The relationship between oxygen consumption rate and viability of in vivo-derived pig embryos vitrified by the micro volume air cooling method.

The aim of this study was to assess the viability of vitrified-warmed in vivo-derived pig embryos after measuring the oxygen consumption rate. Six days after artificial insemination, blastocysts were collected from gilts and vitrified by the micro volume air cooling method. The oxygen consumption rate was measured in 60 vitrified-warmed embryos, which were then cultured for 48h to assess the viability. The survival (re-expansion) rate of embryos after warming was 85.0%. The average oxygen consumption rate of embryos immediately after warming was greater in embryos which could re-expand during subsequent culture (F=0.75±0.04) than that in those which failed to re-expand (F=0.33±0.05). Moreover, the oxygen consumption rate of vitrified-warmed embryos was greater in the hatched (F=0.88±0.06) than that in the not-hatched group (F=0.53±0.04). When the oxygen consumption rate of the vitrified-warmed embryos and the numbers of viable and dead cells in embryos were determined, there was a positive correlation between the oxygen consumption rate and the number of live cells (P<0.01, r=0.538). A total of 29 vitrified embryos after warming and measuring the oxygen consumption rate were surgically transferred into uterine horns of two recipients. Both of the recipients become pregnant and farrowed 12 healthy piglets. These results demonstrate that the oxygen consumption rate of vitrified-warmed pig embryos can be related to the number of live cells and that the measurement of oxygen consumption of embryos after cryopreservation may be useful for estimating embryo survivability.

Plasmon modes in single gold nanodiscs.

Optical properties of single gold nanodiscs were studied by scanning near-field optical microscopy. Near-field transmission spectra of a single nanodisc exhibited multiple plasmon resonances in the visible to near-infrared region. Near-field transmission images observed at these resonance wavelengths show wavy spatial features depending on the wavelength of observation. To clarify physical pictures of the images, theoretical simulations based on spatial correlation between electromagnetic fundamental modes inside and outside of the disc were performed. Simulated images reproduced the observed spatial structures excited in the disc. Mode-analysis of the simulated images indicates that the spatial features observed in the transmission images originate mainly from a few fundamental plasmon modes of the disc.

A prospective study of the incidence and outcomes of incidental dural tears in microendoscopic lumbar decompressive surgery.

Little information is available about the incidence and outcome of incidental dural tears associated with microendoscopic lumbar decompressive surgery. We prospectively examined the incidence of dural tears and their influence on the outcome six months post-operatively in 555 consecutive patients (mean age 47.4 years (13 to 89)) who underwent this form of surgery. The incidence of dural tears was 5.05% (28/555). The risk factors were the age of the patient and the procedure of bilateral decompression via a unilateral approach. The rate of recovery of the Japanese Orthopaedic Association score in patients with dural tears was significantly lower than that in those without a tear (77.7% vs. 87.6%; p < 0.02), although there were no significant differences in the improvement of the Oswestry Disability Index between the two groups. Most dural tears were small, managed by taking adequate care of symptoms of low cerebrospinal fluid pressure, and did not require direct dural repair. Routine MRI scans were undertaken six months post-operatively; four patients with a dural tear had recurrent or residual disc herniation and two had further stenosis, possibly because the dural tear prevented adequate decompression and removal of the fragments of disc during surgery; as yet, none of these patients have undergone further surgery.

The natural history of asymptomatic lumbar canal stenosis in patients undergoing surgery for cervical myelopathy.

