The fungal-like family Saprolegniaceae (Oomycota), also called “water mold,” includes mostly aquatic saprophytes as well as notorious aquatic animal pathogens. Most studies on Saprolegniaceae have been biased toward pathogenic species that are important to aquaculture rather than saprotrophic species, despite the latter’s crucial roles in carbon cycling of freshwater ecosystems. Few attempts have been made to study the diversity and ecology of Saprolegniaceae; thus, their ecological role is not well-known. During a survey of oomycetes between 2016 and 2021, we investigated the diversity and distribution of culturable Saprolegniaceae species in freshwater ecosystems of Korea. In the present study, members of Saprolegniaceae were isolated and identified at species level based on their cultural, morphological, and molecular phylogenetic analyses. Furthermore, substrate preference and seasonal dynamics for each were examined. Most of the species were previously reported as animal pathogens; however, in the present study, they were often isolated from other freshwater substrates, such as plant debris, algae, water, and soil sediment. The relative abundance of Saprolegniaceae was higher in the cold to cool season than that in the warm to hot season of Korea. This study enhances our understanding of the diversity and ecological attributes of Saprolegniaceae in freshwater ecosystems.

BACKGROUND: Cell culture is the golden standard method for Herpes simplex virus (HSV) isolation. However, some specimens require many days to develop any cytopathic effect (CPE). We developeda rapid sensitive culture technique for HSV isolations. METHODS: This study included a total of 133 patients with suspected HSV infection. Specimens were centrifuged onto a Vero cell monolayer in a shell vial. The CPE was observed daily during the5-day incubation by inverted-phase microscope. The direct immunofluorescence (DIF) stain with aHSV specific antibody was performed 2 days after sample inoculation. The negative samples in theDIF stain were reinoculated in the new shell vials after extraction of the monolayer. Polymerase chainreaction for HSV detection was performed using the original samples. RESULTS: The CPE was observed 30 (64%), 39 (83%), 43 (92%), 44 (94%), and 46 (98%) cases at1, 2, 3, 4, and 5 days incubation, respectively. The DIF stain detected 46 cases (98%) at 2 days incubation. The CPE was observed in another 7 cases at 1-day incubation after the reinoculation of negative samples. The PCR detected 47 (100%) of 133 cases. CONCLUSIONS: The reinoculation of negative sample in a shell vial culture is a rapid sensitive methodfor HSV isolation.
Cell Culture Techniques ; Culture Techniques ; Fluorescent Antibody Technique, Direct ; Humans ; Polymerase Chain Reaction ; Simplexvirus* ; Vero Cells

No abstract available.
Glioma*

PURPOSE: This study aimed to investigate the effects of oral hygiene care by oral professionals on periodontal health in type 2 diabetes mellitus patients. MATERIALS AND METHODS: Diabetic participants were recruited at a university hospital and matched at a 1:1 ratio by age and gender, and randomly allocated into intervention (40 people) and control groups (35 people). Tooth brushing instruction, oral health education, and supra-gingival scaling were implemented in all patients at baseline. This program was repeatedly conducted in intervention patients every month for 6 months, and twice at baseline and the sixth month in the control. Oral health was measured by decayed, missing, and filled teeth (DMFT), plaque index, calculus index, bleeding index, patient hygiene performance (PHP) index, tooth mobility, Russel's periodontal index, and community periodontal index (CPI). Diabetes-related factors, oral and general health behaviors, and sociodemographic factors were interviewed as other confounding factors. An analysis of covariance (ANCOVA) was used with SPSS for Windows 14.0. RESULTS: At baseline, there were no significant differences between the two groups in average of periodontal health (calculus index, bleeding index, Russel's periodontal index, CPI, and tooth mobility), diabetes-related factors (fasting blood glucose, postprandial blood glucose, and HbA1c), and in distribution of sociodemographic factors and health behaviors. In intervention group, plaque index, dental calculus index, bleeding index, and PHP index were reduced fairly and steadily from the baseline. There were significant differences in plaque index, dental calculus index, bleeding index, PHP index, and Russel's periodontal index between the two groups at sixth month after adjusted for baseline status. CONCLUSION: Intensive oral hygiene care can persistently improve oral inflammation status and could slow periodontal deterioration.
Adult ; Aged ; Dental Plaque Index ; *Diabetes Mellitus, Type 2 ; Female ; Gingivitis/*prevention & control ; Humans ; Male ; Middle Aged ; Oral Hygiene/education/*methods ; Periodontal Diseases/*prevention & control

