PubMed Health⌕ Search

Biomedical subjects

L W Chen

Publications and source records attributed to L W Chen.

At least 19 recordsLinked to original sources
PubMed2002

Effect of neuropsychiatric symptoms of Alzheimer's disease on Chinese and American caregivers.

BACKGROUND: In Chinese culture, extended family support, acceptance of age-related cognitive changes and filial tradition of caring for elders may decrease caregiver burden and distress in the context of dementia. OBJECTIVE: To study cross-regional and cross-cultural differences in symptom-related caregiver distress due to the behavioral problems of Chinese and American patients with Alzheimer's disease. METHOD: Caregivers of patients with Alzheimer's disease at Taipei Veterans General Hospital, Taiwan (n = 89), Chinese University of Hong Kong (n = 31) and the UCLA Alzheimer's Disease Research Center, Los Angeles, California (n = 169) reported the neuropsychiatric symptoms of patients and their corresponding distress on the Neuropsychiatric Inventory. RESULT: Presence or absence of distress due to the neuropsychiatric symptoms of the patients with Alzheimer's disease was assessed. The three centers differed significantly in the proportions of caregivers with distress caused by depression (p < 0.05) and apathy (p < 0.001). UCLA had higher proportions of caregivers with depression-related distress than Taipei. UCLA caregivers were also more stressed by apathy than caregivers in Taipei and Hong Kong. Logistic regression further supported the findings that depression-related and apathy-related caregiver distress differed between Chinese and American caregivers (p < 0.05). CONCLUSIONS: The results were surprising, in that American and Chinese (Taipei and Hong Kong) caregivers exhibited similar distress or lack of distress in response to delusions, hallucinations, agitation, anxiety, euphoria, disinhibition, irritability, aberrant motor behavior, sleep and appetite symptoms of Alzheimer's disease patients. Chinese caregivers were less affected by depression and apathy in patients with Alzheimer's disease than Caucasian caregivers.

Aged↗
PubMed2001

Cholinergic neurons expressing substance P receptor (NK(1)) in the basal forebrain of the rat: a double immunocytochemical study.

Cholinergic neurons expressing substance P receptor (SPR, NK(1)) were examined in the rat brain using double immunofluorescence. The distribution of SPR-like immunoreactive (SPR-LI) neurons completely overlapped with that of choline acetyltransferase (ChAT)-LI neurons in the medial septal nucleus, the nucleus of diagonal band of Broca, the magnocellular preoptic nucleus, the substantia innominata of basal forebrain, the caudate-putamen, and the ventral pallidum of the basal ganglia. In the mesopontine tegmentum and the cranial motor nuclei of the brainstem, the distribution of SPR-LI and ChAT-LI neurons was partially overlapping. Neurons showing both SPR-like and ChAT-like immunoreactivities, however, were predominantly found above basal forebrain regions and 82-90% of these ChAT-LI neurons displayed SPR-like immunoreactivity, in addition to the confirmatory observation that 100% of the ChAT-LI neurons exhibit SPR-like immunoreactivity in the basal ganglia. In contrast, neurons double-labeled for SPR-like and ChAT-like immunoreactivities were hardly detected in aforementioned regions of the brainstem. The present study has provided morphological evidence for direct physiological modulation of cholinergic neurons by tachykinins through substance P receptor in the basal forebrain of the rat.

Acetylcholine↗
PubMed2001

Probing reactive sites for ion-molecule reactions of anthraquinones with dimethyl ether using an external source ion trap tandem mass spectrometer and computational chemistry.

Gas-phase ion-molecule reactions of anthraquinone derivatives with dimethyl ether (DME) were investigated using an external source ion trap mass spectrometer. Semi-empirical calculations were executed to determine possible reactive sites for the product ions. Collision activated dissociation (CAD) was successfully performed for very low abundance of ion-molecule products. Even for product ions with a relative intensity below 1%, CAD experiments can be successfully performed. Significantly more structural information could be elucidated based on this special feature. Importantly, the CAD spectra of very minor ions could be measured by this ion trap instrument, which significantly enhances the future role of the ion trap as a powerful analytical instrument. CAD of all product ions on anthraquinone compounds typically eliminates neutral molecules such as CO or H(2)O. A hydration phenomenon in the CAD processes resulting from the precursor ions incorporating one molecule of H(2)O and then eliminating one molecule of CO was observed in this study.

