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Abstract: We report a first-in-human clinical trial using chimeric antigen receptor (CAR) T cells targeting CD37, an antigen highly expressed in B- and T-cell malignancies. Five patients with relapsed or refractory CD37+ lymphoid malignancies were enrolled and infused with autologous CAR-37 T cells. CAR-37 T cells expanded in the peripheral blood of all patients and, at peak, comprised >94% of the total lymphocytes in 4 of 5 patients. Tumor responses were observed in 4 of 5 patients with 3 complete responses, 1 mixed response, and 1 patient whose disease progressed rapidly and with relative loss of CD37 expression. Three patients experienced prolonged and severe pancytopenia, and in 2 of these patients, efforts to ablate CAR-37 T cells, which were engineered to coexpress truncated epidermal growth factor receptor, with cetuximab were unsuccessful. Hematopoiesis was restored in these 2 patients after allogeneic hematopoietic stem cell transplantation. No other severe, nonhematopoietic toxicities occurred. We investigated the mechanisms of profound pancytopenia and did not observe activation of CAR-37 T cells in response to hematopoietic stem cells in vitro or hematotoxicity in humanized models. Patients with pancytopenia had sustained high levels of interleukin-18 (IL-18) with low levels of IL-18 binding protein in their peripheral blood. IL-18 levels were significantly higher in CAR-37-treated patients than in both cytopenic and noncytopenic cohorts of CAR-19-treated patients. In conclusion, CAR-37 T cells exhibited antitumor activity, with significant CAR expansion and cytokine production. CAR-37 T cells may be an effective therapy in hematologic malignancies as a bridge to hematopoietic stem cell transplant. This trial was registered at www.ClinicalTrials.gov as #NCT04136275., (© 2024 American Society of Hematology. Published by Elsevier Inc. All rights are reserved, including those for text and data mining, AI training, and similar technologies.)

Aim: To determine the tensile load capacity (TLC) and the tearing characteristics for interrupted and vertical mattress sutures with different insertion points from the wound margin, and the effect of the bite size when using vertical mattress sutures., Materials and Methods: A total of 120 gingiva and lining mucosa samples obtained from pig jaws were divided into groups according to the suturing technique (interrupted and vertical mattress sutures), distance of the insertion points from the wound margin (margin, 1, 3, and 5 mm) and bite size (1, 3, and 5 mm). The TLC of the suture and the tearing characteristics were evaluated using a tensile tester device., Results: The TLC was significantly higher for vertical mattress sutures than for interrupted sutures regardless of the distance of the insertion points from the wound margin (intergroup p < .001). This distance significantly influenced the TLC for vertical mattress sutures (p < .05) but not for interrupted sutures (p > .05). Testing the tearing characteristics revealed that no tissue tearing occurred in groups when the insertion points were more than 3 mm from the wound margin., Conclusion: The TLC is higher for vertical mattress sutures than for interrupted sutures, and it increases when the insertion points are farther from the wound margin., (© 2024 The Authors. Clinical Oral Implants Research published by John Wiley & Sons Ltd.)

Chimeric antigen receptor (CAR) modified T cell therapies targeting BCMA have displayed impressive activity in the treatment of multiple myeloma. There are currently two FDA licensed products, ciltacabtagene autoleucel and idecabtagene vicleucel, for treating relapsed and refractory disease. Although correlative analyses performed by product manufacturers have been reported in clinical trials, there are limited options for reliable BCMA CAR T detection assays for physicians and researchers looking to explore it as a biomarker for clinical outcome. Given the known association of CAR T cell expansion kinetics with toxicity and response, being able to quantify BCMA CAR T cells routinely and accurately in the blood of patients can serve as a valuable asset. Here, we optimized an accurate and sensitive flow cytometry test using a PE-conjugated soluble BCMA protein, with a lower limit of quantitation of 0.19% of CD3+ T cells, suitable for use as a routine assay for monitoring the frequency of BCMA CAR T cells in the blood of patients receiving either ciltacabtagene autoleucel or idecabtagene vicleucel., (© 2024 International Clinical Cytometry Society.)

