ABSTRACT
This study developed probabilistic models to predict Salmonella growth in processed meat products formulated with varying concentrations of NaCl and NaNO2. A five-strain mixture of Salmonella was inoculated in nutrient broth supplemented with NaCl (0%, 0.25%, 0.5%, 0.75%, 0.5%, 1.0%, 1.25%, and 1.75%) and NaNO2 (0, 15, 30, 45, 60, 75, 90, 105, and 120 ppm). The inoculated samples were then incubated under aerobic and anaerobic conditions at 4°C, 7°C, 10°C, 12°C, and 15°C for up to 60 days. Growth (assigned the value of 1) or no growth (assigned the value of 0) for each combination was evaluated by turbidity. These growth response data were analyzed with a logistic regression to evaluate the effect of NaCl and NaNO2 on Salmonella growth. The results from the developed model were compared to the observed data obtained from the frankfurters to evaluate the performance of the model. Results from the developed model showed that a single application of NaNO2 at low concentrations did not inhibit Salmonella growth, whereas NaCl significantly (p<0.05) inhibited Salmonella growth at 10°C, 12°C, and 15°C, regardless of the presence of oxygen. At 4°C and 7°C, Salmonella growth was not observed in either aerobic or anaerobic conditions. When NaNO2 was combined with NaCl, the probability of Salmonella growth decreased. The validation value confirmed that the performance of the developed model was appropriate. This study indicates that the developed probabilistic models should be useful for describing the combinational effect of NaNO2 and NaCl on inhibiting Salmonella growth in processed meat products.
ABSTRACT
UNLABELLED: Urinary tract infections (UTIs) are one of the most common diseases by which humans seek medical help and are caused mainly by uropathogenic Escherichia coli (UPEC). Studying the virulence and antibiotic resistance of UPEC with respect to various phylogenetic groups is of utmost importance in developing new therapeutic agents. Thus, in this study, we analysed the virulence factors, antibiotic resistance and phylogenetic groups among various UPEC isolates from children with UTIs. The phylogenetic analysis revealed that majority of the strains responsible for UTIs belonged to the phylogenetic groups B2 and D. Of the 58 E. coli isolates, 79·31% belonged to group B2, 15·51% to group D, 3·44% to group A and 1·72% to B1. Simultaneously, the number of virulence factors and antibiotic resistance exhibited were also significantly high in groups B2 and D compared to other groups. Among the isolates, 44·8% were multidrug resistant and of that 73% belonged to the phylogenetic group B2, indicating the compatibility of antibiotic resistance and certain strains carrying virulence factor genes. The antibiotic resistance profiling of UPEC strains elucidates that the antimicrobial agents such as chloramphenicol, cefoxitin, cefepime, ceftazidime might still be used in the therapy for treating UTIs. SIGNIFICANCE AND IMPACT OF THE STUDY: As the antibiotic resistance pattern of uropathogenic Escherichia coli varies depending on different geographical regions, the antibiotic resistance pattern from this study will help the physicians to effectively administer antibiotic therapy for urinary tract infections. In addition, the frequency of virulence factors and antibiotic resistance genes among various phylogenic groups could be effectively used to draw new targets for uropathogenic Escherichia coli antibiotic-independent therapies. The study emphasizes need of public awareness on multidrug resistance and for more prudent use of antimicrobials.
