Coxsackie and adenovirus receptor is a critical regulator for the survival and growth of oral squamous carcinoma cells.

Coxsackie and adenovirus receptor (CAR) is essential for adenovirus infection to target cells, and its constitutive expression in various cancerous and normal tissues has been reported. Recently, the biological role of CAR in human cancers of several different origins has been investigated with respect to tumor progression, metastasis and tumorigenesis. However, its biological function in tumor cells remains controversial. Here we report the critical role of CAR in growth regulation of oral squamous cell carcinomas (SCCs) in vitro and in vivo via the specific interaction with Rho-associated protein kinase (ROCK). Loss of endogenous CAR expression by knockdown using specific small interfering RNA (siRNA) against CAR facilitates growth suppression of SCC cells due to cell dissociation, followed by apoptosis. The consequent morphological reaction was reminiscent of anoikis, rather than epithelial-mesenchymal transition, and the dissociation of oral SCC cells was triggered not by lack of contact with extracellular matrix, but by loss of cell-to-cell contact caused by abnormal translocation of E-cadherin from surface membrane to cytoplasm. Immunoprecipitation assays of the CAR-transfected oral SCC cell line, HSC-2, with or without ROCK inhibitor (Y-27632) revealed that CAR directly associates with ROCKI and ROCKII, which results in inhibition of ROCK activity and contributes to maintenance of cell-to-cell adhesion for their growth and survival. Based on these findings, in vivo behavior of CAR-downregulated HSC-2 cells from siRNA knockdown was compared with that of normally CAR-expressing cells in intraperitoneally xenografted mouse models. The mice engrafted with CAR siRNA-pretreated HSC-2 cells showed poor formation of metastatic foci in contrast to those implanted with the control siRNA-pretreated cells. Thus, CAR substantially has an impact on growth and survival of oral SCC cells as a negative regulator of ROCK in vitro and in vivo.

Diffuse dermal angiomatosis.

Diffuse dermal angiomatosis (DDA) is characterized clinically by painful erythematous lesions with ulcers and histologically by a benign, diffuse, and self-limited proliferation of tiny blood vessels in the superficial layers of the reticular dermis. Here we describe a case of DDA with leg ulcer. Erythematous lesions presented around the ulcer and angiogram revealed an occlusion of the superficial femoral artery. The erythematous lesions disappeared after revascularization. Although DDA is extremely rare, early correction of the ischemia in the peripheral artery should be taken into consideration.

Effects of gassericins A and T, bacteriocins produced by Lactobacillus gasseri, with glycine on custard cream preservation.

Lactobacillus gasseri LA39 and LA158 isolated from human-infant feces produce bacteriocins named gassericins A and T, respectively. Both gassericins have high heat stability (121 degrees C, 10 min), good pH tolerance (pH 2-11), and strong bactericidality against many gram-positive bacteria, especially lactic acid bacteria, and thus are expected to be effective food preservatives. A microwell plate assay against 12 strains of custard cream spoilage bacteria showed that the gassericins had broader antibacterial spectra than nisin A. Although the gassericins allowed gram-negative isolates to grow, they successfully inhibited the growth of all tested bacterial strains in microwells with the addition of glycine. Glycine was bacteriostatic against many strains except lactic acid bacteria. For practical use, gassericin A was efficiently produced by cultivation in a food-grade medium improved using cheese whey, nourishing proteose peptone, and surfactant yolk lecithin. The practical preservative effect of gassericin A and glycine was verified from the viability of 4 isolated strains, Bacillus cereus, Lactococcus lactis ssp. lactis, Achromobacter denitrificans, and Pseudomonas fluorescens, in custard creams. Custard cream containing 123 arbitrary units of gassericin A per milliliter entirely growth-inhibited the 2 gram-positive strains. In custard cream containing an insufficient amount of gassericin A (49 arbitrary units/mL), the gram-positive strains gradually grew but were completely inhibited by the addition of 0.5% (wt/wt) glycine. The 2 gram-negative strains did not multiply even in the additive-free custard cream, probably because of the unsuitable growth environment. This is the first report showing the combined effect of bacteriocin and glycine and their application for food preservation, which may be helpful for future use in the food industry.

Microbial community analysis of food-spoilage bacteria in commercial custard creams using culture-dependent and independent methods.

