Cyclops lesions associated with both bundles and selective bundle repair of the anterior cruciate ligament.

BACKGROUND: The remnant of a ruptured anterior cruciate ligament (ACL) can increase the risk of impingement or a cyclops lesion, which can increase challenges to proper tunnel placement. PURPOSE: To evaluate the prevalence of cyclops lesions after ACL reconstruction and to assess the difference in the incidence of cyclops lesions between single-bundle repair and selective bundle repair of the ACL. MATERIAL AND METHODS: This retrospective study included 151 patients who were diagnosed with an ACL tear after knee joint magnetic resonance imaging (MRI) who underwent ACL reconstruction surgery. MRI diagnosis of cyclops lesion formation was made if a soft-tissue mass was seen anteriorly in the intercondylar notch near the tibial insertion of the reconstructed ACL, based on sagittal T2-weighted (T2W) imaging. The size of the cyclops lesions was recorded as the largest diameter on the sagittal T2W imaging. RESULTS: A cyclops lesion was detected in 74 (38.5%) cases. Cyclops lesions were detected more frequently in cases with single-bundle repair of the ACL, but the results were not statistically significant (P = 0.609). Compared with selective bundle repair, cyclops lesions had a significantly higher prevalence in the posterolateral (PL) bundle repair than in the anteromedial (AM) bundle repair (P = 0.027) based on MR images at 6-12 months after surgery. CONCLUSION: The incidence of cyclops lesions did not differ significantly in single-bundle repair and selective bundle repair of ACL. However, selective PL bundle repair of the ACL showed a significantly increased incidence of cyclops lesions compared with selective AM bundle repair.

BAC cloning and heterologous expression of a giant biosynthetic gene cluster encoding antifungal neotetrafibricin in streptomyces rubrisoli.

Polyene natural products including nystatin A1, amphotericin B, ECO-02301, and mediomycin belong to a large family of valuable antifungal polyketide compounds typically produced by soil actinomycetes. A previous study (Park et al., Front. Bioeng. Biotechnol., 2021, 9, 692340) isolated Streptomyces rubrisoli Inha501 with strong antifungal activity and analyzed a large-sized biosynthetic gene cluster (BGC) of a linear polyene compound named Inha-neotetrafibricin (I-NTF) using whole genome sequencing and bioinformatics. In the present study, an entire I-NTF BGC (∼167 kb) was isolated through construction and screening of Streptomyces BAC library. Overexpression of the cloned I-NTF BGC in the wild-type S. rubrisoli Inha501 and its heterologous expression in S. lividans led to 2.6-fold and 2.8-fold increase in I-NTF yields, respectively. The qRT-PCR confirmed that the transcription levels of I-NTF BGC were significantly increased in both homologous and heterologous hosts containing the BAC integration of I-NTF BGC. In addition, the I-NTF aglycone-producing strains were constructed by a target-specific deletion of glycosyltransferase gene present in I-NTF BGC. A comparison of the in vitro biological activities of I-NTF and I-NTF aglycone confirmed that the rhamnose sugar motif of I-NTF plays a critical role in both antifungal and antibacterial activities. These results suggest that the Streptomyces BAC cloning of a large-sized natural product BGC is a valuable approach for natural product titer improvement and biological activity screening of natural product in actinomycetes.

Streptomyces BAC Cloning of a Large-Sized Biosynthetic Gene Cluster of NPP B1, a Potential SARS-CoV-2 RdRp Inhibitor.