Salt stress is a significant abiotic stress that reduces crop yield and quality globally. In this study, we utilized RNA sequencing (RNA-Seq) to identify differentially expressed genes (DEGs) in response to salt stress induced by gamma-ray irradiation in a salt-tolerant soybean mutant. The total RNA library samples were obtained from the salt-sensitive soybean cultivar Kwangan and the salt-tolerant mutant KA-1285. Samples were taken at three time points (0, 24, and 72 h) from two tissues (leaves and roots) under 200 mM NaCl. A total of 967,719,358 clean reads were generated using the Illumina NovaSeq 6000 platform, and 94.48% of these reads were mapped to 56,044 gene models of the soybean reference genome (Glycine&#95;max&#95;Wm82.a2.v1). The DEGs with expression values were compared at each time point within each tissue between the two soybeans. As a result, 296 DEGs were identified in the leaves, while 170 DEGs were identified in the roots. In the case of the leaves, eight DEGs were related to the phenylpropanoid biosynthesis pathway; however, in the roots, Glyma.03G171700 within GmSalt3 , a major QTL associated with salt tolerance in soybean plants, was differentially expressed. Overall, these differences may explain the mechanisms through which mutants exhibit enhanced tolerance to salt stress, and they may provide a basic understanding of salt tolerance in soybean plants.

Chimeric Antigen Receptor (CAR) T cells directed to B cell maturation antigen (BCMA) mediate profound responses in patients with multiple myeloma, but most patients do not achieve long-term complete remissions. In addition, recent evidence suggests that high-affinity binding to BCMA can result in on-target, off-tumor activity in the basal ganglia and can lead to fatal Parkinsonian-like disease. Here we develop CAR T cells against multiple myeloma using a binder to targeting transmembrane activator and CAML interactor (TACI) in mono and dual-specific formats with anti-BCMA. These CARs have robust, antigen-specific activity in vitro and in vivo. We also show that TACI RNA expression is limited in the basal ganglia, which may circumvent some of the toxicities recently reported with BCMA CARs. Thus, single-targeting TACI CARs may have a safer toxicity profile, whereas dual-specific BCMA-TACI CAR T cells have potential to avoid the antigen escape that can occur with single-antigen targeting., (© 2023. The Author(s).)

Immune evasion is a critical step of cancer progression that remains a major obstacle for current T cell-based immunotherapies. Hence, we investigated whether it is possible to genetically reprogram T cells to exploit a common tumor-intrinsic evasion mechanism whereby cancer cells suppress T-cell function by generating a metabolically unfavorable tumor microenvironment (TME). In an in silico screen, we identified ADA and PDK1 as metabolic regulators. We then showed that overexpression (OE) of these genes enhanced the cytolysis of CD19-specific chimeric antigen receptor (CAR) T cells against cognate leukemia cells, and conversely, ADA or PDK1 deficiency dampened this effect. ADA-OE in CAR T cells improved cancer cytolysis under high concentrations of adenosine, the ADA substrate, and an immunosuppressive metabolite in the TME. High-throughput transcriptomics and metabolomics analysis of these CAR T cells revealed alterations of global gene expression and metabolic signatures in both ADA- and PDK1-engineered CAR T cells. Functional and immunologic analyses demonstrated that ADA-OE increased proliferation and decreased exhaustion in CD19-specific and HER2-specific CAR T cells. ADA-OE improved tumor infiltration and clearance by HER2-specific CAR T cells in an in vivo colorectal cancer model. Collectively, these data unveil systematic knowledge of metabolic reprogramming directly in CAR T cells and reveal potential targets for improving CAR T-cell therapy., (©2023 American Association for Cancer Research.)

Auricularia is one of the broadly cultivated edible mushrooms in Korea. Most of the Korean Auricularia strains used for cultivation and breeding are known as A. auricula-judae . Recently, this species has been reported to belong to a species complex. Therefore, this study was carried out to genetically clarify the bred and cultivated Korean A. auricula-judae strains. The internal transcribed spacer (ITS) and IGS1 rDNA region sequences were determined from 10 A. auricula-judae strains by PCR and sequencing. Variation in the nucleotide sequence and sequence length of the two rDNA regions were found among the seven A. auricula-judae strains. A maximum-likelihood (ML) phylogenetic tree based on the ITS sequences clearly placed all the 10 Korean A. auricula-judae strains in the A. heimuer clade of the A. auricula-judae complex. A. heimuer is diverged from A. auricula-judae . An ML phylogenetic tree based on the IGS1 sequences revealed the close relationship between Korean A. heimuer strains to Chinese A. heimuer strains. But each strain could be distinguishable by the IGS1 sequence. Furthermore, progeny strains in the seven Korean strains could be differentiated from their parental strains by the IGS1 sequence based phylogenetic tree. Our results are expected to be used to complement the distinction of domestic Auricularia cultivars., Competing Interests: No potential conflict of interest was reported by the authors., (© 2023 The Author(s). Published by Informa UK Limited, trading as Taylor & Francis Group on behalf of the Korean Society of Mycology.)

