The association between neutropenia and prognosis in stage III colorectal cancer patients receiving adjuvant chemotherapy.

Neutropenia during chemotherapy has been reported to be a predictor of better survival in patients with several types of cancer, although there are no reports on stage III colorectal cancer (CRC). The purpose of this study was to examine the association between neutropenia and prognosis in stage III CRC patients receiving adjuvant chemotherapy consisting of oral uracil and tegafur (UFT) plus leucovorin (LV). We retrospectively analysed 123 patients with stage III CRC who received UFT/LV as adjuvant chemotherapy. The end-point was disease-free survival (DFS). Survival curves of the two categories (neutropenia absent vs. present) were estimated using the Kaplan-Meier method and compared by the log-rank test. We estimated the hazard ratio (HR) for DFS according to neutropenia after adjustment for covariates by multivariate analyses using Cox's regression analysis. A total of 33 (26.8%) patients experienced neutropenia. Patients without neutropenia showed a significantly lower DFS than those with neutropenia (3-year DFS 57.3% vs. 81.2%, P = 0.0213). By multivariate analysis, neutropenia and histological type were independent prognostic factors, with HR of 0.410 (neutropenia absent vs. present, P = 0.045) and 4.793 (well to moderately differentiated vs. poorly differentiated, P = 0.004) respectively. We demonstrated that neutropenia occurring during adjuvant chemotherapy consisting of UFT/LV may be a prognostic factor of recurrence in stage III CRC patients.

Predictive index for the onset of medication overuse headache in migraine patients.

Migraine patients are particularly prone to develop medication overuse headache (MOH). However, the risk factors for the transformation of migraine to MOH are still not clear. We investigated gene polymorphisms, personality traits, and characteristics of headache and lifestyle in 47 migraine patients (aged 36.4 ± 10.3) and 22 MOH patients (aged 39.6 ± 9.9) who progressed from migraine and made a scoring system for a predictive index (PI) of the onset of MOH in patients with migraine. By multivariate logistic stepwise regression analysis, type of migraine, regular and sufficient dietary intake, and methylenetetrahydrofolate reductase (MTHFR) C677T (rs1801133) and dopamine D2 receptor (DRD2) C939T (rs6275) polymorphisms were selected as significant factors that contribute independently to the development from migraine to MOH (P < 0.05). The regression coefficients (ß) of these four selected factors were approximated and scored. The PI score in MOH patients (7.32 ± 1.60) was significantly higher than that in migraine patients (4.62 ± 1.83, P < 0.001). The proposed scoring system should in the future be the object of larger studies to confirm its validity.

Negative predictors of clinical response to triptans in patients with migraine.

Moderate to severe migraine attacks are treated with triptans. However, about 25% of migraineurs fail to respond to triptans. We investigated the involvement of gene polymorphisms, personality traits and characteristics of headache, and made a scoring system for prediction of clinical response to triptans in patients with migraine. Gene polymorphisms including serotonin (5-HT)(1B) receptor G861C and dopamine receptor 2 (DRD2) C939T, personality traits and characteristics of headache were investigated in 46 consistent responders and 14 inconsistent responders to triptans. The multivariate stepwise logistic regression analysis revealed that age, periorbital/deep orbital pain and C/C genotype carrier at DRD2 C939T were significant factors that contributed independently to the negative response to triptans in patients with migraine. Their odds ratios were 6.329 (40-69 vs. 20-39 years, 95% CI 1.441-27.778), 6.772 (no vs. yes, periorbital/deep orbital pain, 95% CI 1.159-39.580) and 14.085 (non-C/C vs. C/C genotype at DRD2 C939T, 95% CI 1.253-166.667), respectively. The predictive index (PI) of clinical response to triptans in patients with migraine was calculated using these three factors. The score in inconsistent responders (1.6 ± 0.6) was significantly higher than that in consistent responders (0.8 ± 0.7, P < 0.001). Sensibility of low-score (RI = 0) group was 100%, and specificity of high-score (PI ≥ 2) group was 87%. The proposed scoring system should in the future be the object of larger studies to confirm its validity.

