PURPOSE: 99mTc-HMPAO is a radiopharmaceutical for imaging cerebral blood flow. HMPAO (RR, SS)-4,8- diaza-3,6,6,9-tetramethylundecan-2,10- dione bisoxime) has three stereoismers such as, meso-, d-, and l-HMPAO. Techentium complexes of meso-HMPAO and d,l-HMPAO are known to have different in vivo brain uptakes. In this study, enantiomers of HMPAO (d-HMPAO and l-HMPAO) were separated from d,l-HMPAO. These enantiomers were labeled with 99mTc and the biodistribution studies were performed in mice. MATERIALS AND METHODS: An intermediate imine product was produced from 2,3-butanedione monooxime and 2,2-dimethyl- 1,3-propanediamine (54% yield) and was reduced into a mixture of three isomers (35% yield). The meso-isomer was separated from d,l-mixture by repeated fractional crystallization (11% yield). The d- and l-enantiomers were subsequently separated by co-crystallization with optical isomers of tartaric acid (25% and 5% yield, respectively). Each enantiomeric HMPAO was labeled with 99mTc by reacting with SnCl2 2H2O and 99mTc-pertechnetate. Biodistribution study was performed 1 hr after tail vein injection to ICR mice. RESULTS: Radiochemical purities of each compound were over 80%. In biodistribution study, the brain uptakes of d,l- d- and l-form were 1.34, 1.12 and 1.67% ID/g, respectively. In case of l-Isomer the brain uptake was higher (1.5 fold) than d-isomer. CONCLUSION: We successfully purified each enantiomeric HMPAO. In biodistribution study of stereoismers of 99mTc-HMPAO in mice, l-HMPAO may show better brain image than d,l-HMPAO which was supplied in a commercial kit.
Animals ; Brain* ; Crystallization ; Dental Calculus ; Diacetyl ; Mice ; Mice, Inbred ICR ; Stereoisomerism ; Technetium Tc 99m Exametazime* ; Veins

BACKGROUND: Whether or not the excitation-contraction (EC) uncoupler, diacetyl monoxime (DAM) and cytochalacin D (Cyto D) alter the ventricular fibrillation activation patterns is unclear. METHODS: We recorded single cell action potentials and performed optical mapping in isolated perfused swine right ventricles at different concentrations of DAM and cyto D during ventricular fibrillation and dynamic pacing. RESULTS: Increasing concentration of DAM results in progressively shortened action potential duration measured to 90% repolarization (APD90), reduced slope of the action potential duration restitution(APDR) curve, decreased Kolmogorov-Sinai entropy, and reduced number of ventricular fibrillation wavefronts. In all right ventricles, 15 to 20 mmol/l DAM converted ventricular fibrillation to ventricular tachycardia. The ventricular fibrillation could be reinduced after the DAM was washed out. In comparison, cyto D (10 to 40 mol/l) has no effects on APDR curve or the dynamics of ventricular fibrillation. The effects of DAM on ventricular fibrillation are associated with reduced number of wavefronts and dynamic complexities in ventricular fibrillation. CONCLUSION: These results are compatible with Restitution Hypothesis of ventricular fibrillation and suggest that DAM may be unsuitable as an EC uncoupler for optical mapping studies of ventricular fibrillation in the swine right ventricles.
Action Potentials ; Diacetyl ; Entropy ; Heart Ventricles* ; Swine* ; Tachycardia, Ventricular ; Ventricular Fibrillation*