Anthraquinones↗
PubMed2001

Reactivation of hepatitis C virus superinfection in a patient seropositive for hepatitis B e antigen.

During the course of chronic hepatitis B virus (HBV) infection, a patient seropositive for hepatitis B e antigen experienced four episodes of acute hepatic necroinflammation. Serum HBV-DNA concentration elevated immediately before the first and third exacerbations, whereas serum hepatitis C virus (HCV) RNA was detected during the second and fourth exacerbations. The nucleotide sequences of HCV hypervariable region derived from samples of the two exacerbations were identical. Interestingly, "de novo" seroconversion of anti-HCV antibody (Abbott HCV EIA 3.0) followed by reversion occurred in both the second and fourth exacerbations with low sample/cutoff ratios. Immunoblot analysis using a line-immunoassay (Inno-LIA HCV Ab III) revealed a single positive band (C1) developing after the second exacerbation. These data indicate that the second exacerbation in this patient was caused by newly acquired acute HCV superinfection, whereas the fourth exacerbation was likely due to reactivation of the previous HCV infection. Recognition of such a case suggests that the presence of de novo seroconversion of anti-HCV may indicate either reactivation or acute superinfection of HCV in a patient seropositive for hepatitis B e antigen.

Adult↗
PubMed2001

Cholinergic neurons expressing neuromedin K receptor (NK3) in the basal forebrain of the rat: a double immunofluorescence study.

By using a double immunofluorescence method we have examined the distribution of cholinergic neurons expressing neuromedin K receptor (NK3) in the rat brain and spinal cord. The distribution of neuromedin K receptor-like immunoreactive neurons completely overlapped with that of choline acetyltransferase-positive neurons in certain regions of the basal forebrain, e.g. the medial septal nucleus, nucleus of the diagonal band of Broca, magnocellular preoptic nucleus and substantia innominata. Partially overlapping distributions of neuromedin K receptor-like immunoreactive and choline acetyltransferase-positive neurons were found in the basal nucleus of Meynert, globus pallidus, ventral pallidum of the forebrain, tegmental nuclei of the pons and dorsal motor nucleus of the vagus. Neurons showing both neuromedin K receptor-like and choline acetyltransferase immunoreactivities, however, were found predominantly in the medial septal nucleus, nucleus of the diagonal band of Broca and magnocellular preoptic nucleus of the basal forebrain: 66-80% of these choline acetyltransferase-positive neurons displayed neuromedin K receptor-like immunoreactivity. Neurons showing both neuromedin K receptor-like and choline acetyltransferase immunoreactivities were hardly detected in other aforementioned regions of the forebrain, brainstem and spinal cord. The present study has provided morphological evidence for direct physiological modulation or regulation of cholinergic neurons by tachykinins through the neuromedin K receptor in the basal forebrain of rats.

Animals↗
PubMed2001

Differential expression of AMPA receptor subunits in dopamine neurons of the rat brain: a double immunocytochemical study.

We have examined the distribution of dopamine neurons expressing alpha-amino-3-hydroxy-5-methyl-4-isoxazole propionate (AMPA) receptor subunits (glutamate receptors 1, 2/3 and 4) in the A8-A15 regions of the rat brain using double immunofluorescence. The distribution of glutamate receptor 1- or 2/3-like immunoreactive neurons completely overlapped that of tyrosine hydroxylase-like immunoreactive neurons in dopamine cell groups in the retrorubral field (A8), the substantia nigra (A9), the ventral tegmental area and the nucleus raphe linealis (A10), and the rostral hypothalamic periventricular nucleus (A14, A15). In the caudal hypothalamic periventricular nucleus (A11), arcuate nucleus (A12) and zona incerta (A13), the distribution was partially overlapping. Neurons double-labeled for tyrosine hydroxylase and glutamate receptor 1 or 2/3 immunoreactivities were, however, exclusively found in certain dopamine cell regions: in areas A14-A15, 85-88% of tyrosine hydroxylase-containing neurons expressed glutamate receptor 1 and 22-25% expressed glutamate receptor 2/3, while in areas A8-A10, 20-43% expressed glutamate receptor 1 and 63-84% expressed glutamate receptor 2/3. In contrast, the double-labeled neurons were hardly detected in the A11-A13 regions. No tyrosine hydroxylase-positive neurons displayed glutamate receptor 4 immunoreactivity, though a partially overlapping distribution of tyrosine hydroxylase- and glutamate receptor 4-immunopositive neurons was also seen in regions A8-10, A11 and A13. The present study has demonstrated the morphological evidence for direct modulation of dopamine neurons via AMPA receptors in rat mesencephalon and hypothalamus. This distribution may provide the basis for a selective dopamine neuron loss in neurodegenerative disorders, such as Parkinson's disease.