Background: We aimed to compare glymphatic dysfunction between patients with end-stage renal disease (ESRD) and healthy controls and analyze the correlation between the glymphatic function and clinical characteristics using the diffusion tensor image analysis along with the perivascular space (DTI-ALPS) index., Methods: We prospectively enrolled neurologically asymptomatic 49 patients with ESRD undergoing dialysis and 38 healthy controls. Diffusion tensor image was conducted using the same 3T scanner, and the DTI-ALPS index was calculated. We compared the DTI-ALPS index between the patients with ESRD and healthy controls. In addition, we conducted a correlation analysis between the clinical characteristics and DTI-ALPS index in patients with ESRD., Results: There were significant differences in the DTI-ALPS index between patients with ESRD and healthy controls. The DTI-ALPS index in patients with ESRD was lower than that in healthy controls (1.460 vs. 1.632, p = 0.003). In addition, there was a significant positive correlation between the DTI-ALPS index and serum parathyroid hormone levels ( r = 0.357, p = 0.011)., Conclusion: We demonstrated glymphatic dysfunction in patients with ESRD, as revealed by the DTI-ALPS index. This study also reveals the feasibility of the DTI-ALPS method to determine glymphatic function in patients with ESRD, which could be used in future research studies., Competing Interests: BY was employed by DEEPNOID. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Heo, Lee, Park, Lee, Park, Kim, Lee, Yoo and Park.)

Aim: To determine the healing outcome following grafting with deproteinized porcine bone mineral (DPBM) with or without collagen membrane coverage in two-wall (both buccal and lingual)-damaged extraction sockets., Materials and Methods: Distal roots of three mandibular premolars in six beagle dogs were extracted, and the whole buccal and lingual bony walls were surgically removed. Three treatment protocols were then applied according to the following group allocation: no graft (None), grafting DPBM (BG), and grafting DPBM with coverage by a collagen membrane (BG + M). Two observational periods (2 and 8 weeks) were used with the split-mouth design, and quantitative and qualitative analyses were performed by microcomputed tomography and histology., Results: The dimensions of the alveolar ridge at both grafted sites (BG and BG + M) remained similar to those of the pristine ridge in the histologic and radiographic analyses, whereas the ungrafted sites (None) collapsed both vertically and horizontally. Both grafting protocols produced substantial bony regeneration, but the addition of a covering membrane enhanced the proportion of mineralized tissue within the augmented area, and the BG + M group also showed a significantly larger area of regenerated ridge than the None group (p < .05)., Conclusions: Bone grafting with collagen membrane can maintain the alveolar ridge dimensions with substantial bone regeneration in a two-wall-damaged extraction socket., (© 2021 The Authors. Clinical Oral Implants Research published by John Wiley & Sons Ltd.)

Purpose: This study was conducted to determine how the distance of the near insertion points in a vertical mattress suture from the wound margin influences the pattern of primary closure in an in vitro experimental model., Methods: Pairs of 180 porcine gingival and alveolar mucosa samples were harvested from 90 pig jaws and fixed to a specially designed model. A vertical mattress suture was performed with the near insertion point at 3 different distances from the wound margin (1-, 3-, and 5-mm) on both the gingival and mucosal samples (6 groups; n=30 for each group). The margin discrepancy and the presence of epithelium between the wound margins were measured on histologic slides., Results: The margin discrepancy decreased significantly as the near insertion point became closer to the wound margin both in mucosal tissue (0.241±0.169 mm, 0.945±0.497 mm, and 1.306±0.773 mm for the 1-, 3-, and 5-mm groups, respectively) and in gingival tissue (0.373±0.304 mm, 0.698±0.431 mm, and 0.713±0.691 mm, respectively). The frequency of complications of wound margin adaptation reduced as the distance of the near insertion point from the wound margin decreased both in the mucosal and gingival tissues., Conclusions: Placing the near insertion point close to the wound margin enhances the precision of wound margin approximation/adaptation using a vertical mattress suture., Competing Interests: No potential conflict of interest relevant to this article was reported., (Copyright © 2021. Korean Academy of Periodontology.)

