REG Iα is a biomarker for predicting response to chemotherapy with S-1 plus cisplatin in patients with unresectable stage IV gastric cancer.

BACKGROUND: The regenerating gene Iα (REG Iα) is involved in gastric carcinogenesis as an antiapoptotic factor. Therefore, we investigated whether REG Iα confers resistance to chemotherapeutic drugs in gastric cancer (GC) cells and whether REG Iα expression is useful for predicting the response to chemotherapy and outcome in patients with GC. METHODS: A total of 70 patients with unresectable stage IV GC received first-line chemotherapy with S-1 and cisplatin (S-1/CDDP). The expression of REG Iα was evaluated immunohistochemically using biopsy samples obtained before chemotherapy, and its relationship to clinicopathological parameters was analysed statistically. The effects of REG Iα gene induction on resistance to 5-FU or CDDP treatment were examined by cell survival assay and flow cytometry. RESULTS: Of the 70 patients with unresectable stage IV GC, 19 (27%) were positive for REG Iα expression. The expression of REG Iα was independently predictive of poorer progression-free and overall survival in such patients (hazard ratio (HR) 2.46; P=0.002 and HR 1.89; P=0.037, respectively). The gene induction of REG Iα conferred resistance to cell death induced by 5-FU or CDDP in GC cells. CONCLUSION: In patients with stage IV GC, REG Iα, which confers resistance to chemotherapeutic drugs in GC cells, is a potential biomarker for predicting resistance to S-1/CDDP treatment.

Cyclic performance of carbon-coated TiO2 for photocatalytic activity of methylene blue decomposition.

Carbon-coated TiO2 powders were prepared from the mixtures of anatase (ST-01) and carbon precursor poly(vinyl alcohol) in different ratios by carbonization at 900 degrees C in a flow of either Ar or N2. Carbon-coated TiQ2 thus prepared was either fixed on an adhesive tape or formed into a film by using the organic binder poly(tetrafluoroethylene) and used for photodecomposition of methylene blue in water repeatedly. On the samples fixed on adhesive tapes, the rate constant was reproduced with about 10-20% scattering during repeated uses in a fresh methylene blue solution up to 7 times. On the samples formed into films by using the organic binder, the rate constant was reproduced after the 3rd or 4th cycle. The mutual relations in rate constant were almost the same among three different conditions for the determination, suspending the sample particles, fixed on an adhesive tape and formed into a film. The rate constant for the photodecomposition was found to give a maximum on the sample containing about 9 mass% carbon.

A newly developed peripheral anterior chamber depth analysis system: principle, accuracy, and reproducibility.

AIM: To develop a new, non-contact system for measuring anterior chamber depth (ACD) quantitatively, and to investigate its accuracy as well as interobserver and intraobserver reproducibility. METHODS: The system scanned the ACD from the optical axis to the limbus in approximately 0.5 second and took 21 consecutive slit lamp images at 0.4 mm intervals. A computer installed program automatically evaluated the ACD, central corneal thickness (CT), and corneal radius of curvature (CRC) instantly. A dummy eye was used for investigating measurement accuracy. The effects of CT and CRC on the measurement results were examined using a computer simulation model to minimise measurement errors. Three examiners measured the ACD in 10 normal eyes, and interobserver and intraobserver reproducibility was analysed. RESULTS: The ACD values measured by this system were very similar to theoretical values. Increase of CRC and decrease in CT decreased ACD and vice versa. Data calibration using evaluated CT and CRC successfully reduced measurement errors. Intraobserver and interobserver variations were small. Their coefficient variation values were 7.4% (SD 2.3%) and 6.7% (0.7%), and these values tended to increase along the distance from the optical axis. CONCLUSION: The current system can measure ACD with high accuracy as well as high intraobserver and interobserver reproducibility. It has potential use in measuring ACD quantitatively and screening subjects with narrow angle.

Expression and subcellular localization of multifunctional calmodulin-dependent protein kinases-I, -II and -IV are altered in rat hippocampal CA1 neurons after induction of long-term potentiation.