Animals↗
PubMed2001

Effect of the antianginal drug bepridil on intracellular Ca2+ release and extracellular Ca2+ influx in human neutrophils.

To understand more fully the effects of bepridil, an antiarrhythmic and antianginal drug, on myocardial ischemia-reperfusion injury and systemic immune responses, its effect on intracellular Ca2+ levels ([Ca2+]i) in human neutrophils was investigated by using fura-2 as a fluorescent probe. Bepridil (10-200 microM) increased [Ca2+]i in a concentration-dependent fashion. This signal was partly inhibited by removal of extracellular Ca2+. In a Ca(2+)-free medium, pretreatment with bepridil (100 microM) abolished the Ca2+ release induced by thapsigargin (1 microM), an endoplasmic reticulum Ca2+ pump inhibitor, and by carbonylcyanide m-chlorophenylhydrazone (2 microM), a mitochondrial uncoupler. Pretreatment with carbonylcyanide m-chlorophenylhydrazone and thapsigargin, respectively, partly inhibited bepridil-induced Ca2+ release. Addition of Ca2+ (3 mM) increased [Ca2+]i after pretreatment with bepridil (100 microM) in a Ca(2+)-free medium. Bepridil (100 microM)-induced Ca2+ release was not altered when phospholipase C was inhibited by U73122 (2 microM). Both Ca2+ release and Ca2+ entry induced by bepridil (100 microM) were augmented by activating protein kinase C with phorbol 12-myristate 13-acetate (10 nM), and were suppressed by inhibiting protein kinase C with GF 109203X (2 microM). Treatment with bepridil (10-20 microM) for 30 min increased the production of reactive oxygen intermediates (ROI) by more than 50%. Collectively, it was found that bepridil increased [Ca2+]i concentration-dependently in human neutrophils by releasing Ca2+ from the endoplasmic reticulum, mitochondria and, possibly, other compartments in a phospholipase C-independent manner. Bepridil also activated Ca2+ influx. The activity of protein kinase C may regulate bepridil-induced Ca2+ release and Ca2+ entry.

Angina Pectoris↗
PubMed2001

Mechanisms and modulation of formyl-methionyl-leucyl-phenylalanine (fMLP)-induced Ca2+ mobilization in human neutrophils.

The effect of fMLP (N-formyl-methionyl-leucyl-phenylalanine), a neutrophil-stimulating bacterial peptide, on Ca2+ mobilization in human neutrophils was examined using fura-2 as a Ca2+ indicator. fMLP (10 nM-10 microM) increased [Ca2+]i concentration-dependently. The [Ca2+]i signal comprised an initial rise followed by a gradual decay and a sustained phase. External Ca2+ removal partly decreased the signal. La3+ (50 microM) pretreatment mimicked the effect of Ca2+ removal. In Ca(2+)-free medium, pretreatment with 1 microM thapsigargin (an endoplasmic reticulum Ca2+ pump inhibitor) prevented 10 microM fMLP from increasing [Ca2+]i; whereas 1 microM thapsigargin still significantly increased [Ca2+]i after pretreatment with 10 microM fMLP. Addition of 3 mM Ca2+ induced a concentration-dependent [Ca2+]i increase after pretreatment with fMLP in Ca(2+)-free medium. This Ca2+ entry was partly inhibited by econazole (25 microM), SKF96365 (50 microM), and a phospholipase A2 inhibitor (aristolochic acid; 20 microM). The fMLP (10 microM)-induced Ca2+ release was abolished by inhibiting phospholipase C with 2 microM U73122. The fMLP-induced [Ca2+]i increase was inhibited by 25% by pretreatment with 10 nM phorbol ester to activate protein kinase C but was augmented by 27% by pretreatment with 2 microM GF 109203X to inactivate protein kinase C. We found that fMLP increase reactive oxygen intermediate (ROI) production in neutrophils, which can be suppressed by U73122 pretreatment. Collectively, this study shows that in human neutrophils, fMLP increased [Ca2+]i concentration-dependently by releasing Ca2+ from phospholipase C-coupled, thapsigargin-sensitive stores, accompanied by Ca2+ entry. The fMLP-induced [Ca2+]i rise was modulated by protein kinase C, and the fMLP-induced Ca2+ entry was abolished by La3+, and was reduced by econazole, SKF96365 and inhibition of phospholipase A2.

