Membrane associated transporter protein gene (SLC45A2) and the genetic basis of normal human pigmentation variation

This work is concerned with the genetic basis of normal human pigmentation variation. Specifically, the role of polymorphisms within the solute carrier family 45 member 2 (SLC45A2 or membrane associated transporter protein; MATP) gene were investigated with respect to variation in hair, skin and eye colour ― both between and within populations. SLC45A2 is an important regulator of melanin production and mutations in the gene underly the most recently identified form of oculocutaneous albinism. There is evidence to suggest that non-synonymous polymorphisms in SLC45A2 are associated with normal pigmentation variation between populations. Therefore, the underlying hypothesis of this thesis is that polymorphisms in SLC45A2 will alter the function or regulation of the protein, thereby altering the important role it plays in melanogenesis and providing a mechanism for normal pigmentation variation. In order to investigate the role that SLC45A2 polymorphisms play in human pigmentation variation, a DNA database was established which collected pigmentation phenotypic information and blood samples of more than 700 individuals. This database was used as the foundation for two association studies outlined in this thesis, the first of which involved genotyping two previously-described non-synonymous polymorphisms, p.Glu272Lys and p.Phe374Leu, in four different population groups. For both polymorphisms, allele frequencies were significantly different between population groups and the 272Lys and 374Leu alleles were strongly associated with black hair, brown eyes and olive skin colour in Caucasians. This was the first report to show that SLC45A2 polymorphisms were associated with normal human intra-population pigmentation variation. The second association study involved genotyping several SLC45A2 promoter polymorphisms to determine if they also played a role in pigmentation variation. Firstly, the transcription start site (TSS), and hence putative proximal promoter region, was identified using 5' RNA ligase mediated rapid amplification of cDNA ends (RLM-RACE). Two alternate TSSs were identified and the putative promoter region was screened for novel polymorphisms using denaturing high performance liquid chromatography (dHPLC). A novel duplication (c.–1176_–1174dupAAT) was identified along with other previously described single nucleotide polymorphisms (c.–1721C>G and c.–1169G>A). Strong linkage disequilibrium ensured that all three polymorphisms were associated with skin colour such that the –1721G, +dup and –1169A alleles were associated with olive skin in Caucasians. No linkage disequilibrium was observed between the promoter and coding region polymorphisms, suggesting independent effects. The association analyses were complemented with functional data, showing that the –1721G, +dup and –1169A alleles significantly decreased SLC45A2 transcriptional activity. Based on in silico bioinformatic analysis that showed these alleles remove a microphthalmia-associated transcription factor (MITF) binding site, and that MITF is a known regulator of SLC45A2 (Baxter and Pavan, 2002; Du and Fisher, 2002), it was postulated that SLC45A2 promoter polymorphisms could contribute to the regulation of pigmentation by altering MITF binding affinity. Further characterisation of the SLC45A2 promoter was carried out using luciferase reporter assays to determine the transcriptional activity of different regions of the promoter. Five constructs were designed of increasing length and their promoter activity evaluated. Constitutive promoter activity was observed within the first ~200 bp and promoter activity increased as the construct size increased. The functional impact of the –1721G, +dup and –1169A alleles, which removed a MITF consensus binding site, were assessed using electrophoretic mobility shift assays (EMSA) and expression analysis of genotyped melanoblast and melanocyte cell lines. EMSA results confirmed that the promoter polymorphisms affected DNA-protein binding. Interestingly, however, the protein/s involved were not MITF, or at least MITF was not the protein directly binding to the DNA. In an effort to more thoroughly characterise the functional consequences of SLC45A2 promoter polymorphisms, the mRNA expression levels of SLC45A2 and MITF were determined in melanocyte/melanoblast cell lines. Based on SLC45A2’s role in processing and trafficking TYRP1 from the trans-Golgi network to stage 2 melanosmes, the mRNA expression of TYRP1 was also investigated. Expression results suggested a coordinated expression of pigmentation genes. This thesis has substantially contributed to the field of pigmentation by showing that SLC45A2 polymorphisms not only show allele frequency differences between population groups, but also contribute to normal pigmentation variation within a Caucasian population. In addition, promoter polymorphisms have been shown to have functional consequences for SLC45A2 transcription and the expression of other pigmentation genes. Combined, the data presented in this work supports the notion that SLC45A2 is an important contributor to normal pigmentation variation and should be the target of further research to elucidate its role in determining pigmentation phenotypes. Understanding SLC45A2’s function may lead to the development of therapeutic interventions for oculocutaneous albinism and other disorders of pigmentation. It may also help in our understanding of skin cancer susceptibility and evolutionary adaptation to different UV environments, and contribute to the forensic application of pigmentation phenotype prediction.

