungary - Isotoptech Ltd, Debrecen, Hungary and China - C.N.R.I.F.F.I. - China National Institute of Food and Fermentation Industries Limited) EIM Module was connected to TC/EA (High Temperature Conversion Elemental Analyzer). Peripherals in all laboratories were further interfaced with isotope ratio mass spectrometer. Furthermore delta O-18 values in wine water were measured by using Gas Bench II interfaced also with isotope ratio mass spectrometer (one laboratory - US). Obtained results from all 4 laboratories have shown that this new approach which uses delta D-n in wine ethanol is more effective in improving detection of illegal wine production practices (sugar enrichment and water dilution) and origin of ethanol, and also detecting the addition of corn or beet sugar, sugar syrup to wine, or dilution of grape must with water prior to alcoholic fermentation

Meningeal rupture caused by gunshot in a dog – a case report

Ozljede kralježnice, a time i kralježnične moždine, u pasa su najčešće izazvane vanjskim uzrocima, primjerice traumom u prometu, padom s visine, nastrijelom ili protruzijom intervetrebralnih diskova. Za otkrivanje višestrukih povreda i određivanje prognoze neophodno je obaviti neurološki pregled. U nekim slučajevima radiografija će biti dovoljna za postavljanje dijagnoze. Međutim, uz osnovno snimanje bez upotrebe kontrastnih sredstava, daljnje pretrage poput mijelografije, epidurografije, stres radiografije ili segmentalnog snimanja će pomoći u razumijevanju uzroka i ozbiljnosti ozljeda kralježnice. Općenito se tijek radioloških pretraga pacijenata s traumom kralježnice može podijeliti na pretrage u budnom stanju i pretrage u anesteziji. Jedna od mogućnosti nastanka traume kralježnice u ljudi i životinja jesu i strijelne ozljede. Vrlo malo kliničkih istraživanja bavi se ozljedama u području vrata i vratne kralježnice, a istraživanja usmjerena na isključivo strijelne ozljede u području vrata, u veterinarskoj medicini baziraju se na prikaze slučajeva ili serije prikaza, do 3 slučaja. Najopsežnije istraživanje o prijelomima vratnih kralježaka obuhvaća 56 slučajeva pasa, u razdoblju od 23 godine, u kojem je u samo 2 slučaja (3,5%) povreda uzrok bio nastrijel. U svim dosada poznatim istraživanjima prekida kontinuiteta mozgovnih ovojnica kod pasa i mačaka nije zabilježen pozitivan ishod liječenja, odnosno preživljavanje, neovisno o etiologiji povreda. Na Klinike Veterinarskog fakulteta zaprimljeno je štene, križanac, oko 5 mjeseci staro. Ležalo je na boku, pri svijesti, tetraplegično. U području vrata i skapularnog podučja vidljivo je nekoliko starijih rana, temeljito sumnjivih na prostrijelne rane. Neurološkim pregledom ustvrđeni su znatni neurološki deficiti. Rendgenskim snimanjem u standardnim projekcijama utvrđeno je otečenje mekih tkiva desne strane vrata i potvrđen prostrijel s desne na lijevu stranu vrata u području šestog (C6) i sedmog (C7) vratnog kralješka. Mijelografski je u visini C7 utvrđen izljev kontrastnog sredstva u okolna tkiva, naročito dorzalno i lateralno, u desno, što dokazuje prekid kontinuiteta spinalnih ovojnica. Nalaz odgovara prostrijelnoj rupturi moždanih ovojnica (tvrde spinalne ovojnice - durae mater i paučinaste ovojnice – arachnoideae) dorzalno u visini od C5 do C7. Zbog težine stanja psa i neizlječivosti životinja je eutanazirana.Spine and spinal cord injuries in dogs are most commonly caused by external trauma, such as vehicular trauma, high rise syndrome, gunshot or intervertebral disc extrusion. Neurological examination is required to diagnose multiple spine injuries and prognosis. In some cases, survey radiography is useful in proper diagnosing. However, myelography, stress radiography, and sectional radiography are of value in addition to routine non-contrast studies in order to completely understand the various causes and severity of cord injuries. The concept of spinal radiography can be conveniently divided in procedures performed on the conscious patient and those performed on the anesthetized patient. In human and veterinary medicine, spinal lesions can be caused by gunshots. There are only a few publications outlining clinical studies of the neck and cervical spine injuries. Furthermore, gunshot injuries in the cervical region have been described in several case reports, though with a maximum of three cases. Only one comprehensive study investigated 56 cases of cervical vertebral fractures in dogs over a 23- year period. This study described only 2 dogs (3.5%) with cervical spine fractures caused by gunshots. No evidence of full recovery or survival was found in any of the publications studding meningeal tear in dogs and cats, regardless of the etiology of the rupture. A mixed breed male puppy, about 5 months old, was admitted to the small animal clinic at the Faculty of Veterinary Medicine. He was conscious, in lateral recumbency, showing tetraplegia. Several chronic wounds were visible in the caudal cervical and scapular region with suspicion of a gunshot wounds. Neurological examination identified significant neurological deficits. X-rays findings included soft tissue swelling on the right side of the neck and gunshot from the right to the left side of the neck in the area of the sixth (C6) and seventh (C7) cervical vertebrae. Myelography finding showed contrast media leakage in the area of C7, especially dorsally and laterally on the right, which confirmed the meningeal tear. This finding was related to the gunshot-caused rupture of the dural and arachnoideal meninges dorsally in the area C5 to C7. Due to the dubious prognosis and lack of any possibility for treatment, the dog was euthanized