We retrospectively examined the prevalence and natural history of asymptomatic lumbar canal stenosis in patients treated surgically for cervical compressive myelopathy in order to assess the influence of latent lumbar canal stenosis on the recovery after surgery. Of 214 patients who had undergone cervical laminoplasty for cervical myelopathy, we identified 69 (32%) with myelographically documented lumbar canal stenosis. Of these, 28 (13%) patients with symptomatic lumbar canal stenosis underwent simultaneous cervical and lumbar decompression. Of the remaining 41 (19%) patients with asymptomatic lumbar canal stenosis who underwent only cervical surgery, 39 were followed up for ≥ 1 year (mean 4.9 years (1 to 12)) and were included in the analysis (study group). Patients without myelographic evidence of lumbar canal stenosis, who had been followed up for ≥ 1 year after the cervical surgery, served as controls (135 patients; mean follow-up period 6.5 years (1 to 17)). Among the 39 patients with asymptomatic lumbar canal stenosis, seven had lumbar-related leg symptoms after the cervical surgery. Kaplan-Meier analysis showed that 89.6% (95% confidence interval (CI) 75.3 to 96.0) and 76.7% (95% CI 53.7 to 90.3) of the patients with asymptomatic lumbar canal stenosis were free from leg symptoms for three and five years, respectively. There were no significant differences between the study and control groups in the recovery rate measured by the Japanese Orthopaedic Association score or improvement in the Nurick score at one year after surgery or at the final follow-up. These results suggest that latent lumbar canal stenosis does not influence recovery following surgery for cervical myelopathy; moreover, prophylactic lumbar decompression does not appear to be warranted as a routine procedure for coexistent asymptomatic lumbar canal stenosis in patients with cervical myelopathy, when planning cervical surgery.

A 5-HT(4)-receptor activation-induced neural plasticity enhances in vivo reconstructs of enteric nerve circuit insult.

BACKGROUND: It was recently reported that some 5-HT(4)-receptor agonists increased neuronal numbers and length of neurites in enteric neurons developing in vitro from immunoselected neural crest-derived precursors. We aimed to explore a novel approach in vivo to reconstruct the enteric neural circuitry that mediates a fundamental distal gut reflex. METHODS: The neural circuit insult was performed in guinea pigs by rectal transection and subsequent end-to-end one layer anastomosis. A 5-HT(4)-receptor agonist, mosapride citrate (10-100 micromol L(-1)) (applied for a patent) was applied locally at the anastomotic site. KEY RESULTS: Mosapride promoted the regeneration of the neural circuit in the impaired myenteric plexus and the recovery of the defecation reflex in the distal gut. Furthermore, mosapride generated neurofilament (NF)-, 5-HT(4)-receptor- and 5-bromo-2'-deoxyuridine (BrdU)-positive cells and surprisingly formed neural network in the newly formed granulation tissue at the anastomotic site 2 weeks after enteric nerve circuit insult. Possible neural stem cell markers, anti-distal less homeobox 2 (DLX2)- and p75-positive and NF-positive cells increased during the same time period. All actions by mosapride were inhibited by the specific 5-HT(4)-receptor antagonist, GR113808 (10 micromol L(-1)). CONCLUSIONS & INFERENCES: These results indicate that activation of enteric neural 5-HT(4)-receptors promotes reconstruction of an enteric neural circuit leading to the recovery of the defecation reflex in the distal gut, and that this reconstruction involves possibly neural stem cells. These findings indicate that treatment with 5-HT(4) agonists could be a novel therapy for generating new enteric neurons to rescue aganglionic gut disorders.

Vibrations of microspheres probed with ultrashort optical pulses.

We use ultrashort optical pulses to excite and detect vibrations of single silica spheres with a diameter of 5 microm placed at the surface of an acoustically mismatched substrate. In addition to the photoelastic detection of picosecond longitudinal acoustic pulses propagating inside the bulk, we detect gigahertz acoustic resonances of the sphere through probe beam defocusing. The mode frequencies are in close accord with those calculated from the elastic vibrations of a free sphere. We also record a resonant enhancement in the amplitude of specific modes of two touching spheres.

Inverse silica opal photonic crystals for optical sensing applications.

This work reports fabrication of inverse silica opal photonic crystal structures from direct polystyrene micro sphere opals using low-temperature sol-gel infiltration of silica, and examines performance of these photonic crystals as environmental refractive index sensors. Sensitivity of the spectral position and optical attenuation of photonic stop gaps is found to allow detection of the index changes by the amount of ~10(-3). The high value of sensitivity, which is comparable with those of other optical sensing techniques, along with simplicity of the optical detection setup required for sensing, and the low-temperature, energy-efficient fabrication process make inverse silica opals attractive systems for optical sensing applications.

