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We prospectively examined influence of androgen deprivation therapy (ADT) on lipid and glucose metabolisms in Japanese patients with prostate cancer.Patients with prostate cancer who were hormone-naive and scheduled to receive long-term ADT were recruited between 2011 and 2013. Body weight, abdominal circumference and blood testing associated with lipid and glucose metabolism were recorded every 3 months during 1 year of ADT. Computed tomography (CT) was performed to measure areas of subcutaneous and visceral fat before and after 1 year of ADT. ADT was limited to a luteinizing hormone-releasing hormone (LHRH) agonist with or without bicalutamide.Of 218 patients registered, data were available from 177 patients who completed 1 year of ADT. Of these, CT was performed before and after 1 year of ADT in 88 patients. Median age was 75 years (range, 49-85 years). Median PSA before ADT was 16.7 ng ml(-1) (range, 0.3-3316). Clinical stage was B (54.2%), C (23.2%) and D (20.9%). Mean increases in body weight and abdominal circumference after 1 year of ADT were 2.9 and 3.0%, respectively. Mean increases in total, low-density lipoprotein and high-density lipoprotein cholesterol and triglycerides were 10.6, 14.3, 7.8 and 16.2%, respectively. Mean increases in fasting blood sugar and hemoglobin A1c (HbA1c) were 3.9 and 2.7%, respectively. Lipid alterations were noted in patients without comorbidities, whereas changes in HbA1c were noted in patients with diabetes mellitus at baseline. These lipid and glucose alterations were prominent in the early ADT period. Both visceral and subcutaneous fat, as measured by CT, increased by20%. The increase in subcutaneous fat was significantly greater than that in visceral fat (P=0.028).One year of ADT significantly changed lipid and glucose metabolism in Japanese patients with prostate cancer. Patient characteristics or comorbidities at baseline may be associated with ADT-induced metabolic changes.

Relationships between surface state of indium tin oxide (ITO) and luminescent properties, and aging characteristics of organic light emitting diodes (OLEDs) were investigated. Polycrystalline ITO surface used as the substrate of the thin film OLED was changed by chemical etching and ultraviolet (UV) irradiation. Initial properties of luminescent characteristics showed no significant difference between different ITO surface states. On the other hand, the device fabricated on UV irradiated ITO showed longer lifetime than that of without UV irradiation. We found that UV irradiation was effective for the aging characteristics of thin film OLED.

Renal cell carcinomas are immunohistochemically positive for oligosaccharides with the Le(x) determinant (Gal beta 1----4[Fuc alpha 1----3]GlcNAc) and its derivatives, as oncofetal antigens, and their expression is closely related to a better prognosis of the patients. This study was designed to clarify the difference in antigen localization at the ultrastructural level between renal cell carcinoma and normal tissues. In normal kidneys, Le(x) detected by monoclonal antibody (MAb) FH 2 and sialylated extended Le(x) (sialyl Le(x)-i) by MAb FH 6 were identified along the plasma membrane of microvilli of proximal tubule epithelial cells, with occasional immunoreactivity along the basolateral plasma membranes. Intracellular localization was very sparse. Renal cell carcinoma showed localization of Le(x) and sialyl Le(x)-i antigens along the cell membrane and in the cytosol as aggregates or filaments. Immunoreactive materials were also observed in the lumen formed among carcinoma cells. The cytosolic immunoreactivity, not observed in the normal kidney, was regarded as "abnormal cytosolic accumulation" of the antigens. This pattern was more pronounced in clear-cell carcinoma. Pretreatment of specimens with chloroform-methanol, which extracts glycolipids, decreased immunoreactivity in carcinoma tissues, particularly that in the cytosol. The extracts contained substances immunoreactive for MAb FH6. Our study has demonstrated that (a) remarkable changes occur in the ultrastructural localization patterns of sialyl Le(x)-i and Le(x) in renal cell carcinoma and (b) considerable amounts of glycolipids are contained in the substances with sialyl Le(x)-i deposited in the cytosol of clear-cell carcinoma.

This paper presents a new dual color FISH technique that will allow the use of cell specimens isolated from formaldehyde-fixed and paraffin-embedded (F-P) tissues. The new FISH method was used to examine numerical aberration in chromosome 17 and c-erbB2 gene amplification in 26 F-P epithelial ovarian cancer tissues. Numerical aberration of chromosome 17 appears frequently in clear cell adenocarcinoma. The frequency of amplification of the c-erbB2 gene in stage III and IV cases was significantly higher than in stage I cases. The FISH technique as established here may serve as a molecular tool for examination of clinico-pathological significance in F-P tissues.

