A KRAS mutation status-stratified randomized phase II trial of gemcitabine and oxaliplatin alone or in combination with cetuximab in advanced biliary tract cancer.

BACKGROUND: Previous clinical trials have not proved that adding epidermal growth factor receptor inhibitors to chemotherapy confers a survival benefit for patients with advanced biliary tract cancer (ABTC). Whether the KRAS mutation status of tumor cells confounded the results of past studies is unknown. PATIENTS AND METHODS: ABTC patients stratified by KRAS status, Eastern Cooperative Oncology Group performance status, and primary tumor location were randomized 1 : 1 to receive GEMOX (800 mg/m(2) gemcitabine and 85 mg/m(2) oxaliplatin) or C-GEMOX (500 mg/m(2) cetuximab plus GEMOX) every 2 weeks. The primary end point was objective response rate (ORR). RESULTS: The study enrolled 122 patients between December 2010 and May 2012 (62 treated with C-GEMOX and 60 with GEMOX). Compared with GEMOX alone, C-GEMOX was associated with trend to better ORR (27% versus 15%; P = 0.12) and progression-free survival (PFS, 6.7 versus 4.1 months; P = 0.05), but not overall survival (OS, 10.6 versus 9.8 months; P = 0.91). KRAS mutations, which were detected in 36% of tumor samples, did not affect the trends of difference in ORR and PFS between C-GEMOX and GEMOX. The two treatment arms had similar adverse events, except that more patients had skin rashes, allergic reactions, and neutropenia in the C-GEMOX arm. Of patients with C-GEMOX, the presence of a grade 2 or 3 skin rash was associated with significantly better ORR, PFS, and OS. CONCLUSIONS: Addition of cetuximab did not significantly improve the ORR of GEMOX chemotherapy in ABTC, although a trend of PFS improvement was observed. The trend of improvement did not correlate with KRAS mutation status. CLINICAL TRIALS NUMBER: This study is registered at ClinicalTrials.gov (NCT01267344). All patients gave written informed consent.

Osteopontin regulates anabolic effect in human menopausal osteoporosis with intermittent parathyroid hormone treatment.

UNLABELLED: In this pilot study, we demonstrated that women with osteopontin (OPN) over-expression show less resistance to postmenopausal osteoporosis than women with normal OPN levels. We hypothesized that the levels of plasma OPN could be used as a treatment indicator for intermittent parathyroid hormone (PTH)-treated menopausal osteoporosis. We demonstrated that plasma OPN levels could be used as a biomarker for early treatment response. INTRODUCTION: Animal studies indicate that OPN-deficient mice are resistant to ovariectomy induced osteoporosis. Our pilot study also demonstrated women with OPN over expression may show less resistance to postmenopausal osteoporosis. The role of plasma OPN in PTH1-34-treated osteoporosis remains unclear. METHODS: From September 2005 to September 2006, 31 menopausal women over 45 years of age with severe osteoporosis were enrolled in our study. Subjects were treated with PTH1-34 subcutaneously at a dose of 20 µg/day. Plasma OPN levels and BMD of the lumbar spine and hip were measured using ELISA and dual-energy X-ray absorptiometry at baseline, 3, 6, and 9 months. Response to the treatment was assessed by the sequential change in bone mineral density and OPN expression using a general linear mixed model. RESULTS: The plasma OPN decreased sequentially and significantly throughout the 9-month treatment course from 20.75 ± 5.36 to 11.2 ± 4.37 ng/ml (p < 0.001). The sequential improvement in the T-score and Z-score was significant in the lumbar spine but not in the hip area. In the lumbar spine, when the plasma OPN decreased by 1 ng/ml the T-score increased by 0.0406 and the Z-score increased by 0.0572 of lumbar spine. CONCLUSION: OPN levels are related to the anabolic effect of PTH in human postmenopausal osteoporosis. Plasma OPN levels could be used as a biomarker for early treatment response.

Increased serum osteopontin is a risk factor for osteoporosis in menopausal women.