Laser manipulation of a smectic liquid-crystal droplet.

The laser trapping of a smectic-A liquid-crystal micro-droplet was spatially traced during its transient into the trapped position. The lateral and angular orientation of the droplet were determined and followed in time during the axial descent of the micro-droplet into the stationary trapped position using the analysis of polarization changes of the light passed through the droplet with temporal resolution of a video refresh rate of 30 ms. The spatial resolution of 0.1-1 microm has been achieved for typical laser trapping powers of 2-600 mW. The axial profile of a laser trapping force (an ellipticity of the focal spot) has been determined. The laser trapping mechanism of smectic micro-droplets is discussed in terms of minimization of a light-droplet interaction.

Laser-induced microexplosion confined in the bulk of a sapphire crystal: evidence of multimegabar pressures.

Extremely high pressures (approximately 10 TPa) and temperatures (5 x 10(5) K) have been produced using a single laser pulse (100 nJ, 800 nm, 200 fs) focused inside a sapphire crystal. The laser pulse creates an intensity over 10(14) W/cm2 converting material within the absorbing volume of approximately 0.2 microm3 into plasma in a few fs. A pressure of approximately 10 TPa, far exceeding the strength of any material, is created generating strong shock and rarefaction waves. This results in the formation of a nanovoid surrounded by a shell of shock-affected material inside undamaged crystal. Analysis of the size of the void and the shock-affected zone versus the deposited energy shows that the experimental results can be understood on the basis of conservation laws and be modeled by plasma hydrodynamics. Matter subjected to record heating and cooling rates of 10(18) K/s can, thus, be studied in a well-controlled laboratory environment.

Comparative study of gene expression of cholinergic system-related molecules in the human spinal cord and term placenta.

By reverse transcription-polymerase chain reaction, Southern blot analysis, direct sequencing, and immunohistochemistry, we studied the expression of cholinergic neuronal markers (choline acetyltransferase [ChAT], vesicular acetylcholine transporter [VAChT], and a high-affinity choline transporter [CHT1]), and gene regulatory molecules (repressor element-1 silencing transcription factor/neuron-restrictive silencer factor [REST/NRSF] and CoREST) in the human spinal cord and term placenta, both of which are well known to contain cells synthesizing acetylcholine. H-type, M-type, N2-type, and R-type ChAT mRNAs, VAChT mRNA, and CHT1 mRNA were detected in the spinal cord, but only H-type, M-type, and N2-type ChAT mRNAs, in the term placenta. REST/NRSF and CoREST were detected in the spinal cord and the placenta, but the amounts of both mRNAs were greater in the placenta than in the spinal cord. Further microdissection analyses revealed that the placental trophoblastic cells contained more REST/NRSF and CoREST transcripts than the spinal large motor neurons. Large motor neurons in the anterior horn of the spinal cord were immunohistochemically stained for ChAT and VAChT. In the placenta, stromal fibroblasts, endothelial cells, and trophoblastic cells of the chorionic villi were positively stained with anti-ChAT antibody but not with anti-VAChT antibody. These findings suggest that transcriptions of the R-type ChAT and VAChT mRNAs are coordinately suppressed in the human term placenta, which might be regulated in part by a REST/NRSF complex that binds to a consensus sequence of repressor element 1/neuron-restrictive silencer element (RE1/NRSE) in the 5' region upstream from exon R, whereas transcriptions of the H-type, M-type, and N2-type ChAT mRNAs might be independent of control by RE1/NRSE. It is possible that at least two separate regulatory mechanisms of gene expression are present for the human cholinergic gene locus, which might be selected by different combinations of DNA motifs and binding proteins to function in neuronal and non-neuronal cells.

c-ret regulates cholinergic properties in mouse sympathetic neurons: evidence from mutant mice.