In order to determine the clinical significance of numerical aberrations on chromosome 17 in uterine cervical and endometrial neoplasias, we investigated 140 cell specimens obtained from the uterine cervix and endometrium using the fluorescence in situ hybridization (FISH) method. These specimens consisted of ten normal cervical epithelium (NCE), 16 cervical intraepithelial neoplasias (CIN 1), 15 CIN 2, 35 CIN 3, 11 early invasive squamous cell carcinoma of the uterine cervix (early invasive SCC), 11 invasive SCC, 13 normal endometrium (NE), 17 endometrial hyperplasias (EH), and 12 endometrial ademocarcinomas (EA). After Papanicolaou staining on these specimens was decolorized, FISH was performed using chromosome 17 specific repetitive DNA probes. Signals of centromere of chromosome 17 in marked atypical cells were counted using a fluorescence microscope. There was a significant difference in the rate of cells with one signal on chromosome 17 between CIN 1 (7.1+/-0.7%) or 2 (7.0+/-0.5%) and CIN 3 (12.6+/-0.9%) (p0.01), and also between CIN 3 and early invasive SCC (19.6+/-1.0%) (p0.01). The rate of cells with three signals was significantly increased when the uterine cervical lesions were progressive to CIN 3 (p0.01). Cells with five or more signals occurred only in early invasive SCC and invasive SCC. There was a significant difference in the rate of cells with three signals between EH (4.8+/-0.6%) and EA (11.4+/-2.1%) (p0.05). Cells with five or more signals occurred only in EA. Examination of the numerical aberrations on chromosome 17 in uterine cervical and endometrial neoplasias has been suggested to be useful as an additional method for the differential diagnosis of these lesions.

Acetylcholine (ACh) caused repetitive transient Cl- currents activated by intracellular Ca2+ in single rat submandibular grand acinar cells. As the concentration of ACh increased the amplitude and the frequency of the transient Cl- currents increased. These responses occurred also in the absence of extracellular Ca2+ but disappeared after several minutes. Repetitive transient Cl- currents were restored by readmission of Ca2+ to the extracellular solution. The higher the concentration of extracellular Ca2+ readmitted, the larger the amplitude of the transient Cl- currents. Ca2+ entry through a store-coupled pathway was detected by application of Ca2+ to the extracellular solution during a brief cessation of stimulation with ACh. In these experiments too, the higher the concentration of Ca2+, the larger the transient Cl- currents activated by Ca2+ released from the stores. The time course of decrease in total charge movements of repetitive transient responses to ACh with removal of extracellular Ca2+ depended on a decrease in charge movements of each transient event rather than a decrease in frequency of the repetitive events. The decrease of charge movements of each transient event was due to a decrease in its amplitude rather than its duration. The results suggest that in this cell type and amplitude-modulated mechanism is involved in repetitive Ca2+ release and that Ca2+ entry is essential to maintain the repetitive release of Ca2+. The results further suggest that the magnitude of Ca2+ entry determines the number of unitary stores filled with Ca2+ which can synchronously respond to ACh.

In 33 patients with primary ovarian cancer who had not received any chemotherapy before surgery, expression of glutathione S-transferase pi (GST-pi) was studied immunohistochemically in relation to the response to chemotherapy with CDDP, and furthermore it was examined whether numerical aberration of chromosome 11 was available as a prognostic indicator. The obtained results were as follows: 1. Of 33 ovarian cancer tissue samples, 19 (57.6%) showed positive staining and 14 (42.4%) negative staining for GST-pi. 2. The 5 year survival rate of the patients with GST-pi positive tumors was significantly lower than that of those with GST-pi negative tumors (p0.01). 3. The frequency of detection of cells showing numerical aberration of chromosome 11 in tissues was significantly higher in ovarian cancers than in benign ovarian tumors and normal ovaries. 4. The 5 year survival rate of the patients with ovarian cancer in which the cells showing numerical aberration of chromosome 11 at 20% or more was significantly lower than that of those in which such cells were under 20% (p0.02). These results show that expression of GST-pi and numerical aberration of chromosome 11 in the tissues of human ovarian cancers are useful as prognostic indicators.

Filled skutterudite compounds attracted many researchers' interest as strongly correlated electron systems. The skutterudite family includes binary, unfilled skutterudite compounds. We have studied the powder x-ray diffraction of unfilled skutterudite compounds TSb3 (T=Rh and Ir) under high pressure up to around 40 GPa, using synchrotron radiation. Pressure-induced irreversible structural changes of TSb3 (T=Rh and Ir) have been observed. Structural instability of the unfilled skutterudite compounds TSb3 (T=Co, Rh and Ir) under high pressure is discussed.

Although chemotherapy is indispensable for the treatment of ovarian cancer, secondary acute leukemia has become increasingly important as one of the most unfavorable late effects according to widespread long-term chemotherapy. We report a patient suffering from acute nonlymphocytic leukemia (ANLL) 3 years after treatment for stage IV ovarian cancer began.

