A comparison of gastric emptying of soluble solid meals and clear fluids matched for volume and energy content: a pilot crossover study.

We previously demonstrated that the gastric emptying time of different liquids with the same volume mainly depended on their energy content, regardless of differences in composition. In this crossover study, we investigated whether the same applies when soluble solid foods are ingested with water. Ten healthy volunteers ingested one of five test diets consisting of two test meals (Calorie Mate® 100 and 200 kcal) and three test solutions (water and glucose solutions of 100 and 200 kcal), each given in a volume of 400 ml, and then underwent ultrasonography to measure the gastric antral cross-sectional area every 10 min for 120 min. The gastric emptying time was defined as the time for the antral cross-sectional area to revert to its initial value. When test diets with the same energy content were ingested, the gastric emptying curves were nearly identical, regardless of whether the original form was solid or liquid. The median (IQR[range]) gastric emptying times of Calorie Mate® of 100 kcal with water vs. isocaloric glucose solution were 65 (60-78 [50-80]) vs. 65 (60-70 [50-80]) min (p = 0.58), and for Calorie Mate® of 200 kcal with water vs. isocaloric glucose solution they were 100 (93-108 [90-120]) vs. 105 (90-110 [90-120]) min (p = 0.54). The median (IQR [range]) for water was 40 (30-40 [30-50]) min. Energy content may be a critical determinant of the gastric emptying time when ingesting soluble solid diets with water.

Determinants of liquid gastric emptying: comparisons between milk and isocalorically adjusted clear fluids.

BACKGROUND: Although current preoperative fasting guidelines apply restrictions to drinks containing milk because of delayed gastric emptying, the safe volume of milk that can be consumed up to 2 h before surgery on a theoretical basis has not yet been defined. We aimed to determine whether delayed gastric emptying depended mainly on the total amount of calories irrespective of compositional differences between milk and clear fluids. METHODS: We prepared five beverages with a uniform volume (500 ml) and step-wise increments in calories (0, 220, and 330 kcal), comprised mainly of non-human milk, pulpless orange juice, water, and gum syrup. The gastric emptying rate of each beverage was determined by ultrasound measurements of the gastric antral cross-sectional area after their ingestion by eight healthy fasting volunteers. RESULTS: The emptying rates of 500 ml of orange juice and 330 ml of non-human milk with 170 ml of water (both were 220 kcal) from the stomach were similar. Furthermore, 450 ml of orange juice with 50 ml of gum syrup and 500 ml of non-human milk (both were 330 kcal) left the stomach at similar rates. The 220 kcal beverages emptied faster than the 330 kcal beverages. CONCLUSIONS: There were no significant differences in liquid gastric emptying after drinking equal volumes of either orange juice or milk as long as both had the same amount of calories. Liquid gastric emptying depends chiefly on the total caloric content. CLINICAL TRIAL REGISTRATION: UMIN000012537.

Dimensional-crossover-driven metal-insulator transition in SrVO3 ultrathin films.

We have investigated the changes occurring in the electronic structure of digitally controlled SrVO(3) ultrathin films across the metal-insulator transition (MIT) by the film thickness using in situ photoemission spectroscopy. With decreasing film thickness, a pseudogap is formed at E(F) through spectral weight transfer from the coherent part to the incoherent part. The pseudogap finally evolves into an energy gap that is indicative of the MIT in a SrVO(3) ultrathin film. The observed spectral behavior is reproduced by layer dynamical-mean-field-theory calculations, and it indicates that the observed MIT is caused by the reduction in the bandwidth due to the dimensional crossover.

Coagulation/fibrinolysis and inflammation markers are associated with disease activity in patients with chronic urticaria.

