ABSTRACT
Precise identification of causative variants from whole-genome sequencing data, including both coding and noncoding variants, is challenging. The Critical Assessment of Genome Interpretation 5 SickKids clinical genome challenge provided an opportunity to assess our ability to extract such information. Participants in the challenge were required to match each of the 24 whole-genome sequences to the correct phenotypic profile and to identify the disease class of each genome. These are all rare disease cases that have resisted genetic diagnosis in a state-of-the-art pipeline. The patients have a range of eye, neurological, and connective-tissue disorders. We used a gene-centric approach to address this problem, assigning each gene a multiphenotype-matching score. Mutations in the top-scoring genes for each phenotype profile were ranked on a 6-point scale of pathogenicity probability, resulting in an approximately equal number of top-ranked coding and noncoding candidate variants overall. We were able to assign the correct disease class for 12 cases and the correct genome to a clinical profile for five cases. The challenge assessor found genes in three of these five cases as likely appropriate. In the postsubmission phase, after careful screening of the genes in the correct genome, we identified additional potential diagnostic variants, a high proportion of which are noncoding.
Subject(s)
Genetic Association Studies/methods , Genetic Diseases, Inborn/diagnosis , Genetic Diseases, Inborn/genetics , Genetic Predisposition to Disease , Genome, Human , Genomics/methods , Rare Diseases , Algorithms , Alleles , Genetic Variation , Genome-Wide Association Study/methods , Genotype , Humans , Models, Theoretical , Phenotype , Whole Genome Sequencing , WorkflowABSTRACT
Thermodynamic stability is a fundamental property shared by all proteins. Changes in stability due to mutation are a widespread molecular mechanism in genetic diseases. Methods for the prediction of mutation-induced stability change have typically been developed and evaluated on incomplete and/or biased data sets. As part of the Critical Assessment of Genome Interpretation, we explored the utility of high-throughput variant stability profiling (VSP) assay data as an alternative for the assessment of computational methods and evaluated state-of-the-art predictors against over 7,000 nonsynonymous variants from two proteins. We found that predictions were modestly correlated with actual experimental values. Predictors fared better when evaluated as classifiers of extreme stability effects. While different methods emerging as top performers depending on the metric, it is nontrivial to draw conclusions on their adoption or improvement. Our analyses revealed that only 16% of all variants in VSP assays could be confidently defined as stability-affecting. Furthermore, it is unclear as to what extent VSP abundance scores were reasonable proxies for the stability-related quantities that participating methods were designed to predict. Overall, our observations underscore the need for clearly defined objectives when developing and using both computational and experimental methods in the context of measuring variant impact.
Subject(s)
Computational Biology/methods , Methyltransferases/chemistry , Mutation , PTEN Phosphohydrolase/chemistry , High-Throughput Nucleotide Sequencing , Humans , Methyltransferases/genetics , PTEN Phosphohydrolase/genetics , Protein StabilityABSTRACT
The Critical Assessment of Genome Interpretation-5 intellectual disability challenge asked to use computational methods to predict patient clinical phenotypes and the causal variant(s) based on an analysis of their gene panel sequence data. Sequence data for 74 genes associated with intellectual disability (ID) and/or autism spectrum disorders (ASD) from a cohort of 150 patients with a range of neurodevelopmental manifestations (i.e. ID, autism, epilepsy, microcephaly, macrocephaly, hypotonia, ataxia) have been made available for this challenge. For each patient, predictors had to report the causative variants and which of the seven phenotypes were present. Since neurodevelopmental disorders are characterized by strong comorbidity, tested individuals often present more than one pathological condition. Considering the overall clinical manifestation of each patient, the correct phenotype has been predicted by at least one group for 93 individuals (62%). ID and ASD were the best predicted among the seven phenotypic traits. Also, causative or potentially pathogenic variants were predicted correctly by at least one group. However, the prediction of the correct causative variant seems to be insufficient to predict the correct phenotype. In some cases, the correct prediction has been supported by rare or common variants in genes different from the causative one.