Nanoparticles and nanofluids have been implemented in energy harvesting devices, and energy harvesting based on magnetic nanofluid flow was recently achieved by using a layer-built magnet and microbubble injection to induce a voltage on the order of 10-1 mV. However, this is not yet suitable for some commercial purpose. The air bubbles must be segmented in the base fluid, and the magnetic flux of the ferrofluids should change over time to increase the amount of electric voltage and current from energy harvesting. In this study, we proposed a novel technique to achieve segmented flow of the ferrofluids and the air layers. This segmented ferrofluid flow linear generator can increase the magnitude of the induced voltage from the energy harvesting system. In our experiments, a ferrofluid-filled capsule produced time-dependent changes in the magnetic flux through a multi-turn coil, and the induced voltage was generated on the order of about 101 mV at a low frequency of 2 Hz. A finite element analysis was used to describe the time-dependent change of the magnetic flux through the coil according to the motion of the segmented flow of the ferrofluid and the air-layer, and the induced voltage was generated to the order of 102 mV at a high frequency of 12.5 Hz.

Metal nanoparticles are used in applications ranging from bio-diagnostics to catalysis due to the expectation to improve attributes or the performance of specific products or processes. The electric arc can be used to produce metal nanoparticles by evaporating the anode and forming the anode vapor. In order to synthesize the nanoparticles of the desired properties, the influence of various input parameters on the growth kinetics has to be fully understood. In this study, we presented two and three dimensional results of numerical simulation of the transferred electric arc taking into account the interaction model between an electric arc and two electrodes. It was found that the predicted temperature of the arc column with two electrodes was in good agreement with the measured data, and the main advantage of this model over our previous one was to predict the temperature distribution of the arc column with two electrodes by two- and three-dimensional computations.

A numerical simulation was developed for magnetic nanoparticles in a liquid dielectric to investigate the AC breakdown voltage of the magnetic nanofluids according to the volume concentration of the magnetic nanoparticles. In prior research, we found that the dielectric breakdown voltage of the transformer oil-based magnetic nanofluids was positively or negatively affected according to the amount of magnetic nanoparticles under a testing condition of dielectric fluids, and the trajectory of the magnetic nanoparticles in a fabricated chip was visualized to verify the related phenomena via measurements and computations. In this study, a numerical simulation of magnetic nanoparticles in an insulating fluid was developed to model particle tracing for AC breakdown mechanisms happened to a sphere-sphere electrode configuration and to propose a possible mechanism regarding the change in the breakdown strength due to the behavior of the magnetic nanoparticles with different applied voltages.

Nanoparticles and nanofluids have been implemented in energy harvesting devices, and energy harvesting based on magnetic nanofluid flow was recently achieved by using a layer-built magnet and micro-bubble injection to induce a voltage on the order of 10-1 mV. However, this is not yet suitable for some commercial purpose. In order to further increase the amount of electric voltage and current from this energy harvesting the air bubbles must be segmented in the base fluid, and the magnetic flux of the segmented flow should be materially altered over time. The focus of this research is on the development of a segmented ferrofluid flow linear generator that would scavenge electrical power from waste heat. Experiments were conducted to obtain the induced voltage, which was generated by moving a ferrofluid-filled capsule inside a multi-turn coil. Computations were then performed to explain the fundamental physical basis of the motion of the segmented flow of the ferrofluids and the air-layers.

ZnO nanoparticles (NPs) with monolayer structures were prepared by atomic layer deposition (ALD) to use for a charge-trap layer (CTL) for nonvolatile memory thin-film transistors (MTFTs). The optimum ALD temperature of the NP formation was demonstrated to be 160 °C. The size and areal density of the ZnO NPs was estimated to be approximately 33 nm and 4.8 × 10 9 cm -2 , respectively, when the number of ALD cycles was controlled to be 20. The fabricated MTFTs using a ZnO-NP CTL exhibited typical memory window properties, which are generated by charge-trap/de-trap processes, in their transfer characteristics and the width of the memory window (MW) increased from 0.6 to 18.0 V when the number of ALD cycles increased from 5 to 30. The program characteristics of the MTFT were markedly enhanced by the post-annealing process performed at 180 °C in an oxygen ambient due to the improvements in the interface and bulk qualities of the ZnO NPs. The program/erase (P/E) speed was estimated to be 10 ms at P/E voltages of -14 and 17 V. The memory margin showed no degradation with the lapse in retention time for 2 × 10 4 s and after the repetitive P/E operations of 7 × 10 3 cycles.

