Extremely low-frequency electromagnetic field induces neural differentiation of hBM-MSCs through regulation of (Zn)-metallothionein-3.

Extremely low-frequency electromagnetic field (ELFEMF) can stimulate neural differentiation in human bone marrow-derived mesenchymal cells (hBM-MSCs), and this provides an opportunity for research on neurodegenerative diseases such as Alzheimer's disease (AD). Metallothionein-3 (MT3), an isoform of the metal-binding proteins, metallothioneins, involved in maintaining intracellular zinc (Zn) homeostasis and the deregulation of zinc homeostasis, has separately been implicated in AD. Here, we investigated the effect of ELFEMF-induced neural differentiation of hBM-MSCs on Zn-MT3 homeostatic interaction. Exposure to ELFEMF induced neural differentiation of hBM-MSCs, which was characterized by decreased proliferation and enhanced neural-like morphology. We observed expression of neuronal markers such as ß-tubulin3, pleiotrophin, and neurofilament-M at the mRNA level and MAP2 at the protein level. ELFEMF-induced neural differentiation correlated with decreased expression of metal-response element-transcription factor 1 and MT3, as well as decreased intracellular Zn concentration. In addition, upregulation of dihydropyrimidinase-related protein 2 was observed, but there was no change in γ-enolase expression. These data indicate a possible regulatory mechanism for MT3 during neural differentiation. Our findings provide considerable insight into molecular mechanisms involved in neural differentiation, which is useful for developing new treatments for neurodegenerative diseases. Bioelectromagnetics. 38:364-373, 2017. © 2017 Wiley Periodicals, Inc.

Divergent C-H Annulation for Multifused N-Heterocycles: Regio- and Stereospecific Cyclizations of N-Alkynylindoles.

N-Alkynylindoles were divergently cyclized for the synthesis of multifused N-heterocycles. An ortho-aryl palladium species was added to the α position of an ynamine to generate (Z)-6-alkylidene/benzylidene-6H-isoindolo[2,1-a]indoles, while Pt-catalyzed ß-addition through π-activation gave 5-alkyl/arylindolo[2,1-a]isoquinolines. Double cyclizations using PdCl2 and oxidant afforded bright yellow benzo[7,8]indolizino[2,3,4,5-ija]quinolines, the synthesis of which was also demonstrated in a different synthetic route.

Safe administration of sotrovimab to a COVID-19 patient with acute phase type 1 diabetes mellitus.

Type 1 diabetes mellitus (T1DM) and poor glycemic control are risk factors for severe coronavirus disease 2019 (COVID-19). Sotrovimab can treat mild-to-moderate COVID-19 in patients at a high risk of progression to severe COVID-19. However, its safety and efficacy in T1DM patients remain to be elucidated. We report the case of a 12-yr-old patient who was treated with sotrovimab for COVID-19 immediately after treatment for diabetic ketoacidosis (DKA) due to new-onset T1DM. He presented with nausea and sore throat and was diagnosed with severe DKA and COVID-19. A productive cough and sputum developed after admission. On the 3rd day of admission, the DKA resolved, and sotrovimab was administered to prevent exacerbation of COVID-19. Although the blood glucose levels increased after the administration of sotrobimab, there was no recurrence of DKA. Hyperglycemia may be a sotrovimab-related adverse event in T1DM patients. Nevertheless, the benefits of sotrovimab treatment may far outweigh the potential risks. Thus, sotrovimab was considered safe for patients with T1DM immediately after treatment of severe DKA.

Genome-wide screening antifungal genes in Streptomyces griseus S4-7, a Fusarium wilt disease suppressive microbial agent.

Streptomyces is a widely studied bacterial genus, particularly with regard to secondary metabolites and antibiotics production. Streptomyces griseus S4-7 was isolated from a strawberry Fusarium wilt disease suppressive soil, and its biological control ability has been well established. However, the antifungal mechanism of strain S4-7 is not yet fully understood at the molecular and biochemical level. Therefore, in this study we created a random mutant library for strain S4-7 with the Tn5 transposon element to investigate antifungal traits on a genome-wide scale. In total 4646 individual mutant strains were created and 13 mutants were selected based on loss of antifungal activity. The knockout genes were identified as electron transfer oxidoreductase (eto),sigma factor-70(sig70) and nrps by Inverse PCR (I-PCR). eto regulates the geranylgeranyl reductase gene, which is involved in terpenoid-quinone biosynthesis, an important factor in cell fitness. In the △eto strain, expression of wbl, a master regulator of the production of secondary metabolites, was significantly reduced. sig70 is responsible for the cell differentiation sensing mechanism in genus Streptomyces. △nrps showed decreased production of hybrid peptide-polyketide siderophores. These results suggest that S. griseus S4-7 may have various antifungal mechanisms, and each mechanism is essential to maximal antifungal activity.