As valuable antibiotics, microbial natural products have been in use for decades in various fields. Among them are polyene compounds including nystatin, amphotericin, and nystatin-like Pseudonocardia polyenes (NPPs). Polyene macrolides are known to possess various biological effects, such as antifungal and antiviral activities. NPP A1, which is produced by Pseudonocardia autotrophica, contains a unique disaccharide moiety in the tetraene macrolide backbone. NPP B1, with a heptane structure and improved antifungal activity, was then developed via genetic manipulation of the NPP A1 biosynthetic gene cluster (BGC). Here, we generated a Streptomyces artificial chromosomal DNA library to isolate a large-sized NPP B1 BGC. The NPP B1 BGC was successfully isolated from P. autotrophica chromosome through the construction and screening of a bacterial artificial chromosome (BAC) library, even though the isolated 140-kb BAC clone (named pNPPB1s) lacked approximately 8 kb of the right-end portion of the NPP B1 BGC. The additional introduction of the pNPPB1s as well as co-expression of the 32-kb portion including the missing 8 kb led to a 7.3-fold increase in the production level of NPP B1 in P. autotrophica. The qRT-PCR confirmed that the transcription level of NPP B1 BGC was significantly increased in the P. autotrophica strain containing two copies of the NPP B1 BGCs. Interestingly, the NPP B1 exhibited a previously unidentified SARS-CoV-2 RNA-dependent RNA polymerase (RdRp) inhibition activity in vitro. These results suggest that the Streptomyces BAC cloning of a large-sized, natural product BGC is a valuable approach for titer improvement and biological activity screening of natural products in actinomycetes.

Protein, Amino Acid, Oil, Fatty Acid, Sugar, Anthocyanin, Isoflavone, Lutein, and Antioxidant Variations in Colored Seed-Coated Soybeans.

Different physiological and genetic studies show that the variations in the accumulation of pigment-stimulating metabolites result in color differences in soybean seed coats. The objective of this study was to analyze the nutrient contents and antioxidant potential in black, brown, and green seed-coated soybeans. Significant variations in protein (38.9-43.3%), oil (13.9-20.4%), total sugar (63.5-97.0 mg/g seed), total anthocyanin (3826.0-21,856.0 µg/g seed coat), total isoflavone (709.5-3394.3 µg/g seed), lutein (1.9-14.8 µg/g), total polyphenol (123.0-385.8 mg gallic acid/100 g seed), total flavonoid (22.1-208.5 mg catechin/100 g seed), 2,2'-azino-bis(3-ethylbenzthiazoline-6-sulphonic acid (ABTS; 275.0-818.8 mg Trolox/100 g seed), and 2,2-diphenyl-1-picrylhydrazyl (DPPH; 96.3-579.7 mg Trolox/100 g seed) were found among the soybean genotypes. Ilpumgeomjeong2 contained the lowest protein but the highest oil and total sugar. The lowest oil-containing Wonheug had the highest protein content. Socheong2 was rich in all four variables of antioxidants. Anthocyanins were detected only in black soybeans but not in brown and green soybeans. The variation in isoflavone content was up to 5-fold among the soybean genotypes. This study could be a valuable resource for the selection and improvement of soybean because an understanding of the nutrient content and antioxidant potentials is useful to develop effective strategies for improving the economic traits; for example, the major emphasis of soybean breeding for fatty acids is to enhance the oleic and linoleic acid contents and to decrease linolenic acid content.

Quantitative Trait Locus Mapping for Drought Tolerance in Soybean Recombinant Inbred Line Population.

Improving drought stress tolerance of soybean could be an effective way to minimize the yield reduction in the drought prevailing regions. Identification of drought tolerance-related quantitative trait loci (QTLs) is useful to facilitate the development of stress-tolerant varieties. This study aimed to identify the QTLs for drought tolerance in soybean using a recombinant inbred line (RIL) population developed from the cross between a drought-tolerant 'PI416937' and a susceptible 'Cheonsang' cultivar. Phenotyping was done with a weighted drought coefficient derived from the vegetative and reproductive traits. The genetic map was constructed using 2648 polymorphic SNP markers that distributed on 20 chromosomes with a mean genetic distance of 1.36 cM between markers. A total of 10 QTLs with 3.52-4.7 logarithm of odds value accounting for up to 12.9% phenotypic variance were identified on seven chromosomes. Five chromosomes-2, 7, 10, 14, and 20-contained one QTL each, and chromosomes 1 and 19 harbored two and three QTLs, respectively. The chromosomal locations of seven QTLs overlapped or located close to the related QTLs and/or potential candidate genes reported earlier. The QTLs and closely linked markers could be utilized in maker-assisted selection to accelerate the breeding for drought tolerance in soybean.

Pathological Mechanism of a Constitutively Active Form of Stromal Interaction Molecule 1 in Skeletal Muscle.

