Linear and nonlinear optical parameters of biodegradable chitosan/polyvinyl alcohol/sodium montmorillonite nanocomposite films for potential optoelectronic applications.

Innovations in sophisticated optoelectronic devices have increased interest in high-refractive index polymers. Herein, we report innovative nanocomposite films with high linear and nonlinear refractive indices prepared by casting chitosan (Cs) with polyvinyl alcohol (PVA) (50:50 wt%) along with different concentrations (10-50 wt%) of sodium montmorillonite (NaMMT) nanoclay. The refractive indices in addition to other optical parameters of homopolymers and hybrid materials were investigated by UV-Vis. spectroscopy and optical modeling to assess their potential applications in optics. Besides, the structure, morphology, and thermal stability of the prepared films were investigated by a multitude of experimental techniques including X-ray diffraction (XRD), attenuated total reflectance-Fourier transform infrared spectroscopy (ATR-FTIR), scanning electron microscopy (SEM), atomic force microscopy (AFM), and thermogravimetric analysis (TGA/DTG). The ATR-FTIR, XRD, SEM, and AFM measurements confirmed the complete exfoliation of NaMMT nanolayers in the Cs/PVA matrix. The TGA/DTG revealed an increase in the thermal stability of Cs/PVA film with increasing clay content. The UV-Vis. measurements revealed a decrease in the optical energy gap (Eg) and a substantial increase in the linear (nD) and nonlinear (n2) refractive indices as clay content increased. Additionally, the nanohybrids displayed low UV transmission and reflected about 80 % of UV rays, making them excellent candidates for UV protection. For the first time, the dissipation factor (tanδ) in the UV/Vis. region has been calculated and fitted with the Drude-Lorentz model to predict the plasma frequency (ωp), resonance frequency (ω0), and electron lifetime (τ) of pristine polymers and nanocomposites.

Hydrogen peroxide pretreatment assisted phytoremediation of sodium dodecyl sulfate by Juncus acutus L.

BACKGROUND: Sodium Dodecyl Sulfate (SDS) an anionic surfactant pollutant has emerged as a serious hazard to the aquatic and terrestrial environment. Due to physical and chemical methodological difficulties for SDS removal, phytoremediation techniques are efficient alternative strategies to tackle such adversities. Juncus acutus L. (J. acutus) is a pioneer wetland species that has been recently exploited for phytoremediation purposes. To our knowledge, the role of exogenous hydrogen peroxide (H2O2), in improving the phytoextraction of SDS has not been examined yet. In this study, pretreatment foliar spray of H2O2 (15 mM) combined with two levels of SDS (50 and 100 ppm) in water culture was evaluated to remove SDS contamination and add value to the phytoremediation process. RESULTS: The outcomes revealed that J. acutus has considerable translocation and bioaccumulation abilities for SDS and can be utilized as an appropriate hyperaccumulator in SDS-contaminated sites. However, the involvement of H2O2 extended phytoremediation capacity and successive removal of SDS. H2O2 significantly assisted in increasing SDS remediation via more accumulation in J. acutus tissues by 29.9 and 112.4% and decreasing SDS concentration in culture media by 33.3 and 27.3% at 50 and 100 ppm SDS, respectively. Bioaccumulation factor (BCF) increased by 13.8 and 13.2%, while translocation factor (TCF) positively maximized by 82.4 and 76.2% by H2O2 application at 50 and 100 ppm SDS, respectively. H2O2 pretreatment could drive the decline in biochemical attributes in SDS-affected plants by modulating stress tolerance indices, pigments, water relations, proline content, enzymatic activities, and further, reduced oxidative stress in terms of electrolyte leakage, cellular H2O2, malondialdehyde (MDA) accumulation. CONCLUSIONS: H2O2 could play a potential role in maximizing phytoremediation capacity of SDS by J. acutus in polluted sites.

H-Shaped Copolymer of Polyethylene and Poly(ethylene oxide) under Severe Confinement: Phase State and Dynamics.