BACKGROUND: The evaluation of disease severity and activity of chronic urticaria (CU) is essential for the adequate treatment of patients. However, there is no reliable biomarker for such evaluations. Recently, markers of blood coagulation and fibrinolysis have been revealed to be elevated in severe cases of CU. In this article, we studied the coagulation/fibrinolysis and inflammation markers and their relationship to disease activity in patients with CU. METHODS: Plasma fibrin degradation products (FDP), d-dimer and serum C-reactive protein (CRP) were measured with the assessment of disease severity and skin reaction to autologous serum in 82 patients with CU and 37 patients with acute urticaria, idiopathic angioedema (AE) or inducible types of urticaria (IU). RESULTS: The levels of FDP in patients with CU were significantly higher than those in patients with IU, but no other differences in FDP, d-dimer and CRP were observed among patients with different types of urticaria. These markers of patients with CU were well correlated with each other and significantly associated with disease severity of CU, but not with skin reactions to autologous serum. In 37 patients with CU, levels of all these parameters reduced as their disease condition improved, while they increased when the disease became aggravated. Regarding FDP, this relationship was observed even if FDP concentrations were within normal range throughout the study. CONCLUSIONS: The measurement of plasma FDP, d-dimer and serum CRP may be useful for the assessment of disease activity of CU.

Increased angiotensin-converting enzyme in peripheral blood monocytes from patients with sarcoidosis.

Angiotensin-converting enzyme (ACE) activity was measured in isolated peripheral blood monocytes and culture medium from 28 patients with sarcoidosis and compared with values obtained from monocytes of 25 normal control subjects. ACE activity was determined by radioimmunoassay of angiotensin II produced from angiotensin I. While there was no measurable ACE activity in monocytes or culture medium from normal controls under the conditions of our study, monocytes from patients with sarcoidosis all showed activity both in cells and culture medium. The mean ACE activity of monocytes from patients with sarcoidosis was 2.0 pg angiotensin II formed/min per 10(5) cells, and that released into medium over a 24-h interval was 30.4 pg angiotensin II/min per 10(5) cells. The monocyte ACE from patients with sarcoidosis was activated by chloride ions and inhibited by EDTA, captopril, and rabbit antiserum to purified human plasma ACE, indicating that enzymatic activity was effected specifically by ACE. Thus, our studies show a significant elevation and release of ACE by peripheral blood monocytes of patients with sarcoidosis under conditions where monocytes of normal control subjects do not demonstrate ACE activity.

Production of bone-resorbing activity and colony-stimulating activity in vivo and in vitro by a human squamous cell carcinoma associated with hypercalcemia and leukocytosis.

A squamous cell carcinoma of 33-yr-old patient who developed marked leukocytosis and hypercalcemia was transplanted into nude mice in which more marked leukocytosis and hypercalcemia also developed. This tumor (LJC-1-JCK) produced a colony-stimulating factor (CSF) and formed a cyst in the tumor from which a CSF-producing cell line (T3M-1) was established. The CSF causes predominantly formation of granulocytic colonies in addition to macrophage colonies. Bone-resorbing activity (BRA) was detected in the cystic fluid and was eluted as two separate peaks with proteins of an apparent molecular weight of 30,000-50,000 and 10,000-20,000. Colony-stimulating activity (CSA) was eluted at an apparent 30,000 mol wt. The conditioned medium of the T3M-1 cells also contained a BRA with an apparent 14,000 mol wt, whereas CSA eluted at an apparent 30,000 mol wt. PTH, epidermal growth factor, transforming growth factor-alpha, prostaglandin Es, and vitamin D could not account for the powerful BRA. In contrast to CSA, BRA was not inactivated by trypsin and more stable at 70 degrees C. When T3M-1 cells were transplanted into nude mice, marked hypercalcemia developed in addition to granulocytosis. Our findings suggest that the tumor produces and secretes a powerful BRA in vivo and in vitro, which is different from CSA in terms of molecular weight, heat stability, and trypsin treatment. We speculate that the synergistic action of CSF that stimulates macrophage colony formation and recruits osteoclast precursors, and BRA, which stimulates mononuclear phagocytes and/or osteoclasts were responsible for a marked increase in osteoclastic bone resorption and humoral hypercalcemia in the patient.

Hard tissue formation in subcutaneously transplanted rat dental pulp.