Calcium phosphate ceramics combined with rhBMP6 within autologous blood coagulum promote posterolateral lumbar fusion in sheep

Posterolateral spinal fusion (PLF) is a procedure used for the treatment of degenerative spine disease. In this study we evaluated Osteogrow-C, a novel osteoinductive device comprised of recombinant human Bone morphogenetic protein 6 (rhBMP6) dispersed in autologous blood coagulum with synthetic ceramic particles, in the sheep PLF model. Osteogrow-C implants containing 74-420 or 1000-1700 µm ceramic particles (TCP/HA 80/20) were implanted between L4-L5 transverse processes in sheep (Ovis Aries, Merinolaandschaf breed). In the first experiment (n = 9 sheep; rhBMP6 dose 800 µg) the follow-up period was 27 weeks while in the second experiment (n = 12 sheep; rhBMP6 dose 500 µg) spinal fusion was assessed by in vivo CT after 9 weeks and at the end of the experiment after 14 (n = 6 sheep) and 40 (n = 6 sheep) weeks. Methods of evaluation included microCT, histological analyses and biomechanical testing. Osteogrow-C implants containing both 74-420 and 1000-1700 µm ceramic particles induced radiographic solid fusion 9 weeks following implantation. Ex-vivo microCT and histological analyses revealed complete osseointegration of newly formed bone with adjacent transverse processes. Biomechanical testing confirmed that fusion between transverse processes was complete and successful. Osteogrow-C implants induced spinal fusion in sheep PLF model and therefore represent a novel therapeutic solution for patients with degenerative disc disease

A novel autologous bone graft substitute containing rhBMP6 in autologous blood coagulum with synthetic ceramics for reconstruction of a large humerus segmental gunshot defect in a dog: The first veterinary patient to receive a novel osteoinductive therapy

Background: Management of large segmental defects is one of the most challenging issues in bone repair biology. Autologous bone graft substitute (ABGS) containing rhBMP6 within autologous blood coagulum (ABC) with synthetic ceramics is a novel biocompatible therapeutic solution for bone regeneration. ----- Case presentation: A 2-year old dog was brought to the veterinary clinics due to pain and bleeding from the right front leg after being unintendedly hit by a gunshot. Radiological examination revealed a large, 3 cm long multisegmental defect of the humerus on the right front leg with a loss of anatomical structure in the distal portion of the bone. The defect was treated surgically and an external fixator was inserted to ensure immobilization. Complete lack of bone formation 3 months following surgery required a full reconstruction of the defect site with a novel ABGS (rhBMP6 in ABC with ceramic particles) to avoid front leg amputation. The healing was then followed for the next 16 months. The callus formation was observed on x-ray images 2 months following ABGS implantation. The bone segments progressively fused together leading to the defect rebridgment allowing removal of the external fixator by 4 months after the reconstruction surgery. At the end of the observation period, the function of the leg was almost fully restored while analyses of the humeral CT sections revealed restoration and cortices rebridgment with a renewal of uniform medullary canal including structural reconstruction of the distal humerus. ----- Conclusion: This large humeral gunshot segmental defect of the front leg in a dog was saved from amputation via inducing bone regeneration using a novel ABGS osteoinductive device containing BMP6 in ABC

