Enhanced optical and nonlinear optical responses in a polyelectrolyte templated Langmuir-Blodgett film.

Optical and nonlinear optical properties like fluorescence and second harmonic generation (SHG) of molecular materials can be strongly influenced by the mode of assembly of the molecules. The Langmuir-Blodgett (LB) technique is an elegant route to the controlled assembly of molecules in ultrathin films, and complexation of ionic amphiphiles in the Langmuir film by polyions introduced in the aqueous subphase provides a simple and efficient access to further control, stabilization, and optimization. The monolayer LB film of the hemicyanine-based amphiphile, N-n-octadecyl-4-[2-(4-(N,N-ethyloctadecylamino)phenyl)ethenyl]pyridinium possessing a "tail-head-tail" structure, shows fluorescence as well as SHG response. The concomitant enhancement of both of these linear and nonlinear optical attributes is achieved through templating with the polyanion of carboxymethylcellulose. Brewster angle and atomic force microscopy reveal the influence of polyelectrolyte templating on the morphology of the Langmuir and LB films. Polarized absorption and fluorescence spectroscopy provide insight into the impact of complexation with the polyelectrolyte on the orientation and deaggregation of the hemicyanine headgroup leading to fluorescence and SHG enhancement in the LB film.

Alteration of X and Y chromosomes in human esophageal squamous cell carcinoma.

The incidence of human esophageal squamous cell carcinoma in males is well-known to be higher than in females and its biological action in male patients is generally much more aggressive than that of the female. Recently, aberrations and/or other abnormalities of the sex chromosomes, especially the Y chromosome, have been postulated to be involved in some of the differences in the incidence and/or biological action of human malignancies between male and female patients. Therefore, in this study, we examined abnormalities of the sex chromosomes in cell smears obtained from 30 male patients diagnosed with esophageal squamous cell carcinoma. In addition, TE series cell lines, derived from esophageal squamous cell carcinomas, were studied for sex chromosome abnormalities by utilizing a simultaneous double color fluorescent in situ hybridization (FISH) and these findings were correlated with various clinicopathological parameters in order to examine its likely biological significance. In esophageal squamous cell carcinoma, Y chromosome loss was detected in all cases studied (1.6-86.9%, mean 22.98 +/- 22.04%), but the loss of the X chromosome was encountered in only 6 of the cases (7.1-40.6%, mean 15.90 +/- 12.46%). There was no significant association between the rate of Y chromosome loss in carcinoma cells and any of the clinicopathological parameters examined including age and stage of the cancer. Loss of the Y chromosome was observed in only two cases of adjacent non-pathological esophageal squamous cell epithelium. Among the TE series examined, the cell lines derived from male patients demonstrated loss of the Y chromosome in all cell lines (1.4-92.9%, mean 44.92 +/- 42.55%), but the great majority of cell lines derived from female patients were associated with the karyotype of XX. These results indicated that the loss of the Y chromosome is associated with the malignant phenotype in human esophageal squamous epithelium, but possibly not with biological behavior. These results also suggested that at least one X chromosome is indispensable for the survival of esophageal squamous cell carcinoma.

Cell cycle inhibitory protein p27 in esophageal squamous cell carcinoma.

BACKGROUND: p27 protein is one of the cdk inhibitors which regulates the progression from G1 to S phase of the cell cycle. Reduced expression of p27 protein has been reported to be correlated with poor clinical outcome in patients with various cancers. MATERIALS AND METHODS: We performed immunohistochemistry of both p27 and Ki67 in 136 cases of resected human esophageal squamous cell carcinoma, and evaluated the association between p27 immunoreactivity and clinicopathological features including clinical outcome in these patients. We also examined the correlation between labeling index or the percentage of positive tumor cells for p27 and Ki67 in serial tissue sections in these cases. RESULTS: Cases with invasion of the muscularis propria or adventitia had significantly (p < 0.05) higher p27 LI (65.0 +/- 23.7) than those with invasion limited to mucosa or submucosa and those with carcinoma in situ (58.9 +/- 18.3). There were no significant correlations between p27 and other clinicopathological factors such as sex, age, tumor size, differentiation type, nodal status and histological stage. The cases with p27 LI below 40% tended to have a worse prognosis than those with p27 LI above 40%. There was no significant correlation between Ki67 and p27 LIs. CONCLUSIONS: Reduced expression of p27 may be correlated with the biological behavior of esophageal squamous cell carcinoma and may play an important role in the early stages of cancer.