Long-term potentiation (LTP) is considered to be associated with an increase in expression as well as activity of Ca(2+)/calmodulin-dependent protein kinases (CaMKs). LTP-induced and control hippocampal slices were studied by immunohistochemical and electronmicroscopic analyses using anti-CaMK-I, -II and -IV antibodies. All three kinases were demonstrated to increase their expression in CA1 neurons. CaMK-I was shown to mainly localize in the cytoplasm of the control and LTP-induced neurons, and a significant increase of immunoreactivity was observed in the latter neurons. A part of CaMK-I was found to translocate to the nuclei of LTP-induced hippocampal CA1 neurons. Direct evidence of the translocation of CaMK-II from cytoplasm to nuclei in LTP was demonstrated by immuno-electronmicroscopy. A significant increase in expression of CaMK-IV in the nuclei was also observed. Our data suggest that all the three CaMKs were actively involved in nuclear Ca(2+)-signaling in LTP.

Receptor-mediated facilitation of cell volume regulation by swelling-induced ATP release in human epithelial cells.

Osmotic swelling induces the release of intracellular ATP in a number of cell types. In the immediate vicinity of the cell surface, released ATP has been shown to reach a concentration high enough to stimulate P2-purinergic receptors in a human epithelial cell line, Intestine 407. The role of released ATP in the regulatory volume decrease (RVD) after cell swelling was thus studied in Intestine 407 cells. The RVD was suppressed by an ATP hydrolyzing enzyme, apyrase, or by a purinergic receptor antagonist, suramin. Extracellular application of ATP accelerated the RVD rate in a concentration-dependent manner. An increase in the cytosolic free-Ca(2+) concentration was induced by a hypotonic challenge, and the swelling-induced Ca(2+) response was partially suppressed by apyrase or suramin. A rise in cytosolic Ca(2+) was also induced by extracellular application of ATP or UTP, but not ADP, 2-methylthio-ATP or alpha,beta-methylene ATP. The ATP-induced Ca(2+) response was blocked by suramin. Therefore, it is concluded that RVD is facilitated by ATP, which is released upon cell swelling, by augmenting intracellular Ca(2+) rise via the stimulation of purinergic (P2Y(2)) receptors in the human epithelial cell.

Circadian rhythm of autonomic nervous function in patients with normal-tension glaucoma compared with normal subjects using ambulatory electrocardiography.

PURPOSE: To compare circadian rhythm of autonomic nervous function in patients with normal-tension glaucoma with subjects with normal eyes. METHODS: Thirty-two patients with normal-tension glaucoma and 32 age-matched normal subjects who had no history of systemic disorders and no currently treated systemic disorders, especially diseases of the autonomic nervous system, were studied. An ambulatory electrocardiogram was installed that recorded heartbeats for 48 hours. Low-frequency and high-frequency values were calculated as markers of the autonomic nervous system status based on heart-rate variability using a power-spectrum analysis. RESULTS: The low-frequency values of patients with normal-tension glaucoma during the spans of an active day and a resting night were significantly greater than those of normal subjects, and this difference was emphasized during the night resting span. However, the high-frequency values of patients with normal-tension glaucoma were similar to those of normal subjects. The normal subjects showed a significant age-related decrease in all investigated parameters except the low-frequency values during the resting span. However, the patients with normal-tension glaucoma showed a significant age-related decrease only in low-frequency values during the active day. Patients with normal-tension glaucoma with progressive visual field defects showed much greater values than other cases, although the values were not significantly different. CONCLUSION: These results indicate that a disturbance of the circadian rhythm of the autonomic nervous system may exist in patients with normal-tension glaucoma.

Ultrasound biomicroscopic study of ciliary body thickness in eyes with narrow angles.

PURPOSE: To determine the ciliary body thickness and other biometric findings in eyes with narrow angles. METHODS: Eighteen otherwise normal eyes with narrow angles in 18 Japanese patients and 18 normal control eyes with open angles in 18 age-matched and sex-matched Japanese patients were studied. A-scan ultrasonography was performed to measure anterior chamber depth, lens thickness, axial length, and relative lens position. Ultrasound biomicroscopy was also performed to obtain measurements of the anterior ocular structures, including anterior chamber depth and ciliary body thickness at sites 1 mm and 2 mm posterior to the scleral spur (positions 1 and 2, respectively). RESULTS: Compared with normal control eyes, the narrow-angle eyes showed a shallower anterior chamber (narrow angle, 1.87 +/- 0.27 mm; control, 2.69 +/- 0.26 mm; P <.0001), a thicker lens (4.97 +/- 0.49 mm, 4.26 +/- 0.53 mm; P <.0001), a more anteriorly located lens (2. 21 +/- 0.13, 2.35 +/- 0.14; P <.0001), a shorter axial length (22.70 +/- 0.97 mm, 23.41 +/- 0.86 mm; P =.012), and a thinner ciliary body (position 1: 454 +/- 107 microm, 602 +/- 86 microm; P <.0001; position 2: 203 +/- 50 microm, 321 +/- 68 microm; P <.0001). Lens thickness was significantly correlated with ciliary body thickness at positions 1 (R(2) = 0.34; P =.0001) and 2 (R(2) = 0.43; P <.0001). Anterior chamber depth was significantly correlated with ciliary body thickness at positions 1 (R(2) = 0.48; P <.0001) and 2 (R(2) = 0.56; P <.0001). CONCLUSION: Thinning of the ciliary body may be one of the important factors associated with the anterior location of the lens, the increased lens thickness, and the decreased anterior chamber depth in eyes with a narrow angle.