The self-assembly and the dynamics of an H-shaped copolymer composed of a polyethylene midblock and four poly(ethylene oxide) arms (PE-b-4PEO) are investigated in the bulk and under severe confinement into nanometer-spaced LAPONITE clay particles by means of small- and wide-angle X-ray diffraction (SAXS, WAXS), differential scanning calorimetry (DSC), polarizing optical microscopy (POM), rheology, and dielectric spectroscopy (DS). Because of the H-shaped architecture, the PE midblock is topologically frustrated and thus unable to crystallize. The superstructure formation in the bulk is dictated solely by the PEO arms as inferred by the crystallization/melting temperature relative to the PEO homopolymer. Confinement produced remarkable changes in the interlayer distance and PEO crystallinity but left the local segmental dynamics unaltered. To reconcile all structural, thermodynamic, and dynamic effects, a novel morphological picture is proposed with interest in emulsions. Key parameters that stabilize the final morphology are the severe chain confinement with the associated entropy loss and the presence of interactions (hydrophobic/hydrophilic) between the LAPONITE and the PEO/PE blocks.

pH and Salt Response of Mixed Brushes Made of Oppositely Charged Polyelectrolytes Studied by in Situ AFM Force Measurements and Imaging.

The response of mixed brushes made of poly(acrylic acid) and poly(2-vinyl pyridine) with a mixing ratio of about 60:40 was studied using atomic force microscopy (AFM) force measurements with colloidal probes and AFM imaging with a sharp tip in the pH range between 2.5 and 8 and at varying KCl concentrations up to 1 M. It was found that under all conditions a dense polyelectrolyte complex layer coexists with excess polyelectrolyte chains in varying swelling states depending on pH and salt concentration. The mixed brush thus combines typical features of polyelectrolyte brushes and complexes. So, the increase of the salt concentration not only led to a transition from osmotic to salted brush regime but also to salt-induced softening or partial decomposition of the complex layer. Attractive forces at high salt concentrations indicated the presence of P2VP chains in the swollen layer even at high pH values.

Pharyngeal stimulus-induced reflexes are impaired in infants with perinatal asphyxia: Does maturation modify?

BACKGROUND: Development of pharyngo-esophageal protective reflexes among infants with hypoxic ischemic encephalopathy (HIE) is unclear. Our aim was to distinguish these reflexes from controls and examine the maturational changes in HIE infants. METHODS: We evaluated 14 HIE infants (seven males) at 41.4±0.6 (HIE Time-1) and 46.5±0.6 (HIE Time-2) weeks postmenstrual age (PMA). Seven controls (three males) were evaluated at 43.5±1.3 weeks PMA. Graded pharyngeal stimulation with liquids (0.1, 0.3, 0.5 mL in triplicate) concurrent with high-resolution manometry was used to analyze sensory-motor components of pharyngeal reflexive swallowing (PRS), upper esophageal sphincter (UES), contractile reflex (PUCR), and esophageal body characteristics. Linear mixed and generalized estimating equation models were used for comparison among groups. KEY RESULTS: Compared to controls, HIE infants (Time-1 and Time-2) exhibited decreased number of pharyngeal peaks and latency to terminal swallow. HIE Time-1 infants showed increased UES resting tone and distal latency, compared to controls and HIE Time-2. Contractile vigor was increasingly abnormal during maturation, compared to healthy controls. Threshold volumes and frequency distribution of primary responses (PRS: PUCR: None) were not significant among all groups. CONCLUSIONS & INFERENCES: Compared to controls, HIE infants display significant hypertonicity of skeletal muscle components, impairment of pharyngeal provocation-induced reflexes and smooth muscle contractile vigor, reflecting poor propagation with maturation. These mechanisms may be responsible for inadequate clearance of secretions, ascending refluxate, and oropharyngeal bolus in HIE infants.

Swelling and Surface Interactions of End-Grafted Poly(2-vinylpyridine) Layers in Acidic Solution: Influence of Grafting Density and Salt Concentration.

In previous studies, the authors found that end-grafted layers of the weak polybase poly(2-vinylpyridine) (P2VP) in aqueous solutions do not only swell and collapse if the pH value and salt concentration are varied but also exhibit a pH- and salinity-dependent adhesion to microsized silica spheres. For a better understanding of these effects, in situ force measurements using the AFM colloidal probe technique were applied to end-grafted P2VP layers of different grafting densities in NaCl solutions at pH 2.5. Although a mushroom-to-brush transition could be seen in the dry state, the layers were in the brush regime in aqueous solutions at all NaCl concentrations and grafting densities. We observed an increase of the brush height with increasing grafting density and a salinity-dependent collapse and reswelling of the brushes. The adhesion between the P2VP layer and a silica sphere depended on both grafting density and salinity. At low salt concentrations, the adhesion reached its highest value at the intermediate grafting density and disappeared with denser brushes. Maximum adhesion was obtained for high NaCl concentrations and the lowest grafting density. From a detailed analysis of the experiments, we gained insight into chain stretching and density profiles under complex ionic conditions and into the mechanism of adhesion of polyelectrolytes to solid surfaces.