Predictors of restenosis after coronary artery stenting.

AIM: The aim of this study was to analyze restenosis after percutaneous coronary intervention, factors related to restenosis after coronary artery stenting and the degree of the risk of restenosis were evaluated. METHODS: The study enrolled 181 patients (249 lesions) who underwent the first coronary artery stenting. Multivariate analysis was performed, and the restenotic index (RI) was calculated by combining the extracted predictors. RESULTS: Among the 181 patients (249 lesions), restenosis occurred in 89 (111 lesions) and did not occur in 92 (138 lesions). Vascular revasculation was performed in 95 restenosed target lesions in 68 patients. The mean period of follow-up angiography after the procedures was 206 days in the restenosis group and 271 days in the non-restenosis group, i.e. significantly shorter in the restenosis group. As a result of multivariate analysis, diabetes mellitus, Cr level, amount of the contrast medium used and stent diameter were selected as significant factors that independently contributed to the restenosis after coronary artery stenting. By combining these factors, the RI was calculated by the following formula for the prediction of restenosis: RI=exp (1.088xCr+0.909xdiabetes mellitus+0.871xcontrast medium+0.591xstent diameter). CONCLUSION: The risk of restenosis after coronary artery stenting can be predicted to an extent according to the RI devised in this study.

Evaluation of paraumbilical vein as a prognostic index of severe liver cirrhotic patients with portal-systemic shunts.

AIM: The aim of this study was to predict the outcome in severe liver cirrhotic patients with portal-systemic shunts. METHODS: One-hundred and sixteen patients with liver cirrhosis diagnosed as Child-Pugh class B and C with portal-systemic shunts confirmed by abdominal ultrasonography, computed tomography and magnetic resonance imaging were enrolled in this study. Twenty-three factors were evaluated concerning clinical laboratory parameters and extracted prognostic factors using the Cox proportional hazards model, and the prognostic index (PI) was prepared by combining these factors. RESULTS: The cumulative survival rates after admission were 64.6%, 35.6% and 25% after 1, 3 and 5 years, respectively. Using multivariate analysis, age, the presence of hepatocellular carcinoma (HCC), portal vein tumor thrombosis (PVTT) and paraumbilical vein (PUV) shunt were selected as significant prognostic factors that contributed independently to the prognosis of severe liver cirrhotic patients with portal-systemic shunts. The PI was calculated with the following formula using these 4 factors. PI = 0.042 x Age + 0.913 x HCC + 0.989 x PVTT + 1.079 x PUV shunt. The group with a high score for PI was found to die with significantly higher frequency than the group with a low score. CONCLUSIONS: It was found that tumor related factors and PUV shunt were the most important factors for severe liver cirrhotic patients with portal-systemic shunts. The PI is suggested to be an appropriate index to predict the prognosis for these patients.

Prognostic factors of primary biliary cirrhosis detected by health screening.

AIM: The liver cirrhosis is likely to differ in the Japanese and Western populations. Thus, we performed a retrospective cohort analysis by a review of clinical records to clarify prognostic factors after the onset of primary biliary cirrhosis (PBC) detected by health screening. METHODS: The subjects were 52 patients with PBC. Thirty-nine factors were evaluated concerning clinical data and extracted prognostic factors using the Cox proportional hazard model. RESULTS: The mean duration of the follow-up period was 5.1 years, during which 6 (11.5%) of the patients died. The cumulative survival rate after the onset of PBC was 93.4% after 5 year, and 67.8% after 10 years. Multivariate analysis indicated 2 factors, i.e. the body mass index (BMI), and IgG, as independent prognostic factors. Their hazard ratios were 0.399 (per 1 kg/m2 of BMI) and 1.282 (per 100 mg/dL of IgG). The prognostic index (PI) was calculated by the following formula using these 2 factors. PI = 0.919 x BMI+0.249 x IgG. CONCLUSIONS: The prediction of the outcome using PI based on the 2 factors provides additional information for the determination of the therapeutic approach in PBC after health screening.

Membrane modification during secretory granule formation in rat somatotrophs.