Super pulse CO2 laser for bracket bonding and debonding.

A super pulse and a normal pulse CO2 laser were used to carry out enamel etching and bracket debonding in vitro and in vivo. The shear bond strength of the orthodontic brackets attached to laser-etched and conventional chemically-etched extracted premolars was measured. The pulp cavity temperature was also measured using the same laser irradiation conditions as the shear test. Both super pulse and normal pulse CO2 laser etching resulted in a lower shear bond strength (super pulse: 6.9 +/- 3.4 kg, normal pulse: 9.7 +/- 5.2 kg) than that of chemical etching (15.3 +/- 2.8 kg). Furthermore, the super pulse CO2 laser was able to create debonding at 2 watts within a period of less than 4 seconds (2.9 +/- 0.9 seconds). The super pulse, when irradiating the ceramic brackets from above, during debonding showed a 1.4 degrees C temperature increase in the dental pulp at 2 watts and an increase of 2.1 degrees C at 3 watts. While etching, directly irradiating the enamel surface at 3 watts, the dental pulp showed a temperature increase of 3.5 degrees C. These temperature increases were within the physiologically acceptable limits of the pulp. These results indicate that, in orthodontic treatments, super pulse CO2 laser debonding is more useful than laser etching.

The expression of Ca2+/calmodulin-dependent protein kinase I in rat retina is regulated by light stimulation.

Ca2+/calmodulin-dependent protein kinase I (CaM-kinase I) in rat retina was analyzed by immunohistochemical analysis, Western blot analysis and kinase activity assay. Western blot analysis revealed two immunoreactive bands similar to those detected in the brain. Developmental studies revealed that CaM-kinase I expression increased in accordance with postnatal development. Expression of CaM-kinase I in the retinas of rats raised in the complete darkness markedly decreased. CaM-kinase I activity assay supported these findings. Synapsin I was shown to be a possible intrinsic substrate of CaM-kinase I in rat retina. These results elucidated that CaM-kinase I is expressed in the retina and may play an important role in the retinal functions and that the expression of CaM-kinase I is regulated by light stimulation.

Alteration of mucin in human conjunctival epithelia in dry eye.

PURPOSE: The objective of this study was to determine whether alteration in mucins could be detected in patients with dry eye symptoms by using the monoclonal antibody H185, which recognizes carbohydrate epitopes on mucin molecules. METHODS: Immunohistochemistry and immunoelectron microscopy was used to examine binding of H185 antibody to conjunctival cells obtained by nitrocellulose filter paper stripping (impression cytology). Two study populations were examined. Study I included 22 patients with dry eye symptoms and 13 normal volunteers. Study II included 16 aqueous-deficient dry eye patients and 14 age-matched control subjects. RESULTS: Results of the studies demonstrated significant differences in binding patterns of H185 to conjunctival cells in normal eyes compared with those of patients with dry eye symptoms. In normal eyes, the antibody bound to apical cells in a mosaic pattern, with cells exhibiting either light, medium, or intense binding. A predominant pattern in patients with dry eye symptoms was loss of the mosaic pattern with replacement by a "starry sky" pattern in which there was a lack of apical cell binding (hence, dark sky) but increased binding to goblet cells (hence, stars in the sky). The starry sky pattern correlated with rose bengal staining. CONCLUSIONS: From these studies it is concluded that there is an alteration either in mucin distribution or mucin glycosylation on the surfaces of apical conjunctival cells in dry eye and that glycosylation of goblet cell mucins changes with the disease.

Activation of cAMP-dependent C1- currents in guinea-pig paneth cells without relevant evidence for CFTR expression.

