We examined the action of mastoparan on beta cell exocytosis. Mastoparan stimulated GABA and insulin release from MIN6 beta cells. On the other hand, mastopraran-induced GABA release was decreased by expressing the tetanus toxin C1 light chain in MIN6 cells. We have then investigated the relationship between SNARE proteins and mastoparan action using adenovirus-mediated gene transfer system. Overexpression of t-SNAREs, syntaxin 1A, and SNAP-25 inhibited the mastoparan-induced insulin release by approximately half-fold of control levels, however, the mastoparan-induced GABA release was not affected by these t-SNAREs overexpression. The overexpression of mutant alpha-SNAP (1-285), which inhibits the wild-type alpha-SNAP function in a dominant negative manner, did not influence either mastoparan-induced GABA or insulin release in spite of its marked inhibition of glucose-stimulated insulin release. Our data indicate that mastoparan stimulates GABA exocytosis via vesicular transport; however, SNARE proteins are differently involved in the exocytosis of insulin and GABA.

Animals↗

PubMed2001

Negative regulation of LPS-stimulated expression of inducible nitric oxide synthase by AP-1 in macrophage cell line J774A.1.

The level of NOS II mRNA was markedly increased during 24 h lipopolysaccharide (LPS) stimulation, but showed no further increase thereafter. On the other hand, the level of NOS II mRNA in J774A.1 cells transfected with an expression vector containing the rat csk cDNA (J.Csk) was significantly increased during 3 h LPS stimulation, but rather decreased thereafter. Although no significant difference was observed in the activation of NF-kappaB by LPS among parental J774A.1, J774A.1 transfected with promoterless vector (J.pBK), and J.Csk cells, activity of c-Jun N-terminal kinase (JNK) and nuclear translocation of nuclear factor activator protein-1 (AP-1) were markedly upregulated in the J.Csk cells. Then luciferase reporter vectors containing NOS II promoter with mutations in two AP-1-like sites (U site, -1126 approximately -1120; L site, -524 approximately -518) were transiently transfected in J774A.1 cells. The promoter activity following LPS stimulation for 24 h was significantly increased by mutation at the L site, but not by mutation at the U site, suggesting that NOS II expression is negatively regulated, at least in part, through the AP-1-like L site in response to LPS.

Animals↗

PubMed2001

Morpholinium 2-chloro-4-nitrobenzoate, 2-chloro-5-nitrobenzoate and 4-chloro-3-nitrobenzoate.

Morpholinium 2-chloro-4-nitrobenzoate, C4H10NO+*C7H3ClNO4-, (I), crystallizes in a non-centrosymmetric space group. The cations and anions are connected by N-H...O hydrogen bonds to afford a 2(1) helical chain. Morpholinium 2-chloro-5-nitrobenzoate, C4H10NO+*C7H3ClNO4-, (II), and morpholinium 4-chloro-3-nitrobenzoate, C4H10NO+*C7H3ClNO4-, (III), both crystallize in a centrosymmetric space group. In (II) and (III), two cations and two anions are held together by N-H.O hydrogen bonds to form a centrosymmetric ring with graph-set descriptor R4(4)(12).

Journal Article↗

PubMed2001

Adenovirus-mediated preproinsulin gene transfer into adipose tissues ameliorates hyperglycemia in obese diabetic KKA(y) mice.

We investigated whether adenovirus-mediated preproinsulin gene transfer into insulin target tissues (adipocytes) ameliorates hyperglycemia in diabetic mice. KKA(y) mice, a genetically obese type 2 diabetic animal model, were treated with a single subcutaneous injection of recombinant adenovirus, Adex1CA-human preproinsulin (Adex1CA-pchi), into the epididymal fat pads. pchi mRNA was expressed only in adipose tissue in which mature insulin was produced. Three days after virus injection these mice showed a marked decrease of blood glucose levels (from about 400 to 200 mg/dl), and an intraperitoneal glucose tolerance test revealed the markedly improved glucose tolerance. There was no significant difference in serum insulin levels between control and recombinant adenovirus-treated KKA(y) mice. The normalized glucose levels in diabetic mice were maintained for at least 2 weeks after the virus injection. This strategy could provide a novel and, most importantly, a simple and convenient gene therapy for obese type 2 diabetes patients.

Adenoviridae↗

PubMed2001

Ammonium hydrogen tartronate at 240 and 20 K.

The low-temperature structure determination of the title compound, alternatively called ammonium hydrogen hydroxypropanedioate, NH(4)(+) x C(3)H(3)O(5)(-), has revealed that the H atom involved in a very short asymmetric O--H...O hydrogen bond [O...O = 2.448 (2) A at 240 K and 2.4393 (10) A at 20 K] is disordered.