Somatotrophs from male rat anterior pituitary were used to investigate the formation of secretory granules. When enzymatically dispersed cells were incubated with cationized ferritin (CF) for 15 min, CF labeled immature secretory granules, but not mature granules of somatotrophs. Most immature granules labeled by CF transformed to the mature types within 120 min. This indicates that the fusion of endocytic vesicles with the immature granules occurs during the maturation process of secretory granules. The internalized CF was distributed not only in the immature secretory granules, but also in the peripheral region of trans Golgi cisternae or GERL. Enzyme cytochemistry revealed that acid phosphatase-positive cisternae (GERL) were the main site for secretory granule formation, and was devoid of thiamine pyrophosphatase (TPPase) activity. A small number of secretory granules were also present in the peripheral regions of TPPase-positive Golgi cisternae. The granule-forming sites, however, lacked TPPase activity, while the remaining region of the same cisterna showed the positive enzyme activity. This indicates that the granule-forming region at the periphery of Golgi cisterna is different from the remaining part of the same cisterna in terms of cytochemical properties. This probably results from the insertion of endocytic vesicle membrane, since the same granule-forming sites preferentially fused with CF-labeled small vesicles which lacked cytochemical TPPase activity. Taken together. Our results suggest that the membrane of secretory granules is modified during the granule formation, at least partly by the fusion of endocytic small vesicles with Golgi cisternae (or GERL), and with immature secretory granules.

Stimulation of tetrahydrobiopterin synthesis induced by insulin: possible involvement of phosphatidylinositol 3-kinase.

Although we recently showed that insulin increases the intracellular concentration of tetrahydrobiopterin (BH4), which is one of the cofactors of nitric oxide (NO) synthase, the mechanism of the effect was not elucidated. In the present study, we examined the signaling pathway of the stimulation of BH4 synthesis by insulin in mouse brain microvascular endothelial cells. Extracellular and intracellular BH4 levels were determined as biopterin by using reversed-phase high performance liquid chromatography with fluorometric detection. Measurement of the level of mRNA for GTP cyclohydrolase I (GTPCH), which is the rate-limiting enzyme for de novo BH4 synthesis, was performed by reverse transcription-polymerase chain reaction (RT-PCR). Addition of insulin to endothelial cells caused an increase of not only the intracellular but also the extracellular BH4 level in a time- and a concentration-dependent manner. Insulin also induced an increase of the level of GTPCH mRNA. Moreover, 2,4-diamino-6-hydroxypyrimidine, an inhibitor of GTPCH, inhibited the insulin-induced enhancement of BH4 synthesis. The increase in the BH4 level and the induction of GTPCH mRNA by insulin were reduced by wortmannin and LY294002, which are both phosphatidylinositol 3-kinase (PI3-kinase) inhibitors. These results suggest that insulin stimulates BH4 synthesis through the de novo synthetic pathway involving induction of GTPCH, and that the signaling pathway involves the activation of PI3-kinase.

Dehydroepiandrosterone decreases serum tumor necrosis factor-alpha and restores insulin sensitivity: independent effect from secondary weight reduction in genetically obese Zucker fatty rats.

Dehydroepiandrosterone (DHEA) and its sulfate ester are the most abundant circulating adrenal steroids in humans. Administration of DHEA has been reported to have beneficial effects on obesity, hyperlipidemia, diabetes, and atherosclerosis in obese rodents, although its effects on insulin resistance have not been fully elucidated. In this study, the effects of DHEA treatment on insulin sensitivity were investigated in genetically obese Zucker rats, an animal model of insulin resistance, using the euglycemic clamp technique. After 0.4% DHEA was administered for 10 days to female obese Zucker rats aged 16 weeks, body weight and plasma insulin decreased and glucose disposal rate (GDR), which was normally reduced in obese rats, rose significantly compared with age- and sex-matched control obese rats. On the other hand, although the pair-fed obese rats also showed levels of weight reduction similar to those of DHEA-treated rats, the increase in GDR of DHEA-treated rats was significantly greater than in pair-fed rats, suggesting a direct ameliorating effect of DHEA on insulin sensitivity of obese rats. Serum concentration of tumor necrosis factor (TNF)-alpha, one of cytokines causing insulin resistance, was also reduced significantly in DHEA-treated, but not in pair-fed obese rats. In conclusion, our results suggest that DHEA treatment reduces body weight and serum TNF-alpha independently, and that both may ameliorate insulin resistance in obese Zucker fatty rats.