Objective: Asthma and chronic obstructive pulmonary disease (COPD) have distinct pathophysiological mechanisms but sometimes share similar clinical manifestations. Distinguishing between these diseases is important. This study compared the profiles of serum biomarkers between patients with asthma and those with COPD., Methods: Serum levels of the chitinase like protein YKL-40, periostin, interleukin (IL)-18, and chemokine (C--C motif) ligand 18 (CCL18) were measured in asthma patients (n = 20), COPD patients (n = 16), and normal controls (n = 20)., Results: Serum levels of YKL-40 were higher in COPD patients [median (range), 55 (17-565) versus 208 (74-922) ng/mL, p < 0.0001], but no differences were observed between asthma and COPD patients after adjusting for age and forced expiratory volume in 1 s (FEV1). No differences in serum levels of periostin, IL-18, or CCL18 were observed between the patient groups. Total IgE and airway hypersensitivity were negatively correlated (r = -0.485, p = 0.007). CCL18 levels were related to patients' age in asthmatic patients (r = -0.562, p = 0.010). Serum levels of CCL18 and IL-18 were positively correlated in patients with COPD (r = 0.696, p = 0.003)., Conclusions: No differences in the serum profiles of periostin, IL-18, or CCL18 were observed between patients with asthma and those with COPD. Serum levels of YKL-40 were not different between asthma and COPD patients after adjusting for age and FEV1. There were negative correlation between CCL18 and age in patients with asthma and positive correlation between IL-18 and CCL18 in patients with COPD.

Persicariae Rhizoma (PR) has been used as an anti-inflammatory and detoxification agent in Korea, and contains the biologically active dyes purple indirubin and blue indigo. Despite synthetic indigo showing genotoxic potential, thorough studies have not been carried out on the genotoxicity of PR. The potential genotoxicity of an aqueous extract of PR containing indigo (0.043%) and indirubin (0.009%) was evaluated using a standard battery of tests for safety assessment. The PR extract did not induce any genotoxic effects under the conditions of this study. The results of a reverse mutation assay in four Salmonella typhimurium strains and one Escherichia coli strain indicated that PR extract did not increase the frequency of revertant colonies in any strain, regardless of whether S9 mix was present or not. The PR extract also did not increase chromosomal aberrations in the presence or absence of S9 mix. Although slight signs of diarrhea were restrictedly detected in the mice treated with 2,000 mg/kg PR extract, no noteworthy changes in the frequency of micronucleated polychromatic erythrocytes were observed at doses ≤2,000 mg/kg in a bone marrow micronucleus test. These results indicate the potential safety of the PR extract, particularly if it is consumed in small amounts compared with the quantities used in the genotoxicity tests.

Temperature in a free-burning arc used for synthesis of nanoparticles and nanostructured materials is generally around 20,000 K just below the cathode, falling to about 15,000 K just above the anode, and decreasing rapidly in the radial direction. Therefore, the electrode erosion is indispensable for these atmospheric plasma systems, as well as for switching devices, due to the high heat flux transferred from high temperature arcs to electrodes, but experimental and theoretical works have not identified the characteristic phenomena because of the complex physical processes. To the previous study, we have focused on the arc self-induced fluid flow in a free-burning arc using the computational fluid dynamics (CFD) technique. At this time, our investigation is concerned with the whole region of free-burning high-intensity arcs including the tungsten cathode, the arc plasma and the anode using a unified numerical model for applying synthesis of nanoparticles and nanostructured materials practically.