Journal Article↗

PubMed2001

Increased insufflation pressure enhances the development of liver metastasis in a mouse laparoscopy model: possible mechanisms.

BACKGROUND: The effect of different insufflation pressures and durations of CO2 pneumoperitoneum on the growth of liver metastasis was investigated in a mouse model. The possible mechanisms involved in the pressure-related enhancement of liver metastasis were also examined. METHODS: Mice inoculated intraportally with colon 26 cells underwent CO2 pneumoperitoneum at different pressures (5,10, or 15 mmHg) for 30 or 60 min, or received no treatment other than tumor cell inoculation (control). The subsequent growth of liver metastases was examined. Mice injected intraportally with 111In-oxine-labeled colon 26 cells underwent pneumoperitoneum at three different pressures or served as controls. The radioactivity of the liver was determined to evaluate tumor accumulation in the liver. Mice received pneumoperitoneum at three different pressures or received trocar placement alone. Changes in plasma interleukin-6 levels were determined. RESULTS: The growth of liver metastases on day 14 was influenced by increased insufflation pressures (p < 0.05) rather than the prolonged duration of pneumoperitoneum without significant interaction. The 15-mmHg pneumoperitoneum group showed a higher (p < 0.05) accumulation of radioactivity in the liver compared with the 5-mmHg pneumoperitoneum group and controls. Pneumoperitoneum groups with 5 and 10 mmHg showed higher (p < 0.05) peak levels of IL-6 compared with controls. CONCLUSIONS: An elevated insufflation pressure plays an important role in the enhancement of liver metastases, and this pressure-related adverse effect may be partly relevant to facilitating accumulation of tumor cells in the liver.

Animals↗

PubMed2001

Opening of mitochondrial K(ATP) channels attenuates the ouabain-induced calcium overload in mitochondria.

We tested whether opening of mitochondrial ATP-sensitive K(+) (mitoK(ATP)) channels depolarizes mitochondrial membrane potential (DeltaPsi(m)) and thereby prevents the mitochondrial Ca(2+) overload. With the use of a Nipkow disk confocal system, the mitochondrial Ca(2+) concentration ([Ca(2+)](m)) and DeltaPsi(m) in rat ventricular myocytes were measured by loading cells with Rhod-2 and JC-1, respectively. Exposure to ouabain (1 mmol/L) for 30 minutes produced mitochondrial Ca(2+) overload, and the intensity of Rhod-2 fluorescence significantly increased to 173+/-16% of baseline (P<0.001). Treatment of myocytes with the mitoK(ATP) channel opener diazoxide (100 micromol/L) blunted the ouabain-induced mitochondrial Ca(2+) overload (131+/-10% of baseline; P<0.001 versus ouabain). Moreover, diazoxide significantly depolarized the DeltaPsi(m) and reduced the intensity of JC-1 fluorescence during application of ouabain to 89+/-2% of baseline (P<0.05). These effects of diazoxide were blocked by the mitoK(ATP) channel blocker 5-hydroxydecanoate (500 micromol/L). These results indicate that opening of mitoK(ATP) channels prevents a mitochondrial Ca(2+) overload in association with DeltaPsi(m) depolarization and thereby protects myocardium against ischemic damage.

Adenosine Triphosphate↗

PubMed2001

Mesenteric lipoma: report of a case with emphasis on US findings.

Mesenteric lipoma (ML) is a relatively rare disease that has been very infrequently reported. We present the clinical data and medical imaging results of an asymptomatic case with ML incidentally detected by sonography (US). On US the lesion was imaged as a well-demarcated hypoechoic mass with multiple linear echoes. The mass also changed location under probe compression. The lesion was imaged as a clearly demarcated non-enhanced homogeneous fat-density mass on CT. Color Doppler US and angiography confirmed the avascular nature of the mass. Knowledge of the US findings in this case is useful for the diagnosis of ML.

Adult↗

PubMed2001

Matrix metalloproteinase-2 and -9 in bile as a marker of liver metastasis in colorectal cancer.

Matrix metallproteinases (MMP)-2 and -9 are associated with cancer invasion and metastasis. MMP-2 and MMP-9 activities have never been assayed in bile. In the present study we investigated whether MMP-2 and -9 activities in the bile could be a marker for evaluation of liver metastasis in colorectal cancer. Fifty-three patients underwent colorectal resection for histologically verified adenocarcinoma. Twenty-six patients had colorectal cancer without liver metastasis and 27 patients had metastatic liver tumor. Six patients were studied as carcinoma-free control. MMP-2 and MMP-9 activities were assayed in bile using gelatin zymography and quantitated. Active MMP-2 activity of colorectal cancer with liver metastasis group (24.1 +/- 2.5 pixel count) was significantly higher than that of colorectal cancer without liver metastasis group (11.4 +/- 1.3 pixel count) (P < 0.001) or of control group (6.4 +/- 1.0 pixel count) (P < 0.001). Active MMP-9 was not detected in bile. ProMMP-9 activity of colorectal cancer with liver metastasis group (530.3 +/- 127.5 pixel count) was significantly higher than that of colorectal cancer without liver metastasis group (213.9 +/- 33.2 pixel count) (P = 0.008). This is the first report showing that the levels of active MMP-2 and proMMP-9 in bile were significantly higher in liver metastasis of colorectal cancer than in metastasis-free colorectal cancer. The results suggest that activities of active MMP-2 and proMMP-9 in the bile may be useful markers for predicting liver metastasis in colorectal cancer.

