JOURNAL OF FOOD BIOCHEMISTRY, Issue 5 2008
MICHAEL R. WEBB
ABSTRACT Anthocyanins and their aglycone anthocyanidins are pigmented flavonoids found in significant amounts in many commonly consumed foods. They exhibit a complex chemistry in aqueous solution, which makes it difficult to study their chemistry under physiologic conditions. Here we used a gel electrophoresis assay employing supercoiled DNA plasmid to examine the ability of these compounds (1) to intercalate DNA; (2) to inhibit human topoisomerase I through both inhibition of plasmid relaxation activity (catalytic inhibition) and stabilization of the cleavable DNA,topoisomerase complex (poisoning); and (3) to inhibit or enhance oxidative single-strand DNA nicking. We found no evidence of DNA intercalation by anthocyan(id)ins in the physiologic pH range for any of the compounds used in this study , cyanidin chloride, cyanidin 3- O -glucoside, cyanidin 3,5- O -diglucoside, malvidin 3- O -glucoside and luteolinidin chloride. The anthocyanins inhibited topoisomerase relaxation activity only at high concentrations (>50 µM), and we could find no evidence of topoisomerase I cleavable complex stabilization by these compounds. However, we observed that all of the anthocyan(id)ins used in this study were capable of inducing significant oxidative DNA strand cleavage (nicking) in the presence of 1 mM dithiothreitol, while the free radical scavenger, dimethyl sulfoxide (DMSO), at concentrations typically used in similar studies, completely inhibited DNA nicking. Finally, we proposed a mechanism to explain the anthocyan(id)in-induced oxidative DNA cleavage observed under our experimental conditions. PRACTICAL APPLICATIONS This study provided improved understanding of the mechanisms by which anthocyan(id)ins interact with DNA. By characterizing the chemistry and solution properties of these important dietary components, we obtained improved information on how the anthocyan(id)ins might function in living systems. [source]

INHIBITION OF HELICOBACTER PYLORI BY PHENOLIC EXTRACTS OF SPROUTED PEAS (PISUM SATIVUM L.)

JOURNAL OF FOOD BIOCHEMISTRY, Issue 1 2006
CHIA-YU HO
ABSTRACT Infection by Helicobacter pylori is associated with gastric and duodenal ulcers. Conventional treatments to eradicate it have side-effects such as diarrhea and dizziness. The excessive use of antibiotics could also lead to antibiotic-resistant bacteria. The use of plant phenolic phytochemicals can be an alternative because of their health benefits due to both antioxidant activity and antimicrobial activity. The pea (Pisum sativum), the world's second most important pulse crop, produces phenolic phytochemicals with antimicrobial potential. Because the synthesis of phenolic compounds increases with stress, we investigated the anti- H. pylori effectiveness of extracts from pea sprouts, germinated in the dark condition following treatment with either distilled water or acetyl salicylic acid. The peas were germinated for 8 days and the sprouted samples were measured for total soluble phenolic content, antioxidant and guaiacol peroxidase activity. Subsequently, the sprout extracts were tested for anti- H. pylori activity using the agar diffusion method and the effective dose was determined based on phenolic content. The results showed that both acetyl salicylic acid-treated and untreated pea sprouts at days 5 and 8 had anti- H. pylori activity. The minimum volume for inhibition was 50 ,L of extracts. The inhibitory effects were dose dependent. From this study, the potential to use natural phenolic phytochemicals from pea sprouts to control H. pylori was found to be promising. This provides a strategy and foundation to design legume phenolics as functional ingredients against H. pylori. [source]

INHIBITION OF GEL WEAKENING OF THREADFIN BREAM SURIMI USING THAI LEGUME SEED PROTEINASE INHIBITORS

JOURNAL OF FOOD BIOCHEMISTRY, Issue 5 2000
SOOTTAWAT BENJAKUL
ABSTRACT Partially purified proteinase inhibitors from cowpea (Vigna unguiculata (L) Wasp), pigeon pea (Cajanus cajan (L.) Millsp.) and bdmbara groundnuts (Voandzeia subterranea (L.) Thou) effectively inhibited sarcoplasmic modori-inducing proteinase extracted from threadfin bream muscle in a concentration dependent manner. Incorporation of these proteinase inhibitors into threadfin bream surimi partially inhibited autolytic degradation and increased the gel force and deformation. Combination of setting and incorporating proteinase inhibitors from cowpea and bambara groundnut var. HY at the level of 30 Kcunits/g resulted in an increase in gel force and deformation by 60% and 26%, respectively. However, the lightness and whiteness of surimi gels decreased slightly when the proteinase inhibitor was added at a level of 30 kunits/g. [source]

INHIBITION OF STALING IN CHAPATI (INDIAN UNLEAVENED FLAT BREAD)

