bluuebunny/crossref_metadata_2025_split huggingface dataset:
Effects of recombinant adenovirus-mediated expression of IL-2 and IL-12 in human B lymphoma cells on co-cultured PBMC
Authors: Ebert Oliver , Wilbert Dorothee , Buttgereit Peter , Ziske Carsten , Flieger Dimitri , Schmidt-Wolf Ingo GH 
Published: October 2004

Abstract
Background
Modulation of the immune system by genetically modified lymphoma cell vaccines is of potential therapeutic value in the treatment of B cell lymphoma. However, the anti-tumor effect of any single immunogene transfer has so far been limited. Combination treatment of recombinant IL-2 and IL-12 has been reported to be synergistic for inducing anti-tumor responses in solid tumors but the potential of IL-2/IL-12 gene modified B cell lymphoma cells has not been explored yet.

Methods
Using three different human B cell lymphoma cell lines and primary samples from patients with B cell neoplasms, expression levels of the coxsackie B-adenovirus receptor (CAR) and alpha (v) integrins were analyzed by fluorescence-activated cell sorter (FACS). Adenoviral transduction efficiencies were determined by GFP expression analysis and IL-2 and IL-12 cytokine production was quantified by enzyme-linked immunosorbent (ELISA) assays. Proliferative activities of peripheral blood mononuclear cells (PBMC) stimulated with either cytokine derived from supernatants of transduced lymphoma cells were measured by cell proliferation (MTT) assays. An EuTDA cytotoxicity assay was used to compare cytotoxic activities of IL-2 and/or IL-12 stimulated PBMC against unmodified lymphoma cells.

Results
We found that B cell lymphoma cell lines could be transduced with much higher efficiency than primary tumor samples, which appeared to correlate with the expression of CAR. Adenoviral-expressed IL-2 and IL-12 similarly led to dose-dependent increases in proliferation rates of PBMC obtained from healthy donors. IL-2 and/or IL-12 transduced lymphoma cells were co-cultured with PBMC, which were assayed for their cytolytic activity against unmodified lymphoma cells. We found that IL-2 stimulated PBMC elicited a significant anti-tumor effect but not the combined effect of IL-2/IL-12 or IL-12 alone.

Conclusion
This study demonstrates that the generation of recombinant adenovirus modified lymphoma cell vaccines based on lymphoma cell lines expressing IL-2 and IL-12 cytokine genes is technically feasible, induces increases in proliferation rates and cytotoxic activity of co-cultured PBMC, and warrants further development for the treatment of lymphoma patients in the future.
The effect of chemotherapy combined with recombination mutant human tumor necrosis factor on advanced cancer
Authors: Liu Xing , Zhang Xiang-fu , Zheng Zhi-weng , Lu Huishan , Wu Xinyuan , Huang Changmin , Wang Chuan , Guang Guoxiang 
Published: October 2004

Abstract
Background
Past studies suggested that tumor necrosis factor (TNF) assisted anti-tumor treatment and intensified the sensitivity of chemotherapy. However its clinical application has been curbed because of its low purity, high dosage, and strong toxicity. This research, through perspective random clinical control experiment, observed the therapeutic effect of the treatment of late malignant tumor through the injection of recombinant mutant human tumor necrosis factor (rmhTNF) combined with general chemotherapy and its adverse reactions.

Methods
105 patients with advanced malignant tumor were randomly divided into trial group, 69 patients, and control group, 36 patients. Injection of rmhTNF 4 × 106u/m2 was given to the trial group, from the 1st to 7th days, the 11th to 17th days combined with chemotherapy course. The chemotherapy plan was as follows: CAP for patients with the NSCLC; FAM for patients with gastric cancer; FC for patients with colorectal cancer. One treatment cycle lasted for 21 days and two cycles were scheduled. The control group was given only the same chemotherapy as the trial group.

Results
In the trial group there was 1 CR case and 12 PR cases, and the response rate is 13/69 (18.84%); in the control group 1 PR case, the response rate 1/36 (2.78%). The response rate of the trial group was significantly higher than that of the control group (P = 0.022). The response rate for NSCLC in the trial group was 8/17 (47.06%), and 1/6 (16.67%) in the control group. The response rates for gastric cancer and colorectal cancer in the trial groups also were higher than those of the control groups. After the treatment the KPS is 89.00 ± 9.92 in the trial group, and 84.17 ± 8.84 in the control group, with a significant difference between the two groups (P = 0.028). The adverse reactions of rmhTNF injection included: pain in the injection area, chill, hardening and swelling and redness in the injection area, fever, ostealgia and myosalgia, and cold-like symptoms. All these adverse reactions were mild and bearable.

