Previous studies have shown that the cytotoxicity of fluoropyrimidines is mediated, in large part, by inhibition of the enzyme
thymidylate synthase (TS). The aim of this study was to determine ...whether the chemosensitivity of human cancer cells to fluoropyrimidines
could be increased by decreasing TS expression with antisense oligodeoxyribonucleotides (ODNs). ODNs (18-mers) targeted at
the AUG translational initiation site of TS mRNA inhibited translation in a sequence- and dose-dependent manner in a rabbit
reticulocyte lysate in vitro translation system. Treatment of human colon cancer HT-29 cells with antisense ODNs decreased
TS catalytic activity in the cells in a dose-dependent manner over a short period, but the longer-term effect of the TS antisense
ODN treatment was actually to increase the amount of TS in the cells and to decrease their sensitivity to 5-fluoro-2'-deoxyuridine
(FdUrd). However, when human nasopharyngeal cancer KB31 cells were transfected with a plasmid (pHaMAGRP) construct containing
the TS antisense fragment (+ 1 to + 422) under the control of a glucose-regulated promoter, the expression of both TS protein
and TS catalytic activity was decreased by nearly 30% (P = 0.014), and sensitivity of these cells to FdUrd was enhanced by
approximately 8-fold (P = 0.021). No changes in the levels of expression of TS protein or FdUrd-associated cytotoxicity were
observed in control, vector-transfected cells. No change was observed in the sensitivity of transfected cells toward either
cisplatin or Adriamycin. These results show that the level of expression of TS in human malignant cells can be down-regulated
with antisense TS RNA, and their sensitivity to fluoropyrimidines can, thereby, be increased.
Objective: To evaluate the safety and immunogenicity of two lots of a heptavalent
Streptococcus pneumoniae conjugate vaccine (PCV) containing seven capsular polysaccharide serotypes (4, 6B, 9V, 14, ...18C, 19F, and 23F) conjugated to the outer membrane complex of
Neisseria meningitidis serogroup B (OMPC) and administered to infants at 2, 4, 6, and 12 months of age.
Methods: One hundred twenty infants were randomly assigned to concurrently receive PCV-OMPC and one of two
Haemophilus influenzae type b (Hib) conjugate-DTwP combination vaccines: (1) Hib with a heterologous protein carrier (CRM
197, TETRAMUNE
®, Group 1) or (2) an experimental Hib-hepatitis b combination vaccine with the homologous carrier (OMPC, Group 2). All infants in Groups 1 and 2 received PCV-OMPC (lot 1) at 12 months of age. Another separate group of 120 infants (Group 3) received a different lot of PCV-OMPC concurrently with Hib-CRM
197 (TETRAMUNE
®) at 2, 4, and 6 months of age and then were randomized to receive either PCV-OMPC or a 23-valent polysaccharide (PS) pneumococcal vaccine at 12 months of age.
Results: Each PCV-OMPC lot was generally well tolerated and no vaccine-related serious adverse events were reported. Following the primary series, serotype-specific anti-pneumococcal geometric mean concentrations (GMC) were highest for serotypes 14, 19F, and 4 and lowest for serotypes 6B and 23F. GMC and seroconversion rates in Group 3 (lot 2) were lower than in Group 1 (lot 1) for serotypes 6B, 14, 18C, and 23F. Antibody responses to serotypes 6B, 14, and 18C were significantly lower in Group 2 compared to Group 1. Following a booster dose of PCV-OMPC at 12 months of age, each lot was immunogenic with at least a 5–10-fold increase in antibody levels, and responses were significantly higher among those who received the PS vaccine.
Conclusions: PCV-OMPC is generally safe in infants, displays variable immune response by serotype, and concomitant receipt of Hib vaccine with homologous carrier may impact on its immunogenicity.