Structure and Dynamics of Asymmetric Poly(styrene-b-1,4-isoprene) Diblock Copolymer under 1D and 2D Nanoconfinement.

The impact of 1- and 2-dimensional (2D) confinement on the structure and dynamics of poly(styrene-b-1,4-isoprene) P(S-b-I) diblock copolymer is investigated by a combination of Scanning Electron Microscopy (SEM), Atomic Force Microscopy (AFM), Grazing-Incidence Small-Angle X-ray Scattering (GISAXS), and Broadband Dielectric Spectroscopy (BDS). 1D confinement is achieved by spin coating the P(S-b-I) to form nanometric thin films on silicon substrates, while in the 2D confinement, the copolymer is infiltrated into cylindrical anodized aluminum oxide (AAO) nanopores. After dissolving the AAO matrix having mean pore diameter of 150 nm, the SEM images of the exposed P(S-b-I) show straight nanorods. For the thin films, GISAXS and AFM reveal hexagonally packed cylinders of PS in a PI matrix. Three dielectrically active relaxation modes assigned to the two segmental modes of the styrene and isoprene blocks and the normal mode of the latter are studied selectively by BDS. The dynamic glass transition, related to the segmental modes of the styrene and isoprene blocks, is independent of the dimensionality and the finite sizes (down to 18 nm) of confinement, but the normal mode is influenced by both factors with 2D geometrical constraints exerting greater impact. This reflects the considerable difference in the length scales on which the two kinds of fluctuations take place.

Helminth infections: therapeutic potential in autoimmune disorders.

Knowledge of immunity enables us to predict that the reactions set in response to infection with helminth would prevent concomitant disease driven by an opposing spectrum of immune events. In another way, the immune response generated to combat the helminth infection could counteract the immunopathological reactions that drive autoimmune diseases. Rodent model systems recapitulate many aspects of human autoimmune diseases and have been enormously useful in defining mechanisms of immunopathology after infection. From this theoretical perspective, many researchers have proved that infection with a variety of helminth can ameliorate disease in murine model systems. Thus, helminth-evoked Th2 events were shown to improve disorders in which Th1 events predominated. This raised the question, 'Can this information be translated into therapies for autoimmune diseases in humans via actual infection, cell delivery or drug intervention?' In this review, we will present some experimental trails to treat autoimmune disorders through establishment of some parasitic infections.

Interaction forces between microsized silica particles and weak polyelectrolyte brushes at varying pH and salt concentration.

The AFM colloidal probe technique was used to measure the interaction between microsized silica spheres and annealed polyelectrolyte brushes made of poly(acrylic acid) (PAA) and poly(2-vinyl pyridine) (P2VP) in KCl solutions of various pH values and salt concentrations. The interaction energy showed a distance dependence that was affected strongly by the swelling and the electric properties of the brushes. Between PAA brushes and silica particles, a repulsive interaction has been observed for all pH values and salt concentrations reflecting the swelling of the brush with varying pH value and the transition from osmotic to salted brush regime with increasing KCl concentration. Force measurements between P2VP brushes and silica particles revealed a much more complex behavior: a steric repulsion by the swollen brush at low pH values, a complex interplay of attractive and repulsive forces at intermediate pH values and a short-ranged attraction between the collapsed brush and the silica particle at basic pH values and high salt concentrations. The results are interpreted in comparison with the Alexander de Gennes model and zeta potential and ellipsometric measurements.

Slow kinetics of phase transformation in a dipole-functionalized discotic liquid crystal.

The kinetics of phase transformation from the liquid crystalline mesophase to the crystalline phase has been studied in a model dipole-functionalized hexabenzocoronene derivative by x-ray diffraction, solid state NMR recoupling techniques, and dielectric spectroscopy. These probes revealed long-lived metastability and an intermediate state involving a change in the unit cell prior to crystallization. The barrier properties were dominant and amount to 1 and 2.5 eV, respectively, for the unit cell transformation and crystallization processes. These effects should be considered in the design of discotic liquid crystals for electronic applications.

Forces between blank surfaces as measured by the colloidal probe technique and by optical tweezers--a comparison.