Custard cream is made from highly nutritive raw materials such as milk and sugar and is easily spoiled by the multiplication of specific microbial contaminants or residents. However, this spoilage microbial community has not been studied. We determined the spoilage microbiota in commercial custard creams using culture-dependent and independent methods. Using the culture-dependent analysis with various agar media, 185 bacterial colonies and 43 eukaryal colonies were isolated from 7 commercial custard cream products. All bacterial isolates were morphologically, physiologically, and genetically identified as bacilli, staphylococci, lactic acid bacteria, and psychrotrophic gram-negative rods. Using culture-independent molecular analysis, the PCR-denaturing gradient gel electrophoresis technique, spoilage of the commercial custard creams was found to be caused by bacilli, staphylococci, lactic acid bacteria, psychrotrophic gram-negative rods, Anoxybacillus sp., Caurobacter sp., and Streptococcus sp. bacteria. The detected spoilage bacteria were the same species as previously detected in spoiled milk products and shown in other reports, suggesting that spoilage bacteria in a raw material easily grow in processed foods made from milk. We determined the spoilage microbial communities in commercial custard creams, and these are the first data concerning spoilage microbiota in nonfermented processed foods using a culture-independent analysis. Our study will be useful for the manufacture and safe preservation of dairy products because the first step toward safe food preservation by food manufacturers is to understand the spoilage microbiota in a target food to select optimal preservatives and to reduce the use of food additives.

Localization and expression of R-cadherin in skeletal muscle following nerve injury.

Rats in which the sciatic nerves were cut were divided into two groups: animals with nerve sutured and animals with nerve not sutured. In the unsutured group, the levels of R-cadherin expression increased and then decreased to values lower than those of controls. In the sutured group, the levels of R-cadherin expression increased and then decreased to almost control values. These results suggest that R-cadherin plays some role in cells of normal and regenerating muscles.

Clinical comparison of the scanning CO2 laser and conventional surgery in the treatment of ingrown nail deformities.

AIM: To compare statistically the clinical effects and postoperative course of the scanning CO(2) laser and conventional surgical method to evaluate the clinical effectiveness of the former for the treatment of ingrown nail deformities. METHODS: We performed vaporization of the nail matrix using the scanning CO(2) laser in 25 cases with ingrown nail deformities. RESULTS: In 21 cases, the recurrence of ingrown nail deformity was not observed during follow-up. All cases were free of postoperative infection. Use of the scanning CO(2) laser reduced postoperative pain, and all patients were able to return to their daily activities by day 3 post-surgery without any problems. CONCLUSIONS: Statistically, the operating time and the duration of postoperative pain were reduced significantly by the scanning CO(2) laser. Furthermore, patients treated with CO(2) laser were able to return to daily life significantly sooner.

[A study on the maternal plasma C21 steroids concentration during pregnancy].

During pregnancy, C21 steroids such as progesterone (P4), and cortisol (F), have been reported to be closely involved in uterine contraction. To clarify the association between changes in steroid hormones and the onset of delivery, we measured the concentrations of six C21 steroids in maternal blood by high performance liquid chromatography. Changes in each steroid hormone were evaluated by analysis of variance during pregnancy. Pregnenolone, a source of C21 steroids, gradually increased during pregnancy. P4 and 17P4, its metabolite, reached a peak 3 weeks before delivery and noticeably decreased thereafter. Accompanying a decrease in P4 and 17P4, 20P4 and F, its metabolite, were noticeably increased. The association among these steroid hormones was also evaluated. The correlation between P4 and F was reversed after 3 weeks before delivery. These results suggest that steroid hormones in the maternal blood begin to change dynamically about 3 weeks before delivery. In particular, a decrease in P4 and an increase in F seem to be closely related to the onset of delivery.

Clinical use of human hepatocyte growth factor in the early detection of HELLP syndrome.

The change in plasma concentration of human hepatocyte growth factor (hHGF) in pregnant women with HELLP (hemolysis, elevated liver enzyme and low platelets) syndrome was investigated, and the following results were obtained. (1) The plasma concentration of hHGF in pregnant women did not change with the gestational stage. (2) The plasma concentration of hHGF in pregnant women with EPH (edema, proteinuria and hypertension) gestosis was 0.19 +/- 0.07 ng/ml and did not differ greatly from that in control pregnant women. (3) The plasma concentration of hHGF in pregnant women with HELLP syndrome was 1.79 +/- 0.35 ng/ml; it was increased prominently compared to control pregnant women. (4) The plasma concentration of hHGF in pregnant women with HELLP syndrome changed parallel to the clinical symptoms of the syndrome.

