Directed evolution and predictive modelling of galactose oxidase towards bulky benzylic and unactivated secondary alcohols

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Global Retinoblastoma Presentation and Analysis by National Income Level

Importance: Early diagnosis of retinoblastoma, the most common intraocular cancer, can save both a child's life and vision. However, anecdotal evidence suggests that many children across the world are diagnosed late. To our knowledge, the clinical presentation of retinoblastoma has never been assessed on a global scale. Objectives: To report the retinoblastoma stage at diagnosis in patients across the world during a single year, to investigate associations between clinical variables and national income level, and to investigate risk factors for advanced disease at diagnosis. Design, Setting, and Participants: A total of 278 retinoblastoma treatment centers were recruited from June 2017 through December 2018 to participate in a cross-sectional analysis of treatment-naive patients with retinoblastoma who were diagnosed in 2017. Main Outcomes and Measures: Age at presentation, proportion of familial history of retinoblastoma, and tumor stage and metastasis. Results: The cohort included 4351 new patients from 153 countries; the median age at diagnosis was 30.5 (interquartile range, 18.3-45.9) months, and 1976 patients (45.4) were female. Most patients (n = 3685 84.7%) were from low-and middle-income countries (LMICs). Globally, the most common indication for referral was leukocoria (n = 2638 62.8%), followed by strabismus (n = 429 10.2%) and proptosis (n = 309 7.4%). Patients from high-income countries (HICs) were diagnosed at a median age of 14.1 months, with 656 of 666 (98.5%) patients having intraocular retinoblastoma and 2 (0.3%) having metastasis. Patients from low-income countries were diagnosed at a median age of 30.5 months, with 256 of 521 (49.1%) having extraocular retinoblastoma and 94 of 498 (18.9%) having metastasis. Lower national income level was associated with older presentation age, higher proportion of locally advanced disease and distant metastasis, and smaller proportion of familial history of retinoblastoma. Advanced disease at diagnosis was more common in LMICs even after adjusting for age (odds ratio for low-income countries vs upper-middle-income countries and HICs, 17.92 95% CI, 12.94-24.80, and for lower-middle-income countries vs upper-middle-income countries and HICs, 5.74 95% CI, 4.30-7.68). Conclusions and Relevance: This study is estimated to have included more than half of all new retinoblastoma cases worldwide in 2017. Children from LMICs, where the main global retinoblastoma burden lies, presented at an older age with more advanced disease and demonstrated a smaller proportion of familial history of retinoblastoma, likely because many do not reach a childbearing age. Given that retinoblastoma is curable, these data are concerning and mandate intervention at national and international levels. Further studies are needed to investigate factors, other than age at presentation, that may be associated with advanced disease in LMICs. © 2020 American Medical Association. All rights reserved

Key issues of a software focused supply chain

10.1109/INDIN.2006.2756552006 IEEE International Conference on Industrial Informatics, INDIN'06747-75

Fingerprinting genomic instability in oral submucous fibrosis

Background: Oral submucous fibrosis (OSF) is a high-risk pre-cancerous condition where 7-13% of these patients develop head and neck squamous cell carcinoma (HNSCC). To date there is no cancer predictive markers for OSF patients. Genomic instability hallmarks early genetic events during malignant transformation causing loss of heterozygosity (LOH) and chromosomal copy number abnormality. However, to date there is no study on genomic instability in OSF. Although this condition is known as a high-risk pre-cancerous condition, there is no data regarding the genomic status of this disease in terms of genetic susceptibility to malignant transformation.\ud \ud Methods: In this study, we investigated the existence of genetic signatures for carcinogenesis in OSF. We employed the high-resolution genome-wide Affymetrix Mapping single nucleotide polymorphism microarray technique to 'fingerprint' global genomic instability in the form of LOH in 15 patient-matched OSF-blood genomic DNA samples.\ud \ud Results: This rapid high-resolution mapping technique has revealed for the first time that a small number of discrete hot-spot LOH loci appeared in 47-53% of the OSF tissues studied. Many of these LOH loci were previously identified regions of genomic instability associated with carcinogenesis of the HNSCC.\ud \ud Conclusion: To our knowledge, this is the first evidence that genomic instability in the form of LOH is present in OSF. We hypothesize that the genomic instability detected in OSF may play an important role in malignant transformation. Further functional association studies on these putative genes may reveal potential predictive oral cancer markers for OSF patients

Upregulation of HIF-1 alpha in malignant transformation of oral submucous fibrosis

Background: Oral submucous fibrosis (OSF) is a precancerous condition showing extensive fibrosis of the submucosa and affects most parts of the oral cavity, including pharynx and upper third of the oesophagus. The molecules involved in the biological pathways of the fibrotic process appeared to be either down- or upregulated at different stages of the disease. Despite the precancerous nature, malignant transformation of the epithelium in the background of fibrosis has not been studied in detail. HIF-1 alpha is a known transcription factor that is induced by hypoxia.\ud \ud Aims: To test the hypothesis that hypoxia plays a role in malignant transformation and progression of OSF.\ud \ud Materials and Methods: We used both formalin-fixed and frozen samples of OSF and normal mucosa to investigate the relationship between HIF-1 alpha and epithelial dysplasia using immunohistochemistry and RT-PCR.\ud \ud Conclusions: Our data indicate that HIF-1 alpha is upregulated at both protein and mRNA levels in OSF and the correlation with epithelial dysplasia is statistically significant (P < 0.001). We propose that HIF-1 alpha may play a role in malignant transformation of OSF. Further, over-expression of HIF-1alpha may contribute to the progression of fibrosis. It may be possible to use HIF-1 alpha as a marker for malignant transformation of OSF

Directed Evolution and Predictive Modelling of Galactose Oxidase towards Bulky Benzylic and Unactivated Secondary Alcohols

The growth of industrial biocatalysis for sustainable chemical manufacturing has been limited by the narrow range of chemistries associated with natural enzymes and experiment-intensive regimes of enzyme engineering. Consequently, there has been deep interest to expand enzyme substrate scopes for broader synthetic utility, and to streamline the enzyme engineering process. In the field of alcohol oxidation, galactose oxidase (GOase) is one of the most established enzymes capable of this important chemical transformation under benign conditions. However, the applicability of GOase towards more complex molecules such as those frequently found in the pharmaceutical, or agrochemical industries remains restricted. Here, by employing a combined approach of directed evolution and predictive modelling, we have identified new GOases with significantly expanded substrate specificity toward both bulky benzylic and unactivated secondary alcohols, showing activity enhancements of up to 2,400-fold compared to the reported benchmark M3-5 mutant. Beneficial mutations conveying relaxed substrate enantioselectivity biases (R/S ratios down to 1.05) and higher thermostabilities (up to 20-fold versus benchmark) have also been identified. We have developed predictive models based on computational tools YASARA, FoldX, SCWRL and Glide that are well correlated with features related to enzyme structure, selectivity, protein stability and catalytic activity. The generated enzyme activity models based on Glide-MM/GBSA (r = -0.85) and YASARA (r = -0.89) have successfully predicted the activity trend of a family of related substrates based on the 1-phenyl-1-alkyl alcohol scaffold with varying alkyl chain lengths. It is envisioned that these in silico models can serve as valuable tools to explore desirable enzyme characteristics, establish enzyme substrate scopes, and accelerate biocatalyst development, thus promoting it as a competitive and competent solution for sustainable chemical manufacturing

MC3R gene polymorphisms are associated with early childhood adiposity gain and infant appetite in an Asian population

10.1111/ijpo.12086Pediatric Obesity116450-458GUSTO (Growing up towards Healthy Outcomes