The well-established atomic force microscopy (AFM)-based colloidal probe technique (CPT) and optical tweezers (OT) are combined to measure the interaction forces between blank SiO(2) surfaces in aqueous ionic solutions (CaCl(2)) of varying concentration at pH 7. Spherical colloids (SiO(2), diameter approximately 4.63 +/- 0.05 microm) taken out of the same batch are used by both methods. In the case of CPT, a single colloid is glued to a cantilever, and the interaction forces with a plain SiO(2) surface are determined in dependence on the concentration of the surrounding medium. For the OT studies, two colloids (one fixed to a micropipet by capillary action, the other held with the optical trap) are approached to each other in nanometer steps, and the resulting forces are measured for the same media as in the CPT experiment. Both techniques fit well to each other and enable one to cover interaction energies ranging from 10(-5) to 1 mN/m. The experimental data are well described by the Derjaguin-Landau-Verwey-Overbeek (DLVO) theory revealing that the effective surface charge density changes slightly with concentration.

Self-assembly, molecular dynamics, and kinetics of structure formation in dipole-functionalized discotic liquid crystals.

The self-assembly, the molecular dynamics, and the kinetics of structure formation are studied in dipole-functionalized hexabenzocoronene (HBC) derivatives. Dipole substitution destabilizes the columnar crystalline phase except for the dimethoxy- and monoethynyl-substituted HBCs that undergo a reversible transformation to the crystalline phase. The disk dynamics are studied by dielectric spectroscopy and site-specific NMR techniques that provide both the time-scale and geometry of motion. Application of pressure results in the thermodynamic phase diagram that shows increasing stability of the crystalline phase at elevated pressures. Long-lived metastability was found during the transformation between the two phases. These results suggest new thermodynamic and kinetic pathways that favor the phase with the highest charge carrier mobility.

The ubiquitin-proteasome system regulates p53-mediated transcription at p21waf1 promoter.

The ubiquitin (Ub)-proteasome system (UPS) promotes the proteasomal degradation of target proteins by decorating them with Ub labels. Emerging evidence indicates a role of UPS in regulating gene transcription. In this study, we provided evidence for the involvement of UPS in the transcriptional activation function of tumor suppressor p53. We showed that both ubiquitylation and proteasomal functions are required for efficient transcription mediated by p53. Disruption of transcription by actinomycin D, 5,6-dichloro-1-beta-D-ribofuranosyl-benzimadazole or alpha-amanitin leads to accumulation of cellular p53 protein. Proteasome inhibition by MG132 increases the occupancy of p53 protein at p53-responsive p21(waf1) promoter. In addition, the Sug-1 component of 19S proteasome physically interacts with p53 in vitro and in vivo. Moreover, in response to ultraviolet-induced DNA damage, both the 19S proteasomal components, Sug1 and S1, are recruited to p21(waf1) promoter region in a kinetic pattern similar to that of p53. These results suggested that UPS positively regulates p53-mediated transcription at p21(waf1) promoter.

Self-assembly and molecular dynamics of oligoindenofluorenes.

The optical properties, self-assembly mechanism, thermal properties, and the associated molecular dynamics of a series of stable blue-emitting oligoindenofluorenes up to the polymer were investigated by photoluminescence, wide-angle X-ray scattering, polarizing optical microscopy, differential scanning calorimetry, and dielectric spectroscopy. Oligomers of indenofluorenes possess high structural order and form smectic mesophases, as opposed to the nematic mesophases formed in the corresponding oligofluorenes. The alpha process associated with the liquid-to-glass transition displays an asymmetric broadening of relaxation times (non-Debye process), with a non-Arrhenius temperature dependence of relaxation times and a strong molecular weight dependence. The low dielectric loss and low ionic conductivity revealed the absence of ketone structural defects, which is consistent with the observed stable blue emission.

Green tea extract attenuates cyclosporine A-induced oxidative stress in rats.

Cyclosporine A (CsA) nephrotoxicity underweighs the therapeutic benefits of such a powerful immunosuppressant. Whether oxidative stress plays a role in such toxicity is not well delineated. We investigated the potential of green tea extract (GTE) to attenuate CsA-induced renal dysfunction in rats. Three main groups of Sprague-Dawley rats were used: CsA, GTE, and GTE plus CsA-receiving animals. Corresponding control groups were also used. CsA was administered in a dose of 20mg kg(-1) day(-1), i.p., for 21 days. In the GTE/CsA groups, the rats received different concentrations of GTE (0.5, 1.0 and 1.5%), as their sole source of drinking water, 4 days before and 21 days concurrently with CsA. The GTE group was treated with 1.5% concentration of GTE only for 25 days. A concomitant administration of GTE, to CsA receiving rats, markedly prevented the generation of thiobarbituric acid-reacting substances (TBARS) and significantly attenuated CsA-induced renal dysfunction as assessed by estimating serum creatinine, blood urea nitrogen, uric acid and urinary excretion of glucose. A considerable improvement in terms of reduced glutathione content and activity of antioxidant enzymes in the kidney homogenate of the GTE/CsA-receiving rats was observed. The activity of lysosomal enzymes, N-acetyl-beta-glucosaminidase, beta-glucuronidase and acid phosphatase was significantly inhibited following GTE co-administration. Our data prove the role of oxidative stress in the pathogenesis of CsA-induced kidney dysfunction. Supplementation of GTE could be useful in reducing CsA nephrotoxicity in rats. However, clinical studies are warranted to investigate such an effect in human subjects.

