Protein composition of human metaphase chromosomes analyzed by two-dimensional electrophoreses.

A large amount of metaphase chromosomes were isolated from synchronized human cell lines by a polyamine procedure. All the chromosomal proteins extracted by an acetic acid extraction method were fully dissolved into the sample solutions for isoelectric focusing (IEF) or radical free and highly reduced (RFHR) two-dimensional electrophoreses (2-DEs). As a result, well-separated and highly reproducible 2-DE patterns were obtained. This could not be attained by an ordinary acetone precipitation method. The 2-DE patterns visualized using Coomassie Brilliant Blue (CBB) staining indicated that more than one hundred proteins were involved in the isolated metaphase chromosomes, although the most abundant proteins, histones, occupied a greater part of the chromosomal proteins. It was also shown that colcemid treatment for cell cycle synchronization had little effect on the 2-DE pattern compared to that obtained without the treatment. Furthermore, no significant differences were observed in the 2-DE patterns among the chromosomal proteins prepared from two different human cell lines, BALL-1 and K562. However, 2-DE analysis of isolated metaphase chromosomes from HeLa cells apparently showed a smaller number of proteins than the BALL-1 and K562 cell lines at a neutral pI range. The present study paves the way for elucidating protein composition of human metaphase chromosomes.

Monitoring of white-rot fungus during bioremediation of polychlorinated dioxin-contaminated fly ash.

Bioremediation is a low-cost treatment alternative for the cleanup of polychlorinated-dioxin-contaminated soils and fly ash when pollution spread is wide-ranging. An interesting fungus, Ceriporia sp. MZ-340, with a high ability to degrade dioxin, was isolated from white rotten wood of a broadleaf tree from Kyushu Island in Japan. We have attempted to use the fungus for bioremediation of polychlorinated-dioxin-contaminated soil on site. However, we have to consider that this trial has the potential problem of introducing a biohazard to a natural ecosystem if this organism is naturalized. We have therefore developed a monitoring system for the introduced fungus as a part of the examination and evaluation of bioremediation in our laboratory. We have also developed a PCR-based assay to reliably detect the fungus at the bioremediation site. DNA isolated from the site was amplified by PCR using a specific primer derived from internal transcribed spacer region (ITS: ITS1, 5.8S rDNA and ITS2) sequences of Ceriporia sp. MZ-340. We successfully monitored Ceriporia sp. MZ-340 down to 100 fg/ micro l DNA and down to 2 mg/g mycelium. We also successfully monitored the fungus specifically at the bioremediation site. The polychlorinated dibenzo- p-dioxin and polychlorinated dibenzofuran content was observed to decrease in response to treatment with the fungus. The species-specific PCR technique developed in the present work is useful in evaluating the possibility of on-site bioremediation using the fungus Ceriporia sp. MZ-340.

Treatment of subungual glomus tumour.

We reviewed 30 patients with subungual glomus tumours of the hand operated on between 1964 and 1997. Seven patients were male and 23 were female. Their age ranged from 16 to 78 years. A transungual approach was selected in 27 patients, and a periungual approach in three. Pre-operative pain subsided in all of the patients, but recurrence of the pain was observed in nine. Nail deformities were observed in nine patients before surgery. After surgery, it disappeared in three patients, persisted in six, and new deformities developed in five. To avoid recurrence of pain, it is important that the accurate pre-operative localisation of the tumour and a complete extirpation should be performed. To avoid nail deformity, it is better to apply a periungual approach for tumours developing in the peripheral region, and a transungual approach followed by meticulous repair of the nail bed for tumours developing in the central region.

A comparison of ON-PUMP vs OFF-PUMP coronary artery bypass surgery among low, intermediate, and high-risk patients: the Hartford Hospital experience.