Platelet aggregation inhibiting activity of human placental chorioepithelial brush border membrane vesicles--the role of alkaline phosphatase.

Human placental chorioepithelial brush border membrane, which is in direct contact with maternal blood flow, has platelet aggregation inhibiting activity. In the present study, the mechanism of this action has been examined in relation to ADP (adenosine diphosphate) degrading activity and alkaline phosphatase activity of brush border membrane vesicles (BBMV). BBMV prepared from human early and term placental villi, inhibited platelet aggregation induced by ADP. BBMV had potent ADP degrading (ADPase) activity. ADP was quickly degraded by BBMV. ADP degrading activity of BBMV was not so different between early and term placenta. Alkaline phosphatase activity of late placental BBMV was about three times greater than that of early placental BBMV. On the other hand, ADP degrading activity of late placental BBMV was almost the same as that of early placental BBMV. Inhibiting activity of platelet aggregation induced by ADP and ADP degrading activity of BBMV, were not inhibited by levamisole (alkaline phosphatase inhibitor).

Changes in blood level of lipid peroxide and vitamin E during pregnancy: clinical significance and relation to the pathogenesis of EPH gestosis.

The present study was undertaken in order to determine the changes in blood levels of lipid peroxide and vitamin E in EPH gestosis. The mean plasma level of lipid peroxide was 0.89 +/- 0.13 nmol/ml for nonpregnant women, while for normal pregnant women it was 2.51 +/- 0.52 nmol/ml, and 4.10 +/- 0.72 nmol/ml for women with EPH gestosis. The mean plasma level of vitamin E was 6.7 +/- 1.0 micrograms/ml for nonpregnant women, while for normal pregnant women it was 14.5 +/- 2.2 micrograms/ml and 12.6 +/- 1.2 micrograms/ml for women with gestosis. The mean red cell level of vitamin E was 3.55 +/- 0.25 micrograms/ml packed red cells in nonpregnant women, while in normal pregnant women it was 2.56 +/- 0.34 micrograms/ml packed cells and 2.90 +/- 0.28 micrograms/ml packed cells for women with EPH gestosis. The mean platelet level of vitamin E was 99 +/- 25 micrograms/g protein for nonpregnant women, while in normal pregnant women it was 232 +/- 24 micrograms/ml protein and 205 +/- 32 micrograms/g protein for women with EPH gestosis. It was shown in this study that in EPH gestosis the plasma level of lipid peroxide increased while the levels of vitamin E which inhibited the formation of lipid peroxide decreased in plasma, red cells and platelets.

Studies on placental inhibition of platelet aggregation: a comparison of human syncytiotrophoblast brush border and basal plasma membranes.

We compared the platelet aggregation inhibiting activity of human placental syncytiotrophoblast brush border membrane vesicles (BBMV) and basal plasma membrane vesicles (BpMV), and obtained the following results. Strong platelet aggregation inhibiting activity is found in placental BBMV. BBMV inhibited platelet aggregation induced by ADP (adenosine diphosphate) and arachidonic acid in a way which depended on the protein concentration of BBMV added. In contrast, BpMV showed no detectable platelet aggregation inhibiting activity. Quite high ADP degrading activity (ADPase activity) was present in the placental BBMV. ADP was quickly degraded by BBMV. In contrast, BpMV did not degrade ADP so quickly. Platelet TXB2 production was almost completely abolished at the protein concentration of 40 micrograms/ml of BBMV. In contrast, BpMV did not significantly inhibit platelet TXA2 (TXB2) production. These results show that syncytiotrophoblast brush border and basal plasma membranes of the human placenta have markedly different properties with respect to platelet aggregation inhibiting activity.

Properties of ADP-degrading activity of human placental syncytiotrophoblast brush border membrane vesicles.