Histone acetyltransferase (HAT) and histone deacetylase (HDAC) activities determine the acetylation status of histones, and have the ability to regulate gene expression through chromatin remodeling. ...Aberrant histone acetylation is known to play a key role in leukemogenesis. To improve the treatment outcome in AML patients, we examined the antiproliferative activity of SK-7041, a novel hybrid synthetic HDACI, in acute myelogenous leukemia (AML) cell lines (KG-1,HL-60 and HEL). This HDACI preferentially inhibited the enzymatic activities of HDAC1 and HDAC2, compared to other HDAC isotypes, indicating that class I HDAC is the major target of SK-7041. SK-7041 effectively induced the time-dependent hyperacetylation of histones, H3 and H4 in AML cell lines. We found that this compound exhibited potent antiproliferative activity against AML cell lines in vitro. The growth inhibitory effect of SK-7041 was related to the induction of apoptosis in AML cell lines. SK-7041(4uM) induced potent cell death, triggering apoptosis in over 60% of cells in 48h by FACS analysis. After 12h of treatment by western analysis, apoptotic cells were increased through activation of caspase-7,-9,-3 and cleaved-PARP (caspase-8 was not affected). Cyclin D1 expression was decreased, and CDK4 expression level was not affected by SK-7041. The level of p21 expression was gradually increased in KG-1 and HL-60 cell lines, whereas treated HEL cell lines showed no change. For further investigation of the genes affected by SK-7041, 20Kb oligonucleotide microarray analysis was performed. The expression of candidate genes, which we selected in chip analysis, was confirmed using real-time PCR. Fifteen genes (RGL1, FYN, CARD9, ABCA7, TNFRSF6B, CASP9, ENPP2, etc.) were consistently up-regulated and twelve genes (PTPN7, CD34, INSIG1, IL16, LHX6, TRIB3, BID, PDCD4, etc.) were down-regulated more than 4-fold by the SK-7041 compound. We suggest that the efficacy of SK-7041 was mainly mediated by the inhibition of proliferation and induction of apoptosis. Taken together, our data suggested that novel HDACI SK-7041 treatment is effective in AML cell lines, and that the further study is warranted.
Traditional mechanisms for rating adherence or fidelity are labor-intensive. We developed and validated a tool to rate adherence to Motivational Enhancement Therapy–-Cognitive Behavioral Treatment ...(MET-CBT) through anonymous client surveys. The instrument was used to survey clients in 3 methadone programs over 2 waves. Explanatory and Confirmatory Factor Analyses were used to establish construct validity for both MET and CBT. Internal consistency based on Cronbach's alpha was within adequate range (α > 0.70) for all but 2 of the subscales in one of the samples. Consensus between clients’ ratings (r wg(j) scores) were in the range of 0.6 and higher, indicating a moderate to strong degree of agreement among clients’ ratings of the same counselor. These results suggest that client surveys could be used to measure adherence to MET-CBT for quality monitoring that is more objective than counselor self-report and less resource-intensive than supervisor review of taped sessions. However, additional work is needed to develop this scale.
We hypothesized that AMPK plays a role in regulation of insulin action in C2C12 myotubes. We expressed constitutively active (Ad‐AMPK‐CA, a generous gift from Ken Walsh) and dominant negative ...(Ad‐AMPK‐DN, produced by Morris Birnbaum) forms of AMPKα by use of adenovirus‐mediated gene transfer. Compared with control (Ad‐GFP), Ad‐AMPK‐CA increased glucose transport by ~50% and increased glucose transport by ~60 and ~70% in the presence of 10 nM insulin and 100 nM insulin, respectively. The increase in insulin sensitivity caused by transduction with Ad‐AMPK‐CA was suppressed by a 3‐h treatment with Compound C (a gift from Merck), an AMPK inhibitor. A 1‐h treatment of myotubes with the AMPK activator 5‐amino‐imidazole‐4‐carboxamide‐1‐β‐D‐ribofuranoside (AICAR) followed by a 2 hour recovery induced increased glucose transport in the presence of insulin (10 nM) in Ad‐GFP myotubes but not in the Ad‐AMPK‐DN group. Our initial western blots suggest that cells transduced with Ad‐AMPK‐CA have decreased serine phosphorylation of IRS‐1 at an mTOR (or S6K) target site, phosphorylation of which is associated with insulin resistance. It appears that there is a synergistic effect of insulin and AMPK on glucose transport in C2C12 myotubes, and our data suggest the possibility that AMPK plays an important physiological role in regulation of insulin signaling.
This work is supported by NIH K01 DK066330.