While dental pulp appears to be able to form mineralized matrices that do not always resemble dentin, the precise characteristics of the hard tissue and the mechanism of its induction remain unknown. Therefore, we evaluated hard tissue induced by transplantation of pulp into subcutaneous tissue. Seven days after transplantation, initial hard tissue was formed at the inner periphery of the pulp. After 14 days, this hard tissue expanded inwardly. Mineralized matrix was immunopositive for osteocalcin, osteopontin, and bone sialoprotein, but negative for dentin sialoprotein. Transplantation of GFP-labeled pulp into wild-type rats showed these formative cells to have been derived from the transplant. TEM observation revealed apatite crystals within necrotic cells and matrix vesicles at the initial stage of calcification. These results indicate that pulp cells possess the ability to form a bone- or cementum-like matrix. Calcification of the matrix may occur in necrotic cells and matrix vesicles, followed by collagenous calcification.

Evaluation of BacT/Alert 3D SA bottles for accurate detection of Mycobacteremia with special reference to Mycobacterium abscessus.

Bacteremia due to Mycobacterium abscessus, a rapid grower, belonging to the Runyon group IV, occurred in an inpatient with fever of unidentified origin in Shinshu University Hospital. To the best of our knowledge, this is the first documented case of M. abscessus bacteremia in Japan. The organism initially grew on Sheep blood agar plates after terminal-subculturing from the BacT/Alert SA aerobic blood culture bottles with no positive signal, and was subsequently identified as M. abscessus using 16S rRNA sequence analysis. We evaluated the BacT/Alert SA bottles for the detection of Mycobacterium species, with special reference to the rapid growers including M. abscessus by seeding experiments and obtained the following findings: 1) The BacT/Alert system shows the positive sign when the bacterial cell counts reach around 10(6) to 10(7) CFU/ml. 2) The System requires around 6 to 7 days of incubation to obtain a sufficient bacterial growth for the positive signal. 3) The System may result in false negative under the 5-day-culture method recommended by American Society for Microbiology in cases of using automated blood culture systems. 4) So-called the blind- or terminal-subcultures from the bottles are inevitable to perform for precluding the false negative cases.

Initial cytotoxicity of novel titanium alloys.

We assessed the biological response to several novel titanium alloys that have promising physical properties for biomedical applications. Four commercial titanium alloys [Super-TIX(R) 800, Super-TIX(R) 51AF, TIMETAL(R) 21SRx, and Ti-6Al-4V (ASTM grade 5)] and three experimental titanium alloys [Ti-13Cr-3Cu, Ti-1.5Si and Ti-1.5Si-5Cu] were tested. Specimens (n = 6; 5.0 x 5.0 x 3.0 mm(3)) were cast in a centrifugal casting machine using a MgO-based investment and polished to 600 grit, removing 250 mum from each surface. Commercially pure titanium (CP Ti: ASTM grade 2) and Teflon (polytetrafluoroethylene) were used as positive controls. The specimens were cleaned and disinfected, and then each cleaned specimen was placed in direct contact with Balb/c 3T3 fibroblasts for 72 h. The cytotoxicity [succinic dehydrogenase (SDH) activity] of the extracts was assessed using the MTT method. Cytotoxicity of the metals tested was not statistically different compared to the CP Ti and Teflon controls (p > 0.05). These novel titanium alloys pose cytotoxic risks no greater than many other commonly used alloys, including commercially pure titanium. The promising short-term biocompatibility of these Ti alloys is probably due to their excellent corrosion resistance under static conditions, even in biological environments.

Expression of cell-cycle-regulating genes in the development of atherosclerosis in Japanese quail (Coturnix japonica).