People called night owls habitually have late bedtimes and late times of arising, sometimes suffering a heritable circadian disturbance called delayed sleep phase syndrome (DSPS). Those with DSPS, those with more severe progressively-late non-24-hour sleep-wake cycles, and those with bipolar disorder may share genetic tendencies for slowed or delayed circadian cycles. We searched for polymorphisms associated with DSPS in a case-control study of DSPS research participants and a separate study of Sleep Center patients undergoing polysomnography. In 45 participants, we resequenced portions of 15 circadian genes to identify unknown polymorphisms that might be associated with DSPS, non-24-hour rhythms, or bipolar comorbidities. We then genotyped single nucleotide polymorphisms (SNPs) in both larger samples, using Illumina Golden Gate assays. Associations of SNPs with the DSPS phenotype and with the morningness-eveningness parametric phenotype were computed for both samples, then combined for meta-analyses. Delayed sleep and "eveningness" were inversely associated with loci in circadian genes NFIL3 (rs2482705) and RORC (rs3828057). A group of haplotypes overlapping BHLHE40 was associated with non-24-hour sleep-wake cycles, and less robustly, with delayed sleep and bipolar disorder (e.g., rs34883305, rs34870629, rs74439275, and rs3750275 were associated with n=37, p=4.58E-09, Bonferroni p=2.95E-06). Bright light and melatonin can palliate circadian disorders, and genetics may clarify the underlying circadian photoperiodic mechanisms. After further replication and identification of the causal polymorphisms, these findings may point to future treatments for DSPS, non-24-hour rhythms, and possibly bipolar disorder or depression.
Bipolar Disorder ; Case-Control Studies ; Comorbidity ; Depression ; Genetics ; Haplotypes ; Humans ; Melatonin ; Phenotype ; Photoperiod ; Polymorphism, Single Nucleotide ; Polysomnography ; Sleep Wake Disorders, Circadian Rhythm ; Strigiformes*

Benzalkonium chloride, diazolidinyl urea, and imidazolidinyl urea are commonly used preservatives in cosmetics. Recent reports suggested that these compounds may have cellular and systemic toxicity in high concentration. In addition, diazolidinyl urea and imidazolidinyl urea are known formaldehyde (FA) releasers, raising concerns for these cosmetic preservatives. In this study, we investigated the effects of benzalkonium chloride, diazolidinyl urea, and imidazolidinyl urea on ROS-dependent apoptosis of rat neural progenitor cells (NPCs) in vitro. Cells were isolated and cultured from embryonic day 14 rat cortices. Cultured cells were treated with 1–1,000 nM benzalkonium chloride, and 1–50 μM diazolidinyl urea or imidazolidinyl urea at various time points to measure the reactive oxygen species (ROS). PI staining, MTT assay, and live-cell imaging were used for cell viability measurements. Western blot was carried out for cleaved caspase-3 and cleaved caspase-8 as apoptotic protein markers. In rat NPCs, ROS production and cleaved caspase-8 expression were increased while the cell viability was decreased in high concentrations of these substances. These results suggest that several cosmetic preservatives at high concentrations can induce neural toxicity in rat brains through ROS induction and apoptosis.
Animals ; Apoptosis* ; Benzalkonium Compounds ; Blotting, Western ; Brain ; Caspase 3 ; Caspase 8 ; Cell Survival ; Cells, Cultured ; Formaldehyde ; In Vitro Techniques ; Rats* ; Reactive Oxygen Species ; Stem Cells* ; Urea