Stromal interaction molecule 1 (STIM1) is the main protein that, along with Orai1, mediates store-operated Ca2+ entry (SOCE) in skeletal muscle. Abnormal SOCE due to mutations in STIM1 is one of the causes of human skeletal muscle diseases. STIM1-R304Q (a constitutively active form of STIM1) has been found in human patients with skeletal muscle phenotypes such as muscle weakness, myalgia, muscle stiffness, and contracture. However, the pathological mechanism(s) of STIM1-R304Q in skeletal muscle have not been well studied. To examine the pathological mechanism(s) of STIM1-R304Q in skeletal muscle, STIM1-R304Q was expressed in mouse primary skeletal myotubes, and the properties of the skeletal myotubes were examined using single-myotube Ca2+ imaging, transmission electron microscopy (TEM), and biochemical approaches. STIM1-R304Q did not interfere with the terminal differentiation of skeletal myoblasts to myotubes and retained the ability of STIM1 to attenuate dihydropyridine receptor (DHPR) activity. STIM1-R304Q induced hyper-SOCE (that exceeded the SOCE by wild-type STIM1) by affecting both the amplitude and the onset rate of SOCE. Unlike that by wild-type STIM1, hyper-SOCE by STIM1-R304Q contributed to a disturbance in Ca2+ distribution between the cytosol and the sarcoplasmic reticulum (SR) (high Ca2+ in the cytosol and low Ca2+ in the SR). Moreover, the hyper-SOCE and the high cytosolic Ca2+ level induced by STIM1-R304Q involve changes in mitochondrial shape. Therefore, a series of these cellular defects induced by STIM1-R304Q could induce deleterious skeletal muscle phenotypes in human patients carrying STIM1-R304Q.

Calsequestrin: a well-known but curious protein in skeletal muscle.

Calsequestrin (CASQ) was discovered in rabbit skeletal muscle tissues in 1971 and has been considered simply a passive Ca2+-buffering protein in the sarcoplasmic reticulum (SR) that provides Ca2+ ions for various Ca2+ signals. For the past three decades, physiologists, biochemists, and structural biologists have examined the roles of the skeletal muscle type of CASQ (CASQ1) in skeletal muscle and revealed that CASQ1 has various important functions as (1) a major Ca2+-buffering protein to maintain the SR with a suitable amount of Ca2+ at each moment, (2) a dynamic Ca2+ sensor in the SR that regulates Ca2+ release from the SR to the cytosol, (3) a structural regulator for the proper formation of terminal cisternae, (4) a reverse-directional regulator of extracellular Ca2+ entries, and (5) a cause of human skeletal muscle diseases. This review is focused on understanding these functions of CASQ1 in the physiological or pathophysiological status of skeletal muscle.

QTL Mapping and Candidate Gene Analysis for Pod Shattering Tolerance in Soybean (Glycine max).

Pod shattering is an important reproductive process in many wild species. However, pod shattering at the maturing stage can result in severe yield loss. The objectives of this study were to discover quantitative trait loci (QTLs) for pod shattering using two recombinant inbred line (RIL) populations derived from an elite cultivar having pod shattering tolerance, namely "Daewonkong", and to predict novel candidate QTL/genes involved in pod shattering based on their allele patterns. We found several QTLs with more than 10% phenotypic variance explained (PVE) on seven different chromosomes and found a novel candidate QTL on chromosome 16 (qPS-DS16-1) from the allele patterns in the QTL region. Out of the 41 annotated genes in the QTL region, six were found to contain SNP (single-nucleotide polymorphism)/indel variations in the coding sequence of the parents compared to the soybean reference genome. Among the six potential candidate genes, Glyma.16g076600, one of the genes with known function, showed a highly differential expression levels between the tolerant and susceptible parents in the growth stages R3 to R6. Further, Glyma.16g076600 is a homolog of AT4G19230 in Arabidopsis, whose function is related to abscisic acid catabolism. The results provide useful information to understand the genetic mechanism of pod shattering and could be used for improving the efficiency of marker-assisted selection for developing varieties of soybeans tolerant to pod shattering.

Study on physical fitness factors affecting race-class of Korea racing cyclists.