The search for signalling systems regulating development of noradrenergic and cholinergic sympathetic neurons is a classical problem of developmental neuroscience. While an essential role of bone morphogenetic proteins for induction of noradrenergic properties is firmly established, factors involved in the development of cholinergic traits in vivo are still enigmatic. Previous studies have shown that the c-ret receptor and cholinergic properties are coexpressed in chick sympathetic neurons. Using in situ hybridization we show now that a loss-of-function mutation of the c-ret receptor in mice dramatically reduces numbers of cells positive for choline acetyltransferase (ChAT) and the vesicular acetylcholine transporter (VAChT) in stellate ganglia of homozygous newborn animals. The number of neurons positive for tyrosine hydroxylase (TH) mRNA, the rate-limiting enzyme of noradrenaline synthesis, is reduced to a smaller degree and expression levels are not detectably altered. Already at embryonic day 16 (E16), ChAT and VAChT-positive cells are affected by the c-ret mutation. At E14, however, ChAT and VAChT mRNAs are detectable at low levels and no difference is observed between wildtype and mutant mice. Our data suggest that c-ret signalling is necessary for the maturation of cholinergic sympathetic neurons but dispensable for de novo induction of ChAT and VAChT expression.

Augmentation effect of postprandial hyperinsulinaemia on growth of human hepatocellular carcinoma.

BACKGROUND: Cirrhotic patients with hepatocellular carcinoma (HCC) frequently have impaired glucose metabolism. AIMS: To investigate whether impaired glucose metabolism affects the growth rate of the tumour. PATIENTS AND METHODS: Tumour doubling time (DT), assessed by ultrasound imaging analysis, was measured in 60 patients with single small HCC (diameter <30 mm). DT was compared with plasma insulin and glucose concentrations following the oral glucose tolerance test (OGTT). The effect of continuous infusion of octreotide (a somatostatin analogue 200 microg/day) for three months on DT in five cases was assessed. RESULTS: The 60 patients were divided into two groups because the median DT was 140 days: rapid growth group (DT 140 days, n=30). Fasting plasma insulin concentration and area under the plasma insulin curve (AUC(ins)) of the OGTT (10.4 (6.2) microU/ml and 262 (152) microU/ml/h, respectively; mean (SD)) in the rapid growth group were significantly higher than those in the slow growth group (7.6 (4.3) and 146 (140), respectively) (p=0.041 and p=0.0006, respectively). In contrast, fasting plasma glucose concentration and area under the plasma glucose curve (AUC(gluc)) in the rapid growth group were significantly lower than those in the slow growth group (p=0.0003 and p=0.0012, respectively). Univariate and multivariate analyses of logistic regression models demonstrated that AUC(ins) was a significant factor contributing to the growth rate of HCC (p=0.001 and p=0.016, respectively). AUC(ins) significantly decreased after octreotide treatment (p<0.02) but AUC(gluc) did not significantly change. DT after treatment increased in three of the five patients and could not be calculated in the remaining two patients because of no change in the diameter of the tumour. CONCLUSIONS: These data suggest that postprandial hyperinsulinaemia is associated with accelerated HCC growth.

Inhibition of proliferation of gastric epithelial cells by a cyclooxygenase 2 inhibitor, JTE522, is also mediated by a PGE2-independent pathway.