BACKGROUND: Off-pump coronary artery bypass (OP-CAB) graft surgery is being used with increasing frequency. This study was designed to compare OP-CAB outcomes with conventional surgical revascularization using cardiopulmonary bypass (CPB) in patients with varying risk categories at a high-volume center. METHODS AND RESULTS: Between 1/1/1999 and 1/31/2001, bypass surgery was performed on 1,312 patients, including 348 OP-CAB cases and 964 CPB cases. Compared to CPB cases, OP-CAB patients were more likely to be female and had a lower incidence of three vessel coronary artery disease, prior percutaneous intervention, and prior bypass surgery. Postoperatively, OP-CAB patients had a lower incidence of renal failure and prolonged ventilatory support, as well as a lower composite endpoint of inhospital mortality, perioperative myocardial infarction, cerebrovascular accident, and/or renal failure. In addition, OP-CAB patients required fewer transfusions and had a shorter total length of hospital stay. In general, morbidity and mortality increased in both OP-CAB and CPB groups with increasing Parsonnet score. CONCLUSIONS: OP-CAB surgery is a safe and effective alternative to conventional coronary artery bypass graft (CABG) surgery, with a lower incidence of major in-hospital adverse clinical events and a decreased requirement for medical resources. Adverse OP-CAB outcomes correlate well with pre-operative Parsonnet Score.

Cloning of the cyclodextrin glucanotransferase gene from alkalophilic Bacillus sp. A2-5a and analysis of the raw starch-binding domain.

The cyclodextrin glucanotransferase (CGTase) gene of alkalophilic Bacillus sp. A2-5a was cloned and expressed in Bacillus subtilis ANA-1 as a host. The DNA region included an open reading frame encoding a 704-amino-acid polypeptide with a typical raw starch-binding motif in its C-terminal region. The CGTase purified from Bacillus sp. A2-5a bound to raw starch as strongly as porcine pancreas alpha-amylase, as expected from the sequence motif. A chromosomal region (a DNA fragment of about 14.1 kbp) including the CGTase gene was also cloned and the nucleotide sequence was determined. Possible cyclodextrinase and putative cyclodextrin-binding protein genes were found in the flanking region of the CGTase gene, which implied that the novel starch-degradation pathway postulated for a gram-negative bacterium [Klebsiella oxytoca; Fiedler et al. (1996) J Mol Biol 256: 279-291] also exists in a gram-positive bacterium i.e. Bacillus.

A hrs binding protein having a Src homology 3 domain is involved in intracellular degradation of growth factors and their receptors.

BACKGROUND: Hrs (hepatocyte growth factor (HGF)-regulated tyrosine kinase substrate) is an early endosomal protein that is rapidly tyrosine-phosphorylated in cells stimulated with growth factors. Hrs is thought to play a regulatory role in the endocytosis of growth factor/receptor complexes through early endosomes. In this study, we searched for Hrs-interacting molecules which may regulate the function of Hrs, using a yeast two-hybrid system. RESULTS: We isolated a cDNA clone encoding a novel Src homology 3 (SH3)-containing protein, and named it 'Hrs binding protein' (Hbp). Hbp was co-immunoprecipitated with Hrs, and its intracellular localization was similar to that of Hrs. The association between Hbp and Hrs was mediated through the coiled coil motifs in Hbp and Hrs. Deletion mutants of Hbp lacking either the SH3 domain or the Hrs binding domain showed dominantly negative effects on the intracellular degradation of a growth factor and its receptor, but not on the internalization of growth factor/receptor complexes. CONCLUSIONS: Hbp is thought to be closely associated with Hrs on early endosomes. Hbp, together with Hrs may play a regulatory role in the vesicular transport of growth factor/receptor complexes through early endosomes, for their degradation.

Rejection and regeneration in peripheral nerve homografts in rats after withdrawal of cyclosporin: morphological and immunohistochemical assessment.

Peripheral nerve homografts 35 mm long were inserted in rats to study rejection and regeneration of nerves after suspension of an immunosuppressant (cyclosporin), and the correlation between the Schwann cells of the host and the donor during the course of regeneration. Sciatic nerve homografts from 36 ACI-RT1a rats were transplanted into Lewis-RT1(1) rats. Isografts were taken from 24 Lewis rats. Cyclosporin 5 mg/kg/day was given subcutaneously for 12 weeks. In the homograft group the myelinated axons that regenerated while the immunosuppressant was being given were covered mainly with Schwann cells from the donor, and after the immunosuppressant was withdrawn both Schwann cells of the donor and axons were rejected. However, while the cyclosporin was being given the Schwann cells of the host migrated into the nerve graft together with a few myelinated axons that escaped rejection, and unmyelinated axons that regenerated after rejection were myelinated by the Schwann cells of host.

[Experimental study of percutaneous hot ethanol injection therapy (PHEIT) by continuous heating device for hepatocellular carcinoma].