Calcium↗
PubMed2001

Inhibition of inducible nitric oxide synthase (iNOS) prevents lung neutrophil deposition and damage in burned rats.

This study was designed to investigate the role of NO and effect of iNOS inhibitor on the lung neutrophil deposition and damage after burn. In Experiment 1, specific pathogen-free (SPF) Sprague-Dawley rats underwent 35% total body surface area (TBSA) burn. On the 4th, 8th, 16th, and 24th h after burn, blood was collected for peroxynitrite-mediated dihydrorhodamine 123 (DHR 123) oxidation assay, and lung tissues were harvested for myeloperoxidase (MPO) test and histologic study. Pulmonary microvascular dysfunction was quantitated by measuring the extravasation of Evans blue dye (EBD). In Experiment 2, S-methylisothiourea (SMT) was given (7.5 mg/kg, intraperitoneal immediately post-burn) to suppress iNOS activity. On the 8th h after burn, the effect of SMT on blood DHR 123 oxidation, lung MPO, lung damage, and lung iNOS expression were evaluated. Lung MPO activity increased up to a maximum of 2-fold 8 h after burn. Blood DHR 123 oxidation increased up to a maximum of 2-fold 8 h after burn. Lung permeability increased up to a maximum of 2.5-fold 4 h after burn. SMT significantly decreased lung MPO activity, blood DHR 123 oxidation, and lung permeability by 31%, 41%, and 54%, respectively. SMT markedly decreased the thermal injury-induced perivascular and interstitial inflammatory cell infiltration and iNOS staining in bronchiolar epithelium, endothelial cells, and perivascular and interstitial inflammatory cells. In conclusion, thermal injury induces blood DHR 123 oxidation, lung neutrophil deposition, lung iNOS expression, and lung damage. Peroxynitrite might play an important role in thermal injury-induced lung neutrophil deposition and damage. Specific inhibition of lung iNOS expression and blood DHR 123 oxidation with SMT on thermal injury not only attenuated the lung neutrophil deposition, but also reduced lung damage.

Animals↗
PubMed2001

Analysis of binding domain and function of chimeric mu/kappa opioid receptors to ohmefentanyl stereoisomers.

AIM: To investigate specific domains in mu opioid receptors that accounted for selective binding of three stereoisomers of ohmefentanyl (Ohm9204, Ohm9202, and Ohm9203) and study the function of chimera II. METHODS: Rat mu and kappa opioid receptors (RMOR, RKOR) and four mu/kappa chimeric receptors (chimeras) I, II, III, and IV were transiently expressed in COS-1 cells. The binding ability and binding domain of receptor to ligands were determined by radioactive ligand and receptor binding experiments. Through measuring cellular cAMP levels, we studied the function of chimera II in mediating signal transduction. RESULTS: Binding affinities of four chimeric receptors were similar to wild type opioid receptors (RMOR and RKOR). The binding affinities of Ohm9204 and Ohm9202 to chimera II were similar to that of RMOR. The binding affinities of Ohm9203 to all six receptors were low. U50488 possessed high binding affinity to chimera I, however dynorphie A(1-9) had some binding affinity to chimera II that was similar to RKOR, which indicated the domains of RKOR accounting for selectively binding to peptide ligand dynorphie A(1-9) and nonpeptide ligand U50488 were different. The efficacy of Ohm9204 and Ohm9203 on inhibiting forskolin-stimulated cAMP accumulation in cells transfected with chimera II was similar to that in cells transfected with RMOR. CONCLUSION: Replacing 194-268 residues of RMOR with 185-262 residues of RKOR does not influence the ability of mu opioid receptor to bind Ohm9204 and Ohm9202 and the receptor mediated inhibition of cellular cAMP level.