Fusing shrinking and expanding active contour models for robust IRIS segmentation

The performance of iris recognition systems is significantly affected by the segmentation accuracy, especially in non- ideal iris images. This paper proposes an improved method to localise non-circular iris images quickly and accurately. Shrinking and expanding active contour methods are consolidated when localising inner and outer iris boundaries. First, the pupil region is roughly estimated based on histogram thresholding and morphological operations. There- after, a shrinking active contour model is used to precisely locate the inner iris boundary. Finally, the estimated inner iris boundary is used as an initial contour for an expanding active contour scheme to find the outer iris boundary. The proposed scheme is robust in finding exact the iris boundaries of non-circular and off-angle irises. In addition, occlusions of the iris images from eyelids and eyelashes are automatically excluded from the detected iris region. Experimental results on CASIA v3.0 iris databases indicate the accuracy of proposed technique.

Focus-score weighted super-resolution for uncooperative iris recognition at a distance and on the move

Uncooperative iris identification systems at a distance and on the move often suffer from poor resolution and poor focus of the captured iris images. The lack of pixel resolution and well-focused images significantly degrades the iris recognition performance. This paper proposes a new approach to incorporate the focus score into a reconstruction-based super-resolution process to generate a high resolution iris image from a low resolution and focus inconsistent video sequence of an eye. A reconstruction-based technique, which can incorporate middle and high frequency components from multiple low resolution frames into one desired super-resolved frame without introducing false high frequency components, is used. A new focus assessment approach is proposed for uncooperative iris at a distance and on the move to improve performance for variations in lighting, size and occlusion. A novel fusion scheme is then proposed to incorporate the proposed focus score into the super-resolution process. The experiments conducted on the The Multiple Biometric Grand Challenge portal database shows that our proposed approach achieves an EER of 2.1%, outperforming the existing state-of-the-art averaging signal-level fusion approach by 19.2% and the robust mean super-resolution approach by 8.7%.

Vitamin D intake needed to maintain target serum 25-Hydroxyvitamin D concentrations in participants with low sun exposure and dark skin pigmentation is substantially higher than current recommendations

Cutaneous cholecalciferol synthesis has not been considered in making recommendations for vitamin D intake. Our objective was to model the effects of sun exposure, vitamin D intake, and skin reflectance (pigmentation) on serum 25-hydroxyvitamin D (25[OH]D) in young adults with a wide range of skin reflectance and sun exposure. Four cohorts of participants (n = 72 total) were studied for 7-8 wk in the fall, winter, spring, and summer in Davis, CA [38.5° N, 121.7° W, Elev. 49 ft (15 m)]. Skin reflectance was measured using a spectrophotometer, vitamin D intake using food records, and sun exposure using polysulfone dosimeter badges. A multiple regression model (R^sup 2^ = 0.55; P < 0.0001) was developed and used to predict the serum 25(OH)D concentration for participants with low [median for African ancestry (AA)] and high [median for European ancestry (EA)] skin reflectance and with low [20th percentile, ~20 min/d, ~18% body surface area (BSA) exposed] and high (80th percentile, ~90 min/d, ~35% BSA exposed) sun exposure, assuming an intake of 200 IU/d (5 ug/d). Predicted serum 25(OH)D concentrations for AA individuals with low and high sun exposure in the winter were 24 and 42 nmol/L and in the summer were 40 and 60 nmol/L. Corresponding values for EA individuals were 35 and 60 nmol/L in the winter and in the summer were 58 and 85 nmol/L. To achieve 25(OH)D ≥75 nmol/L, we estimate that EA individuals with high sun exposure need 1300 IU/d vitamin D intake in the winter and AA individuals with low sun exposure need 2100-3100 IU/d year-round.