OBJECTIVE: Pralidoxime is widely used for the treatment of organophosphate poisoning. Multiple studies have reported its vasoconstrictive property, which may facilitate the restoration of spontaneous circulation (ROSC) after cardiac arrest by increasing the coronary perfusion pressure (CPP). 2,3-Butanedione monoxime, which belongs to the same oxime family, has been shown to facilitate ROSC by reducing left ventricular ischemic contracture. Because pralidoxime and 2,3-butanedione monoxime have several common mechanisms of action, both drugs may have similar effects on ischemic contracture. Thus, we investigated the effects of pralidoxime administration during cardiopulmonary resuscitation in a pig model with a focus on ischemic contracture and CPP.METHODS: After 14 minutes of untreated ventricular fibrillation, followed by 8 minutes of basic life support, 16 pigs randomly received either 80 mg/kg of pralidoxime (pralidoxime group) or an equivalent volume of saline (control group) during advanced cardiovascular life support (ACLS).RESULTS: Mixed-model analyses of left ventricular wall thickness and chamber area during ACLS revealed no significant group effects or group-time interactions, whereas a mixed-model analysis of the CPP during ACLS revealed a significant group effect (P=0.038) and group-time interaction (P<0.001). Post-hoc analyses revealed significant increases in CPP in the pralidoxime group, starting at 5 minutes after pralidoxime administration. No animal, except one in the pralidoxime group, achieved ROSC; thus, the rate of ROSC did not differ between the two groups.CONCLUSION: In a pig model of cardiac arrest, pralidoxime administered during cardiopulmonary resuscitation did not reduce ischemic contracture; however, it significantly improved CPP.
Animals ; Cardiopulmonary Resuscitation ; Diacetyl ; Heart Arrest ; Hemodynamics ; Humans ; Ischemic Contracture ; Organophosphate Poisoning ; Perfusion ; Swine ; Ventricular Fibrillation

Occupational exposure to diacetyl can lead to bronchiolitis obliterans. In this paper, two patients with severe obstructive ventilation disorder who were exposed to diacetyl at a fragrance and flavours factory were analyzed. The clinical manifestations were cough and shortness of breath. One of them showed Mosaic shadows and uneven perfusion in both lungs on CT, while the other was normal. Field investigation found that 4 of the 8 workers in the factory were found to have obstructive ventilation disorder, and 2 had small airway dysfunction. This paper summarizes the diagnostic process of patients in order to improve the understanding of airway dysfunction caused by occupational exposure to diacetyl and promote the development of relevant standards.
Humans ; Diacetyl/adverse effects* ; Occupational Diseases/diagnosis* ; Occupational Exposure/adverse effects* ; Lung ; Bronchiolitis Obliterans/diagnosis*

Saccharomycopsis fibuligera possesses high alpha-amylase and glucoamylase activities that enable it to utilize raw starch as a carbon source. A expression cassette containing the promoter sequence of 3-phosphogylycerate kinase gene (PGK1p), the alpha factor signal sequence from Saccharomyces cerevisiae and the alpha-amylase coding sequence of S. fibuligera was constructed. The alpha-amylase expression cassette was inserted in the ILV2 locus of industrial brewer's yeast strain YSF-5 encoding alpha-acetolactate synthase (AHAS) by homologous recombination. The transformed yeast strain was selected on the media with starch as the sole carbon source and verified by PCR. The transformant exhibited secretive alpha-amylase activity, low AHAS activity and reduced diacetyl production. Effects of temperature, pH, and metal ions on the activity of the alpha-amylase expressed by the transformant were examined. The fermentation performance of host strain YSF-5 and the transformant was also examined.
Beer ; microbiology ; Diacetyl ; metabolism ; Glycogen Synthase Kinase 3 ; genetics ; Recombinant Proteins ; biosynthesis ; genetics ; Recombination, Genetic ; Saccharomyces cerevisiae ; genetics ; metabolism ; Saccharomycopsis ; enzymology ; genetics ; alpha-Amylases ; biosynthesis ; genetics ; metabolism

AIMIn order to elucidate the underlying mechanism of depressed maximal isometric twitch tension normalized by cross sectional area of muscle strip in unloaded soleus.

METHODSThe soleus and extensor digitorum longus (EDL) muscle strips were perfused in vitro and treated by 2,3-Butanedione monoxime (BDM).

RESULTSThe BDM decreased Pt of soleus and EDL in a concentration-dependent manner. The Pt could restored completely to normal level after washing out BDM. The isometric twitch duration was not altered during 1 mmol/L BDM of perfusion, but was shortened at 10 mmol/L dose. The time from maximal to half Pt in EDL was shorter than that in soleus during 10 mmol/L BDM of perfusion. The inhibitory effects of BDM on myosin ATPase activity were higher in EDL than in soleus. The inhibitory extent of BDM on myosin ATPase activity of soleus and EDL was lower than that on Pt.