BACKGROUND: Cyclooxygenase-2 (COX-2) is one of the rate-limiting enzymes for prostaglandin synthesis from arachidonic acid. Although it is known that inhibition of cyclooxygenase activity delays ulcer healing, the regulatory relationship between COX-2 and its metabolites in gastric epithelial cell proliferation is not well known. AIM: To investigate whether COX-2 has an effect on gastric mucosal cell proliferation and further studied whether such effect is mediated only by prostaglandin E2 (PGE2), a representative metabolite of arachidonates in the gastric mucosa. METHODS: Artificial wounds of defined area size were created on complete monolayer cell sheets of isolated rat gastric epithelial cells and rat gastric cell line RGM1 under the addition of arachidonic acid or a COX-2 selective inhibitor, JTE522. Repair of wounds was assessed by monitoring wound size, with cell proliferation detected using 5-bromodeoxyuridine staining. Quantity of secreted PGE2 was measured by enzyme immunoassay. RESULTS: Stimulation of foetal calf serum increased the expression of COX-2 protein and inhibition of COX-2 retarded wound healing with reduction of cell proliferation. Arachidonic acid increased PGE2 production and accelerated restoration. Combination of JTE522 and arachidonic acid resulted in a marked retardation of wound healing compared to the control, but JTE522 did not completely suppress the increase in cellular PGE2 content following the addition of arachidonate. CONCLUSIONS: The difference in the effects of JTE522 on PGE2 production and on wound healing suggest that the involvement of COX-2 in gastric epithelial cell proliferation is not mediated solely by PGE2.

O(2) delivery and the venous P(O(2))-O(2) uptake relationship in pump-perfused canine muscle.

Under the conditions of both an increased red cell affinity for O(2) at a constant rate of O(2) delivery (arterial O(2) content x flow) and a decrease in the rate of O(2) delivery induced by hypoxic hypoxia at constant blood flow, we have obtained a linear relationship between the partial pressure of O(2) in the muscle venous effluent (P(v,)(O(2))) and O(2) uptake (.V(O(2))). The relationship is described by the equation .V(O(2)) = D(a) x P(v,)(O(2)) + .V(O(2)conv)) where D(a) is the apparent O(2) diffusion capacity and .V(O(2)conv)) is O(2) delivery-limited .V(O(2)), and D(a) x P(v,)(O(2)) represents the O(2) diffusion-limited .V(O(2)) .V(O(2)diff)). From these observations, we propose the hypothesis that .V(O(2)) consists of two additive values, .V(O(2)conv)) and .V(O(2)diff)). The mechanism underlying the reduction in .V(O(2)) that is induced by reducing O(2) delivery to markedly below the .V(O(2)conv)) value has only been investigated using a model based on the single compartment of diffusion-limited .V(O(2)), and has not been investigated in terms of this additive .V(O(2)) model. The single compartment analysis appears to overestimate the role of O(2) diffusion in limiting the reduction of .V(O(2)) that occurs in response to a decrease in O(2) diffusion capacity, as reflected by the .V(O(2))/P(v,)(O(2)) ratio. To gain better insight into the mechanism involved, we altered the rate of O(2) delivery by changing arterial P(O(2)) from normoxia (with inhalation of air) to hypoxia (by inhalation of 10-11 % O(2)) and blood flow (with high and low flow rates (n = 7 for both groups), and very low and ischaemic flow rates (n = 4 for both groups)) in pump-perfused dog gastrocnemius preparations during tetanic isometric contractions at 1 Hz. As rates of O(2) delivery were reduced from 23.2 to 10.9 ml min(-1) (100 g)(-1), significant decreases in P(v,)(O(2)) and .V(O(2)) were observed (P < 0.05). From the data of P(v,)(O(2)) and .V(O(2)) values within this range of O(2) delivery rates, we obtained the regression equation .V(O(2)) = 0.22 x P(v,)(O(2)) + 8.14 (r = 0.58). From the equation, the intercept of the .V(O(2))-axis was significantly different from zero (P < 0.05), in accordance with the observation that the .V(O(2)) /P(v,)(O(2)) ratio (ml min(-1) (100 g)(-1) Torr(-1)) increased from 0.54 to 1.35 (P < 0.05). However, at extremely low rates of O(2) delivery (5.6 and 7.3 ml min(-1) (100 g)(-1) the .V(O(2))/P(v,)(O(2)) ratio was 1.51 and 2.80 (P < 0.05), respectively. This indicates a break in the linear .V(O(2))-P(v,)(O(2)) relationship as the rate of O(2) delivery was reduced to below the .V(O(2)conv)) value of the .V(O(2))-axis intercept. These results suggest that the reduction in .V(O(2)) caused by extreme reductions in the rate of O(2) delivery is not attributable to a reduction in O(2) diffusion capacity, as expected from the .V(O(2))/P(v,)(O(2)) ratio, but to a reduction in the O(2) delivery-limited .V(O(2)) component, as evaluated by the .V(O(2))-axis intercept of the linear .V(O(2))-P(v,)(O(2)) relationship.