Conclusions
The administration of rmhTNF injection in combination with general chemotherapy is an effective and secure means in treating advanced malignant tumor.
The J-shape association of ethanol intake with total homocysteine concentrations: the ATTICA study
Authors: Pitsavos Christos , Panagiotakos Demosthenes B , Kontogianni Meropi D , Chrysohoou Christina , Chloptsios Yannis , Zampelas Antonis , Trichopoulou Antonia , Stefanadis Christodoulos 
Published: October 2004

Abstract
Background
Epidemiological studies suggest a non-monotonic effect of alcohol consumption on cardiovascular risk, while there is strong evidence concerning the involvement of homocysteine levels on thrombosis. The aim of this work was to evaluate the association between usual ethanol consumption and homocysteine levels, in cardiovascular disease free adults.

Methods
From May 2001 to December 2002 we randomly enrolled 1514 adult men and 1528 women, without any evidence of cardiovascular disease, stratified by age – gender (census 2001), from the greater area of Athens, Greece. Among the variables ascertained we measured the daily ethanol consumption and plasma homocysteine concentrations.

Results
Data analysis revealed a J-shape association between ethanol intake (none, <12 gr, 12 – 24 gr, 25 – 48 gr, >48 gr per day) and total homocysteine levels (mean ± standard deviation) among males (13 ± 3 vs. 11 ± 3 vs. 14 ± 4 vs. 18 ± 5 vs. 19 ± 3 μmol/L, respectively, p < 0.01) and females (10 ± 4 vs. 9 ± 3 vs. 11 ± 3 vs. 15 ± 4 vs. 17 ± 3 μmol/L, respectively, p < 0.01), after controlling for several potential confounders. The lowest homocysteine concentrations were observed with ethanol intake of < 12 gr/day (Bonferroni α* < 0.05). No differences were observed when we stratified our analysis by type of alcoholic beverage consumed.

Conclusion
We observed a J-shape relationship between homocysteine concentrations and the amount of ethanol usually consumed.
Expression of the largest CD97 and EMR2 isoforms on leukocytes facilitates a specific interaction with chondroitin sulfate on B cells
Authors: Kwakkenbos Mark J , Pouwels Walter , Matmati Mourad , Stacey Martin , Lin Hsi-Hsien , Gordon Siamon , Van Lier René A W , Hamann Jörg 
Published: October 2004

AbstractThe EGF-TM7 receptors CD97 and EMR2 are heptahelical molecules predominantly expressed on leukocytes. A characteristic of these receptors is their ability to interact with cellular ligands via the N-terminal epidermal growth factor (EGF)-like domains. The first two EGF domains of CD97 (but not EMR2) bind CD55 (decay-accelerating factor), while the fourth EGF domain of both CD97 and EMR2 interacts with the glycosaminoglycan chondroitin sulfate (CS). Using fluorescent beads coated with soluble recombinant CD97 and EMR2 protein, and isoform-specific monoclonal antibodies, we have determined the cellular and molecular characteristics of the interaction with CS. The fourth EGF domain of CD97 and EMR2 is expressed on activated lymphocytes and myeloid cells, whereas the ligand is specifically found on B cells within the peripheral blood. The interaction between CD97/EMR2 and CS may therefore play a role in the interaction of activated T cells, dendritic cells, and macrophages with B cells.
Estrogen Acidifies Vaginal pH by Up-Regulation of Proton Secretion via the Apical Membrane of Vaginal-Ectocervical Epithelial Cells
Authors: Gorodeski George I. , Hopfer Ulrich , Liu Chung Chiun , Margles Ellen 
Published: October 2004