The levels of mRNA expression in regulatory genes that are involved in the pathological changes of aortic atherosclerotic and fibroblastic intimal thickening was investigated in Japanese quail. The quail were divided into a control diet group and an atherogenic diet group. The quail were euthanized at 2, 4, 8, and 12 wk after consuming either a control diet or an atherogenic diet. Thereafter, both histological and immunohistochemical studies and mRNA expression analysis of the cell-cycle-regulating genes in aortic atherosclerotic lesions were performed on selected ascending aortas and their large branches. In the atherogenic diet group, aortic lipid-containing intimal and atheromatous lesions were seen mainly at 8 and 12 wk, respectively. Semiquantitative reverse-transcription PCR was used to analyze the alterations of mRNA expression on the development of atherosclerotic lesions. Messenger RNA expression of the c-fos and c-src genes showed peak levels at 8 wk in the atherogenic diet group. However, no significant alteration of c-jun mRNA expression was noted during the entire experimental period. According to the progression of aortic atherosclerotic lesions, c-myc mRNA expression in the atherogenic diet group increased chronologically, and the highest level was observed at 12 wk. Alterations in mRNA expression of proliferating cell nuclear antigen and the p27 gene were similar to that of c-myc. The levels of c-myc, proliferating cell nuclear antigen, and p27 mRNA expression was significantly correlated with the degree of aortic atherosclerotic lesion development at 12 wk in our experiment.

Establishment and characterization of a human colony-stimulating factor-producing cell line from a squamous cell carcinoma of the thyroid gland.

A human colony-stimulating factor (CSF)-producing cell line, T3M-5, has been established in vitro from a squamous cell carcinoma of the thyroid gland transplanted into athymic nude mice [congenitally athymic BALB/c (nu/nu) mice; Central Institute for Experimental Animals, Kawasaki, Japan]. Contaminating fibroblasts derived from a host nude mouse were eliminated by treatment with antiserum raised against nude mouse cells. T3M-5 cells have been continuously propagated during 3 years. The cells grew in a monolayered sheet with about 22 hours of population-doubling time and showed about 40% plating efficiency. The cells exhibited an epithelium-like morphology resembling the structure of the original tumor and showed tumor takes when inoculated into nude mice. Chromosome analysis revealed the cell line to be a human aneuploid line with a hypertriploid mode. The cells possessed the characteristic function of human CSF production in vitro and produced marked neutrophilia in tumor-bearing nude mice that were inoculated with the cultured cells. The molecular weight of the CSF was estimated at about 27,000 and was stable over the pH range 1.0-9.0 at 4 degrees C for 21 hours. The CSF activity was destroyed by either trypsin or chymotrypsin, but it resisted neuraminidase, DNase, and RNase. The cells could be well propagated in roller bottles. About 100 liters of the conditioned medium was obtained with the roller bottle culture method, which formed approximately 500,000,000 colonies of human bone marrow cells. The rate of recovery of CSF activity from the gel-filtration column was high (68.9%). This cell line is therefore expected to aid in the large-scale preparation of human CSF.

Long-term cultivation of a human colony-stimulating factor-producing cell line in a protein-free chemically defined medium.

To examine whether a human colony-stimulating factor (CSF)-producing cell line, T3M-1, can propagate and secrete CSF without protein supplements, long-term cultivation of the cells was carried out in a protein-free chemically defined medium. By the use of stepwise decreases in the fetal bovine serum concentration, continuous growth of T3M-1 cells has been established in a protein-free F-10 medium. The cells designated T3M-1-T2 have been propagated in this medium for 5 years. The population-doubling time of the cells is about 30 hr. Addition of serum stimulated the cell growth (population-doubling time, 17 hr) but did not increase the saturation density. The cells secreted large amounts of CSF (2000 colonies stimulated by 1 ml of the conditioned medium). Addition of serum to the culture increased CSF activity in the conditioned medium (3400 colonies/ml). The results showed that a human cancer cell line, T3M-1-T2, could be propagated in a protein-free chemically defined medium and secrete large amounts of CSF. The cells will serve as an excellent model for better understanding of the cell growth and production of CSF in the absence of any serum contamination.

A human monocyte growth factor produced by lung cancer cells.