PURPOSE: Myelodysplastic syndromes (MDS) are heterogeneous clonal stem cell disorders characterized by morphological and functional abnormal hematopoiesis, resulting in cytopenias in the peripheral blood. MDS in children are very rare and their clinical characteristics, natural history, the most effective treatment and prognostic factors need to be elucidated. In addition, consensus for the classification of pediatric MDS has not been reached yet. This multicenter, retrospective study aimed to describe the characteristics and the disease courses of 75 MDS patients from 11 University Hospitals in Korea, seen between 1991 and 2001. METHODS: Kaplan-Meier probability of leukemic transformation and overall survival were plotted. And the usefulness of prognostic scoring systems, including French-American-British (FAB) classification, Bournemouth scoring system (BSS), and International Prognostic Scoring System (IPSS) in the prediction of transformation to acute myelogenous leukemia (AML) and overall survival was evaluated. RESULTS: The median age was 65 months (2~175 months) and the sex ratio was 2.6: 1 (M: F). Fourteen patients (18.7%) were unable to be allocated into any subtype of FAB. The frequency of FAB subtypes in Korea was similar to that of Western countries except for higher proportion of refractory anemia (RA, 47.5%). Median survival was 54 months with Kaplan-Meier 5-yr survival probability of 31.9% and 2-yr probability of transformation to AML was 23.7%. None of the FAB, BSS, and IPSS was capable of discriminating subgroup of patients for the prediction of survival. However, all of the FAB (P=0.004), BSS (P=0.001), and IPSS (P=0.02) were able to subdivide subgroups for the prediction of transformation to AML. CONCLUSION: The characteristics of pediatric MDS in Korea were different from those of other countries, in light of the higher proportion of RA, the low percentage of inherited diseases, and the low percentage of cytogenetic abnormalities. However, the reasons of the differences were not clear. Moreover, none of the prognostic scoring systems, including IPSS, was reliably predictive of survival, reflecting differences from adult cases. With this multicenter study, we suggest the necessity of a prospective study for the classification and treatment. A newer, effective method should be developed for the prediction of disease progression and survival in pediatric MDS.
Adult ; Anemia, Refractory ; Child ; Chromosome Aberrations ; Classification ; Consensus ; Disease Progression ; Hematopoiesis ; Hospitals, University ; Humans ; Korea* ; Leukemia, Myeloid, Acute ; Myelodysplastic Syndromes* ; Natural History ; Retrospective Studies ; Sex Ratio ; Stem Cells

PURPOSE: Immune pathophysiology of aplastic anemia (AA) has been indirectly inferred from responses to immunosuppressive agents. An association between AA and HLA-A2, or HLA-DR2 (its serologic split, HLA-DR15; or its molecular correspondents, DRB1*1501) has been implicated. The presence of HLA-DR15 (including DR2 or DRB1*15) has been closely associated with a favorable response to immunosuppression in AA. This study was aimed to characterize Korean patients with AA by determining the association with certain HLA alleles, such as HLA-DR2 or HLA-A2, and their implications in terms of the response to immunosuppression. METHODS: One-hundred eighteen children with AA from 10 university hospitals between 1990 and 2001 were enrolled in this multicenter, retrospective study. Among them, HLA data were available from 80 patients. Tests of proportions were used to compare allelic frequencies. RESULTS: The frequency of HLA-A2 (58.8%) or HLA-DR2 (24.7%) in AA was not significantly different from those of the controls. Analysis of the patients treated with immunosuppression (N=86) showed that, 50.0% of patients showed a response, including 16.0% of complete response at 6 months. The presence of DR2 allele did not portend a favorable response to immunosuppressive therapy. CONCLUSION: Unlike Western countries, the association of AA with certain HLA alleles was not documented in the Korean population. Moreover, the presence of HLA-DR2 did not predict a favorable response to immunosuppression. This peculiar characteristics of Korean AA needs to be investigated whether these findings reflect ethnic differences, different contribution of immune-mediated AA, different immune mechanisms, or mere limitation by number of study patients.
Alleles* ; Anemia, Aplastic* ; Child* ; HLA-A2 Antigen ; HLA-DR2 Antigen ; Hospitals, University ; Humans ; Immunosuppression ; Immunosuppressive Agents ; Retrospective Studies

