DNA aptamer-conjugated lipid nanoparticle for targeted PTEN mRNA delivery to prostate cancer cells.

The use of messenger RNA (mRNA) as a cancer vaccine and gene therapy requires targeted vehicle delivery to the site of disease. Here, we designed a mRNA-encapsulating lipid nanoparticle (LNP) conjugated with anti-programmed death-ligand 1 (PD-L1) DNA aptamer that delivers mRNA encoding a tumor suppressor gene, namely phosphatase and tensin homolog (PTEN), to castration-resistant prostate cancer (CRPC) cells expressing PD-L1 on the cell surface. The DNA aptamer-conjugated LNP-based mRNA delivery system (Apt-LNP[PTEN mRNA]) mediated efficient mRNA delivery and transfection in CRPC cells than LNPs without targeting ligands. Cancer-targeted PTEN mRNA delivery using Apt-LNPs achieved significantly higher PTEN expression via aptamer-mediated endocytosis in target cancer cells compared with non-targeted LNP delivery, resulting in significant downregulation of AKT phosphorylation. This enhanced PI3K/AKT pathway regulation, and in turn reduced cell migration after two days along with a 70 % decrease in cell viability, leading to effective apoptotic cell death. In a CRPC xenograft model, Apt-LNP[PTEN mRNA] led to an approximate 60 % reduction in tumor growth, which was attributable to the effective PTEN restoration and PI3K/AKT signaling pathway regulation. PTEN expression was significantly enhanced in CRPC tumor tissues, which abolished cancer cell tumorigenicity. These findings demonstrated the potential of Apt-LNPs for targeted mRNA delivery to cancer cells, thus providing a promising tool for targeted mRNA delivery to a range of cancers and tissues using a conventional LNP systems.

Epigenetic Regulation of the Expression of T Cell Stimulatory and Inhibitory Factors by Histone H3 Lysine Modification Enzymes and Its Prognostic Roles in Glioblastoma

Background@#This study aimed to identify the specific T cell co-stimulatory and co-inhibitory factors that play prognostic roles in patients with glioblastoma. Additionally, the unique histone H3 modification enzymes that regulate the expression levels of these specific costimulatory and co-inhibitory factors were investigated. @*Methods@#The medical records of 84 patients newly diagnosed with glioblastoma at our institution from January 2006 to December 2020 were retrospectively reviewed.Immunohistochemical (IHC) staining for T cell co-stimulatory factors (CD27, CD28, CD137, OX40, and ICOS), T cell co-inhibitory factors (CTLA4, PD1, PD-L1, TIM3, and CD200R), and histone H3 lysine modification enzymes (MLL4, RIZ, EZH1, NSD2, KDM5c, JMJD1a, UTX, and JMJD5) was performed on archived paraffin-embedded tissues obtained by biopsy or resection. Quantitative real time-polymerase chain reaction (qRT-PCR) was performed for specific factors, which demonstrated causal relationships, in order to validate the findings of the IHC examinations. @*Results@#The mean follow-up duration was 27.5 months (range, 4.1–43.5 months). During this period, 76 patients (90.5%) died, and the mean OS was 19.4 months (95% confidence interval, 16.3–20.9 months). Linear positive correlations were observed between the expression levels of CD28 and JMJD1a (R2 linear = 0.982) and those of CD137 and UTX (R2 linear = 1.528). Alternatively, significant negative correlations were observed between the expression levels of CTLA4 and RIZ (R2 linear = −1.746) and those of PD-L1 and EZH1 (R2 linear = −2.118); relationships were confirmed by qRT-PCR. In the multivariate analysis, increased expression levels of CD28 (P = 0.042), and CD137 (P = 0.009), and decreased expression levels of CTLA4 (P = 0.003), PD-L1 (P = 0.020), and EZH1 (P = 0.040) were significantly associated with longer survival. @*Conclusion@#These findings suggest that the expression of certain T cell co-stimulatory factors, such as CD28 and CD 137, and co-inhibitory factors, such as CTLA4 and PD-L1 are associated with prognosis of glioblastoma patients.