Subject(s)
Autism Spectrum Disorder/genetics , Computational Biology/methods , Intellectual Disability/genetics , Sequence Analysis, DNA/methods , Female , Genetic Predisposition to Disease , Humans , Male , Phenotype , Quantitative Trait LociABSTRACT
Whole-genome sequencing (WGS) holds great potential as a diagnostic test. However, the majority of patients currently undergoing WGS lack a molecular diagnosis, largely due to the vast number of undiscovered disease genes and our inability to assess the pathogenicity of most genomic variants. The CAGI SickKids challenges attempted to address this knowledge gap by assessing state-of-the-art methods for clinical phenotype prediction from genomes. CAGI4 and CAGI5 participants were provided with WGS data and clinical descriptions of 25 and 24 undiagnosed patients from the SickKids Genome Clinic Project, respectively. Predictors were asked to identify primary and secondary causal variants. In addition, for CAGI5, groups had to match each genome to one of three disorder categories (neurologic, ophthalmologic, and connective), and separately to each patient. The performance of matching genomes to categories was no better than random but two groups performed significantly better than chance in matching genomes to patients. Two of the ten variants proposed by two groups in CAGI4 were deemed to be diagnostic, and several proposed pathogenic variants in CAGI5 are good candidates for phenotype expansion. We discuss implications for improving in silico assessment of genomic variants and identifying new disease genes.
Subject(s)
Computational Biology/methods , Genetic Variation , Undiagnosed Diseases/diagnosis , Adolescent , Child , Child, Preschool , Computer Simulation , Databases, Genetic , Female , Genetic Predisposition to Disease , Humans , Male , Phenotype , Undiagnosed Diseases/genetics , Whole Genome SequencingABSTRACT
The NAGLU challenge of the fourth edition of the Critical Assessment of Genome Interpretation experiment (CAGI4) in 2016, invited participants to predict the impact of variants of unknown significance (VUS) on the enzymatic activity of the lysosomal hydrolase α-N-acetylglucosaminidase (NAGLU). Deficiencies in NAGLU activity lead to a rare, monogenic, recessive lysosomal storage disorder, Sanfilippo syndrome type B (MPS type IIIB). This challenge attracted 17 submissions from 10 groups. We observed that top models were able to predict the impact of missense mutations on enzymatic activity with Pearson's correlation coefficients of up to .61. We also observed that top methods were significantly more correlated with each other than they were with observed enzymatic activity values, which we believe speaks to the importance of sequence conservation across the different methods. Improved functional predictions on the VUS will help population-scale analysis of disease epidemiology and rare variant association analysis.
Subject(s)
Acetylglucosaminidase/metabolism , Computational Biology/methods , Mutation, Missense , Acetylglucosaminidase/genetics , Humans , Models, Genetic , Regression AnalysisABSTRACT
CAGI (Critical Assessment of Genome Interpretation) conducts community experiments to determine the state of the art in relating genotype to phenotype. Here, we report results obtained using newly developed ensemble methods to address two CAGI4 challenges: enzyme activity for population missense variants found in NAGLU (Human N-acetyl-glucosaminidase) and random missense mutations in Human UBE2I (Human SUMO E2 ligase), assayed in a high-throughput competitive yeast complementation procedure. The ensemble methods are effective, ranked second for SUMO-ligase and third for NAGLU, according to the CAGI independent assessors. However, in common with other methods used in CAGI, there are large discrepancies between predicted and experimental activities for a subset of variants. Analysis of the structural context provides some insight into these. Post-challenge analysis shows that the ensemble methods are also effective at assigning pathogenicity for the NAGLU variants. In the clinic, providing an estimate of the reliability of pathogenic assignments is the key. We have also used the NAGLU dataset to show that ensemble methods have considerable potential for this task, and are already reliable enough for use with a subset of mutations.