Cell population of urinary leucocytes of 22 patients (intestine group) who underwent operations using intestinal segments for the urinary tract was compared with that of 26 complicated UTI patients without surgical intervention (control group). Eosinophils were recognized in 15.5 per cent of urinary leucocytes of the intestine group. However, in the control group, urine eosinophils were recognized only in 0.15 per cent. Although in sterile urines of the intestine group eosinophils were recognized in 30.8 per cent, in infected urines, the percentage of eosinophils decreased. Conversely, the percentage of neutrophils increased to 91.9 per cent. These findings suggest that neutrophils play an important role in infected urines of the intestine group as in urines of the control group. Significant differences were found in the values of urinary secretory IgA, IgG, IgM and urinary osmolarity. To evaluate the influence of these differences on the activity of phagocytosis of urinary leucocytes, the activity of phagocytosis of polymorphonuclear leucocytes (PMN), isolated from the peripheral blood, was investigated in immersion in urines of both groups. The mean rate of phagocytosis of E. coli in urines of both groups showed no statistically significant differences. However, urinary osmolarity of the intestine group was within the suitable range for phagocytosis and the activity of phagocytosis in urine was correlated with the value of IgG, which suggests that IgG has the opsonic effect. In contrast, the activity of phagocytosis in urine of the control group was strongly correlated with the value of urinary osmolarity. The growth of Providencia, Streptococcus, P. aeruginosa, whose frequency of isolation from urine of both groups was different in our previous study, and E. coli was studied in urine of the two groups. No significant difference in the growth of all bacteria was found, however. This finding suggests that the difference in the frequency of isolation of these bacteria from urine possibly depends on the adhesion of bacteria to intestinal epithelium.

The characteristics of urinary tract infections (UTI), secretory IgA and urine composition of 24 patients (intestine group) who underwent operations using intestinal segments in urinary tract were compared with those of 26 complicated UTI patients without surgical intervention (control group). No significant differences were found in the frequency of either mono or polymicrobial infections between both groups. However, the frequency of bacterial isolation was different in the two groups, Streptococcus spp. and Providencia spp. were frequently isolated and P. aeruginosa was rarely isolated from the urine of the intestine group compared with from the urine of the control group. Furthermore, in polymicrobial infections of the intestine group with indwelling catheters, E. faecalis and Providencia spp. were frequently isolated simultaneously. The mean value of urinary secretory IgA was 94.0 micrograms/dl in the intestine group and 25.0 mu/dl in the control group (p less than 0.0001). The mean urinary osmolarity was 370 mOsm/kg in the intestine group and 500 mOsm/kg in the control group (p less than 0.005). The other parameters, including urinary pH and urea N concentration, showed no statistically significant differences. These findings suggest that urinary tract constructed from intestinal segments differs from the urinary tract without surgical intervention in the feature of UTI and anti-bacterial defense mechanisms.

Urinary LMOX concentration was studied in 18 patients with unilateral ureteral obstruction. The concentration of LMOX in the urine from the mild obstructed kidney was 124 to 2,140 micrograms/ml and 10 micrograms/ml in the severely obstructed ones. The difference was probably due to the intensity and the duration of the obstruction. The patient with 99mTc-DMSA renal uptake of less than 3% also had a urinary LMOX concentration of less than 7 micrograms/ml. The above results seem to show that 7 micrograms/ml in urinary LMOX concentration is a significant figure for treatment of UTI. 99mTc-DMSA renal uptake and renal echogram were used to estimate the excretion rate of antibiotics into the urine.

A new fucoganglioside, 6B, which accumulates in human colonic adenocarcinoma but is absent in normal colonic mucosa, was isolated from a monosialoganglioside fraction of colonic adenocarcinomas. The structure of this ganglioside was identified as shown below by methylation analysis, direct probe mass spectrometry, and enzymatic degradation followed by examination of the degradation products with specific monoclonal antibodies. (formula; see text) The hybridoma (FH6) secreting a monoclonal IgM antibody directed to this glycolipid was selected by reactivity of the antibody with this ganglioside and lack of reactivity with other glycolipids having a closely related structure, such as sialosyllactoneotetraosylceramide (IV3NeuAcnLc4), sialosyllactofucopentaosy(III)ceramide (IV3NeuAcIII3FucnLc4), sialosyllactofucopentaosy(II)ceramide (sialosyl-Lea glycolipid; IV3NeuAcIII4FucLc4), and 6C fucoganglioside (sialosyl 2 leads to 6 fucoganglioside; VI6NeuAcIII3FucnLc6). The antibody was highly reactive with a large variety of human cancer cells, but was less reactive or did not react with a variety of normal cells.