The purpose of this study is to compare and analyze the difference of isokinetic muscle functions, anaerobic capabilities, pedaling power and maximum strength according to race-class of Korea racing cyclists. The participants in this study were 57 racing cyclist candidates who graduated from the Korea racing school. One year after graduation, we confirmed race-class of candidates and they were divided into three groups: the first-class racer group (FC, n=14), second-class racer group (SC, n=29), third-class racer group (TC, n=14). The isokinetic muscle strength of trunk and knee flexion/extension was measured using HUMAC NORM and basic physical strength such as squat and bench press was analyzed by Ariel device. As results of this study, % Fat (percentage of body fat) in FC group was the lowest compared to other groups. Isokinetic knee flexion strength was higher in FC group than SC and TC racer groups. Peak and average pedaling power as well as maximum muscle strength were significantly higher in FC group than in other groups. Our findings suggest new evidence that pedaling power and isokinetic muscle strength might be closely associated with race-class of Korea racing cyclists.

Comparison of maximum muscle strength and isokinetic knee and core muscle functions according to pedaling power difference of racing cyclist candidates.

The purpose of this study was to investigate differences of maximum muscle strength and isokinetic knee and core muscle functions according to pedaling power of racing cyclist candidate. Subjects for this study were 200 racing cyclist candidates and divided into four groups: top 10% peak power group (TPP, n=20), low 10% peak power group (LPP, n=20), top 10% average power group (TAP, n=20), and low 10% average power group (LAP, n=20). The maximum muscle strength was consisted of grip strength, bench press and squat measured by Ariel device, and isokinetic knee and core muscle functions were analyzed by Humac Norm device. Significant differences between groups were determined with one-way repeated analysis of variance. As the result of this study, TPP and TAP groups showed significantly decreased body-fat mass and increased free fat mass when compared to LPP and LAP groups. The maximum strength of grip and squat was significantly higher in TPP and TAP than in other groups. Isokinetic knee extension and flexion strength was higher in TPP and TAP groups as well as isokinetic trunk extension and flexion functions were highest in TPP group. Thus, our findings suggest new evidence that muscle mass, maximal muscle strength, and isokinetic muscle functions might be important predictors of racing cyclist performance.

Abrogation of B-RafV600E induced senescence by FoxM1 expression.

B-RafV600E oncogene mutation occurs in various cancers and is associated with tumor initiation. However, genetic modification of B-RafV600E in cells induces MAPK activation and results in oncogene-induced senescence. Overcoming the oncogene-induced senescence by B-RafV600E requires activation of another oncogene pathway, such as AKT signaling. In the present study, we explored the factors involved in overcoming the senescence program in cells activated by B-RafV600E and AKT signaling. B-RafV600E activation caused a feedback inhibition of AKT phosphorylation and resulted in downregulation of FoxM1, one of the AKT downstream components. AKT activation by PTEN downregulation induced FoxM1 expression, and co-expression of B-RafV600E and FoxM1 overcame the cellular senescence. These observations suggested that FoxM1 is critical downstream gene of AKT and functions to overcome B-RafV600E-induced senescence.

Prevalence, toxin gene profile, antibiotic resistance, and molecular characterization of Clostridium perfringens from diarrheic and non-diarrheic dogs in Korea.

Clostridium perfringens causes diarrhea and other diseases in animals and humans. We investigated the prevalence, toxin gene profiles, and antibiotic resistance of C. perfringens isolated from diarrheic dogs (DD) and non-diarrheic dogs (ND) in two animal hospitals in Seoul, Korea. Fecal samples were collected from clinically DD (n = 49) and ND (n = 34). C. perfringens was isolated from 31 of 49 DD (63.3%) and 21 of 34 ND dogs (61.8%). All C. perfringens strains were positive for the α toxin gene, but not for the ß, ε, or ι toxin genes; therefore, all strains were identified as type A C. perfringens. All isolates were cpe-negative, whereas the ß2 toxin gene was identified in 83.9% and 61.9% of isolates from DD and ND, respectively. Most isolates were susceptible to ampicillin (94%), chloramphenicol (92%), metronidazole (100%), moxifloxacin (96%), and imipenem (100%). However, 25.0% and 21.2% of isolates were resistant to tetracycline and clindamycin, respectively. Molecular subtyping of the isolated strains was performed by using pulsed-field gel electrophoresis. Fifty-two isolates were classified into 48 pulsotypes based on more than 90% similarity of banding patterns. No notable differences were observed among the isolates from DD and ND.