The enzymatic properties of ADP (adenosine diphosphate) degradation in human placental syncytiotrophoblast brush border membrane vesicles (BBMV) were explored and the following results were obtained. BBMV had high ADP degrading activity compared to homogenate of placental villi. ADP degrading activity of BBMV; 1.05 +/- 0.05 mumol/mg protein/min placental villi was 21 times higher than that of homogenate of placental villi. Hydrolysis of ADP by BBMV follows Michaelis-Menten saturation kinetics with an apparent Km of 10.9 +/- 0.8 microM and Vmax of 2.10 +/- 0.17 mumol/mg protein/min. The enzyme has a divalent cation requirement. EDTA (2 mM) was found to abolish ADP degrading activity but this could be restored by the addition of either magnesium or calcium ions. Maximum enzyme activity of ADP degradation in BBMV was observed at a pH close to 8.0. The enzyme was insensitive to vanadate, levamisole, oligomycin, ouabain and N-ethylmaleimide (NEM), omeprazole and adenosine (5') pentaphospho (5') adenosine.

Platelet aggregation inhibiting activity of human placental chorioepithelial brush border membrane vesicles.

We investigated the platelet aggregation inhibiting activity of human placental brush border membrane vesicles (BBMV) and obtained the following results. A strong platelet aggregation inhibiting activity existed in placental BBMV. The BBMV inhibited the platelet aggregation induced by ADP, arachidonic acid, collagen and ristocetin in a dose-dependent manner. The protein concentration of BBMV giving 50 per cent inhibition was 52 +/- 6 micrograms/ml for ADP-induced platelet aggregation, 21 +/- 2 micrograms/ml for arachidonic acid-induced platelet aggregation, 19 +/- 2 micrograms/ml for collagen-induced platelet aggregation and 107 +/- 9 micrograms/ml for ristocetin-induced platelet aggregation. There was a high level of ADP degrading activity (ADPase activity) in the placental BBMV. ADP degrading activity of the BBMV: 10.5 +/- 0.5 mumol/mg protein/min was 21 times greater than that of homogenate of the placental villi. The placental BBMV inhibited platelet TXA2 production. In the 40 micrograms/ml protein concentration of placental BBMV, platelet TXA2 production was almost completely inhibited.

Characterization of human placental activity for transport of taurocholate, using brush border (microvillous) membrane vesicles.

The uptake of taurocholate into brush border membrane vesicles prepared from human full term placenta was studied using a rapid filtration technique. The taurocholate uptake into brush border membrane vesicles was sensitive to extravesicular osmolarity, and pre-incubation of the brush border membrane vesicles with the taurocholate increased the uptake of taurocholate into the brush border membrane vesicles. These findings indicate that the uptake of taurocholate by brush border membrane vesicles represents transport into vesicles. The uptake of taurocholate into vesicles was not dependent on Na+ electrochemical gradient (extravesicular > intravesicular). But this uptake was markedly increased when the intravesicular space was rendered electrically more positive by the use of lowly permeant anions or valinomycin-induced K+ diffusion membrane potentials. These findings indicate that the taurocholate transport into brush border membrane vesicles was dependent on membrane potential. The initial rate of taurocholate transport into brush border membrane vesicles exhibited saturation kinetics with respect to the taurocholate concentration, an apparent Km of 67 microM and Vmax of 0.30 nmol/mg protein/20 sec were calculated.

[The platelet aggregation inhibiting activity in human placenta--study on the ADP degrading activity of human placental villi].

We studied the platelet aggregation inhibiting activity and ADP degrading activity of human placental villi (tissue culture supernatant) and brush border membrane vesicles (BBMV) and obtained the following results. 1. There existed a platelet aggregation inhibiting activity in tissue culture supernatant of villi (S-villi) but not in that of decidua or amnion. The S-villi inhibited the platelet aggregation induced by ADP, but not that induced by collagen, arachidonic acid or ristocetin. And, there was also ADP degrading activity (ADPase activity) in the S-villi. ADP was quickly degraded by S-villi. When ADP was preincubated with S-villi, the platelet aggregation induced by ADP was completely lost. 2. There was very strong platelet aggregation inhibiting activity in placental BBMV. The BBMV almost completely inhibited the platelet aggregation induced by ADP, collagen, arachidonic acid and ristocetin. And there was very strong ADP degrading activity in the placental BBMV. ADP was quickly degraded by BBMV. When ADP was preincubated with BBMV, the platelet aggregation induced by ADP was completely lost. 3. The enzymatic character (heat stability, enzymatic kinetics, Ca++ dependency and pH dependency) of ADP degrading activity in BBMV was very similar to that in S-villi. 4. The ADP degrading activity of both S-villi and solubilized BBMV were fractionated by anion exchange column chromatography and gel filtration column chromatography in similar patterns, and it was shown that ADP degrading substance of both S-villi and solubilized BBMV had a molecular weight of about 60K.