Influence of p-coumaric acid on doxorubicin-induced oxidative stress in rat's heart.

The therapeutic value of doxorubicin (DOX) as anticancer antibiotic is limited by its cardiotoxicity. The implication of natural phenolic acids in the prevention of many pathologic diseases has been reported. Herein, the ability of p-coumaric (PC) acid, a member of phenolic acids, to protect rat's heart against DOX-induced oxidative stress was investigated. Three main groups of albino rats were used; DOX, PC, and PC plus DOX-receiving animals. Corresponding control animals were also used. DOX was administered i.p. in a single dose of 15mgkg(-1). PC alone, in a dose of 100mgkg(-1), was orally administered for five consecutive days. In PC/DOX group, rats received PC 5 days prior to DOX. DOX-induced high serum levels of lactic dehydrogenase (LDH) and creatine phosphokinase (CPK), were reduced significantly by PC administration, compared to DOX-receiving rats. Pretreatment with PC ameliorated the cardiac content of glutathione (GSH), and superoxide dismutase (SOD) & catalase (CAT) activities, compared to DOX-receiving rats. On the other hand, accumulation of cardiac content of MDA significantly decreased following PC pretreatment, compared to DOX-treated rats. The data presented here indicate that PC protects rats hearts against DOX-induced oxidative stress in the heart. It may be worthy to consider the usefulness of PC as adjuvant therapy in cancer management.

Modulation of transcriptional activity of p53 by ultraviolet radiation: linkage between p53 pathway and DNA repair through damage recognition.

The increase in the p53 activity in response to DNA damage is thought to be one of the important mechanisms by which p53 contributes to transcriptional activation of p21(wafl), mdm2, and other downstream regulatory genes. To investigate the p53 response to ultraviolet (UV) type of DNA damage, p53 protein level, its transcriptional activity and in vivo ubiquitination were compared in repair-proficient normal human fibroblasts (NHFs) and repair-deficient xeroderma pigmentosum (XP) group A and group C (XP-C) fibroblasts subsequent to irradiation with UV light. Accumulation of p53 protein level was observed with increasing UV doses in all the cell lines; however, discordance between p53 and p21(waf1) and mdm2 levels was observed in NHF and XP-A cells. Induction of p21(waf1) and mdm2 was inhibited by UV irradiation, requiring higher doses in NHF and lower doses in XP-A cells. However, inhibition of p21(waf1) and mdm2 induction was not observed in XP-C cells. Ubiquitin-p53 conjugates could be detected in irradiated or unirradiated NHF and XP-A cells but not in XP-C cells irradiated with 30 and 50 J/m(2) UV light. Using a p53 reporter assay, p53 transcriptional activities were found to be induced by 10 J/m(2) UV exposure and dramatically inhibited with increasing UV doses in NHF cells. Compared with repair-proficient NHF cells, UV inhibition of p53 transcriptional activity was relatively more sensitive in XP-A cells but resistant in XP-C cells. These results indicate that DNA damage by UV, in addition to inducing p53, acts as a trigger for inhibition of p53 transcriptional activity. Overall, recognition of DNA damage links both p53 induction and p53 degradation to DNA repair mechanisms.

Enhanced sensitivity to anti-benzo(a)pyrene-diol-epoxide DNA damage correlates with decreased global genomic repair attributable to abrogated p53 function in human cells.