Subject(s)
Acetylglucosaminidase/genetics , Computational Biology/methods , Mutation, Missense , Ubiquitin-Conjugating Enzymes/genetics , Databases, Genetic , Humans , Machine Learning , Phenotype , ROC Curve , Reproducibility of ResultsABSTRACT
Compared with earlier more restricted sequencing technologies, identification of rare disease variants using whole-genome sequence has the possibility of finding all causative variants, but issues of data quality and an overwhelming level of background variants complicate the analysis. The CAGI4 SickKids clinical genome challenge provided an opportunity to assess the landscape of variants found in a difficult set of 25 unsolved rare disease cases. To address the challenge, we developed a three-stage pipeline, first carefully analyzing data quality, then classifying high-quality gene-specific variants into seven categories, and finally examining each candidate variant for compatibility with the often complex phenotypes of these patients for final prioritization. Variants consistent with the phenotypes were found in 24 out of the 25 cases, and in a number of these, there are prioritized variants in multiple genes. Data quality analysis suggests that some of the selected variants are likely incorrect calls, complicating interpretation. The data providers followed up on three suggested variants with Sanger sequencing, and in one case, a prioritized variant was confirmed as likely causative by the referring physician, providing a diagnosis in a previously intractable case.
Subject(s)
Genetic Variation , Genomics/methods , Rare Diseases/genetics , Child , Genetic Predisposition to Disease , Humans , Sequence Analysis, DNA , SoftwareABSTRACT
Understanding the basis of complex trait disease is a fundamental problem in human genetics. The CAGI Crohn's Exome challenges are providing insight into the adequacy of current disease models by requiring participants to identify which of a set of individuals has been diagnosed with the disease, given exome data. For the CAGI4 round, we developed a method that used the genotypes from exome sequencing data only to impute the status of genome wide association studies marker SNPs. We then used the imputed genotypes as input to several machine learning methods that had been trained to predict disease status from marker SNP information. We achieved the best performance using Naïve Bayes and with a consensus machine learning method, obtaining an area under the curve of 0.72, larger than other methods used in CAGI4. We also developed a model that incorporated the contribution from rare missense variants in the exome data, but this performed less well. Future progress is expected to come from the use of whole genome data rather than exomes.
Subject(s)
Crohn Disease/genetics , Exome Sequencing/methods , Polymorphism, Single Nucleotide , Algorithms , Area Under Curve , Genetic Markers , Genetic Predisposition to Disease , Genome-Wide Association Study , Humans , Machine Learning , PhenotypeABSTRACT
The use of gene panel sequence for diagnostic and prognostic testing is now widespread, but there are so far few objective tests of methods to interpret these data. We describe the design and implementation of a gene panel sequencing data analysis pipeline (VarP) and its assessment in a CAGI4 community experiment. The method was applied to clinical gene panel sequencing data of 106 patients, with the goal of determining which of 14 disease classes each patient has and the corresponding causative variant(s). The disease class was correctly identified for 36 cases, including 10 where the original clinical pipeline did not find causative variants. For a further seven cases, we found strong evidence of an alternative disease to that tested. Many of the potentially causative variants are missense, with no previous association with disease, and these proved the hardest to correctly assign pathogenicity or otherwise. Post analysis showed that three-dimensional structure data could have helped for up to half of these cases. Over-reliance on HGMD annotation led to a number of incorrect disease assignments. We used a largely ad hoc method to assign probabilities of pathogenicity for each variant, and there is much work still to be done in this area.
Subject(s)
Disease/classification , Exome Sequencing/methods , Genetic Variation , High-Throughput Nucleotide Sequencing/methods , Computational Biology , Databases, Genetic , Disease/genetics , Genetic Predisposition to Disease , Humans , Models, Molecular , Mutation, Missense , Phenotype , Proteins/chemistry , Proteins/geneticsABSTRACT
The CAGI-4 Hopkins clinical panel challenge was an attempt to assess state-of-the-art methods for clinical phenotype prediction from DNA sequence. Participants were provided with exonic sequences of 83 genes for 106 patients from the Johns Hopkins DNA Diagnostic Laboratory. Five groups participated in the challenge, predicting both the probability that each patient had each of the 14 possible classes of disease, as well as one or more causal variants. In cases where the Hopkins laboratory reported a variant, at least one predictor correctly identified the disease class in 36 of the 43 patients (84%). Even in cases where the Hopkins laboratory did not find a variant, at least one predictor correctly identified the class in 39 of the 63 patients (62%). Each prediction group correctly diagnosed at least one patient that was not successfully diagnosed by any other group. We discuss the causal variant predictions by different groups and their implications for further development of methods to assess variants of unknown significance. Our results suggest that clinically relevant variants may be missed when physicians order small panels targeted on a specific phenotype. We also quantify the false-positive rate of DNA-guided analysis in the absence of prior phenotypic indication.