1 Effects of various drugs were examined on the mechanical responses of the circular smooth muscle of guinea-pig vas deferens evoked by field stimulation and ACh. 2 During field stimulation (0.1 msec, 3–80 Hz) a contraction consisting of two (initial and second) phases occurred. In addition, another contraction (after-response) occurred after the cessation of the stimulation particularly at higher frequencies (40–80 Hz). 3 Guanethidine (2–20 μM) suppressed both the initial and second phases of the field stimulation-evoked responses. Prazosin (0.1-1 μM) suppressed the second phase but minimally affected the initial phase. 4 Both phases were suppressed by atropine (0.1-1 μM.) and potentiated by physostigmine (5 μM). 5 dTC (0.5–20 μM) did not suppress either phase but potentiated the second phase. 6 ACh (0.28–280 μM) produced a contraction, which was reduced by atropine but was little affected by either dTC or TTX. 7 These results suggest that at the postsynaptic site of the neuromuscular junction in this tissue the sympathetic process activates the smooth muscle through two different types of receptor; one is non-noradrenergic and the other is an α-adrenoreceptor. In addition, a cholinergic process also activates the smooth muscle through muscarinic receptors.

Induction of rat heme oxygenase is mediated by at least two factors: its substrate heme and heat shock (Shibahara, S., Muller, R.M., and Taguchi, H. (1987) J. Biol. Chem. 262, 12889-12892). We have identified the cis-acting element of the rat heme oxygenase gene (HO gene), that was specifically bound by a nuclear protein prepared from rat glioma cells. We have termed this protein as heme oxygenase transcription factor (HOTF), whose estimated molecular weight is about 40,000. The element identified is CCACCACGTGACTCGAG (-51/-35) located just upstream of TATA-like sequence. The functional studies indicate that this sequence is required for the accurate and efficient initiation of transcription from the HO gene promoter. Since the HOTF-binding element is similar to the upstream promoter sequence (UPS) of the adenovirus 2 major late promoter (Ad2MLP), we examined whether rat HOTF is homologous to the upstream stimulatory factor (USF) or major late transcription factor previously identified in HeLa cells, which interacts with the UPS and activates the transcription from the Ad2MLP. HOTF specifically binds to the UPS of Ad2MLP, whereas HOTF failed to form a stable complex with the mutated HOTF-binding element lacking a GTGA sequence (-44/-42 of the HO gene), the sequence of which is identical to a core sequence of the USF-binding sites. Moreover, we show that like USF, HOTF is heat stable. These results suggest that HOTF may be homologous to USF or the major late transcription factor. Since USF is present in uninfected cells, it is conceivable that the expression of the HO gene is regulated at least in part by USF.

A new fucoganglioside, disialosyl Lea, has been found in the disialoganglioside fraction of human colonic adenocarcinoma. The ganglioside has been isolated from four other disialogangliosides by high-performance liquid chromatography followed by preparative high-performance thin-layer chromatography. The structure of the antigen was characterized by its conversion to lactofucopentaosyl(II)ceramide (Lea-active ceramide pentasaccharide), methylation analysis, and high-mass range electron-impact as well as field-desorption mass spectrometry of the permethylated derivative, as shown below: (Formula; see text) Specific removal of alpha 2----3-linked sialosyl residue by influenza virus A2 sialidase, or preferential hydrolysis of the same residue by Clostridium perfringens sialidase in the absence of detergent, resulted in the formation of an intermediate product, monosialosyl LeaII (III4FucIII6NeuAcLc4), which reacted with anti-Lea antibody and with monoclonal antibody FH7 and may have a sialic acid linked at the 6 position of GlcNAc. The IgG3 monoclonal antibody FH7 was established, which reacts specifically with disialosyl Lea and monosialosyl LeaII as above, but does not react with disialosyllactotetraosylceramide (IV3NeuAcIII6Neu-AcLc4), monosialosyl LeaI (IV3NeuAcIII4FucLc4), and other mono- and disialogangliosides isolated from the same cancer tissue. The antibody FH7 may be useful in the detection of human cancer.

A series of glycolipids having the X determinant (Gal beta 1----4 [Fuc alpha----3]GlcNAc) at the terminus and a fucosyl alpha 1----3 residue at the internal GlcNAc residue have been isolated and characterized from tumor tissues (Hakomori, S., Nudelman, E., Levery, S.B., and Kannagi, R. (1984) J. Biol. Chem. 259, 4672-4680. A series of monoclonal antibodies that differentially recognize glycolipids with mono-, di-, and trifucosylated type 2 chain have been isolated and characterized. The antibody FH4 shows a remarkable preferential reactivity towards di-/or trifucosylated type 2 chain, i.e. it does not react with monofucosylated structures, including lactofucopentaosyl (III) ceramide (III3FucnLc4), monofucosyl neolactonorhexaosylceramide (y2, V3FucnLc6), and monofucosyl neolactonoroctaosylceramide (Z1, VII3FucnLc8), but reacts well with di- and trifucosylated type 2 chain structures such as difucosyl neolactonorhexaosylceramide (III3V3Fuc2nLc6) and trifucosyl neolactonoroctaosylceramide (III3V3VII3Fuc3nLc8). Two other monoclonal antibodies, FH5 and ACFH18, preferentially react with trifucosylated type 2 chain structure (III3V3VII3Fuc3nLc8), although cross-reactivity with difucosylated type 2 chain (III3V3Fuc2nLc6) was observed. They showed a minimal cross-reaction with monofucosylated type 2 chain. In contrast, the antibody FH1 does not react with III3FucnLc4 but reacts with V3FucnLc6, III3V3Fuc2nLc6, and III3V3VII3Fuc3nLc8. Two monoclonal antibodies, FH2 and FH3, do not discriminate among various glycolipids having fucosylated type 2 chain, and their reactivities are essentially similar to previously established antibodies directed to the X determinant, such as anti-SSEA-1, WGHS 29, VEP8 and 9, My-1, etc. This series of antibodies will be useful to detect the specific type of glycolipid with fucosylated type 2 chain accumulating in human cancer and in undifferentiated cells.