Subject(s)
Anti-Bacterial Agents/therapeutic use , Drug Resistance, Multiple, Bacterial , Escherichia coli Infections/drug therapy , Urinary Tract Infections/drug therapy , Uropathogenic Escherichia coli , Cefepime , Cefoxitin/therapeutic use , Ceftazidime/therapeutic use , Cephalosporins/therapeutic use , Child , Chloramphenicol/therapeutic use , Escherichia coli Infections/microbiology , Escherichia coli Proteins/genetics , Humans , Microbial Sensitivity Tests , Phylogeny , Republic of Korea , Urinary Tract Infections/microbiology , Uropathogenic Escherichia coli/drug effects , Uropathogenic Escherichia coli/isolation & purification , Uropathogenic Escherichia coli/pathogenicity , Virulence Factors/geneticsABSTRACT
The inhibitory effect of chlorine (50, 100, and 200 mg/kg) was investigated with and without UV radiation (300 mW·s/cm(2)) for the growth of Listeria monocytogenes in chicken breast meat. Using a polynomial model, predictive growth models were also developed as a function of chlorine concentration, UV exposure, and storage temperature (4, 10, and 15°C). A maximum L. monocytogenes reduction (0.8 log cfu, cfu/g) was obtained when combining chlorine at 200 mg/kg and UV at 300 mW·s/cm(2), and a maximum synergistic effect (0.4 log cfu/g) was observed when using chlorine at 100 mg/kg and UV at 300 mW·s/cm(2). Primary models developed for specific growth rate and lag time showed a good fitness (R(2) > 0.91), as determined by the reparameterized Gompertz equation. Secondary polynomial models were obtained using nonlinear regression analysis. The developed models were validated with mean square error, bias factor, and accuracy factor, which were 0.0003, 0.96, and 1.11, respectively, for specific growth rate and 7.69, 0.99, and 1.04, respectively, for lag time. The treatment of chlorine and UV did not change the color and texture of chicken breast after 7 d of storage at 4°C. As a result, the combination of chlorine at 100 mg/kg and UV at 300 mW·s/cm(2) appears to an effective method into inhibit L. monocytogenes growth in broiler carcasses with no negative effects on color and textural quality. Based on the validation results, the predictive models can be used to accurately predict L. monocytogenes growth in chicken breast.
Subject(s)
Chlorine/pharmacology , Food Microbiology , Food Preservation/methods , Listeria monocytogenes/drug effects , Listeria monocytogenes/radiation effects , Meat/microbiology , Ultraviolet Rays , Animals , ChickensABSTRACT
Milk protein is a well-known precursor protein for the generation of bioactive peptides using lactic acid bacteria. This study investigated the antioxidant activity of bovine casein hydrolysate after fermentation with Bifidobacterium longum KACC91563 using the 2,2'-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) assay and total phenolic content (TPC). The antioxidant activities of the 24-h and 48-h hydrolysates were higher than that of the 4-h hydrolysate (2,045.5 and 1,629.3 µM gallic acid equivalents, respectively, vs. 40.3 µM) in the ABTS assay. In contrast, TPC values showed activities of 43.2 and 52.4 µM gallic acid equivalents for the 4-h and 24-h hydrolysates, respectively. Three fractions (≥10 kDa, ≥3 but <10 kDa, and <3 kDa) were separated from the 24-h hydrolysate by ultrafiltration. Among these fractions, the <3 kDa fraction exhibited the highest antioxidant activity (936.7 µM) compared with the other fractions (42.1 and 34.2 µM for >10 kDa and 3-10 kDa fractions, respectively). Through liquid chromatography-electrospray ionization-tandem mass spectrometry analysis, 2 peptides, VLSLSQSKVLPVPQK and VLSLSQSKVLPVPQKAVPYPQRDMPIQA, containing the fragment VLPVPQ that has antioxidant properties, were identified in the <3kDa fraction after 24h of hydrolysis. The present study demonstrates the possibility of antioxidant peptide production from bovine casein using Bifidobacterium longum.
Subject(s)
Bifidobacterium/metabolism , Caseins/metabolism , Animals , Antioxidants/isolation & purification , Antioxidants/pharmacology , Cattle , Chromatography, High Pressure Liquid , Chromatography, Liquid , Chromatography, Reverse-Phase , Electrophoresis, Polyacrylamide Gel , Fermentation , Free Radical Scavengers/pharmacology , Hydrolysis , Peptides/isolation & purification , Peptides/pharmacology , Tandem Mass SpectrometryABSTRACT
Molecular weights (MW) of major proteins in milk of 3 Korean dairy goat breeds were determined by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry, after treatment of milk samples with the reduction buffer used in capillary electrophoresis. The MW of caprine milk proteins were compared with those of Holstein milk counterparts using commercial bovine milk protein standards. The MW of α-lactalbumin, ß-lactoglobulin, and α- and ß-casein standards were 14,197±3.4, 18,326±26.3, 23,591±13.0, and 23,967±12.8 m/z, respectively, whereas those of Holstein milk treated with the reduction buffer were 14,199±8.3, 18,397±25.9, 23,614±64.8, and 23,984±75.6 m/z, respectively. The respective MW of α-lactalbumin in Saanen, Toggenberg, and Alpine milk were 14,194±27.2, 14,266±105.9, and 14,241±13.2 m/z, which were not different from those of the bovine milk. The respective MW of ß- lactoglobulin in corresponding caprine milk were 18,840±31.5, 18,856±26.3, and 18,857±21.3 m/z, which were higher than those in the bovine milk. The MW of ß-casein in corresponding caprine milk were 23,860±27.2, 23,886±12.3, and 23,901±8.4 m/z, which were lower than those in the bovine milk. The results indicated that matrix-assisted laser desorption/ionization time-of-flight mass spectrometry could be used for rapid determination of MW of Korean caprine milk proteins without protein separation steps.