Wistar fatty rat is obese and spontaneously hypertensive.

The purpose of this study was to determine whether genetically obese Wistar fatty rats have higher blood pressure than their lean littermates and if so to elucidate the mechanism of this obesity-related hypertension. We measured blood glucose and plasma insulin levels, blood pressure, and catecholamine and sodium excretions in age-matched female Wistar fatty and lean rats. After 12 weeks of age, the body weight of Wistar fatty rats was significantly greater than that of their lean counterparts. Fasting blood glucose and plasma insulin concentrations were higher in the fatty than the lean rats throughout the observation period (8 to 24 weeks of age). Systolic blood pressure of fatty rats measured by the tail-cuff method was similar to that of lean rats at 8 weeks of age (135 +/- 2 [mean +/- SEM] versus 134 +/- 3 mm Hg) but significantly higher at 16 (158 +/- 2 versus 136 +/- 3 mm Hg, P < .01) and 24 (166 +/- 5 versus 142 +/- 2 mm Hg, P < .01) weeks of age. Urinary norepinephrine excretion was significantly increased in the fatty rats at both 16 (1755 +/- 173 versus 977 +/- 128 ng/24 h, P < .05) and 24 (1907 +/- 283 versus 737 +/- 173 ng/24 h, P < .01) weeks of age. The ratio of urinary norepinephrine excretion to body weight was also significantly increased in the fatty rats. These results show that with increasing body weight Wistar fatty rats develop hypertension, which may be attributable to an increased sympathetic nerve activity.(ABSTRACT TRUNCATED AT 250 WORDS)

cDNA cloning and inducible expression of human multidrug resistance associated protein 3 (MRP3).

Previously, we cloned rat MRP3 as a candidate for an inducible transporter for the biliary excretion of organic anions [Hirohashi et al. (1998) Mol. Pharmacol. 53, 1068-10751. In the present study, we cloned human MRP3 (1527 amino acids) from Caco-2 cells. Human MRP3 is predominantly expressed in liver, small intestine and colon; hepatic expression of MRP3 was observed in humans but not in normal rats. In HepG2 cells, the expression of MRP3 was induced by phenobarbital. These results suggest that MRP3 may act as an inducible transporter in the biliary and intestinal excretion of organic anions.

Role of calcium ions in dopamine release induced by sodium cyanide perfusion in rat striatum.

We have previously reported a transient and remarkable increase in dopamine (DA) release in the rat striatum during application of 2 mM sodium cyanide (NaCN) through a brain microdialysis membrane. In the present study we examined the involvement of extracellular Ca2+ in this response. Rats were divided into 4 groups. In the NaCN group a microdialysis probe inserted into the striatum was perfused with Ringer's solution containing 2 mM NaCN for 60 min. The Ca2+ free + NaCN group was subjected to perfusion with NaCN dissolved in Ca2+ free Ringer's solution, and the CdCl2 + NaCN group with the same plus 0.3 mM CdCl2 (a non-specific Ca2+ channel blocker). In the NaCN and Ca2+ free + NaCN groups DA levels in the dialysates increased to 36- and 44-fold of the control level, respectively, while this was suppressed to only a 16-fold increase in the CdCl2 + NaCN group. In response to a 100 mM KCl perfusion given 3 hr later DA levels were increased (22-fold) in the control group. On the other hand this response was inhibited in the NaCN group (3-fold), but not in the other two groups. An in vitro study with striatal slices showed a gradual increase in intracellular Ca2+ during incubation with 2 mM NaCN. These results suggest that excessive influx of extracellular Ca2+ during NaCN perfusion may contribute partly to the increase in the extracellular DA level in the striatum, and also to the suppression of a DA increase in response to high K+ stimulation observed 3 hr later.