JOURNAL OF FOOD PROCESSING AND PRESERVATION, Issue 3 2008
IRSHAD M. SHAIKH
ABSTRACT Chapati is an unleavened Indian flat bread made from whole wheat flour usually consumed immediately after preparation as it becomes hard on keeping because of staling. Large-scale mechanized preparation of ready-to-eat chapatis will have to address this problem. Investigations were made to study the effect of sodium stearoyl-2-lactylate (SSL), glycerol monostearate, propylene glycol, sorbitol, ,-amylase, xylanase, maltodextrin and guar gum when added to chapati dough, on the inhibition of staling in chapatis stored at 29 ± 1C and 4 ± 1C. Chapatis were prepared from whole wheat flour dough containing (salt, 5% oil, appropriate preservatives and the aforementioned additives. Chapatis were packed in self-sealing low-density polyethylene bags and were stored for 10 days at 29 ± 1C and 4 ± 1C. Stored chapatis were analyzed for various staling parameters such as moisture content, texture, water-soluble starch, in vitro enzyme digestibility, enthalpy change (,H) and sensory quality. Staling of chapatis at 29 ± 1C and at 4 ± 1C of storage was inhibited by all additives to different extents; the extent of staling was less at 4 ± 1C. Maltodextrin at 0.3% (w/w) was found to be the most effective. Several combinations of additives were also studied, and the best combination was (100 ppm) + SSL (0.375%) levels. PRACTICAL APPLICATIONS Chapati, a traditional staple food of Indians, is unleavened flat bread made from whole wheat flour. With rapidly changing lifestyles, changing socio-economic trends and increasing urbanization and consumerism there is a rising demand for convenience foods which require minimum or no preparation time particularly the ready-to-eat (RTE) type of foods. Chapatis are generally baked fresh twice a day for lunch and dinner, and unless eaten immediately after baking, these stale rapidly and become difficult to chew. The most important parameter of chapati quality is its texture. The texture is generally evaluated in terms of tenderness, flexibility and its suitability to be folded into a spoon shape for eating with curried preparation. RTE chapatis are the latest addition to the species of "convenience foods." Keeping this in mind the research was undertaken to improve quality of chapati with the addition of various additives and improvements for the inhibition of staling in chapati. [source]

INHIBITION OF MICROORGANISMS IN SALAD DRESSING BY SUCROSE AND METHYLGLUCOSE FATTY ACID MONOESTERS

JOURNAL OF FOOD PROCESSING AND PRESERVATION, Issue 4 2003
CHING-MIAO YANG
The antimicrobial activity of sucrose and methylglucose esters of medium to long chain fatty acids was studied with two microorganisms involved in the spoilage of salad dressings, Zygosaccharomyces bailii and Lactobacillus fructivorans. The microorganisms were inhibited to various degrees by 0.1, 0.5, and 1.0% synthesized sucrose or methylglucose monoesters using a modified broth dilution method. Sucrose monoesters were most inhibitory when the esterified fatty acid was myristic (C14) or palmitic acid (C16). Methylglucose monoesters with lauric (C12) or myristic acid (C14) exhibited greater inhibition than those with longer chain fatty acids. The least inhibition was generally observed with sucrose and methylglucose oleate (C18:1). Sucrose monoesters were usually more inhibitory than methylglucose monoesters of the same fatty acid, especially for palmitic and stearic (C18) acids. In salad dressing, 1% sucrose monoesters of lauric, myristic, or palmitic acid significantly (P < 0.05) inhibited the growth of Z. bailii and L. fructivorans, and were comparable with or more effective than 0.1% sodium benzoate. Z. bailii growth was nearly completely inhibited by sucrose laurate, myristate and palmitate by 9 days of salad dressing storage. Sucrose monoesters did not delay the lag phase of L. [source]

GROWTH INHIBITION OF CLOSTRIDIUM PERFRINGENS VEGETATIVE CELLS AND SPORES USING CHICKEN IMMUNOGLOBULIN Y

JOURNAL OF FOOD SAFETY, Issue 4 2009
MIN S. SONG
ABSTRACT Egg yolk antibody (IgY) was isolated by the water dilution method from the egg yolk of chickens immunized with Clostridium perfringens vegetative cells and spores. Specific binding activity of IgY against C. perfringens vegetative cells and spores remained relatively high during the immunization period (up to 9 weeks). The titer of specific IgY against C. perfringens spores was 1.4-fold less than that of specific IgY against C. perfringens vegetable cells. The specific IgY powder (10 µg/mL) was found to inhibit the growth of C. perfringens vegetative cells or C. perfringens spores in a liquid medium. The difference of C. perfringens vegetative cell growth between the treatment and control groups was 8.9 × 106 colony forming units (cfu)/mL at 8 h of incubation and 9.95 × 107 cfu/mL at 24 h of incubation. Significant cfu reductions in C. perfringens spores were also observed with specific IgY powder at 24 h of incubation. PRACTICAL APPLICATIONS IgY antibody exerts an antimicrobial activity against pathogens by binding, immobilizing and consequently reducing or inhibiting the growth, replication or colony forming ability of pathogenic bacteria; thus, it is proved to be a viable alternative for antibiotics and preservatives. In this study, IgY against Clostridium perfringens can be used to replace chemical preservatives in food industries. Because IgY functions well at low temperature, it can be used to inhibit the growth of Clostridia which germinate in refrigerated storage conditions, thus preventing foodborne enterotoxicity caused by such bacteria. In practical applications, natural antimicrobial IgY antibody can be applied to meat products for the improvement of food safety. [source]

INHIBITION OF FOODBORNE PATHOGENS AND SPOILING BACTERIA BY ESSENTIAL OIL AND EXTRACTS OF ERIGERON RAMOSUS (WALT.) B.S.P.