Preliminary Study of the Antioxidant Effects of Oenanthe javanica Extract on the Skin of Aged Mice / 대한피부과학회지

No abstract available.

Green discoloration of urine after propofol infusion / 대한마취과학회지

No abstract available.

The mutational landscape of lethal castration-resistant prostate cancer.

Characterization of the prostate cancer transcriptome and genome has identified chromosomal rearrangements and copy number gains and losses, including ETS gene family fusions, PTEN loss and androgen receptor (AR) amplification, which drive prostate cancer development and progression to lethal, metastatic castration-resistant prostate cancer (CRPC). However, less is known about the role of mutations. Here we sequenced the exomes of 50 lethal, heavily pre-treated metastatic CRPCs obtained at rapid autopsy (including three different foci from the same patient) and 11 treatment-naive, high-grade localized prostate cancers. We identified low overall mutation rates even in heavily treated CRPCs (2.00 per megabase) and confirmed the monoclonal origin of lethal CRPC. Integrating exome copy number analysis identified disruptions of CHD1 that define a subtype of ETS gene family fusion-negative prostate cancer. Similarly, we demonstrate that ETS2, which is deleted in approximately one-third of CRPCs (commonly through TMPRSS2:ERG fusions), is also deregulated through mutation. Furthermore, we identified recurrent mutations in multiple chromatin- and histone-modifying genes, including MLL2 (mutated in 8.6% of prostate cancers), and demonstrate interaction of the MLL complex with the AR, which is required for AR-mediated signalling. We also identified novel recurrent mutations in the AR collaborating factor FOXA1, which is mutated in 5 of 147 (3.4%) prostate cancers (both untreated localized prostate cancer and CRPC), and showed that mutated FOXA1 represses androgen signalling and increases tumour growth. Proteins that physically interact with the AR, such as the ERG gene fusion product, FOXA1, MLL2, UTX (also known as KDM6A) and ASXL1 were found to be mutated in CRPC. In summary, we describe the mutational landscape of a heavily treated metastatic cancer, identify novel mechanisms of AR signalling deregulated in prostate cancer, and prioritize candidates for future study.

The Effects of Tetracycline-loaded Silk Fibroin Membrane on Guided Bone Regeneration in a Rabbit Calvarial Defect Model

0.05). Regenerated bone volume (mm3) of 1% TC-loaded SFM, SFM, and control were 36.56+/-8.50, 25.86+/-8.17, and 19.09+/-5.07 at 8 weeks postoperatively, respectively (P<0.05).CONCLUSION: The 1% TC-loaded SFM showed more bone regeneration than the SFM and the uncovered control, in guided bone regeneration.

Quantitative Determination of Plasmodium Parasitemia by Flow Cytometry and Microscopy

The traditional light microscopy has limitations for precise growth assays of malaria parasites in culture or for assessment of new compounds for antimalarial activity; the speed and high reproducibility of flow cytometry can overcome these limitations. A flow cytometric method using PicoGreen, a DNA-binding fluorochrome, was developed with optimal precision suitable for performing growth assays of low-parasitemia field isolates. In addition, intra- and inter-person reproducibility of the flow cytometric and the microscopic method were compared in order to quantitatively demonstrate the improved precision. RNase treatment contributed to the precision of the flow cytometric measurements by enhancing the signal-to-noise ratios. Coefficients of variation of the method were smaller than 10% for 0.1% or higher parasitemia samples. The intra- and inter-person coefficients of variation of the flow cytometric method were three to six times smaller than those of the microscopic method. The flow cytometric method developed in this study yielded substantially more precise results than the microscopic method, allowing determination of parasitemia levels of 0.1% or higher, with coefficients of variation smaller than 10%. Thus, the PicoGreen method could be a reliable high sensitivity assay for analysis of low parasitemia samples and might be applied to a high throughput system testing antimalarial drug activity.