Internationalization is the term that can characterize the current state of the consumer market in many parts of the world. The phenomenon of an internationalized consumer market is noteworthy ...particularly because consumer mobility beyond their national boundaries has increased resulting in a significant growth of diverse immigrant populations in the international marketplace. The theory of intercultural accommodation suggests that consumers who have relocated beyond their national/cultural boundaries adapt to the new market environment developing new consumption patterns. At the same time international consumers do not entirely lose their original ethnic identity that had guided their socialization process. The evolving nature of ethnic identity of international consumers may pose an interesting question about the usefulness of observable ethnicity as a market segmentation basis in international consumer marketing. This study examined how consumers' ethnic identity, specifically the strength of ethnic identification of those who are experiencing intercultural accommodation, influenced consumer apparel shopping orientations and retail format preferences, using a sample of Latino and Asian consumers residing in the U.S. The results of this study showed that Latino and Asian consumers demonstrated significant differences in their retail format preferences but shared similarities in apparel shopping orientations. Some differences in market behavior relative to the strength of ethnic identification were also observed within the Asian sample. Finally, implications and future research directions are discussed.
BACKGROUND.Combination vaccines are urgently needed to reduce the number of injections given to young children. The aim of the study was to evaluate the safety and immunogenicity of a combination ...vaccine that contains diphtheria and tetanus toxoids and acellular pertussis antigens (DTaP), recombinant hepatitis B surface antigen (HepB) and Haemophilus influenzae type b (Hib) polysaccharide conjugated to tetanus toxoid (PRP-T).
METHODS.Four hundred five infants were randomized equally to three groups and immunized at 2, 4 and 6 months of age with(1) DTaP/HepB vaccine used to reconstitute lyophilized PRP-T vaccine and administered as a single injection; (2) DTaP/HepB vaccine and PRP-T vaccine administered as two separate injections; or (3) DTaP, HepB and PRP-T vaccines administered as three separate injections. Safety was closely monitored, and blood specimens were obtained to assess antibody responses to each vaccine antigen.
RESULTS.All study vaccines were well-tolerated, and the rates of systemic and injection site reactions were similar between groups. After the third dose the geometric mean antibody concentrations to Hib were significantly lower in subjects in Group 1 (1.63 μg/ml) compared with subjects in Groups 2 and 3 (6.26 and 6.15 μg/ml, respectively;P < 0.0001). Subjects with antibody concentrations <1.0 μg/ml after the third dose responded well to a booster dose of Hib conjugate vaccine given at 11 to 15 months of age (41 of 44 with anti-PRP ≥1.0 μg/ml). Differences between groups for antibody responses to the other vaccine components were not clinically significant.
CONCLUSIONS.Infants given a combined DTaP/HepB/PRP-T vaccine experienced a significantly lower antibody response to the PRP-T component than infants given PRP-T vaccine as a separate injection. However, the immune response to a booster dose of Hib conjugate vaccine indicated the presence of immunologic memory.
Inhibition of cyclooxygenase-2 (COX-2) pathways may have significant implications for the prevention and treatment of head and neck squamous cell carcinoma (HNSCC). COX-2 is overexpressed in both ...premalignant lesions and invasive HNSCC. We examined COX-2 expression by immunohistochemistry in normal tissues, different stages of premalignant lesions, and carcinoma
in-situ
(CIS). We also evaluated the correlation between COX-2 expression and clinical characteristics of HNSCC patients. Tissue specimens were obtained from: premalignant lesions from 25 subjects enrolled in a biochemoprevention trial; tumor samples collected at diagnosis from 38 HNSCC patients enrolled in an induction chemotherapy trial; and normal control tissues from 10 non-cancer, non-smoking subjects. COX-2 was expressed in early and intermediate stages of premalignant lesions, increasing first in the basal and parabasal layers, then lower spinous, and upper spinous layers. This correlation was noted in normal epithelium (p<0.0001), histologically normal in-field samples (p<0.0001), low-grade dysplasia (p=0.024), and moderate-grade dysplasia (p=0.009), but was lost in the majority of high-grade dysplasia/CIS (p=0.896). COX-2 expression was also noted to increase progressively through the early stages of premalignancy, and to decrease in severe/CIS stage and invasive carcinoma. COX-2 expression in tumors from patients treated with induction chemotherapy was correlated with overall survival after controlling for clinical variables. These findings elucidate the differential expression pattern of COX-2 in stages of head and neck premalignant lesions and invasive carcinoma, supporting the rationale for COX-2 inhibition as an important strategy for cancer chemoprevention. Further validation of COX-2 expression is needed in prospective ongoing chemoprevention trials.
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