JOURNAL OF FOOD SAFETY, Issue 2 2009
ATIQUR RAHMAN
ABSTRACT The antibacterial potential of essential oil and methanolic extracts of Erigeron ramosus (Walt.) B.S.P. was evaluated. Thirty-one components representing 95.3% of the total oil were identified, of which ,-caryophyllene (24.0%), ,-humulene (14.5%), 1,8-cineole (9.0%), eugenol (7.2%), globulol (7.1%), caryophyllene oxide (5.2%), ,-cadinene (5.0%), ,-copaene (4.9%) and widdrol (2.0%) were the major components. The antibacterial activity of essential oil and methanolic extracts of E. ramosus was determined in vitro using the agar diffusion method and minimum inhibitory concentration determination test against 14 (seven gram-positive and seven gram-negative) foodborne bacteria. The essential oil (5 µL/mL, corresponding to 1,000 ppm/disc), methanol extract and its different organic subfractions (7.5 µL/mL, corresponding to 1500 ppm/disc) of E. ramosus displayed a great potential of antibacterial activity against all gram-positive bacteria: Staphylococcus aureus (ATCC 6538 and KCTC 1916), Listeria monocytogenes (ATCC 19116, ATCC 19118, ATCC 19166 and ATCC 15313) and Bacillus subtilis ATCC 6633 and four gram-negative bacteria: Pseudomonas aeruginosa KCTC 2004, Enterobacter aerogenes KCTC 2190 and Escherichia coli (0157:H7 ATCC 43888 and ATCC 8739). The zones of inhibition of different concentrations of essential oil and methanolic extracts against the tested bacteria were found in the range of 10.1,22.3 mm, and MIC values were recorded between 62.5 and 500 µg/mL. PRACTICAL APPLICATIONS The use of essential oil and organic extracts of Erigeron ramosus (Walt.) B.S.P. as antibacterial agents will be suitable for applications on the food industry as natural preservatives or flavoring to control foodborne pathogens. They can be used as growth inhibitors of Listeria monocytogenes, Staphylococcus aureus, Bacillus subtilis, Escherichia coli, Enterobacter aerogenes and Pseudomonas aeruginosa, some important foodborne pathogens and spoiling bacteria. The main reason for their suitability is their natural origin, which consumers find comforting and which is beneficial for the environment, and the very low risk that pathogens will develop resistance to the mixture of components that make up the oil and extracts with their apparent diversity of antibacterial mechanisms. These beneficial characteristics could increase food safety and shelf life. [source]

LISTERIA MONOCYTOGENES AND ESCHERICHIA COLI O157:H7 INHIBITION IN VITRO BY LIPOSOME-ENCAPSULATED NISIN AND ETHYLENE DIAMINETETRAACETIC ACID

JOURNAL OF FOOD SAFETY, Issue 2 2008
T. MATTHEW TAYLOR
ABSTRACT Encapsulation technologies that effectively reduce antimicrobial interaction with food components or protect antimicrobial compounds from food processing measures have the potential to improve the microbiological safety of ready-to-eat foods. Recent application of liposomes for the preservation of cheese has spurred research into their utility in other food matrices. To ascertain the feasibility of encapsulated antimicrobial for the control of Listeria monocytogenes and Escherichia coli O157:H7 growth in a model system, nisin (5.0 and 10.0 µg/mL) and the chelator ethylene diaminetetraacetic acid were entrapped in phospholipid liposomes. While phosphatidylcholine (PC) liposomes did not produce significant inhibition of target pathogens, PC/phosphatidylglycerol 8/2 and 6/4 (mol%) produced significant inhibition of pathogens. Near-complete inhibition of E. coli O157:H7 with liposomal antimicrobials at concentrations below those reported necessary for unencapsulated antimicrobial and chelator suggests that liposomes may represent a powerful technology for the encapsulation of antimicrobials and the control of foodborne pathogens. PRACTICAL APPLICATIONS The activity of many antimicrobials is abolished in many food products for a variety of reasons. Interference and cross-reactions of the antimicrobial and various food constituents, such as protein and fat, are difficult to overcome and often require large amounts of antimicrobial in order to gain significant reductions in the pathogen load in a product. Loss of solubility of some antimicrobials based on pH or ionic strength will negatively affect the antimicrobial potential of a compound like nisin. Liposome encapsulation technologies, such as that reported here, may allow for the maintenance of antimicrobial activity by protecting the antimicrobial against cross-reactions with food components. Additionally, the liposome core represents a microenvironment which can be manipulated by the manufacturer in order to preserve optimal antimicrobial solubility and stability conditions until the time of release. [source]

INHIBITION OF LISTERIA INNOCUA AND L. MONOCYTOGENES IN A LABORATORY MEDIUM AND COLD-SMOKED SALMON CONTAINING LIQUID SMOKE

JOURNAL OF FOOD SAFETY, Issue 2 2001
SUSAN M. VITT
ABSTRACT Five commercial liquid smokes were tested in vitro and the most inhibitory to Listeria monocytogenes ATCC 19115 and L. innocua ATCC 33090 was Charsol Supreme. Chum salmon samples (100-g each) were brined, dipped for 15 s at varying concentrations of liquid smoke, inoculated with L. innocua, cold-processed and analyzed. Liquid smoke concentrations of 60,100% reduced L. innocua by 3-log10/g in the final product. Dwell times of 15 s to 5 min using 60% liquid smoke gradually decreased Listeria survival with an optimum 5-min dip. Isoeugenol was antilisterial in vitro but lacked synergism with liquid smoke in cold-smoked salmon. An immunoassay kit detected low inoculum levels (< 100 CFU/g) of L. innocua in one of three samples that were treated with liquid smoke for two and four minutes. Charsol Supreme was antilisterial but could not be relied on to totally eliminate Listeria in cold-smoked salmon. Panelists found the 0 to 2-min dipped sockeye salmon slightly desirable with no significant (p < 0.05) differences. The 5-min treatment was significantly (p < 0.05) darker, scored lower in desirability and flavor and contained 93 ppm of phenolic compounds. [source]

ACE INHIBITION PRESERVES GLOMERULAR NEPHRIN EXPRESSION IN EXPERIMENTAL DIABETIC NEPHROPATHY

NEPHROLOGY, Issue 1 2002
JR Rumble
[source]