AbstractThe objective of this study was to assess estrogen-dependent cellular mechanisms that could contribute to the acid pH of the vaginal lumen. Cultures of normal human cervical-vaginal epithelial (hECE) cells and endocervical cells were grown on filters, and acidification of the extracellular solutions on the luminal (L-pHo) and contraluminal (CL-pHo) sides was measured. The hECE cells and endocervical cells decreased CL-pHo from 7.40 to 7.25 within 20–30 min of incubation in basic salt solution. Endocervical cells also produced a similar decrease in L-pHo. In contrast, hECE cells acidified L-pHo down to pH 7.05 when grown as monoculture and down to pH 6.05 when grown in coculture with human cervical fibroblasts. This enhanced acid secretion into the luminal compartment was estrogen dependent because removal of endogenous steroid hormones attenuated the effect, whereas treatment with 17β-estradiol restored it. The 17β-estradiol effect was dose dependent (EC50 0.5 nm) and could be mimicked by diethylstilbestrol and in part by estrone and tamoxifen. Preincubation with ICI-182780, but not with progesterone, blocked the estrogen effect. Preincubation of cells with the V-ATPase blocker bafilomycin A1, when administered to the luminal solution, attenuated the baseline and estrogen-dependent acid secretion into the luminal solution. Treatment with EGTA, to abrogate the tight junctional resistance, blocked the decrease in L-pHo and stimulated a decrease in CL-pHo, indicating that the tight junctions are necessary for maintaining luminal acidification. We conclude that vaginal-ectocervical cells acidify the luminal canal by a mechanism of active proton secretion, driven in part by V-H+-ATPase located in the apical plasma membrane and that the baseline active net proton secretion occurs constitutively throughout life and that this acidification is up-regulated by estrogen.
c-Src Regulates Clathrin Adapter Protein 2 Interaction with β-Arrestin and the Angiotensin II Type 1 Receptor during Clathrin- Mediated Internalization
Authors: Fessart Delphine , Simaan May , Laporte Stéphane A. 
Published: October 2004

Abstractβ-Arrestins are multifunctional adapters involved in the internalization and signaling of G protein-coupled receptors (GPCRs). They target receptors to clathrin-coated pits (CCPs) through binding with clathrin and clathrin adapter 2 (AP-2) complex. They also act as transducers of signaling by recruiting c-Src kinase to certain GPCRs. Here we sought to determine whether c-Src regulates the recruitment of AP-2 to β-arrestin and the angiotensin II (Ang II) type 1 receptor (AT1R) during internalization. We show that the agonist stimulation of native AT1R in vascular smooth muscle cells (VSMCs) induces the formation of an endogenous complex containing c-Src, β-arrestins and AP-2. In vitro studies using coimmunoprecipitation experiments and a yeast three-hybrid assay reveal that c-Src stabilizes the agonist-independent association between β-arrestin2 and the β-subunit of AP-2 independently of the kinase activity of c-Src. However, although c-Src expression promoted the rapid dissociation of AP-2 from both β-arrestin and AT1R after receptor stimulation, a kinase-inactive mutant of c-Src failed to induce the dissociation of AP-2 from the agonist-occupied receptor. Thus, the consequence of c-Src in regulating the dissociation of AP-2 from the receptor was also examined on the internalization of AT1R by depleting c-Src in human embryonic kidney (HEK) 293 cells using a small interfering RNA strategy. Experiments in c-Src depleted cells reveal that AT1R remained mostly colocalized with AP-2 at the plasma membrane after Ang II stimulation, consistent with the observed delay in receptor internalization. Moreover, coimmunoprecipitation experiments in c-Src depleted HEK 293 cells and VSMCs showed an increased association of AP-2 to the agonist-occupied AT1R and β-arrestin, respectively. Together, our results support a role for c-Src in regulating the dissociation of AP-2 from agonist-occupied AT1R and β-arrestin during the clathrin-mediated internalization of receptors and suggest a novel function for c-Src kinase in the internalization of AT1R.
Peripheral-Type Benzodiazepine Receptor-Mediated Action of Steroidogenic Acute Regulatory Protein on Cholesterol Entry into Leydig Cell Mitochondria
Authors: Hauet Thierry , Yao Zhi-Xing , Bose Himangshu S. , Wall Christopher T. , Han Zeqiu , Li Wenping , Hales Dale B. , Miller Walter L. , Culty Martine , Papadopoulos Vassilios 
Published: October 2004