Objective: Glucagon in mammals and its homolog (adipokinetic hormone [AKH] in Drosophila melanogaster) are peptide hormones which regulate lipid metabolism by breaking down triglycerides. Although regulatory mechanisms of glucagon and Akh expression have been widely studied, post-transcriptional gene expression of glucagon has not been investigated thoroughly. In this study, we aimed to profile proteins binding with Gcg messenger RNA (mRNA) in mouse and Akh mRNA in Drosophila. Methods: Drosophila Schneider 2 (S2) and mouse 3T3-L1 cell lysates were utilized for affinity pull down of Akh and Gcg mRNA respectively using biotinylated anti-sense DNA oligoes against target mRNAs. Mass spectrometry and computational network analysis revealed mRNA-interacting proteins residing in functional proximity. Results: We observed that 1) 91 proteins interact with Akh mRNA from S2 cell lysates, 2) 34 proteins interact with Gcg mRNA from 3T3-L1 cell lysates. 3) Akh mRNA interactome revealed clusters of ribosomes and known RNA-binding proteins (RBPs). 4) Gcg mRNA interactome revealed mRNA-binding proteins including Plekha7, zinc finger protein, carboxylase, lipase, histone proteins and a cytochrome, Cyp2c44. 5) Levels of Gcg mRNA and its interacting proteins are elevated in skeletal muscles isolated from old mice compared to ones from young mice. Conclusion Akh mRNA in S2 cells are under active translation in a complex of RBPs and ribosomes. Gcg mRNA in mouse precursor adipocyte is in a condition distinct from Akh mRNA due to biochemical interactions with a subset of RBPs and histones. We anticipate that our study contributes to investigating regulatory mechanisms of Gcg and Akh mRNA decay, translation, and localization.

Tolvaptan reduces height-adjusted total kidney volume (htTKV) and renal function decline in autosomal dominant polycystic kidney disease (ADPKD). This study was aimed at investigating the efficacy and safety of tolvaptan in Korean patients with ADPKD during the titration period. Methods: This study is a multicenter, single-arm, open-label phase 4 study. We enrolled 108 patients with ADPKD (age, 19–50 years) with an estimated glomerular filtration rate (eGFR) of >30 mL/min/1.73 m2 and factors defined as indicative of rapid disease progression. After tolvaptan titration, we evaluated efficacy and side effects and assessed factors associated with the effects. Results: After titration for 4 weeks, eGFR and htTKV decreased by 6.4 ± 7.9 mL/min/1.73 m2 and 16 ± 45 mL/m, respectively. No serious adverse drug reactions were observed during the titration period. The greatest eGFR decline was observed in the first week, with a starting tolvaptan dose of 45 mg. Multivariate linear regression for htTKV decline showed that the greater the change in urine osmolality (Uosm), the greater the decrease in htTKV (β, 0.436; p = 0.009) in the 1D group stratified by the Mayo Clinic image classification. Higher baseline eGFR was related to a higher htTKV reduction rate in the 1E group (β, –0.642; p = 0.009). Conclusion: We observed short-term effects and safety during the tolvaptan titration period. The decline of htTKV can be predicted as a short-term effect of tolvaptan by observing Uosm changes from baseline to end of titration in 1D and baseline eGFR in 1E groups.