3,4-Dichloro-N-methyl-N-(2-(1-pyrrolidinyl)-cycloh↗
PubMed2000

Retinal dopaminergic neurons (A17) expressing neuromedin K receptor (NK(3)): a double immunocytochemical study in the rat.

By using a double immunofluorescence method we examined the distribution of dopaminergic neurons (A17) expressing neuromedin K receptor (NKR, NK(3)) in the rat retina. The distribution of NKR-like immunoreactive (-LI) neurons partially overlapped that of tyrosine hydroxylase (TH)-LI neurons in the inner retina of section and flat-mount preparation. Neurons showing both TH- and NKR-like immunoreactivities were found in the retina (A17): 100% of these TH-LI neurons displayed NKR-like immunoreactivity, and they constituted about 3.5% of total NKR-LI neurons. The majority of double-labeled neurons with TH- and NKR-like immunoreactivities were distributed in the proximal inner nuclear layer and the upper part of inner plexiform layer of the retina, and characterized with appearance of amacrine cells. The present study has provided morphological evidence for direct physiological modulation of dopaminergic neurons by tachykinins through NKR in the rat retina (A17).

Animals↗
PubMed2000

Co-localization of NMDA receptors and AMPA receptors in neurons of the vestibular nuclei of rats.

We are interested in studying the co-localization of NMDA glutamate receptor subunits (NR1, NR2A/B) and AMPA glutamate receptor subunits (GluR1, GluR2, GluR2/3 and GluR4) in individual neurons of the rat vestibular nuclei. Immunoreactivity for NR1, NR2A/B, GluR1, GluR2, GluR2/3 and GluR4 was found in the somata and dendrites of neurons in the four major subdivisions (superior, medial, lateral, and spinal vestibular nuclei) and in two minor groups (groups x and y) of the vestibular nuclei. Double immunofluorescence showed that all the NR1-containing neurons exhibited NR2A/B immunoreactivity, indicating that native NMDA receptors are composed of NR1 and NR2A/B in a hetero-oligomeric configuration. Co-expression of NMDA receptor subunits and AMPA receptor subunits was demonstrated by double labeling of NR1/GluR1, NR1/GluR2/3, NR1/GluR4 and NR2A/B/GluR2 in individual vestibular nuclear neurons. All NR1-containing neurons expressed GluR2/3 immunoreactivity, and all NR2A/B-containing neurons expressed GluR2 immunoreactivity. However, only about 52% of NR1-immunoreactive neurons exhibited GluR1 immunoreactivity and 46% of NR1-containing neurons showed GluR4 immunoreactivity. The present data reveal that NMDA receptors are co-localized with variants of AMPA receptors in a large proportion of vestibular nuclear neurons. These results suggest that cross-modulation between NMDA receptors and AMPA receptors may occur in individual neurons of the vestibular nuclei during glutamate-mediated excitatory neurotransmission and may in turn contribute to synaptic plasticity within the vestibular nuclei.

Animals↗
PubMed2000

Noradrenergic neurons expressing substance P receptor (NK1) in the locus coeruleus complex: a double immunofluorescence study in the rat.

By using a double immunofluorescence method we examined the distribution of noradrenergic neurons expressing substance P receptor (NK1) or neuromedin K receptor (NK3) in the rat brainstem. The distribution of SPR-like immunoreactive (-LI) neurons completely overlapped that of tyrosine hydroxylase (TH)-LI neurons in the locus coeruleus (A6), ventrolateral and lateral reticular formation of pons (A5 and A7). Partially overlapping distribution of SPR- and TH-LI neurons were found in certain regions of the medulla oblongata (A1-A4). Neurons showing both SPR- and TH-like immunoreactivities, however, were only found in the locus coeruleus complex (A5-A7): 100% of these TH-LI neurons displayed SPR-like immunoreactivity. Neurons showing both NKR- and TH-like immunoreactivities were not detected in the aforementioned areas of brainstem. The present study has provided morphological evidence for direct physiological modulation of noradrenergic neurons by tachykinins through SPR in locus coeruleus complex (A5-A7).

