Serum Paroxonase 1 level may be an Indicator and Predictor of the Severity of Androgenetic Alopecia.

BACKGROUND: Androgenetic alopecia (AGA) is a common stressful form of hair loss caused by androgen excess, genetic factors, and exposure to oxidative stress (OS) with the formation of reactive oxygen species (ROS). Paraoxonase 1 (PON1) is an enzyme synthesized in the liver bound to high-density lipoproteins to prevent lipid peroxidation. AIM: The aim of our work is to estimate serum PON1 level in patients with AGA and correlate its levels with disease severity which may help in determining if there is a role of ROS in pathogenesis of AGA. SUBJECTS AND METHODS: This study was carried out as a case and control on 40 patients with AGA (diagnosed by typical clinical and dermoscopic finding) versus 40 control subjects. Blood samples were taken from all subjects to assess serum PON1enzyme using enzyme-linked immunosorbent assay kits. RESULTS: There was a significant decrease in serum PON1 concentration level in AGA patients in comparison to controls, in addition, there was a significant decrease correlated with AGA severity (P < 0,001). The study proved that PON1 is considered highly sensitive and specific for AGA cases and a good predictive factor of AGA in healthy subjects. CONCLUSION: This is the first study done to reveal that the level of PON1 significantly decreased in AGA patients, which may give additional proof that OS has role in the pathogenesis of AGA and hence may help in the management of AGA by adding antioxidants in treatment.

TLR4 gene polymorphisms in Egyptian vitiligo patients: insights into emerging association with clinical activity, family history, and response to therapy.

BACKGROUND: Vitiligo is a common pigmentary disorder in which autoimmunity has been suggested to play an important role. Toll-like receptor (TLR) family are recognized different molecular structures expressed on immune cells and have been implicated in a number of autoimmune diseases (AIDs) such as vitiligo. The purpose of this study was to investigate the possible association between TLR4 gene polymorphisms: rs11536858, rs1927911, rs1927914 in Egyptian vitiligo patients and their clinical data, their response to therapy. Using PCR-RFLP for TLR4 gene polymorphisms (rs11536858, rs1927911, and rs1927914), both alleles and genotypes were determined after extraction of DNA in a case-control study of 100 vitiligo Egyptian patients and 100 matched age and sex controls. RESULTS: The distribution of the protective CT genotype of rs1927914 was higher in the control group. After dividing both patients and controls into 2 age groups (below 18 and above 18 years), no significant associations between the genotypes of the selected TLR4 SNPs and the demographic and clinical data of the vitiligo patients in group 1 (below 18 years) were observed. For group 2 (above 18 years), also no significant associations were found except for the association between the CC genotype of rs1927914 and psychiatric trauma, from one side, and between the CT genotype of rs1927911 and alopecia, from the other side. The association between combined genotypes and the risk of vitiligo showed either higher frequency in patients (risky), or controls (protective), and some equal frequencies (non-significant). The association between haplotypes and risk of vitiligo in patients' group revealed the highest frequency for the risky ATT and the least frequency for ATC haplotypes. In control group, the protective GCT haplotype showed the highest frequency while the GTC and GCC showed the least frequency. No significant correlations of haplotypes with clinical and demographic data of selected patients' group were observed apart from that between ACC haplotype and family history of AIDs and between ATT haplotype and remission after phototherapy. CONCLUSIONS: The significant relationship between TLR4 gene polymorphisms and vitiligo patients charcteristics clarify the role of innate immunity in pathogensis of vitiligo and its effect on the used therapies.

Inter-Observer and Intra-Observer Variations in the Assessment of Epithelial Dysplasia in Oral Lichenoid Diseases.

Oral lichen planus (OLP) and oral lichenoid lesions (OLL) can both present with histological dysplasia. Despite the presence of WHO-defined criteria for the evaluation of epithelial dysplasia, its assessment is frequently subjective (inter-observer variability). The lack of reproducibility in the evaluation of dysplasia is even more complex in the presence of a lichenoid inflammation. We evaluated dysplasia in 112 oral biopsies with lichenoid inflammation in order to study the inter-observer and the intra-observer variability.

Ki67 and CD31 Differential Expression in Cutaneous T-Cell Lymphoma and Its Mimickers: Association with Clinicopathological Criteria and Disease Advancement.