Evaluation of marginal fit of 2 CAD-CAM anatomic contour zirconia crown systems and lithium disilicate glass-ceramic crown.

PURPOSE: This study was to evaluate the marginal fit of two CAD-CAM anatomic contour zirconia crown systems compared to lithium disilicate glass-ceramic crowns. MATERIALS AND METHODS: Shoulder and deep chamfer margin were formed on each acrylic resin tooth model of a maxillary first premolar. Two CAD-CAM systems (Prettau®Zirconia and ZENOSTAR®ZR translucent) and lithium disilicate glass ceramic (IPS e.max®press) crowns were made (n=16). Each crown was bonded to stone dies with resin cement (Rely X Unicem). Marginal gap and absolute marginal discrepancy of crowns were measured using a light microscope equipped with a digital camera (Leica DFC295) magnified by a factor of 100. Two-way analysis of variance (ANOVA) and post-hoc Tukey's HSD test were conducted to analyze the significance of crown marginal fit regarding the finish line configuration and the fabrication system. RESULTS: The mean marginal gap of lithium disilicate glass ceramic crowns (IPS e.max®press) was significantly lower than that of the CAD-CAM anatomic contour zirconia crown system (Prettau®Zirconia) (P<.05). Both fabrication systems and finish line configurations significantly influenced the absolute marginal discrepancy (P<.05). CONCLUSION: The lithium disilicate glass ceramic crown (IPS e.max®press) had significantly smaller marginal gap than the CAD-CAM anatomic contour zirconia crown system (Prettau®Zirconia). In terms of absolute marginal discrepancy, the CAD-CAM anatomic contour zirconia crown system (ZENOSTAR®ZR translucent) had under-extended margin, whereas the CAD-CAM anatomic contour zirconia crown system (Prettau®Zirconia) and lithium disilicate glass ceramic crowns (IPS e.max®press) had overextended margins.

Antagonist wear of three CAD/CAM anatomic contour zirconia ceramics.

STATEMENT OF PROBLEM: Little clinical information exists on the antagonist wear of anatomic contour zirconia crowns. PURPOSE: The purpose of this in vitro study was to evaluate the 2-body wear of antagonists for 3 computer-aided design and computer-aided manufacturing (CAD/CAM) anatomic contour zirconia ceramics and veneering porcelain when opposing natural human enamel. MATERIAL AND METHODS: Zirkonzahn Y-TZP (polished zirconia, zirconia with staining, zirconia with staining and glazing), Acucera Y-TZP, Wieland Y-TZP, and Noritake feldspathic ceramic were tested (6 groups). Eight disk-shaped specimens 15 mm in diameter and 5 mm thick were prepared for each group. Forty-eight specimens were fabricated for a wear test against maxillary premolars without caries or previous restorations with 240 000 masticatory cycles in a masticatory simulator. Before the experiment, the surface roughness of each ceramic was measured with a nanosurface 3-dimensional (3D) optical profiler. The surface of the specimens was observed at 50× and 1000× magnification with a field emission scanning electron microscope (FE-SEM) before and after the experiment. The data obtained were statistically analyzed by 1-way ANOVA and the Tukey test for post hoc analysis (α=.05). RESULTS: The SEM observations of each group revealed fine bubbles and porous surfaces in the Noritake feldspathic ceramic group, whereas the polished Zirkonzahn Y-TZP group, Acucera Y-TZP group, and Wieland Y-TZP group had smooth surfaces. The surface roughness of Zirkonzahn Y-TZP after staining and glazing was significantly greater than that of any other groups (P<.01). The tooth opposing the polished Zirkonzahn Y-TZP group demonstrated the least wear (1.11 ± 0.51 mm³), while Zirkonzahn Y-TZP with staining and glazing produced the greatest enamel wear (3.07 ± 0.98 mm³) among the zirconia groups. The Noritake feldspathic ceramic group showed significantly more antagonistic tooth wear than other groups (P<.05). CONCLUSIONS: The antagonist wear of 3 CAD/CAM anatomic contour zirconia ceramics was significantly less than the Noritake veneering ceramic because the surface character of Y-TZP is relatively uniform and homogeneous. Zirkonzahn Y-TZP with staining and glazing was significantly more abrasive than the other zirconia specimens tested. However, it was less abrasive than the Noritake veneering ceramic.