Subject(s)
Milk Proteins/chemistry , Animals , Caseins/chemistry , Cattle , Goats , Lactalbumin/chemistry , Lactoglobulins/chemistry , Molecular Weight , Spectrometry, Mass, Matrix-Assisted Laser Desorption-IonizationABSTRACT
This study investigated the relationship between mast cell numbers and the grade of bladder transitional cell carcinoma (TCC). Bladder TCC biopsies were obtained via transurethral resection and 45 stage T1 specimens were included in the study. Specimens were sorted into two groups, low grade (grade I) and high grade (grades II and III). Samples were stained using haematoxylin and eosin, toluidine blue and immunohistochemical staining for tryptase. Mast cells were examined by light microscopy and cell density was recorded. Mast cell density was significantly higher in high-grade TCC than low-grade TCC. There was also a significant relationship between the number of mast cells identified using toluidine blue staining or immunohistochemical staining for tryptase. Detailed studies of mast cell function will enable the development of more effective antitumour therapies via mast cell manipulation.
Subject(s)
Carcinoma, Transitional Cell/pathology , Mast Cells/pathology , Urinary Bladder Neoplasms/pathology , Adult , Cell Count , Female , Humans , Male , Middle Aged , Neoplasm Grading , Prospective StudiesABSTRACT
Conjugated linoleic acid (CLA) and gamma-linolenic acid (GLA) were encapsulated with hydrated zinc oxide nanoparticles in an effort to improve their time and thermal stability. Encapsulated and nonencapsulated CLA and GLA were stored at 20, 30, 40, and 50 degrees C for 49 d. At various time points, encapsulated CLA and GLA were extracted, methylated, and analyzed using GC-FID. Both encapsulated CLA and control CLA were stable when stored at 20, 30, and 40 degrees C for up to 49 d. However, control CLA was 100% degraded after 28 d at 50 degrees C, whereas encapsulated CLA was stable at 50 degrees C for 49 d. Similarly, both encapsulated GLA and control GLA were stable when stored at 20 degrees C for 49 d, but nonencapsulated GLA was 92% degraded after 49 d at 30 degrees C; encapsulated GLA was stable at 30 degrees C. Therefore, nanoencapsulation improves the time and temperature stability of CLA and GLA.
Subject(s)
Linoleic Acids, Conjugated/administration & dosage , Nanocapsules , Zinc Oxide , gamma-Linolenic Acid/administration & dosage , Drug Stability , Hot Temperature , Time Factors , X-Ray DiffractionABSTRACT
Carcinoid tumor of the pancreas is rare. Moreover, obstructive pancreatitis secondary to a pancreatic carcinoid tumor is extremely rare. We report a case of pancreatic carcinoid tumor in a 50-year-old male who presented with pancreatitis. On multislice helical computed tomography, the main pancreatic duct was obstructed by a small round tumor, and the main pancreatic duct proximal to the tumor was dilated. The correlation between the main pancreatic duct and the tumor was well depicted on minimum intensity projection image. This is the first report of multislice helical computed tomorgraphic and minimum intensity projection image findings of a pancreatic carcinoid tumor presenting with pancreatitis.