Development qRT-PCR Protocol to Predict Strawberry Fusarium Wilt Occurrence.

Strawberry Fusarium wilt disease, caused by Fusarium oxysporum f. sp. fragariae, is the most devastating disease in strawberry production. The pathogen produces chlamydospores which tolerate against harsh environment, fungicide and survive for decades in soil. Development of detection and quantification techniques are regarded significantly in many soilborne pathogens to prevent damage from diseases. In this study, we improved specific-quantitative primers for F. oxysporum f. sp. fragariae to reveal correlation between the pathogen density and the disease severity. Standard curve r2 value of the specific-quantitative primers for qRT-PCR and meting curve were over 0.99 and 80.5°C, respectively. Over pathogen 105 cfu/g of soil was required to cause the disease in both lab and field conditions. With the minimum density to develop the wilt disease, the pathogen affected near 60% in nursery plantation. A biological control microbe agent and soil solarization reduced the pathogen population 2-fold and 1.5-fold in soil, respectively. The developed F. oxysporum f. sp. fragariae specific qRT-PCR protocol may contribute to evaluating soil healthiness and appropriate decision making to control the disease.

Inverse relationship between vitamin D levels and platelet indices in Korean adults.

BACKGROUND AND AIMS: Vitamin D deficiency and increased platelet indices are associated with increased rate or risk of several diseases such as cardiovascular disease and metabolic syndrome, respectively. We investigated whether vitamin D deficiency is associated with increased platelet count (PC) and mean platelet volume (MPV). METHODS AND RESULTS: The study included 3190 subjects older than 20 years. Subjects were divided into three groups based on their vitamin D levels: vitamin D deficiency (<10.0 ng/ml); insufficiency (10-20 ng/ml); and sufficiency (>20.0 ng/ml). The associations between platelet indices and various parameters were analyzed by Pearson's correlation analysis and t-tests. Then, multivariate linear regression analyses were done correcting for associated parameters. PC and MPV showed a negative correlation with vitamin D groups by ANOVA and multiple linear regression. PC was inversely related with vitamin D group after adjusting for sex, age, regular exercise, white blood cell count, total cholesterol, hemoglobin, and creatinine levels (ß ± SE = -3.461 ± 1.512, P = 0.022). MPV was also inversely related with vitamin D group after adjusting for regular exercise, hemoglobin level, and total cholesterol level (ß ± SE = -0.080 ± 0.026, P = 0.002), and this relationship remained statistically significant after adjusting for regular exercise, hemoglobin level, total cholesterol level, diabetes, hypertension, and body mass index (ß ± SE=-0.082 ± 0.026, P = 0.002). CONCLUSION: PC and MPV are inversely associated with vitamin D levels in adults.

CACTA and MITE transposon distributions on a genetic map of rice using F15 RILs derived from Milyang 23 and Gihobyeo hybrids.

Up to 35% of the rice genome consists of various kinds of transposons, and CACTA and MITE are two of the major class 2 DNA transposons in the genome. We have employed the consensus sequences of Rim2/Hipa CACTA, Stowaway MITE Pangrangja, and Tourist MITE Ditto for transposon display (TD) analysis to locate them on a genetic map, with 58 SSR markers used to anchor them. The TD analysis produced a high profile of the polymorphisms between the parental lines, Oryza sativa var. Gihobyeo/O. sativa var. Milyang, in intraspecific F15 RIL lines, locating 368 markers of Rim2/Hipa CACTA, 78 markers of Tourist MITE Ditto, and 22 markers of Stowaway MITE Pangrangja. In the segregation analysis, non-parental segregating bands and segregation distortion bands were observed. The recombinant genetic map spans 3023.9 cM, with 5.7 cM the average distance between markers. The TD markers were distributed unequally on the chromosomes because many TD markers were located in pericentric chromosomal regions except in the cases of chromosomes 2, 3, 6 and 9. Although the number of transposon markers was not sufficient to include all rice class 2 transposons, the current map of CACTA and MITE transposons should provide new insight into the genome organization of rice since no previous DNA transposon map is available.

Genetic diversity and phylogenetic relationship in AA Oryza species as revealed by Rim2/Hipa CACTA transposon display.