Do alpha-adrenergic receptors participate in control of the circadian rhythm of IOP?

The alpha 2-adrenergic antagonists, yohimbine and rauwolscine, and the alpha 1-adrenergic antagonist, bunazosin, were used to explore the role of alpha-adrenergic receptors in the regulation of the circadian rhythms of intraocular pressure and aqueous flow in New Zealand white rabbits. Blockade of alpha 2-adrenergic receptors with yohimbine or rauwolscine produced small decreases in intraocular pressure during both light and dark phases. Rauwolscine had no effect on aqueous flow during the light or dark, but it increased the concentration of norepinephrine in the aqueous during both light and dark. These observations are difficult to reconcile with earlier suggestions that increased sympathetic input to the eye increases intraocular pressure and aqueous flow during the dark. The role of alpha 2-adrenergic receptors in the control of the circadian rhythm of intraocular pressure is unclear. Blockade of alpha 1-adrenergic receptors with bunazosin produced a dose-dependent reduction of IOP during the dark phase of the circadian cycle, a smaller reduction during the light phase, and no reduction during either light or dark in rabbits after superior cervical ganglionectomy or preganglionic section of the cervical sympathetic trunk (decentralization). Bunazosin decreased pupil diameter during the dark phase but had no effect on aqueous flow. Because it is unlikely that alpha 1-adrenergic blockade increased outflow facility or uveoscleral outflow, the mechanism for the role of alpha 1-adrenergic receptors in the control of the circadian rhythm of intraocular pressure in rabbits remains to be identified.

Response and level of beta-adrenergic, vasoactive intestinal peptide, and PACAP receptors during the circadian cycle.

PURPOSE: To determine whether a nocturnal increase of ciliary process beta-adrenergic receptor responsiveness can explain the observation that timolol decreased the aqueous flow rate and intraocular pressure (IOP) during the night but not during the day in rabbits. METHODS: Rabbits were housed in alternating 12-hour periods of light and dark. In vitro stimulation of tissue cyclic adenosine monophosphate (cAMP) levels by isoproterenol (ISO), vasoactive intestinal peptide (VIP), pituitary adenylate cyclase activating polypeptide (PACAP), or a soluble derivative of forskolin (sFSK) was measured in ciliary processes harvested at mid-light phase and early and late dark phase. Inhibition of ISO and VIP stimulation of ciliary process cAMP by an alpha 2-adrenergic agonist and maximal binding of [125I]I-pindolol, [125I]I-VIP, and [125I]I-PACAP in ciliary process membranes were measured at the same three times. RESULTS: Although there may have been a nocturnal increase in the sensitivity of ciliary process cAMP levels to stimulation by ISO, this was not observed consistently, VIP, but not PACAP, stimulation increased at night, but there was no change in the response to sFSK. Inhibition by apraclonidine of elevated ciliary process cAMP levels was constant at all three times. Ligand-binding studies showed little change in ciliary process beta-adrenergic, VIP-, or PACAP-receptor levels at the three times. CONCLUSIONS: There is no convincing evidence for a nocturnal increase in beta-adrenergic receptor sensitivity in rabbit ciliary processes that could explain the difference between day and night effects of timolol on aqueous flow and IOP.

Behavioral stimulation without alteration of beta and 5-HT receptors and adenylate cyclase activity in rat brain after chronic sertraline administration.

Effects of chronic treatment with selective 5-HT reuptake inhibitors (SSRIs) on the monoaminergic functions have not been much investigated in compared with tricyclic antidepressants. Therefore, we compared the effects of 3-week treatment with sertraline, a potent SSRI, to those of imipramine (10 mg/kg, IP, twice a day), on monoamine receptors and adenylate cyclase (AC) activity in rat brain. Two-week treatment with both sertraline and imipramine reduced immobility in the water wheel test to the comparable extent. Sertraline treatment did not affect Kd and Bmax of [3H]CGP12177 and [3H]ketanserin bindings or cAMP, accumulation by norepinephrine, isoproternol, 5'-guanylylimidodiphosphate [Gpp(NH)p] and forskolin in the cortical membrane compared with vehicle-treated rats. On the other hand, imipramine treatment decreased Bmax of both bindings and norepinephrine- or isoproternol-stimulated cAMP accumulation. Treatment with either antidepressant induced no apparent changes in [3H]8-OH-DPAT [2-(N, N-dipropylamino)-8-hydroxy-1,2,3,4-tetrahydronaphthalene] binding in the hippocampal membrane. These results suggested that chronic treatment of sertraline induced little effect on monoamine receptors and AC activity in the brain and that the alteration of these functions may not be primarily involved in antidepressive effects of antidepressants, at least of SSRIs.