REDUCED MATRIX DEGRADATION IN DIABETIC NEPHROPATHY: AMELIORATION WITH ACE INHIBITION

NEPHROLOGY, Issue 3 2000
McLennan Sv
[source]

INHIBITION OF TISSUE FACTOR EXPRESSED BY ISCHAEMIC CARDIOMYOCYTES REDUCES INFARCT SIZE AFTER MYOCARDIAL ISCHEMIA/REPERFUSION INJURY

NEPHROLOGY, Issue 3 2000
Erlich Jh
[source]

INHIBITION OF BRAIN RENIN,ANGIOTENSIN SYSTEM IMPROVES DIASTOLIC CARDIAC FUNCTION FOLLOWING MYOCARDIAL INFARCTION IN RATS

CLINICAL AND EXPERIMENTAL PHARMACOLOGY AND PHYSIOLOGY, Issue 8 2009
IG Araujo
SUMMARY 1Recently, we demonstrated that oral captopril treatment improved diastolic function and attenuated cardiac remodelling after myocardial infarction (MI) in rats. Considering the feasible role of the brain renin,angiotensin system (RAS) in heart failure, in the present study we investigated the role of the captopril injected intracerebroventricularly (i.c.v.) on the progression of cardiac dysfunction. 2Male Wistar rats underwent experimental MI or sham operation. Infarcted animals received daily i.c.v. injections of captopril (approximately 200 mg/kg; MI + Cap) or saline (MI) from 11 to 18 days after infarction. Electro- and echocardiogram assessments were performed before and after i.c.v. treatment (10 and 18 days after MI, respectively). Water and hypertonic saline ingestion were determined daily between 12 and 16 days after MI. 3Electrocardiograms from the MI and MI + Cap groups showed signs that resembled large MI before and after i.c.v. treatment. However, despite similar systolic dysfunction observed in both groups, only captopril-treated rats exhibited reduced left ventricular (LV) dilatation and improved LV filling, as assessed by echocardiograms, and low levels of water ingestion compared with the saline-treated control group. 4The results of the present study suggest that the brain RAS may participate in the development of cardiac dysfunction induced by ischaemia and that inhibition of the brain RAS may provide a new strategy for the prevention of diastolic dysfunction. [source]

AN S296R MUTATION IN THE HUMAN ANDROGEN RECEPTOR CAUSES ACTIVATION OF THE RECEPTOR BY NON-ANDROGENIC STEROIDS AND STRONGER INHIBITION BY THE NUCLEAR RECEPTOR COREPRESSOR N-coR

CLINICAL AND EXPERIMENTAL PHARMACOLOGY AND PHYSIOLOGY, Issue 10 2008
Yi-Dong Li
SUMMARY 1Mutation of the androgen receptor (AR) is believed to contribute to androgen-independent growth of prostate cancer. In the present study, we examined the functional changes associated with the novel somatic mutation S296R in the N-terminal domain of the AR identified from one recurrent prostate cancer sample. 2The results indicate that the S296R mutation does not differ obviously from the wild-type AR in its ability to bind the synthetic androgen methyltrienolone, or in its transcriptional activity induced by dihydrotestosterone (DHT) in the absence or presence of the overexpression of coactivators (steroid receptor coactivator-1, transcription intermediary factor-2, cAMP response element-binding protein-binding protein and p300). However, S296R was found to differ from wild-type AR in that its transcriptional activity could be activated by high concentrations (1 µmol/L) of 17,-oestradiol and progesterone and its transactivity induced by DHT was more obviously inhibited by overexpression of the nuclear receptor corepressor N-coR in CV-1 cells. 3These findings indicate that a point mutation (S296R) in the N-terminal domain of the AR can decrease the ligand specificity of the AR and alter the interaction between S296R and the corepressor N-coR. [source]

INHIBITION OF TOLL-LIKE RECEPTOR-4, NUCLEAR FACTOR-,B AND MITOGEN-ACTIVATED PROTEIN KINASE BY LIGNOCAINE MAY INVOLVE VOLTAGE-SENSITIVE SODIUM CHANNELS

CLINICAL AND EXPERIMENTAL PHARMACOLOGY AND PHYSIOLOGY, Issue 9 2008
Ping-Ying Lee
SUMMARY 1We have shown previously that lignocaine inhibits the upregulation of inducible nitric oxide synthase (iNOS), a crucial factor that initiates the systemic inflammatory response during sepsis, possibly through voltage-sensitive sodium channels (VSSC). Toll-like receptor-4 (TLR-4), nuclear factor (NF)-,B and mitogen activated protein kinases (MAPKs) participate in the upstream regulation of iNOS expression induced by endotoxin. In the present study, we investigated the effects of lignocaine in the regulation of the expression of these enzymes. The role of VSSC in the effects of lignocaine was also investigated. 2Confluent murine macrophages (RAW264.7 cells) were randomized to receive lipopolysaccharide (LPS; 100 ng/mL), LPS + lignocaine (50 µmol/L), LPS + tetrodotoxin (TTX; 1 µmol/L; a VSSC inhibitor), LPS + lignocaine + veratridine (Ver; 50 µmol/L; a VSSC activator) or LPS + TTX + Ver. After reacting with LPS for 0, 15, 30, 45 and 60 min, cell cultures were harvested and enzyme expression was evaluated. 3We found that LPS significantly increased the concentrations of TLR-4, NF-,B and MAPKs, including extracellular regulated kinase (ERK), c-jun N-terminal kinase (JNK) and p38 MAPK, in activated macrophages. Lignocaine and TTX significantly attenuated the effects of LPS on TLR-4, NF-,B, ERK and p38 MAPK expression, but not on JNK. Veratridine mitigated the effects of lignocaine and TTX. 4These data demonstrate that lignocaine has significant inhibitory effects on the activation of TLR-4, NF-,B and MAPKs in activated macrophages. Moreover, these effects involve VSSC. [source]