CACTA is a class 2 transposon, that is very abundantly present in plant genomes. Using Rim2/Hipa CACTA transposon display (hereafter Rim2/Hipa-TD), we analyzed several A-genome diploid Oryza species that have a high distribution of the CACTA motifs. High levels of polymorphism were detected within and between the Oryza species. The African taxa, O. glaberrima and O. barthii, both showed lower levels of polymorphism than the Asian taxa, O. sativa, O. rufipogon, and O. nivara. However, O. longistaminata, another African taxon, showed levels of polymorphism that were similar to the Asian taxa. The Latin American taxon, O. glumaepatula, and the Australian taxon, O. meridionalis, exhibited intermediate levels of polymorphism between those of the Asian and African taxa. The lowest level of polymorphism was observed in O. glaberrima (32.1%) and the highest level of polymorphism was observed in O. rufipogon (95.7%). The phylogenetic tree revealed three major groups at the genetic similarity level of 0.409. The first group consisted of three Asian taxa, O. sativa, O. rufipogon and O. nivara. The second group consisted of three African taxa, O. glaberrima, O. barthii, O. longistaminata, and an American taxon, O. glumaepatula. The third group contained an Australian taxon, O. meridionalis. The clustering patterns of these species matched well with their geographical origins. Rim2/Hipa-TD appears to be a useful marker system for studying the genetic diversity and species relationships among the AA diploid Oryza species.

Bulb Syringe and Pulsed Irrigation: Which Is More Effective to Remove Bacteria in Spine Surgeries?

STUDY DESIGN: Retrospective study. OBJECTIVES: The aim of this study was to investigate the difference of efficacy between bulb syringe irrigation and pulsed irrigation in spinal surgeries. SUMMARY OF BACKGROUND: Pulsed irrigation has been used for more than half a century, especially in trauma surgeries. However, to the best of our knowledge, there has been no study as to the efficacy of pulsed irrigation in spinal surgeries so far. MATERIALS AND METHODS: The consecutive patients who underwent posterior lumbar interbody fusion for 6 months were investigated. Those who underwent surgeries during the first 3 months were irrigated by bulb syringe (group I) and those who underwent surgeries during the next 3 months were irrigated by pulsatile irrigator with 17 psi (group II). Sex, age, surgery time, amount of blood loss, whether associated with diabetes mellitus, smoking, and amount of irrigation solution were compared between the 2 groups to identify homogeneity. Normal saline was used. Intraoperative irrigations were performed 3 times as time elapsed. After the last irrigation, culture specimens were obtained from the muscle layer and intervertebral space. The culture results were compared between the 2 groups. RESULTS: There were 79 cases in group I and 59 cases in group II. In the homogeneity test, surgery time was longer (P=0.011) and amount of irrigation saline was larger (P=0.042) in group II. From the posterior muscle layer, bacteria were cultured in 8/79 cases (10.1%) in group I and 1/59 cases (1.6%) in group II. There was a significant difference between the 2 groups (P=0.046). And from the intervertebral space, it was cultured in 6/79 cases (7.6%) in group I and 5/59 cases (8.5%) in group II. There was no difference between the 2 groups (P=0.546). Staphylococcus epidermidis, Staphylococcus aureus, Staphylococcus hominis, and Staphylococcus saprophyticus were cultured in order of the frequency. CONCLUSIONS: Pulsed irrigation was more effective compared with bulb syringe irrigation in the posterior muscle layer. In the intervertebral space, however, both methods were insufficient to eradicate contaminated bacteria.

High-risk drinking is associated with a higher risk of diabetes mellitus in Korean men, based on the 2010-2012 KNHANES.

We examined the association between alcohol-drinking pattern and diabetes mellitus (DM) in Korean adults. This cross-sectional study included 12,486 participants (5551 men and 6935 women) who participated in the 2010-2012 Korean National Health and Nutrition Examination Survey. We categorized alcohol-drinking pattern into three groups based on the alcohol-use disorders identification test (AUDIT): low-risk (score: 0-7), intermediate-risk (score: 8-14), and high-risk (score: ≥15). DM was defined as having fasting plasma glucose ≥126 mg/dL or taking glucose-lowering medication, including insulin therapy. In the study population, 25.2% of men and 4.7% of women were high-risk drinkers. DM prevalence was 9.2% in men and 5.4% in women. DM prevalence was 9.0% and 5.7% in the low-risk drinking group, 7.6% and 4.1% in the intermediate-risk drinking group, and 11.2% and 3.5% in the high-risk drinking group in men and women, respectively. Compared to the low-risk drinking group, odds ratios (95% confidence intervals) of men and women in the intermediate-risk drinking group for DM were 1.043 (0.779-1.396) and 1.139 (0.712-1.824), respectively, and 1.480 (1.133-1.933) and 0.827 (0.296-2.311) in the high-risk drinking group, after adjusting for age and other confounding factors. In conclusion, high-risk drinking appears to be associated with a higher risk of DM in men, but not in women.