Suppression of cell proliferation and deoxyribonucleic acid synthesis in the cloned rat hepatoma H4-II-E cells overexpressing regucalcin.

The role of endogenous regucalcin (RC) in the regulation of cell proliferation was investigated in the cloned rat hepatoma H4-II-E cells overexpressing RC stably. H4-II-E cells were transfected with RC/pCXN2 vector and the multiple neomycin-resistant clones which overexpress stably RC were selected. The RC content of RC/pCXN2-transfected cells used in this study was 19.7-fold as compared with that of the parental wild type H4-II-E cells. Wild type H4-II-E cells, pCXN2 vector-transfected cells (mock type), and RC/pCXN2-transfected cells (transfectants) were cultured for 24, 48, and 72 h in the presence of fetal bovine serum (10% FBS). Cell numbers of wild and mock type were significantly increased with the time course of culture. Cell numbers of transfectants was significantly suppressed as compared with that of wild and mock type. Deoxyribonucleic acid (DNA) synthesis activity in the nuclear fraction of H4-II-E cells was significantly suppressed in transfectants with culture for 12-48 h. The presence of anti-RC monoclonal antibody (10-50 ng/ml) in the reaction mixture caused a significant increase in DNA synthesis activity in the nuclei of wild type and transfectants; this increase was remarkable in transfectants. The effect of anti-RC monoclonal antibody (50 ng/ml) in increasing DNA synthesis activity in transfectants was completely prevented by the addition of regucalcin (1 microM). This study demonstrates that cell proliferation is suppressed in the cloned rat hepatoma H4-II-E overexpressing RC stably.

Glucose sensing based on interdigitated array microelectrode.

A micro glucose sensor consisting of an interdigitated array gold microelectrode was developed. The interdigitated array structure, which has 10 microns band width and 10 microns band gap, was fabricated in a small region (2.5 x 5 mm2) on a quartz substrate. Glucose oxidase was chemically fixed onto the electrode surface through self-assembled monolayer of 11-mercaptoundecanoic acid; ferroceneacetic acid was used as electron mediator. Electrochemical properties of the glucose oxidase-immobilized microelectrode were investigated by cyclic voltammogram measurements. Results confirmed that the reductive ferroceneacetic acid generated at counter electrode diffuses through a narrow band gap (10 microns) and can reach the working electrode surface.

Stimulatory effect of zinc on insulin-like growth factor-I and transforming growth factor-beta1 production with bone growth of newborn rats.

The effect of zinc, an essential trace element, on insulin-like growth factor-I (IGF-I) and transforming growth factor-beta1 production was investigated to determine the role of this metal in bone growth of newborn rats. Femoral-diaphyseal and metaphyseal tissues were obtained between 1 and 28 days after birth of newborn rats, and cultured for 24 h in a serum-free Dulbecco's modified Eagle's medium containing either vehicle or zinc sulfate (10(-6) - 10(-4) M). Protein concentration in the medium was significantly increased by culture with bone tissues of newborn rats with increasing age (14 and 21 days). Medium IGF-I and TGF-beta1 concentration was gradually reduced with increasing age after birth. The presence of zinc (10(-5) and 10(-4) M) caused a significant increase in protein, IGF-I, and TGF-beta1 concentrations in the medium cultured with the diaphyseal or metaphyseal tissues obtained at 7 and 14 days after birth. The expression of IGF-I and TGF-beta1 mRNA was demonstrated by reverse transcription-polymerase chain reaction (RT-PCR) analysis in the diaphyseal and metaphyseal tissues cultured for 24 h using rat IGF-I or TGF-beta1-specific primers. These expressions were significantly raised in the presence of zinc (10(-4) M) in culture medium. The present study demonstrates that zinc has a stimulatory effect on IGF-I and TGF-beta1 production in the femoral tissues with bone growth of newborn rats.