Human lung cancer cell line, T3M-30, has been shown to produce a growth factor that stimulates proliferation of peripheral blood monocytes. In the presence of this factor, human circulating monocytes were able to proliferate in vitro. Gel exclusion chromatography of the conditioned medium revealed a single peak of monocyte growth-promoting activity at an apparent molecular weight of 16,000. The growth-promoting activity was adsorbed to an anion-exchange column, Mono Q, and eluted with a salt gradient as a single peak of bioactivity at 300 mM NaCl. When the sample was applied to a Vydac C4 column, a reverse-phase high-performance liquid chromatography column, a single peak of activity was observed at a concentration of 76% acetonitrile in 0.1% trifluoroacetic acid. The monocyte growth-promoting activity was heat stable at 56 degrees C. It was partially destroyed by trypsin. The activity was lost after treatment with 2-mercaptoethanol.

Receptor binding of human granulocyte colony-stimulating factor to the blast cells of myeloid leukemia.

Human granulocyte colony-stimulating factor (G-CSF) rapidly loses the biological activity and the receptor binding capacity following radioiodination. We have made a mutein of human G-CSF, KW-2228, in which Thr-1, Leu-3, Gly-4, Pro-5, and Cys-17 were respectively substituted with Ala, Thr, Tyr, Arg, and Ser; showed more potent G-CSF activity; and retained full biological activity and receptor binding capacity at least 2 weeks of radioiodination. G-CSF is an effective growth factor for the blasts of myeloid leukemia. Radioiodinated KW-2228 was prepared using solid-phase glucose oxidase-lactoperoxidase. Human leukemia cell lines and the blast cells from leukemia patients were examined for binding. High affinity binding sites were identified on myeloid cell lines and on the blasts obtained from acute myeloid leukemia patients. Scatchard analysis showed that a single binding site for G-CSF was observed (361-1688 receptors/cell; Kd 128-1400 pM). In contrast, specific binding of 125I-KW-2228 was not demonstrated on lymphoblastic cell lines or the blast cells of acute lymphoid leukemia or lymphoma. This difference was reflected in the effectiveness of G-CSF to stimulate colony formation in acute myeloid leukemia blasts, while G-CSF did not stimulate colony formation of the blast cells from acute lymphoid leukemia.

Superoxide dismutase in various tissues from rabbits bearing the Vx-2 carcinoma in the maxillary sinus.

Distribution profiles of superoxide dismutase isoenzymes in various tissues of rabbits with the Vx-2 carcinoma in the maxillary sinus were compared with those of control rabbits. Copper- and zinc-containing superoxide dismutase (Cu,Zn-SOD) activity in the liver of rabbits decreased significantly 3 weeks after transplantation. Manganese-containing superoxide dismutase (Mn-SOD) activity did not decrease significantly within 5 weeks after transplantation. In other tissues from the tumor-bearing rabbits, Cu,Zn-SOD and Mn-SOD activities were not changed within 5 weeks. No Mn-SOD activity and low Cu,Zn-SOD activity were detected in the Vx-2 carcinoma. These results suggest that the Vx-2 carcinoma has lost most of its ability to defend against oxygen toxicity and this ability decreased only in the liver of rabbits bearing the Vx-2 carcinoma in the maxillary sinus.

Isolation of small cell lung cancer-associated antigen from human brain.

Previous studies have demonstrated that monoclonal antibody TFS-4 recognizes a cell surface antigen with a molecular weight of 124,000 expressed selectively on small-cell lung cancer but not on non-small-cell lung cancers and that it cross-reacts with human brain. The antigenic determinant on small-cell lung cancer and that on brain shared common characteristics, i.e., trypsin sensitivity, heat lability, and neuraminidase resistance, suggesting that they are similar peptides (T. Okabe et al., Cancer Res., 44: 5273-5278, 1984; J-i. Watanabe et al., Cancer Res., 47:826-829, 1987). In order to elucidate the nature of this unique antigen recognized by TFS-4, we have purified the antigen to homogeneity from human brain. The antigen was solubilized from brain with 0.5% Nonidet P-40, precipitated with 50% ammonium sulfate, and subsequently purified by sequential chromatographies, i.e., diethylaminoethyl-Sepharose ion exchange, immunoaffinity, and gel permeation high-pressure liquid chromatography. The antigenic reactivity was assessed by immunoblotting using TFS-4 as a primary antibody. The purified antigen showed a single protein band with a molecular weight of 124,000 on sodium dodecylsulfate-polyacrylamide gel electrophoresis detected by a silver staining technique. The results suggest that the antigen on brain tissues is structurally related to the molecule expressed on small-cell lung cancer.