Subject(s)
Carcinoid Tumor/complications , Carcinoid Tumor/diagnostic imaging , Pancreatic Neoplasms/complications , Pancreatic Neoplasms/diagnostic imaging , Pancreatitis/diagnostic imaging , Pancreatitis/etiology , Tomography, Spiral Computed , Diagnosis, Differential , Humans , Male , Middle AgedABSTRACT
Extrapulmonary small cell carcinoma is a rare neoplasm. It is an aggressive malignant tumour characterized by rapid local progression and early metastasis. We report a case of small cell carcinoma arising in the transverse colon in a 34-year-old man who presented with epigastric pain. On CT, a poorly enhancing bulky mass encircling the transverse colon with extensive regional lymph node metastases was observed. A segmental annular narrowing with thick interhaustral folds of the transverse colon was found by barium enema examination. This is the first report of barium study and CT findings of extrapulmonary small cell carcinoma of the colon.
Subject(s)
Barium Sulfate , Carcinoma, Small Cell/diagnostic imaging , Colonic Neoplasms/diagnostic imaging , Contrast Media , Tomography, X-Ray Computed/methods , Adult , Enema/methods , Humans , MaleABSTRACT
Methanolic extracts of seven herbs (Acorus calamus, Acorus gramineus, Bupleurm facaltum, Dioscorea batatas, Epimedium koreanum, Poria cocos and Zizyphi jujuba) used in traditional Korean medicine for improvement of memory and cognition in old age were tested for cholinesterase inhibitory properties using the Ellman colorimetric method. Significant inhibition of the enzyme at 200 microg/ml was observed for extracts from A. calamus and E. koreanum. The possible bases for the reputation of these and the other herbs tested are discussed in the light of previous investigations into their chemistry and biological activity.
Subject(s)
Cholinesterase Inhibitors/pharmacology , Cognition/drug effects , Phytotherapy , Plant Extracts/pharmacology , Plants, Medicinal , Acetylcholinesterase/drug effects , Acetylcholinesterase/metabolism , Alzheimer Disease/prevention & control , Cholinesterase Inhibitors/administration & dosage , Cholinesterase Inhibitors/therapeutic use , Dose-Response Relationship, Drug , Erythrocytes/enzymology , Humans , Inhibitory Concentration 50 , Korea , Medicine, Traditional , Plant Extracts/administration & dosage , Plant Extracts/therapeutic use , Plant Oils/administration & dosage , Plant Oils/pharmacology , Plant Oils/therapeutic use , Plant RootsABSTRACT
We report a case of gastric pseudoterranoviasis proven by gastrofiberscopy on Dec. 13, 1994. The 34-year-old male patient, residing in Chungju-shi, was admitted to Konkuk University Hospital complaining of prickling epigastric pain. The symptoms suddenly attacked him two days after eating raw marine fish at Chonan-shi. By the gastrofiberscopic examination, a long white-yellowish nematode was found from the fundus region of stomach. The worm was 34.50 x 0.84 mm in size, and was identified as a 3rd stage larva of Pseudoterranova decipiens judging from the position of the intestinal cecum. This is the 12th confirmed case of human pseudoterranoviasis in Korea.
Subject(s)
Anisakiasis/parasitology , Ascaridoidea/isolation & purification , Stomach Diseases/parasitology , Adult , Animals , Gastric Mucosa/parasitology , Gastroscopy , Humans , Korea , Larva , MaleABSTRACT
Interstitial pulmonary emphysema is a well-documented complication of assisted mechanical ventilation in premature infants with respiratory distress syndrome. Localized persistent interstitial pulmonary emphysema (LPIPE) confined to a single lobe was incidentally presented in a 4-day-old female infant. This patient was a normal full-term baby with no respiratory distress symptom and no experience of assisted mechanical ventilation. Chest radiograph showed radiolucent area in right lower lobe zone, which needed differential diagnosis from other congenital lesions such as congenital cystic adenomatoid malformation and congenital lobar emphysema. CT scan showed irregular-shaped air cystic spaces and pathologically, cystic walls primarily consisted of compressed lung parenchyma and loose connective tissue intermittently lined by multinucleated foreign body giant cells.