SUMMARY: Osteopontin (OPN)-deficient mice are resistant to ovariectomy-induced osteoporosis. Therefore, we hypothesized that women with OPN overexpression may show less resistance to postmenopausal osteoporosis. In this study, we first demonstrated that serum OPN levels could be used as a biomarker for the early diagnosis of osteoporosis in postmenopausal women. INTRODUCTION: Animal studies indicate that OPN-deficient mice are resistant to ovariectomy-induced osteoporosis. METHODS: From 2004 to 2006, 124 women over the age of 45 were enrolled in a menopausal group, while another 95 women, from 25 to 45 years of age with regular menstruation, were enrolled into a childbearing age group. The serum concentrations of OPN were calculated using the enzyme-link immunosorbent assay method, and bone mineral densities were determined with dual energy X-ray absorptiometry. RESULTS: Serum OPN levels had a significant positive correlation with age (menopausal group, p < 0.0001) and a negative correlation with body weight, height, hip bone mineral density, and T-scores in the menopausal group. In contrast, there was a positive correlation with the E2 concentration and height, but there was no significant association with the above variables in the childbearing age group. Additionally, high serum OPN levels (>14.7 ng/ml) was a significant risk factor causing menopausal osteoporosis (odds ratio = 2.96, 95% confidence interval, 1.055-8.345). CONCLUSION: Serum OPN levels could be used as a biomarker for the early diagnosis of osteoporosis in postmenopausal women.

High-potentiality preliminary selection criteria and transformation time-dependent factors analysis for establishing Epstein-Barr virus transformed human lymphoblastoid cell lines.

Infection of freshly isolated and cryopreserved lymphocytes with Epstein-Barr virus (EBV) leads to the establishment of human B lymphoblastoid cell lines (LCLs). Techniques for optimal infection of the lymphocytes are vital for the establishment of a human biobank. The present study found that more than half (58-86%) of such established LCLs had transport times of less than 48 h, cell densities exceeding 10(6) cells/ml and cell viabilities greater than 90%. After EBV infection, 3306 freshly isolated lymphocytes required 30.0 +/- 0.1 days to become LCLs. Conversely, 1210 cryopreserved lymphocytes required 36.2 +/- 0.4 days. Cell density and viability of the culture affected transformation time in freshly isolated lymphocytes. On the other hand, blood transport time, cryopreservation time and initial cell viability were major factors in cryopreserved specimens. These results contribute to general information concerning the establishment of a human biobank for EBV infected cells.

Protective effects of partially purified antigens of Actinobacillus pleuropneumoniae on experimentally infected mice.

Three different models of protection experiments in mice using partially purified Actinobacillus pleuropneumoniae antigens such as crude culture supernatant extract (CCSE) and partially purified cell extract (PPCE) were attempted. Biochemical analysis showed that these two immunogens had protein concentration of 0.17-0.2 mg/mL and pentose concentration of 0.012-0.014 mg/mL. In the first model intranasal (IN) vaccination with different doses (from 0.01-10 IN-LD50) against IN challenge with the dose of 20 IN-LD50 containing 1.2 x 10(9) colony forming unit (CFU)/50 microL showed that only those with the dose more than 1 IN-LD50 had slight protection in terms of survival index (SI). In the second model of protection experiment, in which subcutaneous vaccination (s.c.) with the immunogens plus soybean oil against IN challenge with 10 IN-LD50 containing 6 x 10(8) CFU/50 microL, showed that formalin-killed bacteria (bacterin) and CCSE plus PPCE had only a slight protection whereas vaccination with CCSE or PPCE immunogen alone had no protection. In the third model of protection experiment, in which the intramuscular (i.m.) vaccination with the immunogens plus aluminum hydroxide [Al(OH3)] gel against intraperitoneal (i.p.) challenge with the dose of either 2 or 6 i.p.-LD50 containing 1.2-3.6 x 10(8) CFU/0.5 mL of 0.3% mucin saline showed highly effective.

Morphology and function of gill mitochondria-rich cells in fish acclimated to different environments.

The objective of this study is to test the hypothesis that morphologically different mitochondria-rich (MR) cells may be responsible for the uptake of different ions in freshwater-adapted fish. Tilapia (Oreochromis mossambicus) were acclimated to high-Ca, mid-Ca, low-Ca, and low-NaCl artificial freshwater, respectively, for 2 wk. Cell densities of wavy-convex, shallow-basin, and deep-hole types of gill MR cells as well as whole-body Ca(2+), Na(+), and Cl(-) influxes were measured. Low-Ca fish developed more shallow-basin MR cells in the gills and a higher Ca(2+) influx than those acclimated to other media. However, fish acclimated to low-NaCl artificial freshwater predominantly developed wavy-convex cells, and this was accompanied by the highest Na(+) and Cl(-) influxes. Relative abundance of shallow-basin and wavy-convex MR cells appear to be associated with changes in Ca(2+) and Na(+)/Cl(-) influxes, suggesting that shallow-basin and wavy-convex MR cells are mainly responsible for the uptake of Ca(2+) and Na(+)/Cl(-), respectively.