AbstractHormone-induced steroid biosynthesis begins with the transfer of cholesterol from intracellular stores into mitochondria. Steroidogenic acute regulatory protein (StAR) and peripheral-type benzodiazepine receptor (PBR) have been implicated in this rate-determining step of steroidogenesis. MA-10 mouse Leydig tumor cells were treated with and without oligodeoxynucleotides (ODNs) antisense to PBR and StAR followed by treatment with saturating concentrations of human choriogonadotropin. Treatment with ODNs antisense but not missense for both proteins inhibited the respective protein expression and the ability of the cells to synthesize steroids in response to human choriogonadotropin. Treatment of the cells with either ODNs antisense to PBR or a transducible peptide antagonist to PBR resulted in inhibition of the accumulation of the mature mitochondrial 30-kDa StAR protein, suggesting that the presence of PBR is required for StAR import into mitochondria. Addition of in vitro transcribed/translated 37-kDa StAR or a fusion protein of Tom20 (translocase of outer membrane) and StAR (Tom/StAR) to mitochondria isolated from control cells increased pregnenolone formation. Mitochondria isolated from cells treated with ODNs antisense, but not missense, to PBR failed to form pregnenolone and respond to either StAR or Tom/StAR proteins. Reincorporation of in vitro transcribed/translated PBR, but not PBR missing the cholesterol-binding domain, into MA-10 mitochondria rescued the ability of the mitochondria to form steroids and the ability of the mitochondria to respond to StAR and Tom/StAR proteins. These data suggest that both StAR and PBR proteins are indispensable elements of the steroidogenic machinery and function in a coordinated manner to transfer cholesterol into mitochondria.
Hunting for Disease Genes in Multi-Functional Diseases
Authors: Pang Chi-Pui , Baum Larry , Lam Dennis S. C. 
Published: October 2004

Abstract
Disease genes may be identified through functional,
positional, and candidate gene approaches. Although
extensive and often labor-intensive studies such as
family linkage analysis, functional investigation of
gene products and genome database searches are
usually involved, thousands of human disease genes,
especially for monogenic diseases with Mendelian
transmission, have been identified. However, in diseases
caused by more than one gene, or by a combination
of genetic and environmental factors, identification
of the genes is even more difficult. Common
examples include atherosclerosis, cancer, Alzheimer's
disease, asthma, diabetes, glaucoma, and age-related
macular degeneration. There have been conflicting reports
on the roles of associated genes. Even with population-based case-control studies and new statistical
methods such as the sib-ship disequilibrium test and
the discordant alleles test, there is no agreement
on whether α2-macroglobulin (A2M) is a gene for Alzheimer's
disease. Another example is the inconsistent
association between age-related macular degeneration
and ATP-binding cassette transporter (ABCR).
Ethnic variation causes further complications. In our
investigation of LDL-receptor variants in familial hypercholesterolemia,
and the trabecular meshwork inducible
glucocorticoid response protein, or myocillin
(TIGR-MYOC) mutation pattern in primary open angle
glaucoma, we did find dissimilar results in Chinese
compared to Caucasians. New information from the
Human Genome Project and advancements in technologies
will aid the search for and confirm identification
of disease genes despite such challenges.
Gene Amplification as Means for Determining Therapeutic Strategies in Human Cancers
Authors: Tsongalis Gregory J. , Cartun Richard W. , Ricci Andrew 
Published: October 2004

Abstract
Pharmacogenomic analysis aspires to identify individuals
with specific genetic characteristics in order to
predict a positive response or reduce a negative response
to a therapeutic modality. While the search
continues for the many single nucleotide polymorphisms
which will be used in such genetic analyses,
other genetic alterations in specific cell types have
proven useful in determining the potential for response
to therapy. One such genetic alteration is amplification
of entire gene sequences which results in
overexpression of a gene product or protein. Amplification
of the HER2 (neu, erbB-2) oncogene is found in
up to 35% of human breast cancers and is associated
with a poor prognosis. In addition, this genetic alteration
may predict response to various therapeutic
modalities. Assays are available to detect the HER2
protein receptor or copies of the HER2 gene sequence
to determine eligibility for Herceptin treatment or adriamycin
treatment in node positive patients, respectively.
This model represents a somatic event used in
the functional determination of a therapeutic strategy.
Drug-Metabolizing Enzymes, Polymorphisms and Interindividual Response to Environmental Toxicants
Authors: Nebert Daniel W. 
Published: October 2004