Subject(s)
Infant, Newborn, Diseases/pathology , Pulmonary Emphysema/pathology , Diagnosis, Differential , Female , Humans , Infant, Newborn , Infant, Newborn, Diseases/diagnostic imaging , Pulmonary Emphysema/diagnostic imaging , RadiographyABSTRACT
We report a case of lipoleiomyoma which arose in retroperitoneum and presented with progressively distended abdomen in a 56-yr-old woman. The tumor was well encapsulated and consisted of two components, benign smooth muscle cells and mature adipose tissue without any atypia. It is likely to be mistaken as extrarenal angiomyolipoma, well-differentiated liposarcoma and leiomyoma with fatty change. We review the histologic characteristics of previously reported myolipoma and describe essential points of differential diagnosis.
Subject(s)
Angiomyolipoma/pathology , Leiomyoma/pathology , Retroperitoneal Neoplasms/pathology , Diagnosis, Differential , Female , Humans , Liposarcoma/pathology , Middle AgedABSTRACT
UNLABELLED: Meconium aspiration syndrome is related to mechanical obstruction of the airways and subsequent chemical pneumonitis. It has also been suggested that meconium causes inhibition of surfactant function. To assess its inhibitory effect on surfactant function in vitro, we used a stable microbubble (SM) test that was thought to reflect the adequacy of pulmonary surfactant. The mixtures were prepared by adding serial dilutions of human meconium to various concentrations of Surfactant-TA (Surfacten). The SM count at each concentration of surfactant significantly increased with the increasing concentration of surfactant. This shows that the SM test closely reflects the quantified function of surfactant. When various concentrations of meconium were added to the surfactant concentration of 0.05 mg/ml and 0.25 mg/ml, the SM test results were decreased even at low concentrations of meconium. Also the increase in the meconium concentration caused a decrease in the SM test result, which was dependent on the surfactant and the meconium concentration, accordingly. These results suggest that meconium inhibits surfactant function. CONCLUSION: The stable microbubble test is an effective indirect method that tests the changes in surfactant quantity. In the in vitro experiment, we observed an inhibitory effect of meconium on the surfactant activity using the stable microbubble test.
Subject(s)
Biological Products , Meconium Aspiration Syndrome/complications , Meconium/metabolism , Pulmonary Surfactants/metabolism , Humans , In Vitro Techniques , Infant, Newborn , Inhibitory Concentration 50 , Meconium Aspiration Syndrome/metabolismABSTRACT
BRASSINOSTEROID-INSENSITIVE 1 (BRI1) encodes a putative Leucine-rich repeat receptor kinase in Arabidopsis that has been shown by genetic and molecular analysis to be a critical component of brassinosteroid signal transduction. In this study we examined some of the biochemical properties of the BRI1 kinase domain (BRI1-KD) in vitro, which might be important predictors of in vivo function. Recombinant BRI1-KD autophosphorylated on serine (Ser) and threonine (Thr) residues with p-Ser predominating. Matrix-assisted laser desorption/ionization mass spectrometry identified a minimum of 12 sites of autophosphorylation in the cytoplasmic domain of BRI1, including five in the juxtamembrane region (N-terminal to the catalytic KD), five in the KD (one each in sub-domains I and VIa and three in sub-domain VIII), and two in the carboxy terminal region. Five of the sites were uniquely identified (Ser-838, Thr-842, Thr-846, Ser-858, and Thr-872), whereas seven were localized on short peptides but remain ambiguous due to multiple Ser and/or Thr residues within these peptides. The inability of an active BRI1-KD to transphosphorylate an inactive mutant KD suggests that the mechanism of autophosphorylation is intramolecular. It is interesting that recombinant BRI1-KD was also found to phosphorylate certain synthetic peptides in vitro. To identify possible structural elements required for substrate recognition by BRI1-KD, a series of synthetic peptides were evaluated, indicating that optimum phosphorylation of the peptide required R or K residues at P - 3, P - 4, and P + 5 (relative to the phosphorylated Ser at P = 0).