1. To determine whether Paneth cells exhibit functional expression of cAMP-activated Cl- currents and molecular expression of the cystic fibrosis transmembrane conductance regulator (CFTR), we applied whole-cell patch clamp and single-cell mRNA analysis by reverse transcription (RT) followed by polymerase chain reaction (PCR) amplification to single Paneth cells in crypts isolated from the guinea-pig small intestine. 2. Prominent activation of Cl- currents was consistently observed after stimulation with dibutyryl cAMP and forskolin or with vasoactive intestinal polypeptide (VIP). The cAMP-activated Cl- current was inhibited by removal of intracellular ATP or administration of an inhibitor of protein kinase A. 3. Many of the biophysical and pharmacological properties of the currents were phenotypically similar to those of the CFTR Cl- channel, such as the ohmic current-voltage relationship, the anion selectivity with a Type III sequence (Br- > Cl- > I- >> F- >= gluconate-), I--induced blockage, insensitivity to a stilbene-derivative Cl- channel blocker, and sensitivity to a carboxylate analogue Cl- channel blocker. The sensitivity of the current to glibenclamide was, however, much weaker than that reported for the CFTR Cl- channel current. In contrast to the time independence of CFTR currents, the inward component of the Paneth cell Cl- currents exhibited inactivation kinetics. 4. Expression of CFTR mRNA could not be detected by RT-PCR analysis in almost all single Paneth cells, although its expression was consistently detected at the whole-crypt level. The presence of a small number of CFTR-expressing epithelial cells, which were scattered both in villi and crypts but not at the crypt base where Paneth cells were located, was demonstrated by immunocytochemistry. 5. Taken together, it appears that guinea-pig Paneth cells functionally express cAMP-activated Cl- conductance without relevant evidence for molecular expression of CFTR. Functional expression of VIP receptors in the Paneth cells was also demonstrated.

Glibenclamide blocks volume-sensitive Cl- channels by dual mechanisms.

To study the mechanisms of glibenclamide actions on volume-sensitive Cl- channels, whole cell patch-clamp studies were performed at various pH levels in human epithelial Intestine 407 cells. Extracellular application of glibenclamide reversibly suppressed volume-sensitive Cl- currents in the entire range of voltage examined (-100 to +100 mV) and accelerated the depolarization-induced inactivation at pH 7.5. When glibenclamide was applied from the intracellular side, in contrast, no effect was observed. At acidic pH, at which the weak acid glibenclamide exists largely in the uncharged form, the instantaneous current was, in a voltage-independent manner, suppressed by the extracellular drug at micromolar concentrations without significantly affecting the depolarization-induced inactivation. At alkaline pH, at which almost all of the drug is in the charged form, glibenclamide speeded the inactivation time course and induced a leftward shift of the steady-state inactivation curve at much higher concentrations. Thus it is concluded that glibenclamide exerts inhibiting actions on swelling-activated Cl- channels from the extracellular side and that the uncharged form is mainly responsible for voltage-independent inhibition of instantaneous currents, whereas the anionic form facilitates voltage-dependent channel inactivation in human epithelial Intestine 407 cells.

Reticalmin: a novel calcium/calmodulin-dependent protein kinase IV-like protein in rat retina.

Western blot analysis of 100,000 g supernatant of rat retina using a polyclonal anti-Ca2+/ calmodulin-dependent protein kinase IV (CaM-kinase IV) antibody revealed an immunoreactive mass of 35 kDa, termed reticalmin. Lower amount of a isoform of CaM-kinase IV was also expressed in rat retina. Reticalmin did not react with anti-CaM-kinase IV C-terminal peptide antibody which recognized alpha and beta isoforms of CaM-kinase IV and calspermin. Immunohistochemically reticalmin was shown to be localized mainly in the outer segment of photo-receptor cells, and in dendrites of inner plexiform layers and may be in nuclei of ganglion cells and some inner nuclear layer cells.

[Visual field defect in eyes with branch retinal artery occlusion].

There have been few clinical studies that analyzed quantitatively the visual field defects observed in eyes with branch retinal artery occlusion. In the current report we retrospectively studied Humphrey static perimetric results in 7 eyes and Goldmann kinetic perimetric results in 8 eyes with branch retinal occlusion. All but one eye showed altitudinal-like field defects. The static perimetric results demonstrated large difference between sensitivities in each pair of upper and lower measured points adjacent to the horizontal line nasal to Mariott's scotoma, which indicated a steep slope between the defective field and the normal area. The defective field showed either absolute scotoma or sensitivity loss of 20 dB or larger.

Sensitivity of volume-sensitive Cl- conductance in human epithelial cells to extracellular nucleotides.