Personalized oncology through integrative high-throughput sequencing: a pilot study.

Individual cancers harbor a set of genetic aberrations that can be informative for identifying rational therapies currently available or in clinical trials. We implemented a pilot study to explore the practical challenges of applying high-throughput sequencing in clinical oncology. We enrolled patients with advanced or refractory cancer who were eligible for clinical trials. For each patient, we performed whole-genome sequencing of the tumor, targeted whole-exome sequencing of tumor and normal DNA, and transcriptome sequencing (RNA-Seq) of the tumor to identify potentially informative mutations in a clinically relevant time frame of 3 to 4 weeks. With this approach, we detected several classes of cancer mutations including structural rearrangements, copy number alterations, point mutations, and gene expression alterations. A multidisciplinary Sequencing Tumor Board (STB) deliberated on the clinical interpretation of the sequencing results obtained. We tested our sequencing strategy on human prostate cancer xenografts. Next, we enrolled two patients into the clinical protocol and were able to review the results at our STB within 24 days of biopsy. The first patient had metastatic colorectal cancer in which we identified somatic point mutations in NRAS, TP53, AURKA, FAS, and MYH11, plus amplification and overexpression of cyclin-dependent kinase 8 (CDK8). The second patient had malignant melanoma, in which we identified a somatic point mutation in HRAS and a structural rearrangement affecting CDKN2C. The STB identified the CDK8 amplification and Ras mutation as providing a rationale for clinical trials with CDK inhibitors or MEK (mitogen-activated or extracellular signal-regulated protein kinase kinase) and PI3K (phosphatidylinositol 3-kinase) inhibitors, respectively. Integrative high-throughput sequencing of patients with advanced cancer generates a comprehensive, individual mutational landscape to facilitate biomarker-driven clinical trials in oncology.

Deep sequencing reveals distinct patterns of DNA methylation in prostate cancer.

Beginning with precursor lesions, aberrant DNA methylation marks the entire spectrum of prostate cancer progression. We mapped the global DNA methylation patterns in select prostate tissues and cell lines using MethylPlex-next-generation sequencing (M-NGS). Hidden Markov model-based next-generation sequence analysis identified ∼68,000 methylated regions per sample. While global CpG island (CGI) methylation was not differential between benign adjacent and cancer samples, overall promoter CGI methylation significantly increased from ~12.6% in benign samples to 19.3% and 21.8% in localized and metastatic cancer tissues, respectively (P-value < 2 × 10(-16)). We found distinct patterns of promoter methylation around transcription start sites, where methylation occurred not only on the CGIs, but also on flanking regions and CGI sparse promoters. Among the 6691 methylated promoters in prostate tissues, 2481 differentially methylated regions (DMRs) are cancer-specific, including numerous novel DMRs. A novel cancer-specific DMR in the WFDC2 promoter showed frequent methylation in cancer (17/22 tissues, 6/6 cell lines), but not in the benign tissues (0/10) and normal PrEC cells. Integration of LNCaP DNA methylation and H3K4me3 data suggested an epigenetic mechanism for alternate transcription start site utilization, and these modifications segregated into distinct regions when present on the same promoter. Finally, we observed differences in repeat element methylation, particularly LINE-1, between ERG gene fusion-positive and -negative cancers, and we confirmed this observation using pyrosequencing on a tissue panel. This comprehensive methylome map will further our understanding of epigenetic regulation in prostate cancer progression.

A comparison of single molecule and amplification based sequencing of cancer transcriptomes.