Genetically regulated expression of UDP-N-acetylgalactosamine: GM3(NeuGc) N-acetylgalactosaminyltransferase [EC 2.4.1.92] activity in mouse liver.

GM2 containing NeuGc was a major ganglioside in the liver of mouse strains such as BALB/c, DBA/2, C3H/He, and C57BL/10, whereas WHT/Ht mouse liver did not contain GM2(NeuGc) but contained GM3(NeuGc) as a major ganglioside. Since GM3(NeuGc) is a biosynthetic precursor of GM2(NeuGc), WHT/Ht liver was considered to lack the ability to synthesize GM2(NeuGc) from GM3(NeuGc) (Hashimoto, Y., et al. (1983) J. Biochem. 93, 895-901). In this study we measured the activity of UDP-N-acetylgalactosamine : GM3(NeuGc) N-acetylgalactosaminyltransferase in the liver of BALB/c, WHT/Ht, and their progeny. The transferase activity in the microsomal fraction of BALB/c liver was 2.10 +/- 0.32 X 10(-5) units/mg protein (means +/- S.D.), whereas no activity was detected in that of WHT/Ht liver, F1 hybrids between BALB/c and WHT/Ht expressed GM2(NeuGc) as well as the enzyme activity, the level of which was almost half that in BALB/c liver 1.10 +/- 0.12 X 10(-5) units/mg protein). The backcross generation of F1 to WHT/Ht segregated into two groups with respect to expression of GM2(NeuGc) and the transferase activity: 11 of the 21 mice analyzed expressed both GM2(NeuGc) and the transferase activity (1.28 +/- 0.18 X 10(-5) units/mg protein), whereas the rest expressed neither. These results suggest that the expression of GM2(NeuGc) is directly regulated by the activity of UDP-N-acetylgalactosamine: GM3(NeuGc) N-acetylgalactosaminyltransferase in mouse liver.

T lymphopenia in genetically obese-diabetic Wistar fatty rats: effects of body weight reduction on T cells.

Patients with long-standing diabetes may have a propensity for infection-related mortality. In this study, lymphocyte subsets, the proliferative response of splenocytes to mitogens, and circulating levels of tumor necrosis factor alpha (TNF-alpha) in genetically obese-diabetic Wistar fatty (fa/fa) rats (WF) were longitudinally compared versus lean (+/?) litters (WL). Moreover, the effects of weight reduction with voglibose treatment on immunity were evaluated (WFV and WLV). Body weight was significantly increased in WF compared with WL. Hyperglycemia and hyperlipidemia developed, respectively, 11 weeks and 5 weeks thereafter throughout the observation periods. Circulating T cells and T-cell subsets of WF were significantly reduced after 22 weeks. There were also significant decreases in CD4+ and CD8+ thymocytes and the proliferative response of splenocytes. Circulating levels of TNF-alpha were significantly increased in WF. Treatment with voglibose resulted in significantly reduced blood glucose, insulin, cholesterol, triglyceride, and body weight in WFV. After weight reduction, circulating T cells and T-cell subsets were increased and TNF-alpha was decreased significantly in WFV. Our results suggest that the number and function of T cells in WF may be reduced, which may be related at least in part to elevated TNF-alpha levels, although the role of the other factors such as glucose, insulin, cholesterol, and triglycerides on T-cell immunity should be further investigated.

Circadian intraocular pressure management with latanoprost: diurnal and nocturnal intraocular pressure reduction and increased uveoscleral outflow.