Overcoming surfactant inhibition with polymers

ACTA PAEDIATRICA, Issue 12 2000
PA Dargaville
Inhibition of the function of pulmonary surfactant in the alveolar space is an important element of the pathophysiology of many lung diseases, including meconium aspiration syndrome, pneumonia and acute respiratory distress syndrome. The known mechanisms by which surfactant dysfunction occurs are (a) competitive inhibition of phospholipid entry into the surface monolayer (e.g. by plasma proteins), and (b) infiltration and destabilization of the surface film by extraneous lipids (e.g. meconium-derived free fatty acids). Recent data suggest that addition of non-ionic polymers such as dextrano and polyethylene glycol to surfactant mixtures may significantly improve resistance to inhibition. Polymers have been found to neutralize the effects of several different inhibitors, and can produce near-complete restoration of surfactant function. The anti-inhibitory properties of polymers, and their possible role as an adjunct to surfactant therapy, deserve further exploration. [source]

The Potential of Soluble Epoxide Hydrolase Inhibition in the Treatment of Cardiac Hypertrophy

CONGESTIVE HEART FAILURE, Issue 4 2008
Todd R. Harris PhD
First page of article [source]

Potential Role of Type 5 Phosphodiesterase Inhibition in the Treatment of Congestive Heart Failure

CONGESTIVE HEART FAILURE, Issue 1 2003
Stuart D. Katz MD
Endothelial dysfunction is associated with impairment of aerobic capacity in patients with heart failure and may play a role in the progression of disease. Impaired endothelium-dependent vasodilation in patients with heart failure can be attributed to decreased bioavailability of nitric oxide and attenuated responses to nitric oxide in vascular smooth muscle. Impaired vasodilation in response to nitric oxide derived from vascular endothelium or organic nitrates in vascular smooth muscle may be related in part to increased degradation of the second messenger cyclic guanosine monophosphate by type 5 phosphodiesterase. Sildenafil, a specific type 5 phosphodiesterase inhibitor currently approved for the treatment of erectile dysfunction, has been shown to acutely enhance endothelium-dependent vasodilation in patients with heart failure. Further studies are warranted to characterize the safety and efficacy of type 5 phosphodiesterase inhibition in the treatment of chronic heart failure. [source]

Orexins/hypocretins control bistability of hippocampal long-term synaptic plasticity through co-activation of multiple kinases

ACTA PHYSIOLOGICA, Issue 3 2010
O. Selbach
Abstract Aim:, Orexins/hypocretins (OX/Hcrt) are hypothalamic neuropeptides linking sleep,wakefulness, appetite and neuroendocrine control. Their role and mechanisms of action on higher brain functions, such as learning and memory, are not clear. Methods:, We used field recordings of excitatory post-synaptic potentials (fEPSP) in acute mouse brain slice preparations to study the effects of orexins and pharmacological inhibitors of multiple kinases on long-term synaptic plasticity in the hippocampus. Results:, Orexin-A (OX-A) but not orexin-B (OX-B) induces a state-dependent long-term potentiation of synaptic transmission (LTPOX) at Schaffer collateral-CA1 synapses in hippocampal slices from adult (8- to 12-week-old) mice. In contrast, OX-A applied to slices from juvenile (3- to 4-week-old) animals causes a long-term depression (LTDOX) in the same pathway. LTPOX is blocked by pharmacological inhibition of orexin receptor-1 (OX1R) and plasticity-related kinases, including serine/threonine- (CaMKII, PKC, PKA, MAPK), lipid- (PI3K), and receptor tyrosine kinases (Trk). Inhibition of OX1R, CaMKII, PKC, PKA and Trk unmasks LTDOX in adult animals. Conclusion:, Orexins control not only the bistability of arousal states and threshold for appetitive behaviours but, in an age- and kinase-dependent manner, also bidirectional long-term synaptic plasticity in the hippocampus, providing a possible link between behavioural state and memory functions. [source]

The role of inhibitory neurotransmission in locomotor circuits of the developing mammalian spinal cord

ACTA PHYSIOLOGICA, Issue 2 2009
H. Nishimaru
Abstract Neuronal circuits generating the basic coordinated limb movements during walking of terrestrial mammals are localized in the spinal cord. In these neuronal circuits, called central pattern generators (CPGs), inhibitory synaptic transmission plays a crucial part. Inhibitory synaptic transmission mediated by glycine and GABA is thought to be essential in coordinated activation of muscles during locomotion, in particular, controlling temporal and spatial activation patterns of muscles of each joint of each limb on the left and right side of the body. Inhibition is involved in other aspects of locomotion such as control of speed and stability of the rhythm. However, the precise roles of neurotransmitters and their receptors mediating inhibitory synaptic transmission in mammalian spinal CPGs remain unclear. Moreover, many of the inhibitory interneurones essential for output pattern of the CPG are yet to be identified. In this review, recent advances on these issues, mainly from studies in the developing rodent spinal cord utilizing electrophysiology, molecular and genetic approaches are discussed. [source]