The second wave of next generation sequencing technologies, referred to as single-molecule sequencing (SMS), carries the promise of profiling samples directly without employing polymerase chain reaction steps used by amplification-based sequencing (AS) methods. To examine the merits of both technologies, we examine mRNA sequencing results from single-molecule and amplification-based sequencing in a set of human cancer cell lines and tissues. We observe a characteristic coverage bias towards high abundance transcripts in amplification-based sequencing. A larger fraction of AS reads cover highly expressed genes, such as those associated with translational processes and housekeeping genes, resulting in relatively lower coverage of genes at low and mid-level abundance. In contrast, the coverage of high abundance transcripts plateaus off using SMS. Consequently, SMS is able to sequence lower- abundance transcripts more thoroughly, including some that are undetected by AS methods; however, these include many more mapping artifacts. A better understanding of the technical and analytical factors introducing platform specific biases in high throughput transcriptome sequencing applications will be critical in cross platform meta-analytic studies.

Malignant hyperthermia during cesarean section: A case report

Malignant hyperthermia is manifestated in susceptible individuals exposed to triggering drugs, such as depolarizing muscle relaxants and inhalational anesthetics. Various musculoskeletal abnormalities, such as scoliosis, hernias and strabismus, have been associated with malignant hyperthermia susceptibility. During cesarean section of the patient who had undergone scoliosis correction surgery, we experienced malingnant hyperthermia due to succinylcholine and inhalation anesthetics. In our case, as soon as we suspected the episode, all anesthetics were stopped and anesthetic machines were changed to unexposed anesthetic machine for inhalational anesthetics. Dantrolene was given intravenously and the patient was cooled by tepid sponging, cooled fluids. The patient recovered normal temperature and consciousness without any complications.

Protein interaction network underpins concordant prognosis among heterogeneous breast cancer signatures.

Characterizing the biomolecular systems' properties underpinning prognosis signatures derived from gene expression profiles remains a key clinical and biological challenge. In breast cancer, while different "poor-prognosis" sets of genes have predicted patient survival outcome equally well in independent cohorts, these prognostic signatures have surprisingly little genetic overlap. We examine 10 such published expression-based signatures that are predictors or distinct breast cancer phenotypes, uncover their mechanistic interconnectivity through a protein-protein interaction network, and introduce a novel cross-"gene expression signature" analysis method using (i) domain knowledge to constrain multiple comparisons in a mechanistically relevant single-gene network interactions and (ii) scale-free permutation re-sampling to statistically control for hubness (SPAN - Single Protein Analysis of Network with constant node degree per protein). At adjusted p-values<5%, 54-genes thus identified have a significantly greater connectivity than those through meticulous permutation re-sampling of the context-constrained network. More importantly, eight of 10 genetically non-overlapping signatures are connected through well-established mechanisms of breast cancer oncogenesis and progression. Gene Ontology enrichment studies demonstrate common markers of cell cycle regulation. Kaplan-Meier analysis of three independent historical gene expression sets confirms this network-signature's inherent ability to identify "poor outcome" in ER(+) patients without the requirement of machine learning. We provide a novel demonstration that genetically distinct prognosis signatures, developed from independent clinical datasets, occupy overlapping prognostic space of breast cancer via shared mechanisms that are mediated by genetically different yet mechanistically comparable interactions among proteins of differentially expressed genes in the signatures. This is the first study employing a networks' approach to aggregate established gene expression signatures in order to develop a phenotype/pathway-based cancer roadmap with the potential for (i) novel drug development applications and for (ii) facilitating the clinical deployment of prognostic gene signatures with improved mechanistic understanding of biological processes and functions associated with gene expression changes. http://www.lussierlab.org/publication/networksignature/.