1. The effects of various drugs on excitatory junction potentials (EJPs) elicited by field stimulation of the longitudinal smooth muscle of the guinea-pig vas deferens were examined using the sucrose-gap method. 2. Field stimulation with a single pulse (1.0 ms, 20 Vcm-1) produced a depolarization of 3-7 mV. When a single pulse was repetitively applied, the depolarization showed summation, resulting in action potential firing. 3. Tetrodotoxin (1 microM) and guanethidine (1 microM) completely suppressed EJPs whilst propranolol (1 microM) or atropine (0.1 microM) did not affect them. Phentolamine (1 microM) slightly potentiated EJPs. 4. d-Tubocurarine (dTC) at lower concentrations (0.5-2 microM) suppressed EJPs, but at a higher concentration (20 microM) the drug little affected them. 5. Hexamethonium (C6) suppressed EJPs at low concentrations (0.5-5 microM), but potentiated them at a higher concentration (50 microM). 6. Either dTC or C6, each at a concentration of 0.5 microM, did not affect the depolarization induced by exogenously applied ATP or noradrenaline. 7. These results suggest that in this tissue cholinergic nerve activity can contribute to the magnitude of EJPs, and that this effect is mediated through nicotinic receptors.

Effects of various drugs on the mechanical responses of the longitudinal smooth muscle of guinea-pig vas deferens evoked by ACh and field nerve stimulation were examined. ACh (28-280 microM) produced a contraction consisting of two phases. The first phase of the contraction was suppressed by guanethidine and by nicotinic antagonists. The second phase was suppressed only by atropine. Both phases were unaffected by TTX or prazosin. Field stimulation (0.1 msec, 40 Hz) evoked contractions which also consisted of two (early and late) phases. Guanethidine (0.1-1 microM) suppressed both phases whilst prazosin (1 microM) suppressed only the late phase. Atropine (0.1 microM) suppressed both phases whilst physostigmine (5 microM) potentiated both phases of field stimulation-evoked contractions. Pentolinium suppressed both phases of field stimulation-evoked contractions at low concentrations (2-10 microM), but potentiated them at a higher concentration (100 microM). dTC at a low concentration (0.5 microM) suppressed the early phase, but slightly enhanced the late phase of field stimulation responses. At a higher concentration (20 microM), dTC potentiated both phases of the response. Pentolinium and dTC did not affect the contractions induced by 90 mM-K ions, ATP or NA. These results suggest that cholinergic nerves possess an excitatory action not only directly at the smooth muscle but also at the noradrenergic nerve terminals. The role of each receptor is discussed further.

The clinical significance of serum sialyl SSEA-1 antigen was evaluated using the sera of 1261 patients with malignant tumors and 717 patients with non-malignant diseases measured with Otsuka Assay Laboratories' RIA Kits. The results indicate that the antigen was frequently elevated in the sera from the patients with various adenocarcinomas, including lung (45%), pancreas (64%) and ovary (57%). The false positive incidence of antigens in the sera from the patients with non-malignant disorders was as low as 4.9%. No correlation was observed with other tumor markers examined in this study, and the diagnostic efficiency increased significantly by the combined determination of sialyl SSEA-1 antigen level with other markers. The serial determination of the serum sialyl SSEA-1 antigen level has a clinical utility also in monitoring the patients with adenocarcinoma receiving surgical operation and/or chemotherapy as indicated by the results of the longitudinal observation of the patients.

1. Two noradrenergic components of contraction were induced by field nerve stimulation of the circular smooth muscle of guinea-pig vas deferens. One component occurred during the stimulation (former-response) and the other component occurred after stimulation had ceased (after-response), with a distinct tension fall between these two responses. 2. The effects of stimulation frequency and duration upon these components were examined. 3. Prazosin (0.05-0.5 microM) suppressed both responses, whereas nifedipine (0.02 microM) suppressed only the after-response. 4. It is suggested that in this tissue there are two neurogenic alpha-adrenergic contractions which possess different Ca2+ activation mechanisms. The former-response is due to an increase in cytoplasmic Ca2+ concentration through a pathway independent of action potentials, while the after-response is attributed to action potentials arising as a secondary response to the released noradrenaline.