CONCLUSIONThese results suggest that reduction in cross-bridge function of skeletal muscle may be one of reasons induced a decrease in its Pt. BDM is not a specific inhibitor on myosin ATPase activity and can affect multiple parts of excitation-contraction coupling in skeletal muscle.

Animals ; Diacetyl ; analogs & derivatives ; pharmacology ; Isometric Contraction ; drug effects ; physiology ; Male ; Muscle Contraction ; drug effects ; physiology ; Muscle, Skeletal ; drug effects ; physiology ; Myosins ; antagonists & inhibitors ; Rats ; Rats, Sprague-Dawley

Polyamines, putrescine, spermidine and spermine, are ubiquitous in living cells and are essential for eukaryotic cell growth. These polycations interact with negatively charged molecules such as DNA, RNA, acidic proteins and phospholipids and modulate various cellular functions including macromolecular synthesis. Dysregulation of the polyamine pathway leads to pathological conditions including cancer, inflammation, stroke, renal failure and diabetes. Increase in polyamines and polyamine synthesis enzymes is often associated with tumor growth, and urinary and plasma contents of polyamines and their metabolites have been investigated as diagnostic markers for cancers. Of these, diacetylated derivatives of spermidine and spermine are elevated in the urine of cancer patients and present potential markers for early detection. Enhanced catabolism of cellular polyamines by polyamine oxidases (PAO), spermine oxidase (SMO) or acetylpolyamine oxidase (AcPAO), increases cellular oxidative stress and generates hydrogen peroxide and a reactive toxic metabolite, acrolein, which covalently incorporates into lysine residues of cellular proteins. Levels of protein-conjuagated acrolein (PC-Acro) and polyamine oxidizing enzymes were increased in the locus of brain infarction and in plasma in a mouse model of stroke and also in the plasma of stroke patients. When the combined measurements of PC-Acro, interleukin 6 (IL-6), and C-reactive protein (CRP) were evaluated, even silent brain infarction (SBI) was detected with high sensitivity and specificity. Considering that there are no reliable biochemical markers for early stage of stroke, PC-Acro and PAOs present promising markers. Thus the polyamine metabolites in plasma or urine provide useful tools in early diagnosis of cancer and stroke.
Acrolein ; Animals ; Biomarkers ; Brain Infarction ; C-Reactive Protein ; Diacetyl ; DNA ; Early Detection of Cancer ; Eukaryotic Cells ; Humans* ; Hydrogen Peroxide ; Inflammation ; Interleukin-6 ; Lysine ; Metabolism ; Mice ; Oxidative Stress ; Oxidoreductases ; Phospholipids ; Plasma ; Polyamines* ; Putrescine ; Renal Insufficiency ; RNA ; Sensitivity and Specificity ; Spermidine ; Spermine ; Stroke

Recombinant plasmid pICG was constructed by replacing the internal fragment of a-acetohydroxyacid synthase (AHAS) gene (ILV2) with a copy of gamma-glutamylcysteine synthetase gene (GSH1) and copper chelatin gene (CUP1) from the industrial brewing yeast strain YSF31. YSF31 was transformed with plasmid pICG linearized by Kpn I and Pst I. A recombinant strain with high-glutathione and low-diacetyl production was selected. The results of fermentation in 100-L bioreactor showed that the lagering time of beer produced for recombinant strain T2 was shortened by 3 days and the shelf life of the beer was prolonged about 50%. It may be more acceptable for the commercial application, as it does not contain foreign DNA.
Acetolactate Synthase ; genetics ; metabolism ; Beer ; microbiology ; Cloning, Molecular ; Diacetyl ; metabolism ; Fermentation ; Gene Expression Regulation, Fungal ; Glutamate-Cysteine Ligase ; genetics ; metabolism ; Glutathione ; biosynthesis ; Metallothionein ; genetics ; metabolism ; Organisms, Genetically Modified ; genetics ; Saccharomyces cerevisiae ; genetics ; metabolism ; Saccharomyces cerevisiae Proteins ; genetics ; metabolism

OBJECTIVETo investigate the Effect of 2,3-butanedione monoxime (BDM) on calcium paradox-induced heart injury and its underlying mechanisms.