Adenocarcinoma↗

PubMed2001

Asymmetric mono-oxazine: an inevitable product from Mannich reaction of benzoxazine dimers.

The Mannich reaction is detailed, which was carried out on benzoxazine dimers under various conditions, that is, temperature, reaction time, and solvents. Against our expectation, in any condition, instead of generating a disubstitution oxazine compound, the reaction gives a product with only a single oxazine ring, a mono-oxazine benzoxazine dimer, as characterized by FT-IR, 1H NMR, 13C NMR, 2D-NMR (1H-1H COSY, 1H-13C HMQC, and 1H-13C HMBC), and EA. The asymmetrical reaction is found to be based on the original structure of the benzoxazine dimer which has two phenol rings in a different stability as clarified by X-ray structure analysis of the single crystal. All types of benzoxazine dimers indicate the specific structure with a pair of inter- and intramolecular hydrogen bonds. The bond distance indicates that the intramolecular hydrogen bonding is very strong, while the packing structure emphasizes the high stability of the dimer unit and implies the deactivation of one phenol ring in the benzoxazine dimer. In this contribution, we demonstrate one of the quite rare examples, showing how the stereostructure of the reactant molecule is an important factor to control the reaction and give an asymmetric product which we never expected when considering only the chemical formula.

Journal Article↗

PubMed2001

Liver metastases are less established after gasless laparoscopy than after carbon dioxide pneumoperitoneum and laparotomy in a mouse model.

BACKGROUND: Although the liver is the most frequent site of cancer recurrence after conventional open surgery for colorectal cancer, the effect of laparoscopic procedures with or without gas insufflation on the development of liver metastases is largely unknown. METHODS: Male BALB/C mice inoculated intraportally with colon 26 cells were randomized to undergo carbon dioxide pneumoperitoneum (n = 14), abdominal wall lifting (n = 14), or full laparotomy (n = 12), or to serve as control subjects without any procedures other than tumor inoculation (n = 13). RESULTS: The growth of liver metastases 14 days after surgery was enhanced after full laparotomy (p < 0.01) and pneumoperitoneum (p = 0.02), as compared with that in the control subjects, whereas there was no difference in the growth of liver metastases between abdominal wall lifting and the control condition (p = 0.99). CONCLUSIONS: These results suggest that the defense against liver metastasis is better preserved after the gasless procedure than after laparotomy and carbon dioxide pneumoperitoneum in the reported animal model.

Animals↗

PubMed2001

The aglycone of sulfogalactolipids can alter the sulfate ester substitution position required for hsc70 recognition.

3'-Sulfogalactolipids(SGLs), sulfogalactosyl ceramide (SGC), and sulfogalactoglycerolipid (SGG) bind to the N-terminal ATPase-containing domain of members of the heat shock protein 70 family. We have probed this binding specificity using a series of synthetic positional sulfated or phosphorylated glycolipid analogues, containing either a long-chain bisalkyl hydrocarbon-2-(tetradecyl)hexadecane (B30) or C(18) ceramide (SGC(18)) backbone. By TLC overlay and receptor ELISA, recombinant hsc70 bound ceramide-based glycoconjugates having 3'- or 4'-sulfogalactose glycone moieties and the 4'-sulfogalactose positional isomer conjugated to B30. Hsc70 binding was significantly decreased to the 3'-sulfogalactose conjugated to the long-chain branched alkane. 3'-Sulfoglucose conjugated to B30 was not bound, nor were similarly conjugated di-, tri-, and tetra-sulfated or phosphorylated galactolipids. These results highlight the importance of the position, rather than the number of sulfate esters within the galactose ring. This binding selectivity was shared by the sea urchin hsp70-related sperm receptor. A 3'-SGC-based soluble inhibitor, in which the acyl chain was replaced with an adamantyl group, inhibited binding of hsc70 to both 3'- and 4'-SGC species with an IC(50) of 50 and 75 microM, respectively, indicating a shared sulfogalactose binding site. These studies demonstrate the highly specific nature of hsc70/SGL binding and show, for the first time, that the lipid aglycone can alter the substitution position requirement for glycolipid recognition.

Cerebrosides↗