The LewisX (LeX) antigen [characterized by trisaccharide Gal beta 1----4 (Fuc alpha 1----3)N-acetylglucosamine] is an oncodevelopmental antigen in the human colon. Monoclonal antibodies (MoAbs), anti-SSEA-1 and AH8-183, which recognize LeX antigen either on short oligosaccharide side chains or as a terminal immunodeterminant on longer carbohydrate side chains of glycoconjugates, bind to most colon cancer tissues but also to some normal colon mucosae. However, the monoclonal antibodies FH1, FH4, FH6, and IB9, which recognize extended difucosylated and trifucosylated LeX structures or their sialylated derivatives, are more cancer-associated because they rarely bind to normal colon mucosa. In the present study, these MoAbs were used to compare the expression of various LeX-related antigens in premalignant (adenomatous) and nonpremalignant (hyperplastic) colorectal polyps. Antigen expression in polyps was also compared to antigen expressions of normal colon mucosa and colon cancer tissues. The four MoAbs recognizing extended LeX antigens bound to adenomatous polyps (APs) significantly more than to hyperplastic polyps (HPs). In contrast, anti-SSEA-1 and AH8-183 recognizing monofucosyl LeX were less able to distinguish between APs and HPs. In APs, staining with the four MoAbs recognizing extended LeX antigens correlated with the premalignant parameters of larger polyp size, more severe dysplasia, and increased villose component. However, staining with AH8-183 correlated only with polyp size, and anti-SSEA-1 correlated only with polyp size and degree of dysplasia. In general, the staining frequency of HPs was similar to that of normal colon mucosa, although FH6, which did not stain any specimens of normal mucosa, stained a few HPs. The staining frequency of APs was less than that of colon cancer tissues, but these differences were generally not statistically significant. In conclusion, extended LeX antigens and their sialylated derivatives are cancer-associated antigens that are expressed preferentially in premalignant colon polyps, that tend to correlate with malignant potential in these polyps, and that may eventually help to define mechanisms involved in the polyp-to-cancer sequence.

A series of glycolipids having the X determinant (Gal beta 1----4 [Fuc alpha----3]GlcNAc) at the terminus and a fucosyl alpha 1----3 residue at the internal GlcNAc residue have been isolated and characterized from tumor tissues (Hakomori, S., Nudelman, E., Levery, S.B., and Kannagi, R. (1984) J. Biol. Chem. 259, 4672-4680. A series of monoclonal antibodies that differentially recognize glycolipids with mono-, di-, and trifucosylated type 2 chain have been isolated and characterized. The antibody FH4 shows a remarkable preferential reactivity towards di-/or trifucosylated type 2 chain, i.e. it does not react with monofucosylated structures, including lactofucopentaosyl (III) ceramide (III3FucnLc4), monofucosyl neolactonorhexaosylceramide (y2, V3FucnLc6), and monofucosyl neolactonoroctaosylceramide (Z1, VII3FucnLc8), but reacts well with di- and trifucosylated type 2 chain structures such as difucosyl neolactonorhexaosylceramide (III3V3Fuc2nLc6) and trifucosyl neolactonoroctaosylceramide (III3V3VII3Fuc3nLc8). Two other monoclonal antibodies, FH5 and ACFH18, preferentially react with trifucosylated type 2 chain structure (III3V3VII3Fuc3nLc8), although cross-reactivity with difucosylated type 2 chain (III3V3Fuc2nLc6) was observed. They showed a minimal cross-reaction with monofucosylated type 2 chain. In contrast, the antibody FH1 does not react with III3FucnLc4 but reacts with V3FucnLc6, III3V3Fuc2nLc6, and III3V3VII3Fuc3nLc8. Two monoclonal antibodies, FH2 and FH3, do not discriminate among various glycolipids having fucosylated type 2 chain, and their reactivities are essentially similar to previously established antibodies directed to the X determinant, such as anti-SSEA-1, WGHS 29, VEP8 and 9, My-1, etc. This series of antibodies will be useful to detect the specific type of glycolipid with fucosylated type 2 chain accumulating in human cancer and in undifferentiated cells.