Topoisomerase II alpha expression in esophageal squamous cell carcinoma.

BACKGROUND: DNA topoisomerase II alpha (topo II alpha) is associated with active cell proliferation of mammalian cells. Topo II alpha overexpression has been reported in a number of human malignancies and is considered to be related to their biological behaviors. MATERIALS AND METHODS: We examined the expression of topo II alpha immunohistochemically in 136 cases of human esophageal squamous cell carcinoma, 10 foci of squamous dysplasia and 10 non-pathologic squamous epithelium. We calculated the labeling index (LI) or the percentage of immunopositive cells for Topo II alpha and Ki67, and Topo II alpha LI/Ki67 LI (T/K ratio). These findings were then correlated with clinicopathological features of the patients including their clinical outcome. RESULTS: Both topo II alpha and Ki67 immunoreactivity were detected in the nuclei. A significant positive correlation was obtained between Topo II alpha and Ki67 LIs in all the specimens examined. Topo II alpha LI and T/K ratio were 24.5 +/- 8.0% and 1.04 +/- 0.64 for carcinoma, 19.1 +/- 15.2% and 0.68 +/- 0.29 for dysplasia and 14.0 +/- 14.1% and 0.55 +/- 0.17 for non-pathologic epithelium, respectively. Topo II alpha LI and T/K ratio in carcinoma cases were significantly higher than those of normal epithelium. Topo II alpha LI alone did not correlate with any of clinicopathological parameters examined but among carcinoma cases, cases with lymph nodes metastasis or higher histological stages had significantly higher T/K ratio than those without lymph node metastasis or lower histological stages. In addition, carcinoma cases with T/K ratio of greater than 0.8 demonstrated significantly worse prognosis than those with T/K ratio of smaller than 0.8. CONCLUSIONS: The relative overexpression of topo II alpha as compared with Ki67, i.e., increased T/K ratio was detected in esophageal squamous cell carcinoma and is considered to represent a dysregulation or qualitative alteration in topo II alpha, possibly associated with malignances, as reported in other human cancers. In addition, topo II alpha overexpression may also be correlated with the aggressive biological behavior of the patients with esophageal squamous cell carcinoma.

Cyclic ADP-ribose induces Ca2+ release from caffeine-insensitive Ca2+ pools in canine salivary gland cells.

Cyclic ADP-ribose (cADPR), a novel putative messenger of the ryanodine receptor, was examined regarding its ability to mobilize Ca2+ from intracellular Ca2+ stores in isolated cells of parotid and submandibular glands of the dog. cADPR induced a rapid and transient Ca2+ release in the digitonin-permeabilized cells of salivary glands. cADPR-induced Ca2+ release was inhibited by ryanodine receptor antagonists ruthenium red, ryanodine, benzocaine, and imperatoxin inhibitor but not by the inositol 1,4,5-trisphosphate (IP3)-receptor antagonist heparin. Thapsigargin, at a concentration of 3 to 30 microM, inhibited IP3-induced Ca2+ release, while higher concentrations were required to inhibit cADPR-induced Ca2+ release. Cross-potentiation was observed between cADPR and ryanodine or SrCl2, suggesting that cADPR sensitizes the Ca2+-induced Ca2+ release mechanism. Cyclic AMP plays a stimulatory role on cADPR- and IP3-induced Ca2+ release in digitonin-permeabilized cells. Calmodulin also potentiated cADPR-induced Ca2+ release, but inhibited IP3-induced Ca2+ release. Acetylcholine and ryanodine caused the rise in intracellular free Ca2+ concentration ([Ca2+]i) in intact submandibular and parotid cells. Caffeine did not produce any increase in Ca2+ release or [Ca2+]i rise in any preparation. ADP-ribosyl cyclase activity was found in the centrifuged particulate fractions of the salivary glands. These results suggest that cADPR serves as an endogenous modulator of Ca2+ release from Ca2+ pools through a caffeine-insensitive ryanodine receptor channel, which are different from IP3-sensitive pools in canine salivary gland cells. This system is positively regulated by cyclic AMP and calmodulin.