Using sustainable development indicators as a supplementary measure for the integrated management of environmental information system in Taiwan.

Environmental Information System (EIS) transfers environmental data and information to audiences in any field of knowledge or activity interested in environmental problems. Currently, numerous conventional EISs or Environmental Databases (EDs) are available in Taiwan. However, they fail to adequately address the strong correlations among the environmental, social and economic components. Notably, Sustainable Development Indicators (SDIs) combine environmental, social and economic dimensions, as well as illustrate the links among systems. Subsequently, developing a set of SDIs will provide an excellent opportunity to systematically consolidate the information scattered among many archives, as well as promoting an integrated EIS. This work presents an experimental model referred to as the 'Sustainable Development Information System of the Island Taiwan' (SDISIT). This system was developed to assist the integrated management of either EIS or ED. The Factor Analysis Method (FAM) is one tool that may prove useful in easing the developing process. Furthermore, by examining the correlations between the indicators, this method can apply existing SDIs to identify the structure of interrelationships among indicators.

[A study on behavior towards injection among residents and physicians in Kaohsiung & Ping-tung area].

To understand why people in Taiwan preferred iatrogenic injection as their medical therapy, we surveyed the knowledge, attitude, and behavior of iatrogenic injection for people and doctors in Kaohsiung and Ping-tung area by questionnaire. The results of the survey are drawn below: 1. More than 91.7% of the interviewed people would like to receive iatrogenic injection as their medical therapy, and 78.1% of the parents would choose iatrogenic injection for their children. The reason majority were because doctors suggested that and the parents believed iatrogenic injection is a quicker way of recovery. 2. In this study, 72.1% of the people received injection as their usual mode of medical treatment, and 74.9% of them had received injection during the previous month. When they brought their children to the clinics, the percentages of the above two conditions were 60.4% and 59.7% respectively. The factors influencing the reception of injection for people were the efficacy of the injection, education level and medical insurance. The influencing factors for children were the parents' medical behavior and the parents' willingness to accept injection. 3. About 83% of the interviewed doctors preferred iatrogenic injection as treatment for their patients. Almost 60% of them agreed that more patients would receive injection after National Health Insurance. About 72% of the doctors would not change the treatment for their patients, no matter how the system of the medical insurance changes. They would choose the right treatment for the distinct disease.

Capacity of robot handling for Epstein-Barr virus transformation.

OBJECTIVES: Epstein-Barr virus (EBV) transformation has been described as a routine method to establish human B lymphoblastoid cell lines. Each established lymphoblastoid cell line represents one unique genetic information carrier and can produce unlimited quantities of DNA materials available for downstream applications and research. Undoubtedly, it is of great value to human clinical and experimental genetic studies. However, the current process of EBV transformation requires much manpower in the routine renewal of medium, which is time-consuming. This situation can become a serious problem especially when establishing a human B lymphoblastoid cell bank. A modified and cost-effective protocol for EBV transformation should be considered. MATERIALS AND METHODS: In the present study, process in EBV transformation was modified to fit the requirements of robot handling. RESULTS: 1 mL of whole blood was demonstrated to be sufficient to perform EBV transformation. Additionally, EBV transformation can performed in 96-deep-well plates that are directly and widely used with automatic work platforms. CONCLUSIONS: Based on these facts, a process of EBV transformation can be modified to fit the requirements of robot handling.

Light Activation of Mg-dependent Adenosine 5'-Triphosphatase in Isolated Euglena Chloroplasts.

Enhancement of Mg(2+)-dependent ATPase activity in Euglena gracilis chloroplasts by light in the presence of a sulfhydryl compound has been demonstrated. A number of uncouplers and energy transfer inhibitors were studied for their effects on the light enhancement of ATPase activity simultaneously with their effects on photophosphorylation. Results suggest that the light-enhanced ATPase activity in Euglena chloroplasts is an energy-initiated process and that the energy is supplied through electron flow upon illumination of the chloroplasts. However, by studying the difference in their response toward the various uncouplers and inhibitors, it seems that the two processes (photohydrolysis of ATP and photophosphorylation) share only the latter part of their energy-transferring pathway.

Sensitization of mice by inhalation of antigen after infection with Bordetella pertussis.