DNA damage from exposure to environmental chemical carcinogens and failure of repair systems to eliminate these lesions from the genome are considered as the crucial initial steps in the development of various human malignancies. Many cellular proteins are known to play vital roles to overcome the effects of DNA damage. Among such proteins, p53 is known to respond to DNA damage by accumulating in the nucleus and inhibiting cell cycle progression to facilitate DNA repair and the maintenance of genomic stability. In this study, we have investigated the role of p53 protein in modulating nucleotide excision repair of anti-benzo-(a)pyrene-diol-epoxide (BPDE)-DNA adducts and related effects using human fibroblasts with normal (p53-WT) and altered p53 protein (p53Mut and p53-Null). Interestingly, irrespective of the presence or absence of p53, the anti-BPDE dose-dependent p21 protein induction response was qualitatively comparable in all of the three cell lines. However, cells with defective p53 function were deficient for the removal of anti-BPDE-DNA adducts from the overall genome compared to cells with wild-type p53 activity. Strand-specific repair analysis within the individual strands of the p53 gene revealed decreased repair of adducts from the nontranscribed strand in p53-Mut and p53-Null cells. However, the repair of the transcribed strand appeared to be identical in all of the three cell lines. Furthermore, p53-Mut and p53-Null cells were more sensitive than p53-WT cells and displayed increased levels of anti-BPDE-induced apoptosis. Thus, wild-type p53 is required for the efficient global genomic repair of anti-BPDE-induced DNA adducts from the overall genome, but not for transcription-coupled repair of actively transcribed genes. These findings indicate that inefficient DNA repair of potentially cytotoxic and mutagenic lesions from the nontranscribed strand due to the loss of p53, but not the loss of p21, function might be responsible for enhanced cytotoxicity and apoptosis in human cells upon DNA damage.

p53-degradation by HPV-16 E6 preferentially affects the removal of cyclobutane pyrimidine dimers from non-transcribed strand and sensitizes mammary epithelial cells to UV-irradiation.

Nucleotide excision repair (NER), the most versatile and ubiquitous mechanism for DNA repair, operates to remove many types of DNA base lesions. We have studied the role of p53 function in modulating the repair of DNA damage following UV irradiation in normal and p53-compromised human mammary epithelial cells (HMEC). The effect of UV-induced DNA damage on cellular cytotoxicity and apoptosis was determined in conjunction with global, gene- and strand-specific repair. Cytotoxicity studies, using clonogenic survival and MTT assays, showed that HPV-16 E6-expressing HMEC were more UV sensitive than p53-WT cell lines. High apoptotic index obtained with p53-compromised cells was in conformity to both the low clonogenic survival and the low cellular viability. No discernible differences in the formation of initial UV-induced cyclobutane pyrimidine dimers (CPD) were observed in the cell lines of varying p53 functional status. However, the extent and the rate of damage removal from genome overall were highest for p53-WT cells. Further examination of strand-specific repair in the p53 gene revealed that the removal of CPD in the non-transcribed strand (NTS) was slower in p53-compromised cells compared to the normal p53-WT cell lines. These results suggest that loss of p53 function, in the absence of other genetic alterations, decreased both overall amount of CPD repaired and their removal rate from the genome. Additionally, normal function of p53 is required for the repair of the NTS, but not of the transcribed strand (TS) in genomic DNA in human epithelial cells. Thus, failure of quantitative removal of CPD by global genomic repair (GGR), due to loss of p53 function, causes the enhanced UV sensitivity and increased damage-induced apoptosis via a p53-independent pathway. Nevertheless, recovery of cells from UV damage requires normal p53 function and efficient GGR.

Decreased DNA repair efficiency by loss or disruption of p53 function preferentially affects removal of cyclobutane pyrimidine dimers from non-transcribed strand and slow repair sites in transcribed strand.

The tumor suppressor protein p53 plays a central role in modulating the cellular responses to DNA damage. Several recent studies, undertaken with the whole genomic DNA or full-length gene segments, have shown that p53 is involved in nucleotide excision repair and it selectively influences the adduct removal from the non-transcribed strand in the genome. In this study, we have analyzed the damage induction at nucleotide resolution by ligase-mediated polymerase chain reaction and compared the repair of ultraviolet radiation-induced cyclobutane pyrimidine dimers within exon 8 of p53 gene in normal and Li-Fraumeni syndrome fibroblasts as well as in normal and human papillomavirus 16 E6 and E7 protein-expressing human mammary epithelial cells. The results demonstrate that (i) loss or disruption of p53 function decreases efficiency of DNA repair, by preferentially affecting the repair of non-transcribed strand and of intrinsically slow repair sites in transcribed strand; (ii) mutant p53 protein affects DNA repair, at least of non-transcribed strand, in a dominant negative manner; and (iii) pRb does not have an effect on the repair of DNA damage within transcribed or non-transcribed strand. The overall data suggest that p53 could regulate excision repair or related events through direct protein-protein interaction.