Prevalence of Sleep Disorder and Associated Factors in Family Practice / 가정의학회지

BACKGROUND: Sleep is an essential restorative physiologic phenomenon. Impaired sleep results in significant negative effect to the health. Symptoms like sleep initiation difficulty, frequent awakening, severe snoring have related to poor sleep quality. We studied frequency and compared the characteristics of common sleep disorders at family practice. METHODS: We surveyed patients over 18 years of age and their guardians who visited 16 familial practices for 6 days. We investigated sleep characteristics, frequency of sleep disorder and associated factors by questionnaires and analyzed by frequency analysis, Spearman's correlation coefficient, multiple logistic regression. RESULTS: We enrolled 1,117 participants. Older participants were more likely to report early sleep onset and off time, short sleep duration. Mean number of awakening during a typical night is 1.69. Female complained difficulties in initiation and maintenance of sleep more than male. A total of 32.5% had these insomnia symptoms and related to hypertension, stroke, stress, arthralgia, depression, urological disorder. 31.1% had excessive daytime sleepiness, related to stress, arthralgia, depression. Loud snoring and gasp for breath showed positive correlation between male, high BMI. Disrupted sleep over 3 times was related to old age, female, diabetes, hypertension, stroke, stress, arthralgia, depression. Restless leg syndrome were high in elderly, high BMI, stress, arthralgia and depression. CONCLUSION: About one in three who visit in primary medical practice have sleep disorder symptoms like insomnia, daytime fatigue, snoring. 3% of them have gasp for breath, 8% have restless leg syndrome.

Robust methods for accurate diagnosis using pan-microbiological oligonucleotide microarrays.

BACKGROUND: To address the limitations of traditional virus and pathogen detection methodologies in clinical diagnosis, scientists have developed high-throughput oligonucleotide microarrays to rapidly identify infectious agents. However, objectively identifying pathogens from the complex hybridization patterns of these massively multiplexed arrays remains challenging. METHODS: In this study, we conceived an automated method based on the hypergeometric distribution for identifying pathogens in multiplexed arrays and compared it to five other methods. We evaluated these metrics: 1) accurate prediction, whether the top ranked prediction(s) match the real virus(es); 2) four accuracy scores. RESULTS: Though accurate prediction and high specificity and sensitivity can be achieved with several methods, the method based on hypergeometric distribution provides a significant advantage in term of positive predicting value with two to sixty folds the positive predicting values of other methods. CONCLUSION: The proposed multi-specie array analysis based on the hypergeometric distribution addresses shortcomings of previous methods by enhancing signals of positively hybridized probes.

PhenoGO: an integrated resource for the multiscale mining of clinical and biological data.

The evolving complexity of genome-scale experiments has increasingly centralized the role of a highly computable, accurate, and comprehensive resource spanning multiple biological scales and viewpoints. To provide a resource to meet this need, we have significantly extended the PhenoGO database with gene-disease specific annotations and included an additional ten species. This a computationally-derived resource is primarily intended to provide phenotypic context (cell type, tissue, organ, and disease) for mining existing associations between gene products and GO terms specified in the Gene Ontology Databases Automated natural language processing (BioMedLEE) and computational ontology (PhenOS) methods were used to derive these relationships from the literature, expanding the database with information from ten additional species to include over 600,000 phenotypic contexts spanning eleven species from five GO annotation databases. A comprehensive evaluation evaluating the mappings (n = 300) found precision (positive predictive value) at 85%, and recall (sensitivity) at 76%. Phenotypes are encoded in general purpose ontologies such as Cell Ontology, the Unified Medical Language System, and in specialized ontologies such as the Mouse Anatomy and the Mammalian Phenotype Ontology. A web portal has also been developed, allowing for advanced filtering and querying of the database as well as download of the entire dataset http://www.phenogo.org.

Transcriptome sequencing to detect gene fusions in cancer.

Recurrent gene fusions, typically associated with haematological malignancies and rare bone and soft-tissue tumours, have recently been described in common solid tumours. Here we use an integrative analysis of high-throughput long- and short-read transcriptome sequencing of cancer cells to discover novel gene fusions. As a proof of concept, we successfully used integrative transcriptome sequencing to 're-discover' the BCR-ABL1 (ref. 10) gene fusion in a chronic myelogenous leukaemia cell line and the TMPRSS2-ERG gene fusion in a prostate cancer cell line and tissues. Additionally, we nominated, and experimentally validated, novel gene fusions resulting in chimaeric transcripts in cancer cell lines and tumours. Taken together, this study establishes a robust pipeline for the discovery of novel gene chimaeras using high-throughput sequencing, opening up an important class of cancer-related mutations for comprehensive characterization.