BACKGROUND: Cell proliferation and angiogenesis are important in progression of cancerous processes. Differentiating cutaneous T-cell lymphoma (CTCL) from its mimicking dermatoses and prognosticating it are challenging. AIM: This study assesses cell proliferation and angiogenesis in different CTCL subtypes using immunohistochemistry (IHC) for Ki67 and CD31 to testify their usability in differentiating CTCL from mimicking dermatoses and discriminating CTCL subtypes from each other with correlation to clinicopathological parameters and disease advancement. PATIENTS AND METHODS: IHC for Ki67 and CD31 were applied to skin biopsies from 81 patients divided into CTCL (n=59) and dermatoses (n=22) groups. Hot-spot analysis was used to score Ki67 and CD31 microvascular density (MVD) semiquantitatively. Statistical analysis was performed to compare Ki67 index and MVD between CTCL and dermatoses. CTCL subgroups were compared to each other. Ki67 index and CD31 were compared to age, gender, skin and nodal involvement, blood tumor burden and TNMB stage. RESULTS AND CONCLUSION: There were significant differences in proliferation index and MVD between dermatoses and CTCL, and between dermatoses and all CTCL subtypes with exception of Ki67 in early mycosis fungoides (MF) and CD31 in patch lesions. Increased cell proliferation and MVD were significantly associated with older age, T3 and 4 skin involvement, significant nodes (N1-3), positive blood tumor burden (B1,2) in CTCL and TNMB stage of MF. Both markers differentiated significantly late from early MF, classic MF from its variants and non-MF CTCL from total MF, but not from late MF. In conclusion, Ki67 and CD31 expression in skin biopsies using IHC reproduces the role of proliferation and angiogenesis in the differential diagnosis and prognostication of CTCL being expressed at higher levels in aggressive than indolent CTCL. Therapeutic targeting of cell proliferation and angiogenesis may improve patient's outcome in CTCL. Usability of these markers into patient's stratification should be considered in further studies.

Enhanced optical breakdown in KB cells labeled with folate-targeted silver-dendrimer composite nanodevices.

Enhanced optical breakdown of KB tumor cells folate-targeted with silver-dendrimer composite nanodevices (CNDs) is described. CNDs [(Ag(0))(25)-PAMAM_E5.(NH(2))(42)(NGly)(74)(NFA)(2.7)] were fabricated by reactive encapsulation, using a biocompatible template of dendrimer-folic acid (FA) conjugates. Preferential uptake of the folate-targeted CNDs (of various treatment concentrations and surface functionality) by KB cells was visualized with confocal microscopy and transmission electron microscopy. Intracellular laser-induced optical breakdown threshold and dynamics were detected and characterized by high-frequency ultrasonic monitoring of resulting transient bubble events. When irradiated with a near-infrared, femtosecond laser, the CND-targeted KB cells acted as well-confined activators of laser energy, enhancing nonlinear energy absorption, exhibiting a significant reduction in breakdown threshold and thus selectively promoting intracellular laser-induced optical breakdown. FROM THE CLINICAL EDITOR: This study presents a novel method to selectively destroy cancer cells by combining biochemical targeting with topical laser irradiation. A human epidermoid cancer cell line was targeted with folated silver-dendrimer composite nanodevices and the labeled cancer cells were subsequently destroyed by the microbubbles generated due the enhanced energy uptake of the silver nanoparticles from the laser irradiation, as compared to unlabeled cells.

Penetration and precision of subsurface photodisruption in porcine skin tissue with infrared femtosecond laser pulses.

The surgical precision of photodisruption with ultrafast optical pulses depends on the accurate delivery of optical energy to sites of interest. As light penetration is limited in turbid tissues, localization and precision of subsurface breakdown highly depend on the interacting effects of increased power requirements and external focusing conditions. Infrared femtosecond breakdown extent in excised porcine skin tissue was investigated using a high-frequency ultrasonic technique which sensitively detected laser-induced bubbles. Using a focused laser source, optical parameters including laser fluence and focusing numerical aperture (NA) were controlled. Decreasing NA improved penetration, while increasing NA improved precision. At lower NA, penetrations of up to 1 mm could be achieved with a single laser excitation at the cost of an expanded breakdown region which reduced precision. Even at higher NA, however, maximum penetration reduced to 400 microm and precision was still limited with extended breakdown regions for focusing depths greater than 100 microm. Glycerol was used as an index matching material, which helped reduce scattering and improved penetration by 150 to 400 microm for all NAs. Nonetheless, multiple breakdown sites and the corresponding reduction in precision were observed even with glycerol treatment.

Acoustic estimation of thermal distribution in the vicinity of femtosecond laser-induced optical breakdown.

Laser-induced optical breakdown (LIOB), or photo-disruption, can generate individual microbubbles in tissues for biomedical applications. We have previously developed a co-localized high-frequency ultrasound system to detect and characterize these laser-induced microbubbles. Because ultrasound speed varies with temperature, this system can also be used to directly estimate thermal effects in the vicinity of photodisruption. In this study, individual bubbles (sizes 60-100 microm) were created at the bottom of a water tank using a 793-nm, 100-fs Ti:Sapphire laser pulsed at 250 kHz. During and after breakdown, pulse-echoes from the tank bottom in the region surrounding a bubble were recorded with a single-element 85-MHz ultrasonic transducer, and temperature-dependent pulse-echo displacements were calculated using phase-sensitive correlation tracking. These displacements were then fit to a finite-element heat transfer model to estimate the effective thermal distribution. Estimates were calculated for laser exposure times ranging from 6.25 to 312.5 ms (1600 to 78 000 laser pulses), at 1.5 and 4 J/cm2 fluences. Results suggest a minimal temperature increase (<1 degrees C) within 100 microm of a bubble created with <1600 laser pulses at 1.5 J/cm2 fluence. This implies that LIOB can be controlled to be thermally noninvasive in the bubble vicinity.