Robust mean super-resolution for less cooperative NIR iris recognition at a distance and on the move

Less cooperative iris identification systems at a distance and on the move often suffers from poor resolution. The lack of pixel resolution significantly degrades the iris recognition performance. Super-resolution has been considered to enhance resolution of iris images. This paper proposes a pixelwise super-resolution technique to reconstruct a high resolution iris image from a video sequence of an eye. A novel fusion approach is proposed to incorporate information details from multiple frames using robust mean. Experiments on the MBGC NIR portal database show the validity of the proposed approach in comparison with other resolution enhancement techniques.

Feature-domain super-resolution for IRIS recognition

Uncooperative iris identification systems at a distance suffer from poor resolution of the captured iris images, which significantly degrades iris recognition performance. Superresolution techniques have been employed to enhance the resolution of iris images and improve the recognition performance. However, all existing super-resolution approaches proposed for the iris biometric super-resolve pixel intensity values. This paper considers transferring super-resolution of iris images from the intensity domain to the feature domain. By directly super-resolving only the features essential for recognition, and by incorporating domain specific information from iris models, improved recognition performance compared to pixel domain super-resolution can be achieved. This is the first paper to investigate the possibility of feature domain super-resolution for iris recognition, and experiments confirm the validity of the proposed approach.

Feature-domain super-resolution framework for gabor-based face and iris recognition

The low resolution of images has been one of the major limitations in recognising humans from a distance using their biometric traits, such as face and iris. Superresolution has been employed to improve the resolution and the recognition performance simultaneously, however the majority of techniques employed operate in the pixel domain, such that the biometric feature vectors are extracted from a super-resolved input image. Feature-domain superresolution has been proposed for face and iris, and is shown to further improve recognition performance by capitalising on direct super-resolving the features which are used for recognition. However, current feature-domain superresolution approaches are limited to simple linear features such as Principal Component Analysis (PCA) and Linear Discriminant Analysis (LDA), which are not the most discriminant features for biometrics. Gabor-based features have been shown to be one of the most discriminant features for biometrics including face and iris. This paper proposes a framework to conduct super-resolution in the non-linear Gabor feature domain to further improve the recognition performance of biometric systems. Experiments have confirmed the validity of the proposed approach, demonstrating superior performance to existing linear approaches for both face and iris biometrics.

Iris based identity verification robust to sample presentation security attacks

Iris based identity verification is highly reliable but it can also be subject to attacks. Pupil dilation or constriction stimulated by the application of drugs are examples of sample presentation security attacks which can lead to higher false rejection rates. Suspects on a watch list can potentially circumvent the iris based system using such methods. This paper investigates a new approach using multiple parts of the iris (instances) and multiple iris samples in a sequential decision fusion framework that can yield robust performance. Results are presented and compared with the standard full iris based approach for a number of iris degradations. An advantage of the proposed fusion scheme is that the trade-off between detection errors can be controlled by setting parameters such as the number of instances and the number of samples used in the system. The system can then be operated to match security threat levels. It is shown that for optimal values of these parameters, the fused system also has a lower total error rate.

Feature-domain super-resolution for iris recognition

Uncooperative iris identification systems at a distance suffer from poor resolution of the acquired iris images, which significantly degrades iris recognition performance. Super-resolution techniques have been employed to enhance the resolution of iris images and improve the recognition performance. However, most existing super-resolution approaches proposed for the iris biometric super-resolve pixel intensity values, rather than the actual features used for recognition. This paper thoroughly investigates transferring super-resolution of iris images from the intensity domain to the feature domain. By directly super-resolving only the features essential for recognition, and by incorporating domain specific information from iris models, improved recognition performance compared to pixel domain super-resolution can be achieved. A framework for applying super-resolution to nonlinear features in the feature-domain is proposed. Based on this framework, a novel feature-domain super-resolution approach for the iris biometric employing 2D Gabor phase-quadrant features is proposed. The approach is shown to outperform its pixel domain counterpart, as well as other feature domain super-resolution approaches and fusion techniques.

Human identification at a distance using iris and face

This research has successfully applied super-resolution and multiple modality fusion techniques to address the major challenges of human identification at a distance using face and iris. The outcome of the research is useful for security applications.

A Human identification technique using images of the iris and wavelet transform

A new approach for recognizing the iris of the human eye is presented. Zero-crossings of the wavelet transform at various resolution levels are calculated over concentric circles on the iris, and the resulting one-dimensional (1-D) signals are compared with model features using different dissimilarity functions.