Modeling and characterization of disease associated subnetworks in the human interactome using machine learning.

The availability of large-scale, genome-wide data about the molecular interactome of entire organisms has made possible new types of integrative studies, making use of rapidly accumulating knowledge of gene-disease associations. Previous studies have established the presence of functional biomodules in the molecular interaction network of living organisms, a number of which have been associated with the pathogenesis and progression of human disease. While a number of studies have examined the networks and biomodules associated with disease, the properties that contribute to the particular susceptibility of these subnetworks to disruptions leading to disease phenotypes have not been extensively studied. We take a machine learning approach to the characterization of these disease subnetworks associated with complex and single-gene diseases, taking into account both the biological roles of their constituent genes and topological properties of the networks they form.

A case of cardiac arrest following bupivacaine injection for shoulder arthroscopy : A case report

Intra-articular administration of local anesthetics such as bupivacaine can produce short-term postoperative analgesia in patients who are undergoing shoulder arthroscopy. Yet bupivacaine can result in cardiovascular toxicity that can lead to cardiac arrest. We experienced a case of 63-year-old male patient with severe cardiac toxicity that led to ventricular fibrillation 15 minutes after injecting 0.5% bupivacaine into the patient's shoulder joint for shoulder arthroscopy.

Incidental diagnosis of multiple myeloma during the treatment of low back pain: A case report / 대한마취과학회지

A 49-year-old male patient presented at our clinic with back pain due to an insignificant injury that had occurred approximately 7 months earlier. Although the patient had been treated at primary clinics, the pain had gradually become aggravated and characterized by resting and night pain. We initially diagnosed the patient with myofascial pain syndrome and began treatment comprised of trigger point injection (TPI) on the back muscles to control the pain. His symptoms improved after the first treatment (TPI), but he complained of back pain again several days later. At that time he also reported that he had lost 10 kg over the 4 months preceding his initial visit. Plain radiographs of the thoracolumbar spine revealed L1-L5 compression fractures and generalized osteopenia. The patient was then diagnosed with multiple myeloma based on the results of a bone marrow biopsy. This case demonstrates the importance of using comprehensive diagnostic approaches when the patient manifests symptoms that are unresponsive to conventional treatment.

Trend of the Subjects and Participants of the Korean Academy of Family Medicine Conference / 가정의학회지

BACKGROUND: Throughout the past 20 years in the Korean academy of family medicine seasonal conference, on-going study is done to promote overall development and satisfaction of the conference participants and to overlook the trend of the conference subject, the number of classes, the number of participants, etc. METHODS: About 2,132 topics during the conference from the year 1992 to 2007 collected from the Korean academy of family medicine website were categorized by subject based on the standard of the contents of the latest textbook. There were a total of 7 main classifications including 5 categories like 'principles of family medicine', 'disease prevention and health promotion', 'symptoms', 'clinical procedures', 'diseases' and adding 2 categories such as each committee's classes and other subjects. The scope of the changes of the main and sub-titles were categorized as in the 1990s and 21 century. RESULTS: The number of attendees has increased during the past 20 years, especially the residents were the main portion of the participants. On the proportion of the clinical topics, there was a remarkable increase of geriatric medicine, palliative medicine, obesity, exercise, nutrition, gastroscopy, and colonoscopy procedure in the later half rather than the former half period. In the field of the main category, the core principle subjects of family medicine seemed to be decreased in contrast to disease category. CONCLUSION: During the last 20 years, the titles of family medicine conference are changing with the trend of practice. The core knowledge of family medicine should be maintained and balanced for the future of family medicine conference.