Optical and acoustic detection of laser-generated microbubbles in single cells.

Acoustically monitored laser-induced optical breakdown (LIOB) has potential as an important tool to diagnose and treat living cells. Laser-induced intracellular microbubbles are readily detectable using high-frequency ultrasound, and LIOB can be controlled to operate within two distinct regimes. In the nondestructive regime, a single, short-lived bubble can be generated within a cell, without affecting its immediate viability. In the destructive regime, the induced photodisruption quickly can kill a targeted cell. To generate and monitor this range of bioeffects in real time, we have developed a system integrating an ultrafast laser source with optical and acoustic microscopy. Experiments were performed on monolayers of Chinese hamster ovary (CHO) cells. A 793 nm, 100 fs laser pulsed at 3.8 kHz was tightly focused within each cell to produce the photodisruption, and a 50 MHz ultrasonic transducer monitored the resultant bubble via continuous pulse-echo recordings. Photodisruption was also observed using bright field microscopy, and cell viability was assessed following laser exposure with a trypan blue assay. By controlling laser pulse fluence and exposure duration, either nondestructive or destructive LIOB could be produced. The intracellular position of the laser focus was also varied to demonstrate that cell viability was affected by the specific location of material breakdown.

Acoustic detection of controlled laser-induced microbubble creation in gelatin.

A high-frequency (85 MHz) acoustic technique is used to identify system parameters for controlled laser-induced microbubble creation inside tissue-mimicking, gelatin phantoms. Microbubbles are generated at the focus of an ultrafast 793-nm laser source and simultaneously monitored through ultrasonic pulse-echo recordings. Displayed in wavefield form, these recordings illustrate microbubble creation, and integrated backscatter plots provide specifics about microbubble characteristics and dissolution behavior. By varying laser parameters, including pulse fluence (or pulse energy flux, J/cm2), total number of pulses delivered, and the period between pulses, the size, lifetime, and dissolution dynamics of laser-induced microbubbles may be independently controlled. Pulse fluence is the main size-controlling parameter, whereas both increases in pulse fluence and pulse number can lengthen microbubble lifetime from tens to hundreds of milliseconds. In short, a microbubble of particular lifetime does not necessarily have to be of a particular size. Microbubble behavior, furthermore, is independent of pulse periods below a fluence-dependent threshold value, but it exhibits stochastic behavior if pulse repetition is too slow. These results demonstrate that laser pulse fluence, number, and period may be varied to deposit energy in a specific temporal manner, creating and stabilizing microbubbles with particular characteristics and, therefore, potential uses in sensitive acoustic detection and manipulation schemes.

Trapping cavitation bubbles with a self-focused laser beam.

We observed that laser-induced cavitation bubbles in water can be trapped in a self-focused laser beam. Both optical imaging and acoustic detection have been utilized to confirm bubble trapping. Transverse and longitudinal trapping forces were measured to be as large as 87 and 11 pN, respectively. This result is contrary to conventional wisdom, since the mechanism of trapping in conventional optical tweezers implies that a low-index particle (a bubble being the limiting case) should be antitrapped.

Lateral speckle tracking using synthetic lateral phase.

In traditional speckle tracking, lateral displacement (perpendicular to the beam direction) estimates are much less accurate than axial ones (along the beam direction). The accuracy of lateral tracking is very important whenever spatial derivatives of both axial and lateral displacements are required to give a full description of a two-dimensional (2-D) strain field. A number of methods have been proposed to improve lateral tracking by increasing the sampling rate in the lateral direction. We propose an alternate method using synthetic lateral phase (SLP). The algorithm, a direct analog of the phase zero-crossing approach used in axial displacement estimation, synthesizes the lateral phase first, then performs a zero-crossing detection on this synthetic phase to obtain lateral displacement estimates. The SLP is available by simply eliminating either the positive or negative half of the lateral spectrum of the original analytic signal. No new data need to be acquired for this procedure. This new algorithm was tested on both simulations and measurements from a cardiac phantom model. Results show that the method greatly improves the accuracy of lateral tracking, especially for low strain cases (< or =1%). The standard deviation of the estimation error of the lateral normal strain obtained with this approach has an approximate factor of 2-3 improvement for low strain cases. The conceptual and computational simplicity of this new method makes it a practical approach to improve lateral tracking for elasticity imaging.