Based on their mechanism of action, the most frequently used ocular hypertensive agents, the beta-blockers, cannot be assumed to reduce IOP during sleep. The need for drugs that reduce IOP around-the-clock is underscored, however, by the fact that inadequate nocturnal ocular perfusion pressure is considered to be one of the likely causes of glaucomatous optic neuropathy especially in some cases of normal tension glaucoma. The studies reviewed here demonstrate that latanoprost, a new ocular hypotensive prostaglandin F2 alpha analogue, applied once a day at a concentration of 0.005%, maintains a statistically highly significant IOP reduction around-the-clock. The magnitude of this IOP reduction was found to be essentially identical during the day and at night, both in patients maintained on timolol and in those not receiving other glaucoma medication. Latanoprost-induced IOP reduction was also found to be associated with increased uveoscleral outflow in normotensive volunteers, both during the day and at night. These circadian studies suggest that this new ocular hypotensive agent can be expected to be particularly useful for the medical management of some forms of glaucoma, such as normal tension glaucoma, when the cause of the glaucomatous damage cannot be linked specifically to diurnal IOP abnormalities.

Amended structure of side chains in a cell wall mannan from Candida albicans serotype A strain grown in yeast extract-Sabouraud liquid medium under acidic conditions: detection of the branched side chains corresponding to antigenic factor 4.

In a previous study, we reported the excess production of alpha-1,3-linked mannose residues with the complete disappearance of beta-1,2-linked mannose residues in cell wall mannans of Candida albicans serotype A strain cells, which were grown in yeast extract-Sabouraud liquid medium at pH 2.0. In the present study, we examined the immunochemical reactivity of the same mannan of NIH A-207 with an enzyme-linked immunosorbent assay (ELISA) using several antisera to antigenic factors of the genus Candida (FAbs) and the structure of the mannan by two-dimensional homonuclear Hartmann-Hahn analysis. The ELISA showed that the mannan reacts to FAb 4 but not to FAbs 13b and 34, which are reported to be antibody factors against linear side chains containing an alpha-1,3-linked mannose residue. In the Hartmann-Hahn analysis, we found two branched side chains, Man alpha 1-2Man alpha 1-3[Man alpha 1-6]Man alpha 1-(2Man alpha 1-)(2)2Man and Man alpha 1-3[Man alpha 1-6]Man alpha 1-(2Man alpha 1-)(2)2Man, instead of the previously reported linear side chains. The branched side chains are oversynthesized under acidic conditions.

Regulation of dopamine uptake by basic fibroblast growth factor and epidermal growth factor in cultured rat astrocytes.

We examined the characteristics of dopamine (DA) uptake and its regulation by neurotrophic factors such as basic fibroblast growth factor (bFGF) and epidermal growth factor (EGF) in cultured rat astrocytes. In the presence of inhibitors of monoamine oxidase (MAO) and catechol-O-methyl-transferase (COMT), astrocytes took up DA by Na(+)-dependent and Na(+)-independent mechanisms that were sensitive to a reduction in temperature. The Na(+)-dependent and Na(+)-independent components increased linearly with increasing [3H]DA concentration (1-1000 microM), and showed no saturation. Na(+)-dependent DA uptake was significantly inhibited by ouabain, a Na(+)-K+ ATPase inhibitor. In bFGF-treated astrocytes, [3H]DA uptake increased in a time-dependent manner until 48 h, and declined after 72 h in both the presence and absence of Na+. In EGF-treated astrocytes, [3H]DA uptake increased in a time-dependent manner until 72 h in both the presence and absence of Na +. This enhancement of DA uptake induced by EGF or bFGF was significantly inhibited when the cells were cultured with actinomycin D, cycloheximide, or brefeldin A. Actinomycin D and brefeldin A also significantly inhibited the basal uptake of [3H]DA into astrocytes. These results suggest the existence of Na(+)-dependent and Na(+)-independent DA uptake in cultured rat astrocytes, and that EGF or bFGF might stimulate the expression and translocation of the extraneuronal DA transporter.