Proliferative fasciitis of the forearm: case report with immunohistochemical, ultrastructural and DNA ploidy studies and a review of the literature.

A case of proliferative fasciitis arising in the left forearm of a 56-year-old man was examined. The lesion was preceded by blunt trauma, measured 1.5 x 1.3 x 1.0 cm, was poorly circumscribed and appeared white to light gray on the cut surface. Light microscopic examinations revealed that spindle cells and giant cells with one or two nuclei and abundant basophilic cytoplasm were arranged without any organized patterns in collagenous stroma. Ultrastructurally, well-developed rough endoplasmic reticulum separated by varying amounts of fine to course fibrillar materials was detected in the giant cells. Only vimentin immunoreactivity was detected in both spindle and giant cells. The Ki-67 labeling index of spindle cells was 35% but that of giant cells was less than 5%, and this reflects the quiescent or slow-growing features of these giant cells in proliferative fasciitis. DNA content of the cells, which was examined by image cytometry, demonstrated diploidy in both spindle (DNA index=1.01) and giant (DNA index=1.09) cells.

Inhibition of Na+,K(+)-ATPase by platelet-activating factor in dog submandibular glands.

Physiological stimulation of dog submandibular gland has been shown to generate platelet-activating factor (PAF). However, PAF is not released from cells in the tissue. To assess its intracellular activity, the effect of PAF on Na+,K(+)-ATPase was examined in dog submandibular gland cells. PAF inhibited Na+,K(+)-ATPase in membrane preparations, and the inhibitory effect was dependent on the protein concentration in the enzyme preparation. The inhibitory effect of a low concentration of PAF was antagonized by a PAF-receptor antagonist, BN 50,739, but at high concentrations, PAF was not antagonized. Kinetic analysis of PAF inhibition of Na+,K(+)-ATPase suggests that the inhibition of Na+,K(+)-ATPase by PAF is not due to competition by PAF at K(+)- or Na(+)-binding sites on the enzyme, but by complex inhibitory mechanisms. These results suggest that PAF may interact with specific and nonspecific site of action resulting in the inhibition of Na+,K(+)-ATPase. Ouabain increased mucin release from dog submandibular gland cells. Because Na+,K(+)-ATPase and ion exchange pathways are important in the secretory responses of acinar cells, PAF may regulate intracellularly the secretory function of acinar cells by modulating Na+,K(+)-ATPase and ionic homeostasis.

Detection of apoptotic cells in cytology specimens: an application of TdT-mediated dUTP-biotin nick end labeling to cell smears.

We applied TdT-mediated deoxyuridine triphosphate (dUTP)-biotin nick end labeling (TUNEL) to cytologic smears in order to detect the cells undergoing apoptosis. These smears were obtained by scraping the cut surface of 9 cases of carcinoma, including renal-cell carcinoma (3 cases), esophageal squamous-cell carcinoma (3 cases), and gastric adenocarcinoma (3 cases), and were fixed and prepared by different methods. The results were also compared with those of tissue sections. TUNEL in smears was generally associated with higher background nuclear stain than in tissue sections. Smears that were fixed in 4% or 8% paraformaldehyde or absolute methanol exhibited results comparable with those of tissue sections, with minimum background in all cases examined. There were no significant differences in TUNEL labeling index among tissue sections and smears fixed in 4% or 8% paraformaldehyde or in absolute methanol. Smears treated in Carnoy's fixative (3:1 methanol:acetic acid) and air-dried smears demonstrated a higher background. TUNEL positivity could not be detected in slides decolorized from May-Grünwald-Giemsa stain. Markedly high background, which may occur as a result of artifactural DNA breaks, was also observed in slides decolorized from Papanicolaou stain, in which TUNEL-positive cells could be evaluated only in 3/8 cases. Application of the TUNEL method to cytology specimens has disadvantages or limitations compared to its application to histological sections, but the method is considered the most suitable one for detecting cells undergoing apoptosis in cytology materials.