When is high-Ca2+ microdomain required for mitochondrial Ca2+ uptake?,

ACTA PHYSIOLOGICA, Issue 1 2009
A. Spät
Abstract Ca2+ release from IP3 -sensitive stores in the endoplasmic reticulum (ER) induced by Ca2+ -mobilizing agonists generates high-Ca2+ microdomains between ER vesicles and neighbouring mitochondria. Here we present a model that describes when such microdomains are required and when submicromolar [Ca2+] is sufficient for mitochondrial Ca2+ uptake. Mitochondrial Ca2+ uptake rate in angiotensin II-stimulated H295R adrenocortical cells correlates with the proximity between ER vesicles and the mitochondrion, reflecting the uptake promoting effect of high-Ca2+ peri-mitochondrial microdomains. Silencing or inhibition of p38 mitogen-activated protein kinase (MAPK) or inhibition of the novel isoforms of protein kinase C enhances mitochondrial Ca2+ uptake and abolishes the positive correlation between Ca2+ uptake and ER-mitochondrion proximity. Inhibition of protein phosphatases attenuates mitochondrial Ca2+ uptake and also abolishes its positive correlation with ER-mitochondrion proximity. We postulate that during IP3 -induced Ca2+ release, Ca2+ uptake is confined to ER-close mitochondria, because of the simultaneous activation of the protein kinases. Attenuation of Ca2+ uptake prevents Ca2+ overload of mitochondria and thus protects the cell against apoptosis. On the other hand, all the mitochondria accumulate Ca2+ at a non-inhibited rate during physiological Ca2+ influx through the plasma membrane. Membrane potential is higher in ER-distant mitochondria, providing a bigger driving force for Ca2+ uptake. Our model explains why comparable mitochondrial Ca2+ signals are formed in response to K+ and angiotensin II (equipotent in respect to global cytosolic Ca2+ signals), although only the latter generates high-Ca2+ microdomains. [source]

Simultaneous flow cytometric detection of basophil activation marker CD63 and intracellular phosphorylated p38 mitogen-activated protein kinase in birch pollen allergy,

CYTOMETRY, Issue 1 2009
Nicolaas E. Aerts
Abstract Background: Phosphorylation of p38 MAPK is a crucial step in IgE-receptor signaling in basophils. The relation of p38 MAPK to the well-validated diagnostic cell surface marker CD63 has not been evaluated in a clinical allergy model. Methods: Expression of CD63 and phosphorylation of p38 MAPK were analyzed flow cytometrically in anti-IgE-gated basophils from 18 birch pollen allergic patients, five grass pollen allergic patients, and five healthy individuals, after 3 and 20 min of stimulation with recombinant major birch pollen allergen (rBet v 1). Additional time points and the influence of p38 MAPK inhibitor SB203580 were studied in birch pollen allergic patients. Results: Phospho-p38 MAPK and CD63 were expressed dose-dependently in birch pollen allergic patient basophils within 1 minute of rBet v 1 stimulation. P38 MAPK phosphorylation was fastest and subsided gradually while CD63 expression remained elevated for at least 20 min. Inhibition of p38 MAPK significantly inhibited CD63 upregulation. With optimal stimulation of the cells (1 ,g/mL), sensitivity and specificity for the discrimination between patients and a group of control individuals (grass pollen allergic patients and healthy controls) were 94% and 100% for CD63 at 3 and 20 min and for phospho-p38 MAPK at 3 min. Conclusion: Antigen-induced p38 MAPK phosphorylation in human basophils essentially contributes to CD63 upregulation. It is a sensitive and specific intracellular marker for allergy diagnosis and offers new insight into the mechanisms of basophil activation. © 2008 Clinical Cytometry Society [source]

Directional change produced by perpendicularly-oriented microgrooves is microtubule-dependent for fibroblasts and epithelium

CYTOSKELETON, Issue 5 2009
Douglas W. Hamilton
Abstract Anisotropic substrata such as micromachined grooves can control cell shape, orientation, and the direction of cell movement, a phenomena termed topographic guidance. Although many types of cells exhibit topographic guidance, little is known regarding cell responses to conflicting topographic cues. We employed a substratum with intersecting grooves in order to present fibroblasts and epithelial cells with conflicting topographic cues. Using time-lapse and confocal microscopy, we examined cell behavior at groove intersections. Migrating fibroblasts and epithelial cells typically extended a cell process into the intersection ahead of the cell body. After travelling along the "X" groove to enter the intersection, the leading lamellipodia of the cell body encountered the perpendicular "Y" groove, and spread latterly along the "Y" groove. The formation of lateral lamellipodia resulted in cells forming "T" or "L" morphologies, which were characterized by the formation of phosphotyrosine-rich focal adhesions at the leading edges. The "Y" groove did not prove an absolute barrier to cell migration, particularly for epithelial cells. Analysis of cytoskeletal distribution revealed that F-actin bundles did not adapt closely to the groove patterns, but typically did align to either the "X" or "Y" grooves. In contrast microtubules (MT) adapted closely to the walls. Inhibition of microtubule nucleation attenuated fibroblast and epithelial cell orientation within the intersection of the perpendicular grooves. We conclude that MT may be the prime determinant of fibroblast and epithelial cell conformation to conflicting topographies. Cell Motil. Cytoskeleton 2009. © 2009 Wiley-Liss, Inc. [source]

The involvement of phospholipases C and D in the asymmetric division of subsidiary cell mother cells of Zea mays