METHODSThirty-two adult male SD rats were randomized into 4 groups, namely the control group, BDM treatment control group, calcium paradox group, and BDM treatment group. Isolated Sprague Dawley male rat hearts underwent Langendorff perfusion and the left ventricular pressure (LVP) and left ventricular end-diastolic pressure (LVEDP) were monitored. Left ventricular developed pressure (LVDP) was calculated to evaluate the myocardial performance. Lactate dehydrogenase (LDH) content in the coronary flow was determined. Triphenyltetrazolium chloride staining was used to measure the infarct size, and myocardial cell apoptosis was tested with TUNEL method. Western blotting was used to determine the expression of cleaved caspase-3 and cytochrome c.

RESULTSCompared with the control group, BDM at 20 mmol/L had no effect on cardiac performance, cell death, apoptotic index or the content of LDH, cleaved caspase-3 and cytochrome c at the end of perfusion under control conditions (P>0.05). Calcium paradox treatment significantly decreased the cardiac function and the level of LVDP and induced a larger infarct size (P<0.01), an increased myocardial apoptosis index (P<0.01), and up-regulated expressions of cleaved caspase-3 and cytochrome c (P<0.01). BDM (20 mmol/L) significantly attenuated these effects induced by calcium paradox, and markedly down-regulated the levels of LVEDP and LDH (P<0.01), lowered myocardial apoptosis index, decreased the content of cleaved caspase-3 and cytochrome c (P<0.01), increased LVDP, and reduced the infarct size (P<0.01).

CONCLUSIONBDM suppresses cell apoptosis and contracture and improves heart function and cell survival in rat hearts exposed to calcium paradox, suggesting the value of BDM as an potential drug for myocardial ischemia reperfusion injur.

Animals ; Apoptosis ; Calcium ; adverse effects ; Caspase 3 ; metabolism ; Cytochromes c ; metabolism ; Diacetyl ; analogs & derivatives ; pharmacology ; Heart ; drug effects ; physiopathology ; In Vitro Techniques ; L-Lactate Dehydrogenase ; metabolism ; Male ; Myocardial Reperfusion Injury ; chemically induced ; drug therapy ; Rats ; Rats, Sprague-Dawley ; Ventricular Function, Left

Animals integrate various environmental stimuli within the nervous system to generate proper behavioral responses. However, the underlying neural circuits and molecular mechanisms are largely unknown. The insulin-like signaling pathway is known to regulate dauer formation, fat metabolism, and longevity in Caenorhabditis elegans (C. Elegans). Here, we show that this highly conserved signaling pathway also functions in the integrative response to an olfactory diacetyl and a gustatory Cu(2+) stimuli. Worms of wild-type N2 Bristol displayed a strong avoidance to the Cu(2+) barrier in the migration pathway to the attractive diacetyl. Mutants of daf-2 (insulin receptor), daf-18 (PTEN lipid phosphatase), pdk-1 (phosphoinositide-dependent kinase), akt-1/-2 (Akt/PKB kinase) and sgk-1 (serum- and glucocorticoid-inducible kinase) show severe defects in the elusion from the Cu(2+). Mutations in DAF-16, a forkhead-type transcriptional factor, suppress the integrative defects of daf-2 and akt-1/-2 mutants. We further report that neither cGMP nor TGFβ pathways, two other dauer formation regulators, likely plays a role in the integrative learning. These results suggest that the insulin-like signaling pathway constitutes an essential component for sensory integration and decision-making behavior plasticity.
Animals ; Caenorhabditis elegans ; genetics ; physiology ; Caenorhabditis elegans Proteins ; genetics ; physiology ; Chemotaxis ; genetics ; physiology ; Copper ; physiology ; Cyclic GMP ; genetics ; physiology ; Diacetyl ; metabolism ; Insulin ; metabolism ; Longevity ; Signal Transduction ; Smell ; genetics ; physiology ; Taste ; genetics ; physiology ; Transforming Growth Factor beta ; genetics ; physiology