Subject(s)
Computational Biology/methods , Sequence Analysis, DNA/methods , Databases, Genetic , Genetic Predisposition to Disease , Genetic Testing , Humans , PhenotypeABSTRACT
Correct phenotypic interpretation of variants of unknown significance for cancer-associated genes is a diagnostic challenge as genetic screenings gain in popularity in the next-generation sequencing era. The Critical Assessment of Genome Interpretation (CAGI) experiment aims to test and define the state of the art of genotype-phenotype interpretation. Here, we present the assessment of the CAGI p16INK4a challenge. Participants were asked to predict the effect on cellular proliferation of 10 variants for the p16INK4a tumor suppressor, a cyclin-dependent kinase inhibitor encoded by the CDKN2A gene. Twenty-two pathogenicity predictors were assessed with a variety of accuracy measures for reliability in a medical context. Different assessment measures were combined in an overall ranking to provide more robust results. The R scripts used for assessment are publicly available from a GitHub repository for future use in similar assessment exercises. Despite a limited test-set size, our findings show a variety of results, with some methods performing significantly better. Methods combining different strategies frequently outperform simpler approaches. The best predictor, Yang&Zhou lab, uses a machine learning method combining an empirical energy function measuring protein stability with an evolutionary conservation term. The p16INK4a challenge highlights how subtle structural effects can neutralize otherwise deleterious variants.
Subject(s)
Computational Biology/methods , Cyclin-Dependent Kinase Inhibitor p18/genetics , Genetic Variation , Cell Line, Tumor , Cell Proliferation , Computer Simulation , Cyclin-Dependent Kinase Inhibitor p16 , Cyclin-Dependent Kinase Inhibitor p18/chemistry , Databases, Genetic , Genetic Predisposition to Disease , Humans , Machine Learning , Protein StabilityABSTRACT
The advent of next-generation sequencing has dramatically decreased the cost for whole-genome sequencing and increased the viability for its application in research and clinical care. The Personal Genome Project (PGP) provides unrestricted access to genomes of individuals and their associated phenotypes. This resource enabled the Critical Assessment of Genome Interpretation (CAGI) to create a community challenge to assess the bioinformatics community's ability to predict traits from whole genomes. In the CAGI PGP challenge, researchers were asked to predict whether an individual had a particular trait or profile based on their whole genome. Several approaches were used to assess submissions, including ROC AUC (area under receiver operating characteristic curve), probability rankings, the number of correct predictions, and statistical significance simulations. Overall, we found that prediction of individual traits is difficult, relying on a strong knowledge of trait frequency within the general population, whereas matching genomes to trait profiles relies heavily upon a small number of common traits including ancestry, blood type, and eye color. When a rare genetic disorder is present, profiles can be matched when one or more pathogenic variants are identified. Prediction accuracy has improved substantially over the last 6 years due to improved methodology and a better understanding of features.
Subject(s)
High-Throughput Nucleotide Sequencing/methods , Whole Genome Sequencing/methods , Area Under Curve , Genetic Predisposition to Disease , Human Genome Project , Humans , Phenotype , Quantitative Trait LociABSTRACT
Precision medicine aims to predict a patient's disease risk and best therapeutic options by using that individual's genetic sequencing data. The Critical Assessment of Genome Interpretation (CAGI) is a community experiment consisting of genotype-phenotype prediction challenges; participants build models, undergo assessment, and share key findings. For CAGI 4, three challenges involved using exome-sequencing data: Crohn's disease, bipolar disorder, and warfarin dosing. Previous CAGI challenges included prior versions of the Crohn's disease challenge. Here, we discuss the range of techniques used for phenotype prediction as well as the methods used for assessing predictive models. Additionally, we outline some of the difficulties associated with making predictions and evaluating them. The lessons learned from the exome challenges can be applied to both research and clinical efforts to improve phenotype prediction from genotype. In addition, these challenges serve as a vehicle for sharing clinical and research exome data in a secure manner with scientists who have a broad range of expertise, contributing to a collaborative effort to advance our understanding of genotype-phenotype relationships.