The distribution of a novel difucoganglioside (6B ganglioside, NeuAc alpha 2----3Gal beta 1----4[Fuc alpha 1----3]GlcNAc beta 1----3Gal beta 1----4[Fuc alpha 1----3]GlcNAc beta 1----3Gal beta 1----4Glc beta 1----1Cer) in various normal adult and fetal tissues, as well as in cancer tissues, has been studied by immunoperoxidase staining with a specific monoclonal antibody, FH6, directed to this antigen. A large variety of embryonic and fetal tissues (stomach, colon, small intestine, pancreas, esophagus, lung, and heart) showed a diffuse, weakly positive staining, particularly in the epithelial layer, up to the 70th to 80th day of gestation. However, no staining was observed in various normal adult tissues, including gastrointestinal and glandular epithelial tissues which were stained positively by antibody N-19-9 (directed to sialyl-Lea) or CSLEXI (directed to sialyl-Lex). FH6-positive loci were limited to the proximal convoluted tubuli in kidney and granulocytes. In contrast, 44 of 76 cases of cancer tissue tested, including gastric, colonic, lung, breast, and renal cancers, showed clearly positive staining. The intensity of staining in gastric and colonic cancer tissues by FH6 antibody was weaker and less frequent, although the incidence of positive staining for lung (50%) and breast cancer (86%) was significantly higher than that of the antigen stained by monoclonal antibody FH4 (Y. Fukushi, S. Hakomori, and T. Shepard, J. Exp. Med., 159: 506-520, 1984), which is directed to the asialo core of the FH6 antigen. The antigen levels in the serum of patients with various cancers, inflammatory diseases, and normal subjects were determined by radioimmunoassay. The antigen level was found to be significantly higher in the serum of some patients with cancer, particularly lung, liver, and pancreatic cancers, as compared with the serum levels in other types of cancer, noncancerous diseases, and normal subjects.

SSEA-1 antigen (stage-specific embryonic antigen-1) has been shown to be a series of carbohydrate antigens having type-2 chain and X-hapten structures. These carbohydrate antigens are remarkably accumulated in various human cancer tissues, and activity secreted into the blood stream. Frequently SSEA-1 antigens are further modified with fucoses or sialic acid in human cancer tissues, thus forming various subgroups of antigens such as fucosyl SSEA-1, sialyl SSEA-1 or polyfucosylated antigens. Many monoclonal antibodies are established which can discriminate each subgroup of antigens. Assay systems for these antigens in the sera of cancer patients have been developed using these monoclonal antibodies. Sialyl SSEA-1 is especially elevated in the sera of patients with adenocarcinoma of the lung. The antigens detected with these monoclonal antibodies are mucin-like glycoproteins(cancer-associated mucin, CAM). Various types of cancer-associated mucins can be characterized by respective monoclonal antibodies. It is therefore possible to classify cancer-associated mucins according to the structure of their carbohydrate side chains using these monoclonal antibodies.

Interaction between rat bladder epithelium and Escherichia coli was observed using the transmission electron microscope. As a first step, the bacterial cell coat attaches to the cell coat of the luminal membrane. Then, the luminal membrane enfolds to envelope the bacterium and take it within the cytoplasm. The unit membranes covering the bacteria are destroyed and some of these bacteria are found to be deformed and probably dead in the cytoplasm. Finally, the superficial epithelial cells invaded by many bacteria are collapsed and desquamated into urine. However, these epithelia show a rather prompt recovery and form the normal three layers within 24 hr after inoculation. This experiment suggests one of the defense mechanisms of the vesical epithelium againts E. coli. It is likely that the onset of clinical cystitis may be prevented by the same mechanism during the occasional bacterial invasion of the bladder.

The quantity of tumor-associated antigens carrying type 2 chain polylactosamines with four types of fucosyl determinants, LeX (X-hapten), poly-LeX, sialyl LeX, and LeY (Y-hapten), present in sera of patients with various malignant and non-malignant disorders, as well as the qualitative chemical properties of the carrier molecules in sera, have been investigated using four monoclonal antibodies, each of which defines one of these determinants. The following findings are of particular importance: the serum levels of LeX defined by antibody FH2 and poly-LeX defined by ACFH18 in patients with cancer were occasionally high (incidence about 10%); however, the majority of patients did not show elevated levels; the serum level of the antigen, defined by monoclonal antibody FH6 (termed sialyl LeX-i since this determinant is carried by i antigen), was significantly high in patients with cancers originating from organs from which adenocarcinomas often develop. For example, among various types of lung cancer, only adenocarcinoma but not squamous cell carcinoma, small cell carcinoma, or large cell carcinoma showed a high level of sialyl LeX-i antigen in sera. The incidence of high antigen levels in sera of patients with adenocarcinomas of lung was as high as 76% of the observed cases; the serum level of Ley (Y-hapten) was frequently high in patients with hepatoma (incidence, 34%); sialyl LeX-i antigen was separated on gel filtration as a glycoprotein with an average molecular weight greater than 10(6). It was characterized by its susceptibility to basehydrolysis, Pronase digestion, and sialidase and endo-beta-galactosidase treatment and is assumed to be a high molecular weight mucin-type glycoprotein; sialyl LeX-i antigen expressed in sera of patients with cancer was soluble in perchloric acid, while the same antigen in sera of patients with noncancerous diseases and normal subjects was mostly insoluble in perchloric acid. LeX, a poly-LeX, and essentially all LeY antigens in sera of patients with cancer were perchloric acid-insoluble.