Percutaneous ethanol injection therapy (PEIT) is widely used as a local treatment for hepatocellular carcinoma (HCC). However, because only a small amount of ethanol can be used in one PEIT session and because the antitumor effect is limited, this modality is indicated only when there are three or fewer tumors and when the tumor diameter is < or = 3 cm. To obtain a more potent and certain antitumor effect, we have devised a new treatment called percutaneous hot ethanol injection therapy (PHEIT), and developed a Continuous Heating Device with which ethanol can be heated and locally injected at a specified temperature. The continuous Heating Device is composed of three major components: a syringe heater, a needle thermocontroller, and a needle tip thermosensor. A disposable syringe filled with liquid is inserted into the syringe heater, which heats the liquid to a desired temperature by adjusting the voltage. The needle thermocontroller is a puncture guide needle to which a heating device has been attached. The needle-tip thermosensor constantly measures, displays and records the temperature of the liquid at the needle tip during injection. Also, because the Continuous Heating Device is a closed-circuit system, there is no risk of accidental a fire, which ensures procedural safety. It is also possible to use this device to safely heat and inject a variety of other liquids, such as physiological saline and anticancer agents and thus contribute to the widespread development of ultrasound-guided injection therapy.

L* protein of the DA strain of Theiler's murine encephalomyelitis virus is important for virus growth in a murine macrophage-like cell line.

Strain GDVII and other members of the GDVII subgroup of Theiler's murine encephalomyelitis virus (TMEV) are highly virulent and cause acute polioencephalomyelitis in mice. Neither viral persistence nor demyelination is demonstrated in the few surviving mice. On the other hand, strain DA and other members of the TO subgroup of TMEV are less virulent and establish a persistent infection in the spinal cord, which results in a demyelinating disease. We previously reported that GDVII does not actively replicate in a murine macrophage-like cell line, J774-1, whereas DA strain productively infects these cells (M. Obuchi, Y. Ohara, T. Takegami, T. Murayama, H. Takada, and H. Iizuka, J. Virol. 71:729-733, 1997). In the present study, we used recombinant viruses between these strains of the two subgroups to demonstrate that the DA L coding region of DA strain is important for virus growth in J774-1 cells. Additional experiments with a mutant virus indicate that L* protein, which is synthesized out of frame with the polyprotein from an additional alternative initiation codon in the L coding region of TO subgroup strains, is a key determinant responsible for the cell-type-specific restriction of virus growth. L* protein may play a critical role in the DA-induced restricted demyelinating infection by allowing growth in macrophages, a major site for virus persistence.

A better long-term outcome in cardiac transplant recipient with a history of previous open heart operations.

OBJECTIVE: To investigate the effect of previous open heart operations (POHO) on the outcome of heart transplantation (HTX). METHODS: Between November 1984 and May 1996, HTX was performed on 151 patients at Hartford Hospital. Among them, 61 patients had previous open heart operations (POHO) (group A), and 90 did not (group B). The average follow-up period was 1615 +/- 1185 days for group A and 1330 +/- 1125 days for group B. The recipient age was 55 +/- 10 years for group A and 48 +/- 12 years for group B (P < 0.01). There were 17 patients (26%) in group A and 14 (50%) in group B who were over 60 years of age. There was more coronary artery disease (74% versus 37%, P < 0.001) as etiology, and more diabetics in group A (P < 0.02). RESULTS: The time for cardiopulmonary bypass (133 +/- 20 min versus 106 +/- 18 min, P < 0.01) and aortic clamp time (73 +/- 16 min versus 61 +/- 13 min, P < 0.01) were longer in group A. The operative mortality (within 30 days) was 0 and 2.2%, and the cumulative deaths were 16 (26%) and 43 (48%) respectively for group A and group B (P < 0.01). The causes of death were (group A vs group B): infection (31% vs 26%), rejection (13% vs 28%, P < 0.05), malignancy (25% vs 16%), cardiac event (6% vs 14%) and others (25% vs 16%). In patients over 60, there were 4 deaths (24%) in group A and 7 (50%) in group B. The difference was not significant. No patients died of rejection in this subgroup. The actuarial survival rates in group A versus group B were: 1 year, 93% versus 83%; 2 years, 85% versus 74%; 3 years, 81% versus 71%; 5 years, 76% versus 58%; and 10 years, 57% versus 24% (P < 0.01). CONCLUSION: The survival rate in patients who had POHO is much higher than that in patients who had HTX as their primary operation.