Subject(s)
Bipolar Disorder/genetics , Crohn Disease/genetics , Exome Sequencing/methods , Precision Medicine/methods , Warfarin/therapeutic use , Computational Biology/methods , Databases, Genetic , Genetic Predisposition to Disease , Humans , Information Dissemination , Pharmacogenomic Variants , Phenotype , Warfarin/pharmacologyABSTRACT
PURPOSE: To investigate the effect of resolvin D1 (RvD1) on the Nod-like receptor family pyrin domain-containing (NLRP3) inflammasome and the nuclear factor-kappa beta (NF-κB) pathway in streptozotocin (STZ)-induced diabetic retinopathy in rats. METHODS: Ninety-six male rats were divided into four groups: control, STZ, RvD1, and vehicle. The rats with diabetic retinopathy induced by STZ in the RvD1 and vehicle groups were given an intravitreal injection of RvD1 (1,000 ng/kg) or the same dosage of vehicle, respectively. All rats were euthanized 7 days following treatment. Hematoxylin and eosin staining was used to observe the pathological changes in the retinal tissues. The location and expression of the NLRP3 inflammasome components, including NLRP3, caspase-associated recruitment domain (ASC), and caspase-1, in the retinas were detected using immunohistochemistry, real-time PCR, and western blot, respectively. Retinal homogenate of rats were collected for the detection of the downstream molecules interleukin 1 beta (IL-1ß) and IL-18 of the NLRP3 inflammasome with enzyme-linked immunosorbent assay kits. RESULTS: The levels of NLRP3, ASC, cleaved caspase-1, IL-1ß, and IL-18 were upregulated in the retinas of the STZ-induced diabetic rats; however, these changes were partially inhibited by the RvD1 treatment. Furthermore, the administration of RvD1 suppressed activation of NF-kB, which was upregulated in STZ-induced diabetic retinopathy. CONCLUSIONS: RvD1 plays a protective role in STZ-induced diabetic retinopathy by inhibiting the level of activation of the NLRP3 inflammasome and associated cytokine production, suggesting targeting of this pathway might be an effective strategy in treatment of diabetic retinopathy.
Subject(s)
Diabetes Mellitus, Experimental/prevention & control , Diabetic Retinopathy/prevention & control , Docosahexaenoic Acids/pharmacology , Inflammasomes/metabolism , NF-kappa B/metabolism , NLR Family, Pyrin Domain-Containing 3 Protein/metabolism , Signal Transduction/physiology , Animals , Blotting, Western , Caspase 1/metabolism , Cytokines/metabolism , Diabetes Mellitus, Experimental/metabolism , Diabetic Retinopathy/metabolism , Enzyme-Linked Immunosorbent Assay , Inflammasomes/genetics , Inflammation/prevention & control , Intravitreal Injections , Male , NLR Family, Pyrin Domain-Containing 3 Protein/genetics , Rats , Rats, Sprague-Dawley , Real-Time Polymerase Chain Reaction , StreptozocinABSTRACT
Antibodies are fundamental effectors of humoral immunity, and have become a highly successful class of therapeutics. There is increasing evidence that antibodies utilize transient homotypic interactions to enhance function, and elucidation of such interactions can provide insights into their biology and new opportunities for their optimization as drugs. Yet the transitory nature of weak interactions makes them difficult to investigate. Capitalizing on their rich structural data and high conservation, we have characterized all the ways that antibody fragment antigen-binding (Fab) regions interact crystallographically. This approach led to the discovery of previously unrealized interfaces between antibodies. While diverse interactions exist, ß-sheet dimers and variable-constant elbow dimers are recurrent motifs. Disulfide engineering enabled interactions to be trapped and investigated structurally and functionally, providing experimental validation of the interfaces and illustrating their potential for optimization. This work provides first insight into previously undiscovered oligomeric interactions between antibodies, and enables new opportunities for their biotherapeutic optimization.