The sensitivity to extracellular nucleotides of volume-sensitive Cl- channel activity was investigated by whole cell and single-channel patch-clamp recordings in a human small intestinal epithelial cell line (Intestine 407) during steady osmotic swelling. Adenine nucleotides added to the bathing solution suppressed whole cell volume-sensitive Cl- currents with the potency sequence of ATP > ADP > AMP. In contrast, extracellular adenosine 3',5'-cyclic monophosphate (cAMP) at over 0.1 mM increased volume-sensitive Cl- currents in the entire voltage range examined, whereas guanosine 3',5'-cyclic monophosphate was without effect. Neither the single-channel conductance nor the open probability was affected by extracellular cAMP. Extracellular ATP (at over 30 microM), in the Mg(2+)-free form, inhibited the whole cell volume-sensitive Cl- current, preferentially in the outward direction. By exposure to extracellular ATP, the single Cl- channel current became flickery at positive potentials. These results indicate that the volume-sensitive Cl- channel in the human epithelial cell is stimulated voltage independently by extracellular cAMP but blocked voltage dependently by the Mg(2+)-free form of extracellular ATP.

Auditory isochrony: time shrinking and temporal patterns.

It has previously been reported that the duration of short time intervals is conspicuously underestimated if they are preceded by shorter neighbouring time intervals. This illusion was called 'time shrinking' and it was argued that it strongly affects the perception of auditory rhythms. In the present study this supposition has been pursued in three experiments. In the first, temporal patterns consisting of two, three, and four intervals had to be judged for anisochrony, which was invoked by offsetting the last sound from its isochronous position. By a constant method, it was determined that the last sound of fast sequences (50 ms base interval) had to be delayed by about 30 ms in order for isochronous rhythms to be perceived. Another interesting finding was that for sound sequences with base intervals up to 200 ms it was the difference limen, rather than Weber's ratio, that was constant for anisochrony detection. In the second experiment, the temporal patterns comprised two intervals, presented serially or separately. The deviation of isochrony could be on either the first or the second interval. The data, gathered by an adaptive method, showed time shrinking to be effective even up to a base interval of 200 ms. The third experiment involved a constant method and anisochrony was implemented on the first interval of two interval patterns. Time shrinking affected perceived isochrony in sequences with base intervals of 50, 100, and 200 ms. It is argued that the paradoxical results of anisochrony detection can be explained in terms of time shrinking. Some anomalies of rhythm perception and production that are the result of time shrinking are discussed.(ABSTRACT TRUNCATED AT 250 WORDS)

Combined occlusion of the central retinal artery and vein.

A 56-year-old man, complaining of sudden visual loss in his left eye, demonstrated cream-colored retinal edema along a macular branch of the central retinal artery, overall delay of fluorescence in angiography and general depression of the central visual field, which were interpreted as the incomplete form of central retinal artery occlusion (CRAO) mimicking cilioretinal or branch retinal artery occlusion. After receiving paracentesis and fibrinolytic agents, the patient recovered his vision gradually, while the ophthalmoscopic findings progressed to show central retinal vein occlusion (CRVO) changes consisting of dilated and tortuous retinal veins and scattered intraretinal hemorrhages. The condition associated with the retinal edema indicated combined obstruction of the central retinal artery and the central retinal vein (combined CRAO/CRVO). Six months later, both the ocular fundus and the vision returned to normal. The similar cases in Japanese literature in which ophthalmoscopic findings of combined CRAO/CRVO was followed by aggravation of CRVO changes with or without recovery of vision were reviewed.

[Intrathecal morphine for postoperative pain relief after transvaginal hysterectomy].

We studied the effect of a low-dose intrathecal morphine (0.1 or 0.2 mg) in postoperative pain relief and the incidence of side effects. Two hundred and fifteen patients scheduled for transvaginal hysterectomy were divided into 3 groups according to intrathecal morphine doses: M1 (morphine 0.1 mg N = 75), M2 (morphine 0.2 mg N = 69) and C (control N = 71). A standard mid-line lumbar puncture was performed using a 25-gauze needle in the L3/4 interspace. Preservative-free morphine hydrochloride mixed in hyperbaric tetracaine solution was administered intrathecally. Pain relief was significantly greater for the first 24 hrs in groups M1 and M2 compared with group C. Respiratory depression was not seen in any groups. The incidence of vomiting was about 40% in all groups. We conclude that intrathecal morphine 0.1-0.2 mg is useful for pain relief after transvaginal hysterectomy and accompanies no major side effects.