[A clinical study of retrograde cerebral perfusion in long duration].

The safety limit of retrograde cerebral perfusion (RCP) in aortic arch reconstruction was evaluated in comparison with long (> or = 60 min: group A = 28 cases) and short (< 60 min: group B = 88 cases) RCP time (RCPT). RCPT was 60-94 min in group A and 24-59 min in group B. Hospital mortality was 7.1% and 6.8% in group A and B, respectively. There was no case who was died due to cerebral insufficiency. Postoperative neurological complication was observed 3 in group A and 3 in group B. There was no significance between 2 groups. There were no significant differences in the time to awake, ICU stay, electroencephalogram, and cognitive functions between 2 groups. Based on these results, the safety limit of RCP is over than 60 min and, maybe, about 90 min. We monitored the increase of deoxygenated hemoglobin (HbR) during RCP using a near infrared spectroscopy (n = 55). Since HbR increased gradually during RCP, HbR reflects oxygen metabolism non-invasively in the brain. HbR can be one of useful indexes of the brain protection during RCP.

Stimulation of platelet-activating factor synthesis by neurotransmitters in salivary glands.

Platelet-activating factor (PAF), a phospholipid mediator exhibiting potent biological activities, has been shown to stimulate amylase release from the pancreas and salivary glands. The capacity of salivary glands for PAF biosynthesis in response to stimulation has also been demonstrated. To elucidate the role of PAF in salivary glands, we studied the regulation of platelet-activating factor synthesis by the autonomic nervous system in canine salivary glands. Acetylcholine and ionomycin stimulated PAF production in dispersed cells from parotid, submandibular, and sublingual glands of dogs. Norepinephrine and phenylephrine, but not isoproterenol, also stimulated PAF production in submandibular gland cells. Norepinephrine-induced PAF production was blocked by phentolamine but not by propranolol. Acetylcholine and norepinephrine increased both the PAF production and liberation of [14C]arachidonic acid from cells pre-labeled with [14C]arachidonic acid in the presence of Ca2+ in the medium. These stimulants increased [14C]arachidonic acid liberation without the accompanying production of PAF in Ca(2+)-deprived medium. No activators or inhibitors of protein kinase C produced or affected acetylcholine-induced PAF production. Lyso-PAF:acetyl-CoA acetyltransferase was activated in the cells treated with acetylcholine, norepinephrine, isoproterenol, and 8Br-cyclic AMP. Deprivation of Ca2+ in the medium markedly reduced acetylcholine-induced activation of the transferase, but little affected norepinephrine-, isoproterenol-, and 8Br-cyclic AMP-induced activation. Dithiothreitol-insensitive cholinephosphotransferase activity was also increased by acetylcholine, norepinephrine, isoproterenol, and 8Br-cyclic AMP, and the deprivation of Ca2+ in the medium further increased the activation of the enzyme activity by these agents. These results suggest that PAF synthesis in canine salivary glands is under the control of muscarinic cholinergic and alpha-adrenergic systems via Ca(2+)-dependent remodeling pathways, and that the independent activation of either phospholipase A2 or acetyltransferase is insufficient for PAF production in submandibular gland cells, i.e., the concurrent activation of these enzymes is required.

Inhibition of the association with nuclear matrix of pRB, p70 and p40 proteins along with the specific suppression of c-MYC expression by geldanamycin, an inhibitor of Src tyrosine kinase.

Geldanamycin is an antibiotic that preferentially inhibits G1/S transition and causes G2/M arrest in human leukemia HL-60 cells. With it, we selectively inhibited recombinant Src tyrosine kinase without significantly inhibiting protein kinase A. The perturbation of cell cycling by geldanamycin was accompanied by marked suppression of c-MYC expression. In contrast to this, pRB expression was remarkably enhanced by geldanamycin. In the untreated HL-60 cells, c-MYC was apparently enriched in nuclear matrix preparation, and significant amounts of hyperphosphorylated pRB, p70 and p40 proteins were observed to associated with the nuclear matrix. The amounts of these proteins associated with the nuclear matrix, however, were markedly decreased by treatment with geldanamycin. This finding suggests that the association of c-MYC, hyperphosphorylated pRB, p70 and p40 proteins with the nuclear matrix is essential in cell cycling, especially in G1/S and G2/M progressions, and that this association is a part of signal transduction pathway in Src kinase activation.