Characterization of a gene cluster for glycogen biosynthesis and a heterotetrameric ADP-glucose pyrophosphorylase from Bacillus stearothermophilus.

A chromosomal region of Bacillus stearothermophilus TRBE14 which contains genes for glycogen synthesis was cloned and sequenced. This region includes five open reading frames (glgBCDAP). It has already been demonstrated that glgB encodes branching enzyme (EC 2.4.1.18 [H. Takata et al., Appl. Environ. Microbiol. 60:3096-3104, 1994]). The putative GlgC (387 amino acids [aa]) and GlgD (343 aa) proteins are homologous to bacterial ADP-glucose pyrophosphorylase (AGP [EC 2.7.7.27]): the sequences share 42 to 70% and 20 to 30% identities with AGP, respectively. Purification of GlgC and GlgD indicated that AGP is an alpha2beta2-type heterotetrameric enzyme consisting of these two proteins. AGP did not seem to be an allosteric enzyme, although the activities of most bacterial AGPs are known to be allosterically controlled. GlgC protein had AGP activity without GlgD protein, but its activity was lower than that of the heterotetrameric enzyme. The GlgA (485 aa) and GlgP (798 aa) proteins were shown to be glycogen synthase (EC 2.4.1.21) and glycogen phosphorylase (EC 2.4.1.1), respectively. We constructed plasmids harboring these five genes (glgBCDAP) and assayed glycogen production by a strain carrying each of the derivative plasmids on which the genes were mutated one by one. Glycogen metabolism in B. stearothermophilus is discussed on the basis of these results.

Influence of distal nerve segment volume on nerve regeneration in silicone tubes.

The influence of a volume of a distal nerve segment upon nerve regeneration in an 8-mm gap created within a silicone tube was examined. The rats were randomly divided into four groups. Each group had 5 mm, 1 mm, or a half volume of 1-mm nerve segment (a nerve piece of 1 mm transected longitudinally) inserted into the distal end of a silicone tube of 11 mm. The empty group without a nerve segment was used as control. Diameters of regenerated cylindrical structure between the nerve ends in the silicone tube were measured under an operation microscope and myelinated axon diameter, myelinated axon density, myelin sheath width, and ratio of myelinated axon area to total cross sectional area were measured using the transverse sections at the midpoint of the silicone tube at 6 weeks after surgery. Although there was a significant difference in all of those parameters between the control group and any of the remaining three groups, no significant difference was found between any pair of these three groups. The results of this study indicated that the degree of nerve regeneration does not correlate with the volume of a distal nerve segment and even a very small piece can play an important role in supporting regenerating nerve beyond a definitive gap.

Hepatocyte growth factor activator inhibitor, a novel Kunitz-type serine protease inhibitor.

Hepatocyte growth factor (HGF) activator is a serine protease that is produced and secreted by the liver and circulates in the blood as an inactive zymogen. In response to tissue injury, the HGF activator zymogen is converted to the active form by limited proteolysis. The activated HGF activator converts an inactive single chain precursor of HGF to a biologically active heterodimer in injured tissue. The activated HGF may be involved in the regeneration of the injured tissue. In this study, we purified an inhibitor of HGF activator from the conditioned medium of a human MKN45 stomach carcinoma cell line and molecularly cloned its cDNA. The sequence of the cDNA revealed that the inhibitor has two well defined Kunitz domains, suggesting that the inhibitor is a member of the Kunitz family of serine protease inhibitors. The sequence also showed that the primary translation product of the inhibitor has a hydrophobic sequence at the COOH-terminal region. Inhibitory activity toward HGF activator was detected in the membrane fraction as well as in the conditioned medium of MKN45 cells. These results suggest that the inhibitor may be produced as a membrane-associated form and secreted by the producing cells as a proteolytically truncated form.

Identification of alloantisera reacting with HLA-C blank (Cx52) using a mouse L-cell transfected with the HLA-Cw*1201 allele.