Abstract
Individual risk of toxicity or cancer reflects the amount
of exposure to environmental agents, combined with
one's underlying genetic predisposition. More than six
dozen human ecogenetic polymorphisms have been
described; whereas some of these have been demonstrated
to be associated with altered risks of toxicity or
cancer, others presently remain equivocal and require
further study. Thus, genetic differences in the regulation,
expression and activity of “environmental susceptibility
genes” can be decisive in defining susceptibility
to toxicity or cancer.
“Drug-metabolizing enzymes” (DMEs) are regarded
as one class of environmental susceptibility genes.
DME genes have actually existed on this planet for
more than 2.5 billion years, and might more appropriately
be named “effector-metabolizing enzymes.” Receptors
controlling DME levels have been called “DME
receptors.” DMEs have functioned in many critical life
processes in prokaryotes and, more recently, in countless
basic functions in plants and animals—events that
evolved long before the existence of pharmaceutical
companies and apothecaries. DME genes exist in every
eukaryotic cell and probably in all prokaryotes. Virtually
all environmental agents act as either agonists or
antagonists—in competing with endogenous ligands
that bind to DME receptors and/or competing as substrates
for the DMEs. Over the past decade it has become
clear that each of us has our own “individual fingerprint”
of unique alleles coding for DMEs. The
underlying genetic predisposition of each patient will
reflect combinations of poor- and extensive-metabolizer
phenotypes; if these enzymes cooperate in the
same metabolic pathway for any given drug or environmental
agent, such ecogenetic variability might be
synergistic and lead to as much as 30- or >40-fold differences
in activation or degradation. The end result
can be large interindividual differences in risk of environmentally
caused toxicity or cancer.
How to Manage Individualized Drug Therapy: Application of Pharmacogenetic Knowledge of Drug Metabolism and Transport
Authors: Meisel Christian , Roots Ivar , Cascorbi Ingolf , Brinkmann Ulrich , Brockmöller Jürgen 
Published: October 2004

Abstract
Significant fractions of health budgets must be spent
for treatment of drug side effects and for inefficient
drug therapy. Hereditary variants in drug metabolizing
enzymes, drug transporters, and drug targets are important
determinants of drug response and toxicity
and may therefore aid in selection and dosage of
drugs. Today there is extensive knowledge of genetic
polymorphisms of cytochrome P450 (CYP) enzymes
2A6, 2C9, 2C19, and 2D6; of phase-2 enzymes such as
thiopurine S-methyltransferase; and more recently of
drug transporters such as the MDR-1 gene-product P-glycoprotein,
affecting a significant share of currently
used drugs. However, application of pharmacogenetic
knowledge to clinical routine is limited in current practice.
To promote the application of pharmacogenetic
knowledge in clinical routine, research on genotype-based
dose adjustments is still necessary—as is the
promotion of faster and cheaper genotype analyses.
Furthermore, the benefits of CYP genotype-directed
drug therapy should be evaluated in properly designed
prospective studies. Once such steps have been successfully
taken, drug therapy could well become more
prevention-directed and patient-tailored than it is possible
today, replacing the current “one drug in one
dose for one disease” strategy by a more individualized
approach.
P-Glycoprotein and Bioavailability-Implication of Polymorphism
Authors: Liu Yan , Hu Ming 
Published: October 2004

Abstract
P-Glycoprotein (P-gp) may have a significant impact
on systemic and tissue/cellular bioavailability of drugs
because it functions as an “anti-absorption” mechanism
that effluxes drug molecules out of the lipid bilayer
and cytoplasm. The ability to reduce bioavailability
at the tissue/cellular level was first discovered
during the investigation of the causes of multidrug resistance
(MDR) in cancer chemotherapy. Initially, it
was thought that MDR is only caused by P-gp. Recently,
many other transporters such as multidrug resistance-related protein (MRP) have also been identified.
The ability of P-gp to impact systemic drug
bioavailability was only recently recognized. Dr. Alfred
Schinkel's group was first to show a significant improvement
in the systemic bioavailability of several
drugs in the MDR1 knockout mice. The same group
also discovered that the blood-brain barrier (BBB) has
a very high expression level of P-gp, and that this protein
is necessary to restrict the entrance of various
drug molecules into the central nervous system (CNS).
Polymorphism in the normal human cells has not
been reported, but it has been discovered in human
cancer cells. Functional implication of P-gp polymorphism
in changing the tissue bioavailability has been
studied in rodents. These studies strongly support the
role of P-gp in restricting tissue bioavailability of anticancer
drugs. These studies also support the effectiveness
of P-gp in limiting CNS toxicity of the cytotoxic
drugs.
The functional implication of P-gp on systemic
bioavailability is much less well defined in humans, although
it appears to be quite obvious in MDR1 knockout
mice. Pharmacokinetic models clearly suggest that
a change in the absorption rate will have a significant
impact on systemic blood level of a drug. However,
whether functionally significant polymorphisms of P-gp
exist in humans has not been determined. If they do
exist, they will surely impact on both systemic bioavailability
and drug interaction potentials of many drugs.
In the drug development process, several models
may be used to select a lead compound that may or
may not interact with P-gp, depending on whether the
interaction is desirable. Several inhibitors of P-gp are
currently on the clinical trial stage. Natural inhibitors
of P-gp have also been discovered. There is no doubt
that these new developments will have significant impact
on the bioavailability of a variety of anticancer
and CNS drugs in the next decades.