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Current Pharmaceutical
Analysis
ISSN: 1573-4129
Current Pharmaceutical
Analysis
Volume 3, Number 3, August 2007
Contents
Microarrays as a Tool for Gene Expression Profiling:
Application In Ocular and Craniofacial Research Pp.
158-169
Maria Visalli, Mario Venza, Isabella Venza, Claudia Torino
and Diana Teti
[Abstract]
Chromatographic Removal of DNA from Protein Solutions
by Cationic Polymer Beads Pp. 170-179
Masayo Sakata and Masashi Kunitake
[Abstract]
Review of Hiv-1 Protease Inhibitor Assay Methods
Pp. 180-185
Duik Hur, Robin DiFrancesco, Jill Hochreiter, Qing Ma,
Judianne Slish, Chiedza C. Maponga and Gene D. Morse
[Abstract]
Single Molecule Bioanalysis Pp. 186-194
Jan Hesse and Gerhard J. Schütz
[Abstract]
Pharmacological and Clinical Applications of Natriuretic
Peptides: Accepted Knowledges or Changing Views?
Pp. 195-204
Izzi Valerio, Chiurchiù Valerio and Baldini P.
Morena
[Abstract]
Evaluation of Measuring Methods of Human Serum Albumin-Drug
Binding Affinity Pp. 205-212
Toshihiko Hanai
[Abstract]
Therapeutic Effects and Mechanisms of Action of Cyclosporine
A Ophthalmic Solution in the Treatment of Vernal Keratoconjunctivitis
Pp. 213-220
Daisuke Shii and Atsuki Fukushima
[Abstract]
Abstracts
[Back to top]
Microarrays as a Tool for Gene Expression
Profiling: Application In Ocular and Craniofacial Research
Maria Visalli, Mario Venza, Isabella Venza, Claudia Torino
and Diana Teti
Many approaches have been applied in biological and clinical
studies aimed at simultaneously monitoring the expression
of a large number of genes and at identifying the gene expression
patterns associated with specific cell types.
In the last decade, microarrays are emerged as an innovative
tool to detect differentially expressed genes as disease-specific
markers and to cluster the co-expressed genes regulating the
molecular interactions. In fact, microarrays have been widely
used for tumour diagnosis and classification, prediction of
prognoses and treatment, pharmaceutical safety assessment,
and understanding of molecular mechanisms, biochemical pathways,
and gene networks.
Besides the clinical relevance and the wide application of
this technology, several problems remain to be addressed.
This review illustrates the current and future directions
of the microarray platform, and highlights the successes and
strengths, as well as the shortcomings and limitations of
this technology.
An overview of microarray impact on the ocular and craniofacial
diseases is provided.
[Back to top]
Chromatographic Removal of DNA from Protein Solutions
by Cationic Polymer Beads
Masayo Sakata and Masashi Kunitake
Drugs produced using recombinant gene technology often contain
nucleic acids (DNA) as impurities. Such contaminants must
be removed from solutions used for injection, because the
influence of such DNA on the living body has not been fully
clarified. Various techniques for DNA removal described in
the patent literature are not broadly applicable, as they
are tailored to meet specific product requirements. To remove
DNA from a solution of high-molecular weight compounds, such
as proteins, the adsorption method has proven to be most effective.
This review intends to discuss various methods of chromatographic
removal and separation of DNA from protein solutions using
cationic polymer adsorbents. We will first discuss chromatographic
properties of various DNA-specific polymer adsorbents, and
then describe effects of adsorbent’s pore size, its
amino-group content, its surface pKa
value, and buffer’s condition on removal of DNA from
a protein solution. Finally, we will discuss how to optimize
a method of removal DNA without affecting the recovery of
important compounds, such as proteins.
[Back to top]
Review of Hiv-1 Protease Inhibitor Assay Methods
Duik Hur, Robin DiFrancesco, Jill Hochreiter, Qing Ma,
Judianne Slish, Chiedza C. Maponga and Gene D. Morse
In support of individualizing antiretroviral treatment by
combining therapeutic drug monitoring (TDM) and HIV resistance
tests, numerous assays have been reported in the literature
in an attempt to optimize treatment outcomes. These assays
vary with regard to analysis method of separation and detection
such as using high performance liquid chromatography (HPLC)
coupled to mass tandem spectrometry (HPLC/MS or HPLC/MSMS)
or ultraviolet spectrometry (HPLC/UV). To prepare plasma samples
prior to analysis, clinical samples are subject to chemical
procedures including solid phase extraction (SPE), liquid-liquid
extraction (LLE), and protein precipitation (PP). Information
searches were performed using database searching tools such
as PubMed, MEDLINE, Web of Science, and EM-BASE using the
key words protease inhibitors, HIV, assay, quantification,
determination, therapeutic drug monitoring and pharmacokinetics.