The HLA-C locus frequently has a serologically undefined "blank" (CwBL) specificity. A cDNA clone derived from the HLA-C gene with a blank specificity (Cx52) strongly associated with the most common haplotype in a Japanese population, A24-CwBL-B52-DR2-DQ6-DP9, has been recently cloned and sequenced in our laboratory and officially designated Cw*1201 as an allelic name, indicating that the inability to define the HLA-C antigen serologically in this haplotype is not due to an HLA-C antigen gene deletion or mutation, but to the absence of typing sera. In this paper, a mouse L-cell transfectant expressing this Cw*1201 gene product was constructed and employed for screening of alloantisera recognizing the HLA-C antigen with the Cw*1201 specificity. Two alloantisera against Cw*1201 were thus identified and characterized using HLA homozygous B-cell lines and local panel PBL cells, indicating that a transfectant expressing a single HLA provides an efficient screening system for collection of HLA-typing sera, especially reacting with serologically unclassifiable "blank" antigens.

Establishment and characterization of a cell line, KaMi, from human lung large cell carcinoma.

A cell line of human lung large cell carcinoma (LCC) was established directly from the metastatic skin tumor tissue. The clinical course of the patient who carried this carcinoma was peculiar; generalized lymphadenopathy, histologically resembling Hodgkin's disease, was found as the first clinical symptom. The lung tumor was not discovered until the time of autopsy. This cell line (KaMi) grew adherent to culture vessels with the population doubling time of 20.6h, formed colonies in soft agars with efficiency of 22.6%, and formed tumors in athymic nude mice. The authenticity of KaMi was confirmed by chromosomal analysis and isoenzyme patterns. KaMi cells bore a strong resemblance to the original tumor cells which were composed of small spindle cells, large polygonal cells, and multinucleated giant cells. Immunohistochemically, KaMi cells showed a weak tendency to differentiate to squamous cells, and these immunohistochemical reactivities were almost compatible to those of the original tumor cells, but ultrastructurally, KaMi cells were more immature than the original ones. Treatment with several reagents could not augment a differentiation of KaMi cells. Cytokeratin profiles showed a tendency of squamous cell differentiation. KaMi cells may aid in elucidating the pathogenesis and biology of LCC and its relationship to other lung tumors.

[Influence of systolic left ventricular blood flow direction on genesis of senile calcification of the aortic valve].

To assess the factors which may initiate and accelerate degenerative senile calcification of the aortic valve, two-dimensional echocardiograms and the clinical characteristics of 259 consecutive cases with senile calcification of the aortic valve were studied. The results were compared with those of similar studies among 186 consecutive cases with the normal aortic valves. An aortic cusp with an area of increased echo greater than 3 mm in width and with decreased pliability was regarded as calcified. Among patients with calcification of one aortic cusp, 114 exhibited calcification of a noncoronary cusp, 17 calcification of the left coronary cusp and 3 calcification of the right coronary cusp (p < 0.001). Among patients with calcification of 2 aortic cusps, 39 had calcification of a noncoronary and left coronary cusps, 3 calcification of the left and right coronary cusps and 16 calcification of the right and noncoronary cusps (p < 0.001). In patients with calcification of their aortic valves, the end-diastolic angle between the interventricular septum and the ascending aorta was 102 +/- 10 degrees; whereas, it was 89 +/- 10 degrees in the control group (p < 0.001). There were no differences in frequency of aortic root calcification, mitral annular calcification, hypertension, ischemic heart disease, hyperlipidemia, hyperuricemia, or hyperglycemia, between patients with and without calcification of their aortic valves. Of the female patients ranging in age from 65 to 74 years, 88% in those with calcification of 3 cusps and 30% in those with calcification of one cusp (p < 0.05) had mitral annular calcification.(ABSTRACT TRUNCATED AT 250 WORDS)

[Studies on porphyrin related compounds and tumor tissue affinities. I. Synthesis of a carrier for tumor imaging agent, bifunctional chelating agent coupled porphyrin (ATN-2)].

A carrier for a new tumor imaging agent, a bifunctional chelating agent (BCA)-coupled porphyrin (ATN-2), has been synthesized from protoporphyrin dimethyl ester in 4 steps. At first, the hydrobromination of protoporphyrin dimethyl ester is carried out to obtain a monobromo derivative. The derivative is treated with ethylene glycol. The resulting porphyrin has an ether group at the either 7- or 12-position of the ring. Metallation with GaCl3 of the porphyrin having a ethylene glycol residue affords Ga-metalloporphyrin. Final condensation of the metalloporphyrin with diethylenetriaminepentaacetic acid (DT-PA) gave BCA-coupled porphyrin (ATN-2). The chelation of ATN-2 with 111InCl3 easily afforded [111In]ATN-2. This agent was used for imaging transplantable pancreatic carcinoma in Syrian golden hamster at 72 h after postinjection. The efficacy of the new agent was compared with that of [67Ga]citrate. The images with [111In]ATN-2 were found to be clearer than those with [67Ga]citrate. Therefore, ATN-2, a carrier for 111InCl3, seems to be more useful for tumor imaging agents.