CYTOSKELETON, Issue 11 2008
Panagiotis Apostolakos
Abstract In the present study, the involvement of phospholipase C and D (PLC and PLD) pathways in the asymmetric divisions that produce the stomatal complexes of Zea mays was investigated. In particular, the polar organization of microtubules (MTs) and actin filaments (AFs) and the process of asymmetric division were studied in subsidiary cell mother cells (SMCs) treated with PLC and PLD modulators. In SMCs treated with butanol-1 (but-1), which blocks phosphatidic acid (PA) production via PLDs, AF-patch formation laterally to the inducing guard cell mother cell (GMC) and the subsequent asymmetric division were inhibited. In these SMCs, cell division plane determination, as expressed by MT preprophase band (MT-PPB) formation, was not disturbed. Exogenously applied PA partially relieved the but-1 effects on SMCs. In contrast to SMCs, but-1 did not affect the symmetric GMC division. Inhibition of the PLC catalytic activity by neomycin or U73122 resulted in inhibition of asymmetric SMC division, while AF-patch and MT-PPB were organized as in control SMCs. These data show that the PLC and PLD signaling pathways are involved in the transduction and/or perception of the inductive stimulus that is emitted by the GMCs and induces the polar AF organization and asymmetric SMC division. In contrast, division plane determination in SMCs, as expressed by MT-PPB formation, does not depend on PLC and PLD signaling pathways. Cell Motil. Cytoskeleton 2008. © 2008 Wiley-Liss, Inc. [source]

In vivo phosphorylation of regulatory light chain of myosin II in sea urchin eggs and its role in controlling myosin localization and function during cytokinesis

CYTOSKELETON, Issue 2 2008
Ryota Uehara
Abstract Phosphorylation of myosin regulatory light chain (RLC) at Ser19 (mono-phosphorylation) promotes filament assembly and enhances actin-activated ATPase activity of non-muscle myosin, while phosphorylation at both Ser19 and Thr18 (di-phosphorylation) further enhances the ATPase activity. However, it has not well been addressed which type of phosphorylation is important in regulating myosin during cytokinesis. Here, we investigated subcellular localization in sea urchin eggs of mono-phosphorylated and di-phosphorylated RLC by both quantitative biochemical and spatiotemporal cytological approaches. Mono-phosphorylated RLC was dominant in the equatorial cortex throughout the whole process of cytokinesis. Inhibition of myosin light chain kinase (MLCK) decreased mono-phosphorylated RLC both in the cortex and in the cleavage furrow, and blocked both formation and contraction of the contractile ring. Two different types of ROCK inhibitor gave inconsistent results: H1152 blocked both RLC mono-phosphorylation in the cleavage furrow and contraction of the contractile ring, while Y27632 affected neither the mono-phosphorylation nor cell division. These results suggest that there may be other targets of H1152 than ROCK, which is involved in the RLC phosphorylation in the cleavage furrow. Furthermore, it was revealed that localization of myosin heavy chain in the cleavage furrow, but not in the cortex, was perturbed by inhibition of RLC mono-phosphorylation. These results suggested that RLC mono-phosphorylation by more than two RLC kinases play a main role in regulation and localization of myosin in the dividing sea urchin eggs. Cell Motil. Cytoskeleton 2007. © 2007 Wiley-Liss, Inc. [source]

Na+/Ca2+ exchanger modulates the flagellar wave pattern for the regulation of motility activation and chemotaxis in the ascidian spermatozoa

CYTOSKELETON, Issue 10 2006
Kogiku Shiba
Abstract Ion channels and ion exchangers are known to be important participants in various aspects of sperm physiology, e.g. motility activation, chemotaxis, the maintenance of motility and the acrosome reaction in the sperm. We report here on a role of the K+ -independent Na+/Ca2+ exchanger (NCX) on ascidian sperm. Reverse-transcriptase PCR reveals that the NCX is expressed in the testis while immunoblotting and immunolocalization demonstrate that the NCX exists on the sperm in the ascidian Ciona savignyi and C. intestinalis. A potent blocker of the NCX, KB-R7943 was found to block sperm-activating and -attracting factor (SAAF)-induced motility activation, sperm motility and sperm chemotaxis. We further analyzed the effects of this blocker on motility parameters such as the flagellar waveform, curvature, beat frequency, amplitude and wavelength of the sperm flagella. Inhibition of the NCX caused two distinct effects: a low concentration of KB-R7943 induced symmetric bending, whereas a high concentration of KB-R7943 resulted in asymmetric flagellar bending. These findings suggest that the NCX plays important roles in the regulation of SAAF-induced sperm chemotaxis, motility activation and motility maintenance in the ascidian. This study provides new information toward an understanding of Ca2+ transport systems in sperm motility and chemotaxis. Cell Motil. Cytoskeleton 2006. © 2006 Wiley-Liss, Inc. [source]

Myoblast attachment and spreading are regulated by different patterns by ubiquitous calpains

CYTOSKELETON, Issue 4 2006
Germain Mazères
Abstract The calcium-dependent proteolytic system is a large family of well-conserved ubiquitous and tissue-specific proteases, known as calpains, and an endogenous inhibitor, calpastatin. Ubiquitous calpains are involved in many physiological phenomena, such as the cell cycle, muscle cell differentiation, and cell migration. This study investigates the regulation of crucial steps of cell motility, myoblast adhesion and spreading, by calpains. Inhibition of each ubiquitous calpain isoform by antisense strategy pinpointed the involvement of each of these proteases in myoblast adhesion and spreading. Moreover, the actin cytoskeleton and microtubules were observed in transfected cells, demonstrating that each ubiquitous calpain could be involved in the actin fiber organization. C2C12 cells with reduced ,- or m-calpain levels have a rounded morphology and disorganized stress fibers, but no modification in the microtubule cytoskeleton. Antisense strategy directed against MARCKS, a calpain substrate during C2C12 migration, showed that this protein could play a role in stress fiber polymerization. A complementary proteomic analysis using C2C12 cells over-expressing calpastatin indicated that two proteins were under-expressed, while six, which are involved in the studied phenomena, were overexpressed after calpain inhibition. The possible role of these proteins in adhesion, spreading, and migration was discussed. Cell Motil. Cytoskeleton 63: 2006. © 2006 Wiley-Liss, Inc. [source]