An evaluation of screening for lung cancer in Niigata Prefecture, Japan: a population-based case-control study.

Although an annual screening programme for lung cancer has been carried out widely in Japan since 1987, there is insufficient evidence to confirm its efficacy in terms of reducing mortality. In order to evaluate the efficacy of the lung cancer screening which has been widely carried out in Japan since 1987, a case-control study was conducted in Niigata Prefecture, Japan. In the study area, chest X-ray examinations for all participants and sputum cytology for high-risk participants were offered annually. Case subjects, who had died from lung cancer (174), and control subjects matched by sex, year of birth, residence and smoking status (801), who had been alive at the time of diagnosis of the corresponding case, were selected from the National Health Insurance holders. Screening histories of the subjects were compared between cases and matched controls for the identical calendar period before the time of diagnosis of the cases. The odds ratio of death from lung cancer for those screened within 12 months vs those not screened was 0.401 (95% CI: 0.272-0.591) with adjustment by smoking index. Our results suggest that annual lung cancer screening might reduce mortality from lung cancer by approximately 60%.

Clarithromycin resistance, but not CYP2C-19 polymorphism, has a major impact on treatment success in 7-day treatment regimen for cure of H. pylori infection: a multiple logistic regression analysis.

Mutations in the gene encoding the CYP2C-19 enzyme for PPI metabolism have been shown to enhance the chance for a cure in a H. pylori-positive patients using a two-week dual-therapy regimen involving omeprazole and amoxicillin. However, the impact of CYP2C-19 genetic polymorphism on eradication rates of a one-week triple-therapy regimen has not been examined. In this cohort study, 156 H. pylori-positive peptic ulcer or NUD patients who presented to our university hospital were recruited. They were treated by one-week omeprazole-amoxicillin-clarithromycin therapy. Host and bacterial predictive factors including H. pylori susceptibility and CYP2C-19 genotyping, as well as cure rate for H. pylori infection, were studied. Cure rate was 85.9% (95% CI: 79-91%) on an intent to treat (ITT) basis. By multiple logistic regression analysis, only clarithromycin resistance had a significant impact on treatment success (odds ratio 28.7: 95% CI: 6-172). CYP2C-19 genetic polymorphism was not associated with a significant change in cure rate. These observations indicate only clarithromycin susceptibility, not CYP2C-19 polymorphism, has a major impact on the treatment success when using a seven-day OAC H. pylori treatment regimen.

Molecular cloning and sequencing of the cDNA coding for a novel regucalcin gene promoter region-related protein in rat, mouse and human liver.

The molecular cloning and sequencing of the cDNA coding for a novel regucalcin gene promoter region-related protein (RGPR) was investigated using rat, mouse and human liver cDNA library with a yeast one-hybrid system and a rapid amplification of cDNA ends (RACE) method. The clone coding an unknown protein was isolated, and a novel protein was identified. This protein was termed as RGPR-p117. RGPR-p117 in rat, mouse and human liver consisted of 1058, 1051 and 1060 amino acid residues with calculated molecular mass of 117, 115 and 117 kDa and estimated pI of 5.69, 5.70 and 5.71, respectively. The homologies of amino acids among rat, mouse and human RGPR-p117 were at least 70%. RGPR-p117 had a leucine zipper motif. The expression of RGPR-p117 mRNA was found in the liver, kidney, heart, spleen, and brain of rats. The database search of the human RGPR-p117 showed that its gene consisted of at least 26 exons spanning approximately 4.1 kbp and localized on human chromosome 1q25.2. Furthermore, we found a cDNA clone which was highly identical to a front half part of the human RGPR-p117 cDNA, using the BLAST search of human RGPR-p117. This cDNA clone was a splicing variant of human RGPR-p117, which derived from human placental choriocarcinoma. Our study demonstrates that a novel gene coding RGPR-p117 is present in rat, mouse and human.