The Survivorship and Clinical Results of Minimally Invasive Unicompartmental Knee Arthroplasty at 10-Year Follow-up.

BACKGROUND: In this study, we investigated the long-term clinical results and survival rate of minimally invasive unicompartmental knee arthroplasty (UKA) by collecting cases that had been implanted more than 10 years ago. METHODS: One hundred and twenty-eight patients (166 cases) who underwent Oxford phase 3 medial UKA using the minimally invasive surgery from January 2002 to December 2002 were selected. The mean age of the patients at the time of surgery was 61 years, and the duration of the follow-up was minimum 10 years. Clinical and radiographic assessments were performed using the Knee Society clinical rating system, and the survival analysis was done by the Kaplan-Meier method with 95% confidence interval (CI). RESULTS: The mean Knee Society knee and function scores improved significantly from 53.8 points (range, 25 to 70 points) and 56.1 points (range, 35 to 80 points) preoperatively to 85.4 points (range, 58 to 100 points) and 80.5 points (range, 50 to 100 points) at 10-year follow-up, respectively (p < 0.001). Failures following the UKA occurred in 16 cases (9.6%), and the mean time of the occurrence of the failure was 6.2 years after the surgery. The 10-year survival rate was 90.5% (95% CI, 85.9 to 95.0) when failure was defined as all the reoperations, whereas the 10-year survival rate was 93.4% (95% CI, 89.6 to 97.1) when the cases in which only revision total knee arthroplasty was defined as failure. CONCLUSIONS: The results of this study show outstanding functions of the knee joint and satisfactory 10-year survival rate after minimally invasive UKA. Therefore, minimally invasive UKA could be a useful method in the treatment of osteoarthritis in one compartment of knee joint.

Isolation of callus-specific mRNAs from differentiating embryogenic somatic calli of Pimpinella brachycarpa by cDNA-AFLP.

We used cDNA-AFLP to analyze gene expression profiles during somatic embryogenesis and to isolate stage-specific genes in Pimpinella brachycarpa. Ninety-six combinations of AFLP primers produced 3,785 fragments, of which 478, 403, and 436 were specific to calli, somatic embryos, and plants, respectively. 262 fragments were shared by callus and somatic embryo, 324 by somatic embryo and plant, and 222 by callus and plant. The number of fragments common to all three stages was 1,660. Seventy-five of the 478 callus-specific fragments were cloned and sequenced and their functions sought in the GenBank database. Thirty-three of them showed significant homology to GenBank data base entries and twelve of these were homologous to genes with products of known function such as ATPases, tubulins, kinases, transcription factors, ubiquitins, and ankyrin. Callus-specific expression was confirmed by performing RT-PCR with six of the twelve fragments. Since the somatic embryo is induced by auxin (Thidiazuron), possible down regulation of these genes by auxin is also discussed.

Demineralized bone matrix, as a graft enhancer of auto-local bone in posterior lumbar interbody fusion.

STUDY DESIGN: A case controlled study with prospective data collection. PURPOSE: To evaluate the early influence and the final consequence of demineralized bone matrix (DBM) on auto-local bone as a graft enhancer in posterior lumbar interbody fusion (PLIF). OVERVIEW OF LITERATURE: DBM is known as an osteoinductive material; however, it has not been clearly recognized to enhance auto-local bone with a small amount. METHODS: Patients who had a PLIF were allocated into two groups. Group I (70 cases) used auto-local bone chips and group II (44 cases) used DBM as an additive to auto-local bone, 1 mL per a segment. Group selection was alternated. Early assessment was performed by computed tomography at 6 months and final assessment was done by simple radiography after 24 months at least. The degree of bone formation was assessed by 4 grade scale. RESULTS: The subjects of both groups were homogenous and had similar Oswestry Disability Index at final assessment. The ratio of auto-local bone chips and DBM was 6:1. The degree of bone formation at 6 months after surgery was superior in group II. However, there was no significant difference between the two groups at the final assessment. CONCLUSIONS: DBM was not recognized to enhance auto-local bone with small amount.