Intranasal infection of mice with Bordetella pertussis or injection of pertussis vaccine previous to administration of an albumin aerosol augments sensitivity toward albumin. Sensitization was demonstrated by provocation of anaphylactic reactions following intravenous injection of antigen.

Enhancement of acousto-optic filter resolution using birefringence dispersion in CdS.

A CdS acousto-optic tunable filter has been constructed and operated near the band gap. The filter exhibits a large enhancement of spectral resolution that is due to birefringence dispersions. Over the wavelength range 530 to 545 nm, a half-power bandwidth of 20 A was obtained with an angular aperture of 38 degrees.

Expression of growth-related genes and drug-resistance genes in HTLV-I-positive and HTLV-I-negative post-thymic T-cell malignancies.

This study was designed to investigate the biologic and molecular basis of the aggressive behavior of high-grade post-thymic T-cell malignancies. Freshly frozen tumor tissues from (1) human T-cell leukemia/lymphoma virus type I (HTLV-I)-positive adult T-cell lymphoma (ATL) (7 cases), (2) HTLV-I-negative aggressive T-cell lymphoma (12 cases), and (3) HTLV-I-negative nonaggressive T-cell lymphoma (11 cases) were studied for the expression of several growth-related genes or proliferation antigens including interleukin-2 receptor (IL-2R), Ki-67, transforming growth factor-beta (TGF-beta), topoisomerase, and the multidrug resistance (MDR) gene by immunohistochemistry and Northern blot hybridization. Our results showed that tumor cells associated with HTLV-I and anaplastic morphology had an enhanced expression of Ki-67, TGF-beta, and topoisomerase, as compared to nonaggressive T-cell lymphoma. The expression of IL-2R was limited to ATL and one Ki-1 lymphoma. The MDR gene was frequently expressed in ATL, but only infrequently in other, HTLV-I-negative, malignancies. Clinical progression or relapse was associated with the expression of MDR, in addition to an increased expression of Ki-67. We therefore conclude that the aggressive clinical behavior of high-grade T-cell lymphoma may result mainly from the high proliferative activity of tumor cells, but the association with HTLV-I and clinical relapse is further complicated by the development of drug resistance.

Semi-commercial-scale production of Japanese B encephalitis virus vaccines from tissue culture.

This study was undertaken to modify and develop procedures for tissue culture-inactivated Japanese B encephalitis (JBE) virus vaccine production in large quantities. Various types of glass bottles were tried and, considering many advantages, long cylindrical roller (CR) bottles were selected. Several variables were investigated including number and volume of trypsinized cells to be seeded, volume of growth medium required for optimum cell growth, amount of calf serum, and volume of harvest medium for a high-titer virus yield. A good confluent cell sheet in CR bottles was obtained within a week by increasing the calf serum from 4 to 10% and when such tissue in a CR bottle was inoculated with 45,000 viral mean tissue culture infective doses directly into the medium, the cytopathological effects (CPE) appeared on day 5. High-titer virus yields were obtained when the harvests were made at 4(+) CPE using medium 199 with 2% human albumin at pH 8.3 to 8.5. No appreciable gain in titer was found from such harvests by blending to release intracellular virions. The production methods finally adopted gave consistently good results, and several inactivated JBE virus vaccine lots with minimum immunizing doses, ranging from 0.005 to 0.017 ml, were prepared using a large number of CR bottles in a simulated commercial-scale production system.

Development of ceruloplasmin in pigs during the neonatal period.

Colostrum-free newborn pigs were raised artificially to investigate the appearance of ceruloplasmin in the serum of piglets. The sensitivity of the p-phenylenediamine oxidase activity assay, commonly used as a measure of ceruloplasmin, has been increased 10-fold. Based on this activity, no evidence was found that this copper protein was present in the serum of baby pigs at birth, although some evidence was obtained that the protein moiety (apoceruloplasmin) was present. Holoceruloplasmin usually became detectable in the serum 10-15 hours after birth. Its concentration increased slowly at first and then at an accelerated rate. The ceruloplasmin in the sera of 2-3-day-old piglets was isolated and compared with that isolated from the serum of adults. Contrary to a previous report, the physical and chemical properties of ceruloplasmin synthesized during the first 3 days of life of the piglet showed no significant difference from that synthesized by the adult.

Allograft reconstruction for the treatment of long bone defects: report of 9 cases.