Effect of far-infrared irradiation on catechins and nitrite scavenging activity of green tea.

The processed green tea leaves were irradiated by far-infrared (FIR) at eight temperatures (80, 90, 100, 110, 120, 130, 140, and 150 degrees C) for 10 min. After FIR irradiation, green teas were prepared by soaking the leaves in boiling water, and the physicochemical characteristics of the green tea were determined. FIR irradiation at 90 degrees C increased total phenol contents of green tea from 244.7 to 368.5 mg/g and total flavanol contents from 122.0 to 178.7 mg/g, compared with non-irradiated control. FIR irradiation also significantly affected the amounts of epigallocatechin and epigallocatechin gallate. Nitrite scavenging activity also increased with increasing FIR irradiation until the temperature reached 110 degrees C. However, the overall color changes of green tea irradiated with FIR at 90 and 100 degrees C were negligible. These results indicate that the chemical quality of green tea is significantly affected by FIR irradiation temperature of the green tea leaves.

Encapsulated Bifidobacterium bifidum potentiates intestinal IgA production.

We asked whether Bifidobacterium bifidum regulates the synthesis of IgA by mucosal lymphoid cells. B. bifidum alone, but not Clostridium perfringens, significantly induced total IgA and IgM synthesis by both mesenteric lymph nodes (MLN) and Peyer's patch (PP) cells. We, further, investigated the mucosal antibody production following peroral administration of B. bifidum to mice. Ingested B. bifidum significantly increased the number of Ig (IgM, IgG, and IgA) secreting cells in the culture of both MLN and spleen cells. Nonetheless, B. bifidum itself does not induce the own specific antibody responses, implying that B. bifidum does not provoke unnecessary immune reaction. Subsequently, it was found that encapsulation of B. bifidum further augments the total IgA production in the culture of both MLN and spleen cells. Finally, we found that the immuno-stimulating activity of B. bifidum is due to its cellular components but not due to any actively secreting component(s) from bacteria.

Regulation of TNF-alpha-mediated hyperplasia through TNF receptors, TRAFs, and NF-kappaB in synoviocytes obtained from patients with rheumatoid arthritis.

Although the etiology of rheumatoid arthritis (RA) has not been clearly understood to date, the hyperplasia of the synovial membrane imposed by pro-inflammatory cytokines has been suggested to play a crucial role in the progression of this disease. TNF-alpha, a potent pro-inflammatory cytokine, was detected at highly enhanced concentrations in the blood and synovial fluids of patients with RA relative to those of patients with osteoarthritis and normal subjects. To evaluate the role of TNF-alpha in the synovial hyperplasia during the pathogenic state, we investigated cellular outcomes and molecular mechanisms of synoviocytes in response to TNF-alpha. Following TNF-alpha treatment, fibroblast-like synoviocytes (FLS) obtained from patients with RA proliferated, unlike the cells from a normal subject that were unaffected. This TNF-alpha induced proliferation of synoviocytes obtained from RA patients coincided with down-regulation of TNFR1 and up-regulation of TNFR2 and TRAF1-6, as well as NF-kappaB activation. TNF-alpha-induced proliferation of synoviocytes was inhibited by transfection with a dominant negative mutant form of I-kappaBalpha cDNA (I-kappaBalphadN). Moreover, following TNF-alpha treatment, transfectants with I-kappaBalphadN underwent apoptosis, whereas mock-transfectants did not. Taken together, these results suggest that high levels of TNF-alpha present in RA synovium play an important role in the synovial hyperplasia of RA by suppressing apoptosis and promoting proliferation of synoviocytes through NF-kappaB-dependent signaling pathways mediated by up-regulated TNFR2 and TRAF1-6 molecules.