ABSTRACT
Purpose: To investigate the effects of Resolvin E1 (RvE1) on corneal allograft rejection in a high -risk corneal allograft transplantation model. Methods: High-risk corneal beds were created via placement of intrastromal sutures in the corneas of BALB/c mice for 2 weeks. Allogeneic corneal transplantation was performed by transplanting corneas of C57BL/6 mice onto BALB/c hosts. RvE1 or normal saline (control) was subconjunctivally injected. Allograft survival was observed by slit lamp biomicroscope, and inflammatory cell infiltration was detected by hematoxylin and eosin and immunohistochemistry. The percentage of Th1, Th17, and Treg cells in draining lymph nodes (DLNs) were evaluated by flow cytometric analysis. The levels of Th1, Th2, and Th17-associated cytokines in the grafts were measured by cytometric bead array and real-time PCR. Results: RvE1 treatment significantly improved allograft survival compared to the control group. After RvE1 treatment, the infiltration of neutrophils and CD4+ T (Th1/Th17) cells were decreased in corneal grafts, and the percentage of Th1/Th17 cells in DLNs were reduced. In addition, RvE1 treatment significantly reduced the mRNA expression of proinflammatory cytokines in the graft including IL-1α, IL-1ß, TNF-α, IL-2, IL-6, IFN-γ, IL-17A, IL-17F, IL-21, and IL-22 as well as the protein level of the proinflammatory cytokines, including IL-2, TNF, IL-6, IFN-γ, and IL-17. However, RvE1 treatment did not alter the percentage of Treg cells in DLNs and the expression of IL-4, IL-5, and IL-10. Conclusions: RvE1 treatment improves allogeneic corneal graft survival in a high-risk corneal transplantation model via inhibiting the Th1/Th17-related inflammation.
Subject(s)
Corneal Transplantation/methods , Eicosapentaenoic Acid/analogs & derivatives , Graft Rejection/prevention & control , Graft Survival/drug effects , Animals , Cornea/metabolism , Cytokines/metabolism , Eicosapentaenoic Acid/pharmacology , Immunohistochemistry , Mice , Mice, Inbred BALB C , Mice, Inbred C57BL , RNA, Messenger/metabolism , T-Lymphocytes, Regulatory/cytology , Th1 Cells/cytology , Th17 Cells/cytology , Transplantation, HomologousABSTRACT
Neutrophil extracellular traps (NETs), the product of NETosis, is found to localize pathogens and crystals in immune response. Recent studies have found that excessive NETs lead to disease conditions such as diabetes and its complications like diabetic retinopathy (DR). However, the correlation between NETs and high glucose or DR remains unclear. Here, we found NETs level was significantly increased in the serum of diabetic patients, especially in proliferation diabetic retinopathy (PDR) patients. High glucose dramatically increased NETs production in diabetic individuals with time prolonging. The activation of NADPH oxidase was involved in the NETs process which is triggered by high glucose. Moreover, we verified the infiltration of neutrophils in the eyes and adhesion to vascular endothelial cells in diabetic rat models. NETs formation was observed in the vitreous bodies and retinas of diabetic individuals, which indicates NETs may play a role in the pathogenesis of diabetic retinopathy. Furthermore, anti-VEGF therapy downregulates NETs production indicating that NADPH oxidase-derived ROS may be another signaling pathway involved in anti-VEGF therapy.