Subject(s)
Arabidopsis Proteins , Arabidopsis/enzymology , Protein Kinases/metabolism , Amino Acid Motifs , Amino Acid Sequence , Arabidopsis/genetics , Molecular Sequence Data , Phosphorylation , Phytosterols/metabolism , Protein Kinases/chemistry , Protein Kinases/genetics , Recombinant Proteins/chemistry , Recombinant Proteins/genetics , Recombinant Proteins/metabolism , Serine/chemistry , Signal Transduction , Threonine/chemistryABSTRACT
BACKGROUND: The morphological definition of atrial chambers, and the determination of atrial laterality, are based on analysis of the structure of the atrial appendages. The systemic and pulmonary venous connections to the heart, nonetheless, are important in the management of patients having isomeric appendages. In this study, therefore, we analysed the morphology of the postero-superior walls of the atrial chambers so as to provide evidence concerning the morphogenetic background of those hearts, and to improve operative management. METHODS: We reviewed 15 autopsied specimens with isomeric right appendages, and 10 with isomeric left appendages, paying particular attention to the morphology of the systemic and pulmonary venous connections. The postero-superior walls of the atrial chambers can be made up of the atrial body, the systemic venous components, or the pulmonary venous component. We analysed the contributions made by each of these components. RESULTS: The postero-superior walls of the atrial chambers were markedly variable, but could be grouped into five patterns. Bilaterally well-developed systemic venous components and absence of the pulmonary venous component within the hypoplastic atrial body were present in 9 hearts with extracardiac pulmonary venous connections in the setting of right isomerism. Bilaterally well-developed systemic venous components, and a hypoplastic pulmonary venous component within the hypoplastic atrial body, were present in 5 hearts with intracardiac pulmonary venous connections in right isomerism. Bilaterally well-developed systemic venous components, and a hypoplastic pulmonary venous component within the sizable atrial body, were present in 1 heart with an intracardiac pulmonary venous connection in right isomerism. A well-developed pulmonary venous component within the atrial body, and hypoplasia of one systemic venous component, were present in 7 hearts with left isomerism. A well-developed pulmonary venous component within the atrial body, and hypoplasia of bilateral systemic venous components, were present in 3 hearts with left isomerism. CONCLUSIONS: The postero-superior walls of the atrial chambers in hearts with isomeric atrial appendages can be analysed on the basis of a compound structure made of bilateral systemic venous components, a central pulmonary venous component, and the body of the atrium. Hearts with isomeric right appendages have absence or hypoplasia of the pulmonary venous component, while hearts with isomeric left appendages have hypoplastic systemic venous components.
Subject(s)
Heart Atria/pathology , Levocardia/pathology , Pulmonary Veins/pathology , Autopsy , Bronchi/embryology , Bronchi/pathology , Echocardiography , Female , Fetus/embryology , Fetus/pathology , Heart Atria/diagnostic imaging , Heart Atria/embryology , Humans , Infant, Newborn , Levocardia/diagnostic imaging , Levocardia/embryology , Male , Pulmonary Veins/diagnostic imaging , Pulmonary Veins/embryology , Situs Inversus/embryology , Situs Inversus/pathology , Spleen/embryology , Spleen/pathologyABSTRACT
Sulfotransferase (ST) is considered to be generally not induced by xenobiotics. However, it has been reported that steroids such as dexamethasone (DEX) and pregnenolone-16a-carbonitrile (PCN) are effective ST inducers in rats, and sulfation of xenobiotics is quite different in rats and mice. The present study is primarily aimed at determining the effect of sulfate and steroids on the metabolism of acetaminophen (AA) in vitro using monolayer cultured hepatocytes of Sprague-Dawley rats and ICR mice. Hepatocytes of rats and mice were incubated with inorganic sulfate (0.25, 0.5, 1.0, 2.0, 4.0 mM) and AA in SO4-depleted media. AA sulfation rates increased as the sulfate concentration was raised to 1.0 mM in rats, whereas the addition of inorganic sulfate to the media had a lesser effect in mice hepatocytes. After pretreatment with DEX (0.1, 1.0, 10, 100 microM) and PCN (0.1, 1.0, 10 microM) for 3 d, hepatocytes were incubated with AA in media containing 4.0 mM SO4-. Pretreatment of the hepatocytes with DEX caused an increase in the glucuronidation and sulfation of AA by 2-3 folds in rats, but to a lesser extent in mice. PCN significantly enhanced the formation of AA-glucuronide and AA-sulfate in mice, but had a minimal effect on rat hepatocytes. In summary, sulfate and DEX markedly enhanced the formation of AA-sulfate in rats hepatocytes, and DEX and PCN increased the sulfation of AA in mice hepatocytes. These results partially support the claim that DEX and PCN are effective ST and uridine diphosphate-glucuronyltransferase inducers in vivo.