Animals↗
PubMed2000

Human mu-opioid receptor overexpressed in Sf9 insect cells functionally coupled to endogenous Gi/o proteins.

Human mu-opioid receptor (HmuOR) with a tag of six consecutive histidines at its carboxyl terminus had been expressed in recombinant baculovirus infected Sf9 insect cells. The maximal binding capacity for the [3H] diprenorphine and [3H]ohmefentanyl (Ohm) were 9.1 +/- 0.7 and 6.52 +/- 0.23 nmol/g protein, respectively. The [3H] diprenorphine or [3H] Ohm binding to the receptor expressed in Sf9 cells was strongly inhibited by mu-selective agonists [D-Ala2, N-methyl-Phe4, glyol5]enkephalin (DAGO), Ohm, and morphine, but neither by delta nor by kappa selective agonist. Na+ (100 mM) and GTP (50 microM) could reduce HmuOR agonists etorphine and Ohm affinity binding to the overexpressed HmuOR. mu-selective agonists DAGO and Ohm effectively stimulated [35S]GTP-gammaS binding (EC50 = 2.7 nM and 6.9 nM) and inhibited forskolin- stimulated cAMP accumulation (IC50 = 0.9 nM and 0.3 nM). The agonist-dependent effects could be blocked by opioid antagonist naloxone or by pretreatment of cells with pertussis toxin (PTX). These results demonstrated that HmuOR overexpressed in Sf9 insect cells functionally coupled to endogenous G(i/o) proteins.

Animals↗
PubMed2000

Differential regulation of Ca2+ influx by fMLP and PAF in human neutrophils: possible involvement of store-operated Ca2+ channel.

Calcium (Ca2+) influx into human polymorphonuclear cells (PMNs) in response to N-formyl-Met-Leu-Phe (fMLP) and platelet-activating factor (PAF) stimulation was studied. Whole blood was taken by venous puncture from healthy human volunteers. PMNs were isolated, diluted, and incubated with 2 microM fura-2 AM. The cytosolic free calcium concentration, [Ca2+]i, in human neutrophils was determined by microfluorometry. We found that the net area under the fMLP- or PAF-induced [Ca2+]i rise curve in Ca2+-free medium decreased to 75% or 30% of the area under the curve in Ca2+ medium. Treatment of PMNs with phorbol myristate acetate (PMA), a protein kinase C activator, completely abolished the intracellular Ca2+ level stimulated by PAF, but not the intracellular Ca2+ level stimulated by fMLP. Treatment of PMNs with PAF did not abolish the intracellular Ca2+ level elevation stimulated by fMLP. In addition, treatment of PMNs with fMLP did not abolish intracellular Ca2+ level elevation stimulated by PAF. Loperamide, a positive modulator for store-operated calcium (SOC) channels, elicited an increase in intracellular calcium after the activation of SOC channels stimulated by fMLP or PAF. After the addition of guanosine 3',5'-cyclic monophosphate, N2,2'-O-Dibutyryl-, sodium salt (db-cGMP), the initial increase of PAF- or fMLP-induced PMNs intracellular Ca2+ fluorescences was well preserved, but the slope and the peak height of fluorescence curves declined compared with the curves without db-cGMP. In conclusion, we found that PAF and fMLP regulate the Ca2+ influx of PMNs in different ways. Most of the PAF-induced [Ca2+]i rise resulted from Ca2+ influx, and most of the fMLP-induced [Ca2+]i elevation resulted from intracellular stores release. The initial mobilization of intracellular Ca2+ stores in PAF-stimulated signals is mediated by protein kinase C (PKC) phosphorylation, but not in fMLP-stimulated route. SOC channels are present and important in the fMLP- or PAF-induced PMNs Ca2+ influx. There was no apparent cross-regulation between PAF- and fMLP-stimulated intracellular Ca2+ influx.