Long-term posterolateral spinal fusion in rabbits induced by rhBMP6 applied in autologous blood coagulum with synthetic ceramics

Autologous bone graft substitute (ABGS) containing rhBMP6 in autologous blood coagulum (Osteogrow) is a novel therapeutic solution for bone regeneration. This study is aimed to investigate the long-term outcome of ABGS with synthetic ceramics (Osteogrow-C) in rabbit posterolateral spinal fusion (PLF) model. Osteogrow-C implants were implanted bilaterally between rabbit lumbar transverse processes. We compared the outcome following implantation of ABGS with ceramic particles of different chemical composition (TCP and biphasic ceramics containing both TCP and HA) and size (500-1700 µm and 74-420 µm). Outcome was analyzed after 14 and 27 weeks by microCT, histology, and biomechanical analyses. Successful bilateral spinal fusion was observed in all animals at the end of observation period. Chemical composition of ceramic particles has impact on the PLF outcome via resorption of TCP ceramics, while ceramics containing HA were only partially resorbed. Moreover, persistence of ceramic particles subsequently resulted with an increased bone volume in implants with small particles containing high proportion of HA. ABGS (rhBMP6/ABC) with various synthetic ceramic particles promoted spinal fusion in rabbits. This is the first presentation of BMP-mediated ectopic bone formation in rabbit PLF model with radiological, histological, and biomechanical features over a time course of up to 27 weeks

Evaluation of synthetic ceramics as compression resistant matrix to promote osteogenesis of autologous blood coagulum containing recombinant human bone morphogenetic protein 6 in rabbit posterolateral lumbar fusion model

Posterolateral lumbar fusion (PLF) is a commonly performed surgical procedure for the treatment of pathological conditions of the lumbosacral spine. In the present study, we evaluated an autologous bone graft substitute (ABGS) containing rhBMP6 in autologous blood coagulum (ABC) and synthetic ceramics used as compression resistant matrix (CRM) in the rabbit PLF model. In the pilot PLF rabbit experiment, we tested four different CRMs (BCP 500-1700 μm, BCP 1700-2500 μm and two different TCP in the form of slabs) which were selected based on achieving uniform ABC distribution. Next, ABGS implants composed of 2.5 mL ABC with 0.5 g ceramic particles (TCP or BCP (TCP/HA 80/20) of particle size 500-1700 μm) and 125 μg rhBMP6 (added to blood or lyophilized on ceramics) were placed bilaterally between transverse processes of the lumbar vertebrae (L5-L6) following exposition and decortication in 12 New Zealand White Rabbits observed for 7 weeks following surgery. Spinal fusion outcome was analysed by μCT, palpatory segmental mobility testing and selected specimens were either tested biomechanically (three-point bending test) and/or processed histologically. The total fusion success rate was 90.9% by both μCT analyses and by palpatory segmental mobility testing. The volume of newly formed bone between experimental groups with TCP or BCP ceramics and the different method of rhBMP6 application was comparable. The newly formed bone and ceramic particles integrated with the transverse processes on histological sections resulting in superior biomechanical properties. The results were retrospectively found superior to allograft devitalized mineralized bone as a CRM as reported previously in rabbit PLF. Overall, this novel ABGS containing rhBMP6, ABC and the specific 500-1700 μm synthetic ceramic particles supported new bone formation for the first time and successfully promoted posterolateral lumbar fusion in rabbits