Binding site of omeprazole in hog gastric H+,K(+)-ATPase.

Omeprazole transforms into an active compound in an acidic environment, which is able to modify a sulfhydryl group of gastric H+,K(+)-ATPase. Omeprazole was transformed into a strongly fluorescent molecule by UV-light irradiation (excitation wavelength = 290 nm, emission wavelength = 335 nm). The omeprazole-modified residue of hog H+,K(+)-ATPase was estimated by the fluorescence of the omeprazole moiety and limited tryptic digestion of the enzyme. Among the four main tryptic digested subfragments, omeprazole was bound to the 67, 42 and 32-kDa subfragments, but not to the 52-kDa subfragment. Taking the amino acid sequence of this ATPase into consideration, we propose that omeprazole specifically binds with Cys322 in hog H+,K(+)-ATPase (Cys321 in rat).

The potency of substituted benzimidazoles such as E3810, omeprazole, Ro 18-5364 to inhibit gastric H+, K(+)-ATPase is correlatedwith the rate of acid-activation of the inhibitor.

The half maximal inhibitory concentrations (IC50) of substituted benzimidazoles for the H+, K(+)-ATPase in hog gastric vesicles were measured by using the pyruvate kinase-lactate dehydrogenase-linked system in which hydrolysis of ATP was coupled with the oxidation of NADH. The vesicles were incubated in a solution containing a high concentration of KCl, valinomycin and Mg-ATP, and the intravesicular medium was acidified. The inhibitor was activated in the acidic medium and reacted with SH groups on the luminal (intravesicular) side of the ATPase. The active compound formed in the extravesicular medium (pH 6.11) was quenched by GSH. Under these conditions, IC50 of new compound E3810, 2[(4-(3-methoxypropoxy)-3-methylpyridine-2-yl)methyl-sulfinyl]-1H- benzimidazole sodium salt, was 0.072 microM and that of omeprazole was 0.47 microM at 25 degrees. On the other hand, the rates of formation of active compounds, tetracyclic sulfenamide derivatives, from original substituted benzimidazoles in 0.1 N HCl (k) were determined by measuring optical density at the characteristic wavelengths of the active compounds. There was a good correlation between IC50 and k for various substituted benzimidazoles including E3810, methoxy derivative of E3810, omeprazole, Ro 18-5364, H compound, picoprazole and timoprazole. This fact suggest that the rate of the formation of the acid-activated compound is a main factor determining the potency of the inhibitor.

Acid activation of omeprazole in isolated gastric vesicles, oxyntic cells, and gastric glands.

Omeprazole, a potent inhibitor of gastric hydrogen ion transporting, potassium-stimulated adenosine triphosphatase, was found to be transformed into an SH-reactive strong fluorescent molecule (excitation and emission wavelengths of 370 and 560 nm, respectively) in an acidic medium. The addition of glutathione- or protein-containing sulfhydryl groups such as pepsin to the medium decreased the fluorescence. Also, the increase in the pH of the medium decreased the fluorescence. The fluorescent molecule was identified to be an acid-activated planar cyclic sulfenamide derivative of omeprazole. The transformation was studied in H+-preaccumulated hog gastric vesicles, which contain the hydrogen ion transporting, potassium-stimulated adenosine triphosphatase. The addition of omeprazole to the vesicle suspension induced a rapid increase in the fluorescence intensity, indicating that omeprazole was activated in the intravesicular space. Then, the intensity biphasically decreased with time. The slower small decrease was due to the reaction of the sulfenamide with sulfhydryl group(s) located on the acid secretory side of the hydrogen ion transporting, potassium-stimulated adenosine triphosphatase. Omeprazole was also activated in the acidic lumina of isolated rabbit gastric glands that were stimulated with histamine. Furthermore, direct evidence was obtained from the imaging of the fluorescence that omeprazole was activated in the acidic compartments of the isolated Xenopus oxyntic cell.