The CA 19-9 antigen is a monosialosyl Lea blood group antigen which has been shown to be a useful tumor-associated antigen for the diagnosis of gastrointestinal cancers. Recently, a sialylated derivative of this antigen, disialosyl Lea, was isolated from a colon cancer liver metastasis and a monoclonal antibody (FH7) recognizing this novel determinant was developed. The present study simultaneously compared the expression of Lea, monosialosyl Lea, and disialosyl Lea antigens in a variety of nonmalignant, premalignant, and malignant tissues of the colorectum and pancreas with an aim toward elucidating whether disialosyl Lea is expressed as a tumor-associated antigen. In normal colonic mucosa, disialosyl Lea expression closely resembled Lea expression in overall frequency, segmental distribution, and cellular localization whereas monosialosyl Lea (CA 19-9) was essentially absent. Along the crypt axis, Lea was more often expressed in goblet cells of the upper crypt whereas disialosyl Lea was found in goblet cells along the entire crypt. Fetal colonic mucosa expressed all three antigens, as did most colorectal cancers regardless of location within the colon or degree of differentiation. The majority of hyperplastic polyps and practically all adenomatous polyps also expressed these three antigens, and in adenomas, antigen expression was independent of polyp size, villous morphology, or degree of dysplasia. In the normal pancreas, the three antigens were expressed on ductal, ductular and centroacinar cells of all specimens. The majority of pancreatic cancers expressed all three antigens. Thus, in the normal colon, the absence of monosialosyl Lea (CA 19-9) in the presence of disialosyl Lea suggests that an alpha 2,6 sialyltransferase is active, which results in the masking of CA 19-9 antigen expression. These results further support the concept that specific sialyltransferases play a role in regulating the expression of tumor-associated antigens.

Biochemical studies have revealed that some normal cells express the LeX trisaccharide Gal beta 1----4(Fuc alpha 1----3)GlcNAc either on short-chain fucolipids or as a single immunodeterminant on glycolipid oligosaccharide side chains. Cancer cells, including those from colonic adenocarcinomas, express this antigen on longer type 2 blood group side chains as difucosylated or trifucosylated fucolipids. Moreover, sialylated forms of difucosylated LeX also accumulate in colon cancer but not in normal colonic mucosa. In the present study, six monoclonal antibodies which selectively recognize the various LeX-related antigens were used for immunohistochemical examination of these antigens in serial sections of human colonic tissue. All of these antigens were oncodevelopmental in human colon. Monoclonal antibodies anti-SSEA-1 and AH8-183, directed against short-chain, monofucosylated LeX, were unable to discriminate well between normal and malignant colonic tissue. However, the other four antibodies were much better at distinguishing cancer from normal tissue. FH6 was the most specific in that no normal tissues bound this antibody. However, FH6 failed to stain poorly differentiated cancers and some colloid-type carcinomas. FH4, which was also highly specific, stained almost all cancers, regardless of the degree of differentiation. FH4 primarily stained cancer cell cytoplasm, whereas the sialylated antigen defined by FH6 predominantly stained cell membranes. Differences were noted between the expression of LeX-related antigens in autopsied normal mucosa compared to mucosa of benign colonic diseases. Monoclonal antibodies recognizing long-chain polyfucosylated and sialylated LeX-related antigens appear to be useful tools for detection of colon cancer.

The present study was undertaken in order to examine whether dilazep (1,4-bis-[3-(3,4,5-trimethoxybenzoyloxy)propyl]perhydro-1, 4-diazepine dihydrochloride monohydrate) attenuates myocardial acidosis induced by coronary artery occlusion in dogs. In dogs with nonischemic normal heart, dilazep (300 or 500 micrograms/kg i.v.) increased blood flow in the left anterior descending coronary artery (LAD) with a decrease in heart rate and diastolic blood pressure. In other dogs, LAD flow was reduced by an occluder by 57 to 68% (partial occlusion) for 90 min. Partial occlusion for 30 min decreased myocardial pH by 0.67 to 0.87 pH units, increased ST segment of the surface electrocardiogram, and decreased regional myocardial contractile force. Dilazep was injected i.v. 30 min after partial occlusion. The decrease in myocardial pH induced by partial occlusion was attenuated by the injection of 300 micrograms/kg of dilazep insignificantly and by that of 500 micrograms/kg of dilazep significantly. Restoration of myocardial [H+] induced by dilazep was calculated from the myocardial pH data. Dilazep (500 micrograms/kg) restored myocardial [H+] induced by partial occlusion by 56.7%, and saline solution restored it by 27.4% 60 min after the drug injection, the actual restoration induced by dilazep being 29.3%. Dilazep, however, did not restore the ST segment elevation and contractile force decrease. It is concluded that dilazep attenuates myocardial acidosis during ischemia.