Monoclonal antibody that distinguishes small-cell lung cancer from non-small-cell lung cancer.

To examine whether a monoclonal antibody, TFS-4, can distinguish small-cell lung cancer from non-small-cell lung cancers, an extensive survey of fresh lung tumors, cancers from other organs, and normal tissue specimens has been carried out. The antibody has been shown to react specifically with small-cell lung cancer (15 of 15) but not with squamous cell carcinoma (0 of 20), adenocarcinoma (0 of 20) of the lung, or large-cell lung cancer (0 of 2). It reacted neither with other malignancies, including colorectal cancer, gastric cancer, and malignant lymphoma, nor with such normal tissues as trachea, lung, liver, pancreas, colon, kidney, spleen, skin, striated muscle, bone marrow, or peripheral blood cells. Interestingly, the antibody cross-reacted with central nervous tissues. The antigenic determinant on small-cell lung cancer and that on human brain were both heat labile and trypsin sensitive, but resisted treatment with neuraminidase, suggesting that they represent similar peptides. TFS-4 may be of clinical use in the diagnosis of small-cell lung cancer, while the antigen may help investigate the nature and origin of small-cell lung cancer.

Establishment of different clonal strains from a human sarcoma of the stomach: tumorigenic heterogeneity in athymic nude mice.

Twenty five clonal strains have been isolated from a single human sarcoma of the stomach. Two different types of clones have been recognized by their morphology and behavior in vitro. Type I clones were characterized by criss-crossed arrays and multilayers with high terminal density. Type II clones grew in a well-organized monolayer with lower saturation density. Although both types of clones exhibited fibroblastic appearance, type I clones showed a more rounded, refractile shape. The cells of this type showed multiple regions of criss-crossed arrays and multilayers throughout the culture vessels. Saturation density of this type of clone was 2- to 3-fold (1.7 to 2.1 X 10(5) cells/sq cm) higher than that of type II clones. Chromosomal analysis revealed that type I clones were human aneuploid ones with modal chromosome numbers ranging from 51 to 61. With the exception of clones 11 and 19, type I clones were able to produce tumors in athymic nude mice when injected s.c. Type II clones exhibited a more flattened and elongated appearance. The cells grew in a well-organized monolayer resembling fingerprint whorls. They showed lower saturation density (0.7 to 0.9 X 10(5) cells/sq cm). Chromosomal examinations revealed the clones to be human aneuploid ones with modal numbers from 47 to 54. Tumor formation was not observed in nude mice given injections of this type of cell. Both types of clones did not bear antigens cross-reacting with the antiserum against mouse spleen cells but had surface antigens which were affected by the antibody against HL-60 cells and complement. These results suggested that this human sarcoma was heterologous and that cells with widely different tumorigenic potential preexisted in the parental cell population.

Immune elimination of host fibroblasts for the cultivation of human tumors transplanted into nude mice.

We report here a useful method for the isolation and cultivation of human tumor cells in vitro from human tumors grown in nude mice. A rabbit was immunized with spleen cells obtained from adult nude mice. The rabbit antiserum in the presence of complement effectively killed cultured cells derived from various mouse tissues, but it was not cytotoxic to cultured cells from human tissues including tumors. When mixed cultures consisting of human tumor cells and nude mouse fibroblasts were treated with the antiserum and complement, the nude mouse fibroblasts were completely removed from the cultures, and the human tumor cells could be propagated without noticeable changes in morphological features. Primary cultures of heterotransplanted human tumors grown in nude mice were also successfully treated, resulting in the ultimate elimination of fibroblastic cells derived from the stroma of the tumor. The functional properties of the tumor cells (production of human chorionic gonadotropin by choriocarcinoma cells and production of carcinoembryonic antigens by pancreas carcinoma cells) were also maintained after the antiserum treatment.