Subject(s)
Acetaminophen/metabolism , Dexamethasone/pharmacology , Glucocorticoids/pharmacology , Liver/drug effects , Pregnenolone Carbonitrile/pharmacology , Sulfates/pharmacology , Animals , Cells, Cultured , Liver/cytology , Liver/metabolism , Male , Mice , Mice, Inbred ICR , Rats , Rats, Sprague-Dawley , Species SpecificityABSTRACT
We have analyzed the control exerted by the pathways of oxidative phosphorylation, gluconeogenesis, ureagenesis, and maintenance ATP consumption over each other's rates in isolated, perfused rat liver using top-down metabolic control analysis. The livers from fasted rats were perfused with 3-hydroxybutyrate as respiratory substrate, lactate as substrate for gluconeogenesis, and ammonium as substrate for urea synthesis, in conditions where these pathways were only linked by their common intermediates: ATP, ADP, and Pi. The rates of oxygen consumption, glucose and urea synthesis were measured continuously. The pathways were perturbed either by adding specific inhibitors or by adding new pathways that consumed ATP, and the relative changes in pathway rates were used to calculate the flux control coefficients of each pathway over all pathway rates. When the liver was in a relatively inactive metabolic state, where ATP was only being used by the maintenance ATP-consuming pathways, then essentially all the control over ATP production and consumption was located in the maintenance ATP consumers with ATP production having no control. Whereas, when the liver was in a highly active state using extra ATP for both glucose and urea synthesis, then ATP production (from oxidative phosphorylation) had strong control over its own rate and the rates of glucose and urea synthesis, but gluconeogenesis and ureagenesis still had strong control over their own rates and negative control over each others rates, i.e. they competed for the limited ATP supply. The rate of the maintenance ATP consumers is remarkably insensitive to changes in ATP production and consumption, but exerts considerable control over all other pathways. These results indicate that the general assumption that the rates of ATP production and consumption are controlled exclusively by ATP consumers is false under conditions where a significant amount of ATP is used for biosynthetic processes, such as glucose and urea synthesis, and indicate that the latter processes may be partly controlled by regulators of ATP production and by other ATP-consuming pathways.
Subject(s)
Adenosine Triphosphate/metabolism , Gluconeogenesis/physiology , Liver/metabolism , Oxidative Phosphorylation , Urea/metabolism , 3-Hydroxybutyric Acid , Acetoacetates/metabolism , Ammonia/metabolism , Amobarbital/pharmacology , Animals , Glucose/metabolism , Hydroxybutyrates/metabolism , Lactic Acid/metabolism , Oxygen Consumption/physiology , Perfusion , Pyruvic Acid/metabolism , Quinolinic Acid/pharmacology , Rats , Valine/analogs & derivatives , Valine/pharmacologyABSTRACT
Brassinosteroids are known to promote cell elongation in a wide range of plant species but their effect on cell division has not been as extensively studied. We examined the effect of brassinolide on the kinetics and final division frequencies of regenerating leaf mesophyll protoplasts of Petunia hybrida Vilm v. Comanche. Under optimal auxin and cytokinin conditions, 10-100 nM brassinolide accelerated the time of first cell division by 12 h but had little effect on the final division frequencies after 72-120 h of culture. One micromolar brassinolide showed the same acceleration of first cell division but inhibited the final division frequency by approximately 20%. Under sub-optimal auxin conditions, 10-100 nM brassinolide both accelerated the time of first cell division and dramatically increased the 72- to 120-h final division frequencies. Isolated protoplasts may provide a useful model system to investigate the molecular mechanisms of brassinosteroid action on cell proliferation.