[History of the syndromes of Yin (retention of body fluids) - the relationship between the syndromes of Yin in Chinese medicine and those of Yin in Buddhist medicine].

We traced the history of the term "yin" by means of Chinese medical books and classical Chinese Buddhist literature. As a result of our study, the following points became clear. The syndromes of yin described in the Shan han za bing lun are divided into two groups. Those of the first group are characterized by short retention of body fluids. The yu yin is representative of this group. On the other hand, those of the second group are distinguished by long retention of body fluids. The tan yin is typical of this group. It was deduced that the former group originated in Chinese traditional medicine because it is found in the Su wen and the Ling shu. The latter is supposedly derived from Buddhist medicine because it is similar to the syndromes of yin in Buddhist medicine.

Antitumor activity of a spicamycin derivative, KRN5500, and its active metabolite in tumor cells.

KRN5500, (6-[4-Deoxy-4-(2E,4E)-tetradecadienoylglycyl]amino-L-glycero - beta-L-mannoheptopyranosyl]amino-9H-purine), was semi-synthesized in an attempt to increase the therapeutic effects of spicamycin analogues. The present study evaluated the antitumor activity of KRN5500 against murine tumors and human tumor xenografts. KRN5500 prolonged the survival of P388 leukemia- and B16 melanoma-bearing mice but was marginally effective on colon adenocarcinoma 26. The antitumor activity of KRN5500 (4 mg/kg/day x 5, IV) against xenografts of 10 human stomach, 14 colon and 2 esophageal cancers was evaluated with two parameters: the tumor growth inhibition rate (TGIR) and the tumor mass reduction by comparison with mitomycin C (MMC, 6.7 mg/kg/day x 1,IV). KRN5500 showed a marked efficacy in the human tumor xenograft model. The overall response rate of 26 cancers to KRN5500, evaluated by TGIR, was approximately equal to their response rate to MMC (72% vs. 73%). However, more tumors were reduced by KRN5500 than by MMC (52% vs. 39%). It is notable that the response rates of 14 colon cancers to KRN5500 were significantly higher than those to MMC, both in TGIR (69% vs. 58%) and in tumor mass reduction (46% vs. 23%). Among the tumors sensitive to KRN5500, COL-1 showed a marked response (TGIR 93%) and a significant reduction in tumor mass (0.22-fold the starting volume). In the mode of action, KRN5500 was found to show an inhibitory effect on protein synthesis in P388 cells (IC50 1.5 microM). However, KRN5500 was ineffective even at 170 microM in inhibition of protein synthesis in rabbit reticulocyte lysates.(ABSTRACT TRUNCATED AT 250 WORDS)

[History of the term "tan" (phlegm) (II) -- studies of the term "tan" found in medical books before the Liang dynasty].

The first description of the term "tan" is found in the Shan Han Za Bing Lun published at the end of the later Han dynasty. This book refers to this term as a disorder of the cold fluid in the body. The meaning of this term was revised to a viscous fluid as the years went on. The term "tan" from the Zhou Hou Bai Yi Fang published in the Liang dynasty was the same sense of phlegm as that found in present works. Our studies showed that changes of the words used for the concept "phlegm" in Chinese medical books coincided with those in Chinese Buddhist literature. We suggested that the concept "phlegm" in Buddhist medicine influenced Chinese tradition medicine.

[History of the term "tan" (phlegm) (I) - Studies of the term "tan" described in classical Chinese translations of Buddhist literature].

We traced the origin of the term "tan" (phlegm) in Chinese traditional medicine. This term itself is not found in early works but is represented by the terms of tuo (saliva) and xian (drivel). However, in the theory of tri-dosa of Indian traditional medicine, phlegm is an important element. Our studies of classical Chinese translations of Buddhist literature showed that the concept of tan in Buddhist medicine was introduced into China through Buddhism.