Subject(s)
Diabetes Mellitus, Type 2/complications , Diabetic Retinopathy/etiology , Diabetic Retinopathy/metabolism , Extracellular Traps/metabolism , Hyperglycemia/metabolism , NADPH Oxidases/metabolism , Analysis of Variance , Animals , Cell Adhesion , Chi-Square Distribution , Diabetes Mellitus, Type 2/drug therapy , Diabetic Retinopathy/blood , Diabetic Retinopathy/drug therapy , Disease Models, Animal , Endothelial Cells/physiology , Extracellular Traps/drug effects , Humans , Intravitreal Injections , Neutrophil Infiltration/physiology , Rats , Reactive Oxygen Species/metabolism , Recombinant Fusion Proteins/administration & dosage , Recombinant Fusion Proteins/pharmacology , Recombinant Fusion Proteins/therapeutic use , Signal Transduction/drug effects , Statistics, Nonparametric , Vascular Endothelial Growth Factor A/antagonists & inhibitors , Vascular Endothelial Growth Factor A/metabolism , Vitreous Body/metabolismABSTRACT
Monoclonal antibodies (mAbs) have become a major class of protein therapeutics that target a spectrum of diseases ranging from cancers to infectious diseases. Similar to any protein molecule, mAbs are susceptible to chemical modifications during the manufacturing process, long-term storage, and in vivo circulation that can impair their potency. One such modification is the oxidation of methionine residues. Chemical modifications that occur in the complementarity-determining regions (CDRs) of mAbs can lead to the abrogation of antigen binding and reduce the drug's potency and efficacy. Thus, it is highly desirable to identify and eliminate any chemically unstable residues in the CDRs during the therapeutic antibody discovery process. To provide increased throughput over experimental methods, we extracted features from the mAbs' sequences, structures, and dynamics, used random forests to identify important features and develop a quantitative and highly predictive in silico methionine oxidation model.
Subject(s)
Antibodies, Monoclonal/chemistry , Complementarity Determining Regions/chemistry , Machine Learning , Methionine/chemistry , Antibodies, Monoclonal/administration & dosage , Antibodies, Monoclonal/metabolism , Antigens/metabolism , Antineoplastic Agents, Immunological/administration & dosage , Antineoplastic Agents, Immunological/chemistry , Antineoplastic Agents, Immunological/metabolism , Complementarity Determining Regions/metabolism , Computer Simulation , Humans , Kinetics , Oxidation-Reduction , Protein Binding , Treatment OutcomeABSTRACT
The objective of this paper is to review the natural products and the pharmacological functions of Ganodermataceae family. Presently, studies on the bioactive components of Lingzhi are focused on polysaccharides and triterpenes/triterpenoids compounds. New Ganoderma polysaccharides, including their molecular weights, glycosyl residue compositions, glycosyl linkage and branches, are summarized in this paper. Also presented are new types of triterpenes and their characteristics from Lingzhi. Taking Ganoderma lucidum as an example, we reviewed its pharmacological functions in anti-tumor and immune-modulating activities for treating hypoglycemosis, hepatoprotection, and the effect on blood vessel system. Based on the advances in Lingzhi research in the past few decades, both G. lucidum and G. sinense are considered as the representative species of medicinal mushroom Lingzhi in China. Until 2001, G. tsugae was only advised to be used as the materials of the health products. The biologically-active components related to pharmacological functions of these three species were studied more than other Ganodermataceae family species; however, which have been used in less modern folk medicine.
Subject(s)
Drugs, Chinese Herbal/therapeutic use , Ganoderma , Phytotherapy/methods , Antineoplastic Agents, Phytogenic/therapeutic use , Drugs, Chinese Herbal/pharmacology , Humans , Hypoglycemic Agents/therapeutic use , Immunotherapy/methods , ReishiABSTRACT
A novel course, "Participation in Research Program (PRP)" in life sciences is open for 1st to 3rd year undergraduates. PRP introduces the principles of a variety of biological methods and techniques and also offers an opportunity to explore some specific knowledge in more detail prior to thesis research. In addition, the PRP introduces some methodologies that have been proven to be successful at each institution to participants. Through disciplines crossing, students were trained theoretically and practically about modern techniques, facilitating the efficient commutation of general laboratory skills and modern laboratory skills, and the possession of higher research ability. Therefore, during some basic training (e.g., usage and maintenance of equipments, designing and completing experiments, analyzing data and reporting results, etc.), a series of capabilities are strengthened, such as basic experimental skills, searching appropriate methods, explaining unknown biological phenomena, and the capacity of solving problems. To determine the efficiency of these strategies, we carefully examined students' performance and demonstrated the progress in students' basic abilities of scientific research in their training.