Calcium↗
PubMed2000

Nitric oxide synthase inhibitor ameliorates oral total parenteral nutrition-induced barrier dysfunction.

The expression of inducible nitric oxide synthase (iNOS) is increased in the intestine and results in mucosal damage after endotoxin challenge. Although the oral administration of total parenteral nutrition (TPN) solution promotes bacterial translocation (BT) and increases the intestinal permeability, the role of NO in the nutrition-induced loss of mucosal barrier function remains unclear. The distribution of fluorescein isothiocyanate-dextran (FITC-dextran, 4400) across the lumen of small intestine in rat was examined to investigate the role of NOS activity on the intestinal permeability under oral TPN feeding. Fifty-one rats were randomly divided into 4 groups. Group I (control group) was fed with rat chow, group II received TPN solution orally. Groups III and IV received TPN solution supplemented with NOS inhibitors. On day 9, FITC-dextran was injected into the intestinal lumen. After 30 min, blood samples were taken from portal vein and analyzed for plasma FITC-dextran level by fluorescence spectrophotometry. Samples of small intestine were frozen and sectioned in a cryostat for morphological and NOS histochemical studies. Homogenates of small intestine were used for NOS activity measurement. The plasma level of FITC-dextran showed a significant increase (P < 0.05) in rats fed with oral TPN compared with the control ones. Supplement with NOS inhibitors significantly decreased the intestinal permeability in groups III and IV compared with group II. Similarly, the total NOS activities showed a significant 2-fold increase (P< 0.05) in group II, and NOS inhibitors decreased the elevated NOS activity. These data suggest that oral TPN feeding for 9 days leads to an increase in permeability to dextran and the total NOS activity of small intestine, and both induction of the intestinal permeability and NOS activity were inhibited by treatment with NOS inhibitors. Addition of S-methylisothiourea (SMT), an iNOS selective inhibitor, profoundly inhibited 66% of the induced iNOS activity (P < 0.05) and reduced 74% of the diet-induced increase in intestinal permeability (P < 0.05) in group II. The induced permeability change in rats receiving oral TPN is mainly due to the activity of intestinal mucosal iNOS. The induction of iNOS is an important mediator for intestinal barrier dysfunction. Administration of SMT, which specifically decreases iNOS activity, is useful in the prevention of diet-induced barrier failure.

Administration, Oral↗
PubMed2000

Mechanisms underlying ATP-induced Ca2+ mobilization in human neutrophils.

The effect of ATP on Ca2+ mobilization in human neutrophils was examined by using fura-2 as a Ca2+ indicator. ATP (0.1-100 microM) caused a significant [Ca2+]i increase in a concentration-dependent manner. The [Ca2+]i signal comprised an initial rise followed by a plateau. Removal of external Ca2+ diminished the peak value of the [Ca2+]i signal. In Ca2+-free medium, pretreatment with an endoplasmic reticulum Ca2+ pump inhibitor, thapsigargin, prevented ATP from releasing Ca2+. In contrast, thapsigargin still increased [Ca2+], after pretreatment with 10 microM ATP. These results indicate that 10 microM ATP released Ca2+ mainly from thapsigargin-sensitive stores. Adding 3 mM Ca2+ induced a concentration-dependent increase in [Ca2+]i after pretreatment with ATP or thapsigargin in Ca2+-free medium, suggesting ATP induced Ca2+ influx via capacitative Ca2+ entry. ATP (10 microM)-induced Ca2+ release was abolished by inhibiting phospholipase C with 2 microM U73122, indicating that inositol-1,4,5-trisphosphate (IP3) mediates ATP-induced Ca2+ release. Conversely, ATP-induced [Ca2+]i increase was abolished by activating protein kinase C (PKC) with 10 nM phorbol myristate acetate (PMA), but was not altered by inhibiting PKC with 2 microM GF 109203X. This implies ATP-induced [Ca2+]i increase is a PMA-linked event. Together, the results suggest ATP increases [Ca2+]i in human neutrophils by releasing Ca2+ from IP3-coupled, thapsigargin-sensitive Ca2+ stores, and inducing Ca2+ influx via the process of capacitative Ca2+ entry. The ATP-induced Ca2+ signal is a PMA-linked event.

Adenosine Triphosphate↗