Sustained increase in arterial blood pressure and vascular resistance induced by infusion of arachidonic acid in rats

ACTA PHYSIOLOGICA, Issue 1 2000
Kirkebø
The haemodynamic responses to arachidonic acid (AA) have been investigated in seven groups of anaesthetized rats. Sodium arachidonate was infused intravenously for 4 or 20 min, and arterial blood pressure was recorded continuously. Cardiac output and organ blood flow were measured by microspheres. Infusion of arachidonate caused first a fast drop in arterial blood pressure, thereafter it increased steadily for 5,15 min towards a pressure about 25 mmHg above control level. The high pressure was maintained for at least 1 h. Repeated infusions of arachidonate gave similar responses. Inhibition of cyclo-oxygenase by indomethacin prevented the initial pressure drop to arachidonate, but not the sustained increase in pressure. Arterial pressure, total vascular resistance and blood flow in the kidneys, adrenals and spleen were significantly reduced, whereas cardiac output was not changed 4 min after start infusion of arachidonate. However, average blood pressure was significantly increased 22 and 35 min after start infusion (from 103.9 ± 2.9 to 128.1 ± 6.1 and 135.8 ± 4.6 mmHg). Mean vascular resistance increased simultaneously (from 3.5 ± 0.2 to 4.7 ± 0.4 and 5.2 ± 0.4 mmHg 100 mL,1), while cardiac output, stroke volume and heart rate were maintained or slightly reduced. The renal blood flow was significantly lowered (from average 4.9 ± 0.1 to 3.3 ± 0.2 and 4.0 ± 0.2 mL min,1). Indomethacin did not prevent the changes in vascular resistance or organ blood flow recorded after 20,35 min. On the other hand, inhibition of both cyclo-oxygenase, lipoxygenase and the cytochrome P450 pathways by eicosatetrayonic acid (ETYA) normalized all haemodynamic parameters. Likewise, the rise in pressure was prevented by 17-octadecynoic acid (17-ODYA), an inhibitor of the cytochrome P450 enzyme activity. Thus, arachidonate infusion caused a transient decrease, and then a sustained increase in arterial pressure and vascular resistance, and a long-lasting reduction in renal blood flow, possibly owing to release of a cytochrome P450 dependent vasoconstrictor metabolite of AA. [source]

Remodeling of the actin cytoskeleton of target hepatocytes and NK cells during induction of apoptosis

CYTOSKELETON, Issue 2 2001
W. Marty Blom
Abstract Natural Killer cells are immune cells that recognize and eliminate altered and non-self cells from the circulation. To study the interaction between NK cells and target cells, we set up an experimental system consisting of rat Interleukin-2 activated Natural Killer cells (A-NK cells) and rat hepatocytes with a masked Major Histocompatibility Complex (MHC). The masking of the MHC induces recognition of the hepatocytes by the NK cells as non-self. We showed that in vitro apoptosis is rapidly induced in the hepatocytes [Blom et al., 1999] after co-incubation with A-NK cells. Now we describe the morphological changes that occur during and after interaction of A-NK cells with hepatocytes. Confocal laser scanning microscopy showed that the actin cytoskeleton of the NK cells was remodeled during attack of hepatocytes. Some NK cells were in close contact with the hepatocytes while others had formed actin-containing dendrites of varying length that made contact with the hepatocytes. However, dendrite formation is not obligatory for induction of apoptosis because cells that were unable to form these did induce FAS-dependent apoptosis in hepatocytes. Apparently both direct as well as distant contact resulted in apoptosis. Formation of the dendrites was calcium-dependent as EGTA largely prevented it. Importantly, chelation of the calcium also suppressed killing of the hepatocytes. Within 1 h after addition of the A-NK cells, morphological changes in hepatocytes that are characteristic of apoptosis, such as the formation of apoptotic bodies and fragmented nuclei, became apparent. Specifically, the actin cytoskeleton of the hepatocytes was remodeled resulting in the formation of the apoptotic bodies. Inhibition of caspase activity by z-Val-Ala-DL-Asp-fluoromethylketone (100 ,M) partly protected against the rearrangement of the actin filaments in the hepatocytes. Cell Motil. Cytoskeleton 49:78,92, 2001. © 2001 Wiley-Liss, Inc. [source]

Paruresis (Psychogenic Inhibition of Micturition): cognitive behavioral formulation and treatment

DEPRESSION AND ANXIETY, Issue 11 2008
Mark J. Boschen Ph.D.
Abstract Paruresis is a condition characterized by difficulty or inability to urinate in situations where others are present, or may soon be present. Despite knowledge that paruresis can significantly impact on occupational functioning, social functioning, and quality of life, there exists a paucity of research into effective treatments. Although cognitive conceptualizations have been advanced for other anxiety disorders, there has not been a comprehensive cognitive behavioral model of paruresis. This article presents a revised cognitive and behavioral conceptualization of paruresis, drawing on empirical evidence from other anxiety disorders. Using this conceptualization, a cognitive-behavioral intervention strategy is outlined, with clear targets for cognitive and behavioral strategies. Depression and Anxiety, 2008. © 2007 Wiley-Liss, Inc. [source]