From April 1980 to December 1987, we experienced 18 cases of frozen intercalary allografts including 13 cases of bone tumor, 3 cases of congenital pseudoarthrosis of the tibia and 2 cases of traumatic bone defect. Nine cases were followed up for more than 2 years. The average allograft of length was 10.5cm (ranging from 3.5cm to 22cm). All cases had supplemental autografts in combination with intercalary allografts. The average time of radiographic union was 5 months (ranging from 4 months to 8 months) between allograft and autogenous cancellous bone graft, and 11.3 months (ranging from 7 months to 18 months) between allograft and host bone. We use the bone scan for early determination of the fate of allografts. The bone scan usually becomes positive at about 6 months after transplantation. The functional result was 89% "excellent" and "good" according to Henry J. Mankin's grading. One non-union and 3 leg length discrepancies of more than 2cm were encountered. In this study, we found that the deep frozen allograft for the treatment of large bone defects is very encouraging.

Controlling factors in the development of ceruloplasmin in pigs during the neonatal growth period.

The sequence of ceruloplasmin development and limiting factors controlling this process in neonatal piglets was studied in a randomized block design. Guided by the change in serum ceruloplasmin levels, the liver copper concentrations of piglets fed copper-deficient and copper-supplemented diets were compared in three different periods. The plasma of piglets was devoid of both apo- and holo-ceruloplasmin at birth. The system responsible for the synthesis of apoceruloplasmin developed shortly after birth as indicated by a steady increase in serum ceruloplasmin activity during the first week without being affected by the diet. Copper apparently was not the limiting factor at this stage since very high concentrations of copper were found in the livers of newborn piglets. However, the liver copper was depleted quickly within 1 to 2 weeks and became the limiting factor if the diet was deficient in copper. The livers of these piglets were fractionated by differential centrifugation to compare the liver copper distribution in the two dietary groups with and without copper supplementation. The treatments did not affect the copper distribution on a percentage of total liver copper basis. However, fractionation study revealed that, unlike adult pig livers, about 70% to 80% of copper was present in the heavier particle fractions of piglet livers.

Attack rate of dengue-like illness among teachers in Kaohsiung City, 1988.

The present study is based on a questionnaire evaluating 2,770 school teachers, including those from elementary, junior-high, senior-high, and occupational schools, in Kaohsiung city. The response rate was 91.1% (2,626/2,770). The self-reported cumulative incidence of dengue-like illness was 7.9% (207/2,626) and the physician-diagnosed cumulative incidence was 5.6% (148/2,626). There is no significant difference between distribution of infections among subjects' gender, educational background, and the associated school regardless of self-reported cases or diagnosed cases. Among those self-reported dengue infected teachers, 60.1% (119/207) required sick leaves. The difference of sick leaves among subjects by either education background or associated school was not significant. However, more females stayed home because of the illness than males did.

Pulmonary function and respiratory symptoms of Portland cement workers in southern Taiwan.

Respiratory symptoms and pulmonary function were studied in a cross-sectional survey of Portland cement workers in southern Taiwan. The study cohort consisted of 661 cement workers who were classified into low, medium, and high exposure groups with mean exposure values of 0.22, 0.55 and 1.24 mg/m3 of respirable dust concentrations, respectively. The prevalence of respiratory symptoms was not found to be significantly related to exposure, with the exception of a higher prevalence of coughing found among the heavy exposure group (OR = 1.55, P < 0.05). Both non-smoking and smoking cement workers showed no significant differences in the pulmonary function indices among the three exposure groups.

[The study of Apgar score and infant birth weight in the central Taiwan].

The objective of this study was to assess the relationship between birth weight and the Apgar Score. We collected data on the birth weights and the 1 min and 5 min Apgar Score of new born infants between 1982 and 1987 at a teaching hospital in Central Taiwan. Compared to babies with normal Apgar Score, infants with low Apgar Scores were found to born with low and very low birth weights. In the 1 min of life test, the relative risks of low birth weights among infants with Apgar Scores of 0 to 3 and 4 to 6 were 115.0 and 5.9 times higher than those of normal infants, respectively. In the very low birth weight category, the relative risks of the above score were 252.5 and 51.1, in this order. In the 5 min of life test, the relative risks of the above scores were 16.2 and 12.1 in the low birth weight category, respectively. However, among babies of very low birth weight, the relative risks of the same scores were 121.2 and 84.9, in this order. In conclusion, the 5 min Apgar Score might be a useful prognostic index for the relationship between health and birth weight of new born infants.