[Studies of the fifth section in the introduction of the Qian jin yao fang - Writings quoted from the Shen nong ben cao jing and the Yao dui].

The fifth section in the introduction of the Qian jin yao fang is composed of three paragraphs. The first paragraph is the same as that of the Shen nong ben cao jing. The second and the third ones begin with the sentences of "Lei gong say ..." and "Yao dui says ..." respectively. We inquired into the origin of these three paragraphs. As a result of our investigation, we inferred that the first and the second paragraphs were part of the writings of the Lei gong ji zhu shen nong ben cao and that the third one was part of the Lei gong yao dui. We also pointed out that the substance of these paragraphs was influenced directly and indirectly by Buddhist medicine.

Mechanism of action of an antifungal antibiotic, RI-331, (S) 2-amino-4-oxo-5-hydroxypentanoic acid; kinetics of inactivation of homoserine dehydrogenase from Saccharomyces cerevisiae.

An antifungal antibiotic (S) 2-amino-4-oxo-5-hydroxypentanoic acid, inhibited the biosynthesis of the aspartate family of amino acids (methionine, isoleucine and threonine) followed by the inhibition of protein biosynthesis in Saccharomyces cerevisiae. This inhibition was effected by impeding the biosynthesis of their common intermediate precursor, homoserine. The inhibition of biosynthesis of homoserine by the antibiotic was attributable to inactivation of homoserine dehydrogenase [EC 1.1.1.3], which is involved in the conversion of aspartate semialdehyde to homoserine in the metabolic pathway leading to threonine, methionine and isoleucine. Since such enzymic activity is not present in animal cells, the selective antifungal activity of the antibiotic is thus explained.

Calcium dependency of adrenergic and muscarinic cholinergic stimulation of mucin release from dog submandibular gland cells.

Stimulation of muscarinic cholinergic, alpha-adrenergic and beta-adrenergic receptors elicited mucin release from dispersed dog submandibular cells. The secretory response to acetylcholine was much more pronounced than to adrenergic agonists, and largely dependent on the presence of extracellular Ca2+, but the dependency on extracellular Na+ was slight. Ionomycin also stimulated mucin release. In rat submandibular cells, neither muscarinic cholinergic agonists nor ionomycin were as effective mucosecretagogues as beta-adrenergic agonists. alpha-Adrenoceptor-mediated release was decreased by chelating extracellular Ca2+ with EGTA. The beta-adrenoceptor-mediated response was diminished by extensive exposure of cells to EGTA, due at least in part to the requirement of Ca2+ for beta-adrenoceptor stimulation of cAMP formation. 8-br-cAMP stimulated 45Ca2+ release from cells preloaded with 45Ca2+. The 8-br-cAMP-induced mucin release was eliminated in ionomycin-pretreated cells, but not inhibited by chelating extracellular Ca2+ and by the treatment of the cells with TMB-8 or in the cells loaded with BAPTA. These results suggest that not only the adrenergic system but also the muscarinic cholinergic system may participate in the regulation of mucin release in dog submandibular gland, and also provide the possibility that, in addition to a cAMP-mediated mechanism, Ca(2+)-dependent mechanisms may be involved in mucosecretion in dog submandibular acini.

The mechanism of antifungal action of (S)-2-amino-4-oxo-5-hydroxypentanoic acid, RI-331: the inhibition of homoserine dehydrogenase in Saccharomyces cerevisiae.

We have explored the mechanism by which an antifungal antibiotic, (S)-2-amino-4-oxo-5-hydroxypentanoic acid, RI-331, preferentially inhibits protein biosynthesis in Saccharomyces cerevisiae, by inhibiting the biosynthesis of the aspartate family of amino acids, methionine, isoleucine and threonine. This inhibition was effected by inhibiting the biosynthesis of their common intermediate precursor homoserine. The target enzyme of RI-331 was homoserine dehydrogenase (EC.1.1.1.3) which is involved in converting aspartate semialdehyde to homoserine in the pathway from aspartate to homoserine. The enzyme is lacking in animals. So the antibiotic is selectively toxic to prototrophic fungi.