Publications were reviewed and the data categorized by the
specific protease inhibitor, the limit of quantification (LOQ),
the type of assay, chromatographic conditions, sample preparation,
and total assay run time. The analytical methods summarized
in the review reflect the timeframe since the PI were introduced,
their eventual combination with ritonavir, transitioning toward
once or twice daily dosing and more recently their use in
simplified regimes for chronic dosing. As antiretrovirals
are introduced into developing countries additional considerations
related to drug quality and potency may require that laboratories
adapt these assays in support of clinical pharmacology research
and patient monitoring.
[Back to top]
Single Molecule Bioanalysis
Jan Hesse and Gerhard J. Schütz
Beyond doubt, nanobioscience represents one of the
most innovative concepts in current basic and applied research.
Technological advances provided the essentials for this ongoing
scientific revolution, by making possible the observation,
investigation and manipulation of biomaterials down to single
molecules. Time-resolved single molecule detection has become
the method of choice to characterize structural transitions
between different functional states of isolated biomolecules.
The development of ultra-sensitive detection schemes also
has a strong impact on bioanalysis, as the sensitivity of
biochemical assays could be dramatically increased. Whenever
the available amount of sample is the limiting factor for
unambiguous diagnosis e.g. in medical diagnostics, bioanalytics
with single molecule sensitivity can be expected to become
even an enabling technology. In this review, we describe methodological
requirements for ultra-sensitive detection, and point out
major advantages, in particular the novel information content.
[Back to top]
Pharmacological and Clinical Applications of Natriuretic
Peptides: Accepted Knowledges or Changing Views?
Izzi Valerio, Chiurchiù Valerio and Baldini P.
Morena
Natriuretic peptides (NPs) are endogenous low molecular weight
peptides known for their hypotensive activities. Due to their
ability to induce both natriuresis (Na+
secretion) and diuresis, in fact, these peptides can profoundly
decrease the body haemodynamic load, an effect which represents
a main goal for all therapies aiming at improving the health
of those subjects affected by different forms of heart and
kidney failure. Over the years, many evidences have been provided
for the involvement of NPs in the control of different cardiovascular
parameters both in physiological and pathological conditions.
This has led many researchers to test the possible therapeutic
effects of different NPs and the pharmacological industries
to develop exogenous NPs forms with a higher activity. This
review aims then at summarizing the current knowledge about
the therapeutic and clinical applications of NPs, also considering
“atypical” NPs and the problems yet to be solved.
[Back to top]
Evaluation of Measuring Methods of Human Serum Albumin-Drug
Binding Affinity
Toshihiko Hanai
This study evaluated the feasibility of measuring HSA-drug
binding affinity by ordinary (free solution) method. NMR is
used to identify where a drug binds. An HSA-immobilized column
and mimic ion-exchange columns were developed to measure HSA-drug
binding affinity. The correlation between capacity ratios
(log k) measured using these columns and HSA-drug
binding affinity (log nK) was not perfect. The log
k values measured using the mimic ion-exchangers
well correlated with log nK values measured by a
modified Hummer-Dreyer method with purified HSA. The log k
(log KHSA) of basic drugs
correlated well with the log nK values measured by
the modified Hummer-Dreyer method, but not for the acidic
drugs. Simple measurement as the percent concentration of
bound drug is another matter. Computational chemical prediction
methods were proposed. One uses the molecular properties of
drugs. Another involves the direct calculation of binding
energy required using model phases. Further study is necessary
to develop quick measurement and prediction method to accelerate
the drug discovery process.
[Back to top]
Therapeutic Effects and Mechanisms of Action of Cyclosporine
A Ophthalmic Solution in the Treatment of Vernal Keratoconjunctivitis
Daisuke Shii and Atsuki Fukushima
Cyclosporine A ophthalmic solutions have recently emerged
as an effective treatment for vernal keratoconjunctivitis
(VKC). Cyclosporine A is known to have multiple inhibitory
effects on T-cells, and allergic conjunctivitis models have
shown that cyclosporine A eye drops can effectively inhibit
T-cell-mediated eosinophil and neutrophil migration. Inhibitory
effects of cyclosporine A on other inflammatory cells have
also been documented. Release of histamine and cytokines from
mast cells was shown to be inhibited by cyclosporine A and
cyclosporine A eye drops were able to inhibit histamine release
and migration of inflammatory cells in allergic conjunctivitis
models mediated by mast cells. Cyclosporine A has also been
reported to induce eosinophil apoptosis and block the ability
of these cells to release eosinophil cationic protein (ECP).
In neutrophils, cyclosporine A can inhibit the release of
myeloperoxidase (MPO) and reactive oxygen species. In fibroblasts,
cyclosporine A has been demonstrated to inhibit proliferation
and pro-collagen production. In vitro, cyclosporine
A has variable effects on inflammatory cells, underscoring
the need for further experimentation to elucidate the full
range of its cytologic effects and their relation to therapeutic
outcome. The diverse inhibitory effects of cyclosporine A
on inflammatory cells clearly underlie the ability of this
drug to alleviate the symptoms of VKC.
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