Sanguinarine is a benzophenanthridine alkaloid, derived from the root of Sanguinaria canadensis and other poppy Fumaria species, which is known to have antimicrobial, antiinflammatory and antioxidant properties. Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) is known to induce apoptosis in cancer cells but spare most normal cells. However, its effects are limited in some types of cancer cells, including AGS human gastric adenocarcinoma cells. In the present study, we showed that treatment with TRAIL in combination with subtoxic concentrations of sanguinarine sensitized TRAIL-mediated apoptosis in AGS cells. Combined treatment with sanguinarine and TRAIL effectively induced Bid cleavage and loss of mitochondrial membrane potential, leading to the activation of caspases, and cleavage of poly(ADP-ribose) polymerase and beta-catenin. The cytotoxic effects of the combined treatment were significantly inhibited by z-DEVD-fmk, a caspase-3 inhibitor, which demonstrates the important role of caspase-3 in the observed cytotoxic effect. In addition, the levels of Akt protein were markedly reduced in cells co-treated with sanguinarine and TRAIL. Apoptosis induced by the combined treatment was markedly increased by the phosphatidylinositol-3'-kinase inhibitor, LY294002 (Akt-upstream inhibitor), through the mitochondrial amplification step and caspase activation, suggesting that interactions of the synergistic effect were at least partially mediated through the Akt-dependent pathway.

Methionine sulphoxide reductases (Msr) catalyse the reduction of oxidized methionine to methionine. These enzymes are divided into two classes, MsrA and MsrB, according to substrate specificity. Although most MsrA and MsrB exist as separate enzymes, in some bacteria these two enzymes are fused to form a single polypeptide (MsrAB). Here, we report the first crystal structure of MsrAB from Streptococcus pneumoniae (SpMsrAB) at 2.4 A resolution. SpMsrAB consists of an N-terminal MsrA domain, a C-terminal MsrB domain and a linker. The linker is composed of 13 residues and contains one 3(10)-helix and several hydrogen bonds interacting with both MsrA and MsrB domains. Interestingly, our structure includes the MsrB domain complexed with an SHMAEI hexa-peptide that is the N-terminal region of neighbouring MsrA domain. A kinetic analysis showed that the apparent K(m) of SpMsrAB for the R-form-substrate was 20-fold lower than that for the S-form substrate, indicating that the MsrB domain had a much higher affinity for the substrate than the MsrA domain. Our study reveals the first structure of the MsrAB by providing insights into the formation of a disulphide bridge in the MsrB, the structure of the linker region, and the distinct structural nature of active site of each MsrA and MsrB domain.

Piceatannol is a polyphenol that is found in abundant quantities in grapes and wine. Although recent experimental data revealed the proapoptotic potency of piceatannol, the molecular mechanisms underlying the anti-leukemic activity have not yet been studied in detail. This study examined the effects of piceatannol on the growth of the human leukemia cell line U937. The results showed that piceatannol inhibits the viability of U937 cells by inducing apoptosis, as evidenced by the formation of apoptotic bodies, DNA fragmentation and the accumulation of the sub-G1 phase. RT-PCR and immunoblotting data showed that treating the cells with piceatannol caused the down-regulation of anti-apoptotic Bcl-2 and cIAP-2 expression. Piceatannol-induced apoptosis was also associated with the proteolytic activation of caspase-3, and the degradation/cleavage of poly (ADP-ribose) polymerase protein. z-DEVD-fmk, a caspase-3-specific inhibitor, blocked the activation of caspase-3 and increased the survival of the piceatannol-treated U937 cells, suggesting that caspase-3 activation is essential for piceatannol-induced apoptosis.

Aim: To investigate whether subtoxic concentration of beta-sitosterol (SITO) combined with TNF-related apoptosis-inducing ligand (TRAIL) induces apoptosis in TRAIL-resistant MDA-MB-231 breast cancer cells., Methods: Cell viability and growth were assessed by 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphnyl-2H-tetrazolim bromide assays, chromatin condensation, release of lactate dehydrogenase (LDH), and Annexin V+ cells. The apoptosis-related proteins were detected by Western blotting., Results: Treatment with TRAIL in combination with subtoxic concentrations of SITO sensitized MDA-MB-231 breast cancer cells to TRAIL-mediated apoptosis. The synergistic treatment induced chromatin condensation, DNA fragmentation, the release of LDH, and Annexin V+ cells. The indicators of apoptosis are correlated to the induction of caspase activities, which results in the cleavage of poly(ADP-ribose)polymerase. Both the cytotoxic effects and apoptotic characteristics induced by the synergistic treatment were significantly inhibited by a pan-caspase inhibitor z-VAD-fmk, demonstrating the important role of caspases. These results indicate that caspases are crucial regulators of apoptosis induced by the combined treatment of SITO and TRAIL in MDA-MB-231 cells., Conclusion: The synergistic treatment of SITO and TRAIL induces apoptosis, which can serve as a potential preventive and therapeutic agent.

In archaea, RNA endonucleases that act specifically on RNA with bulge-helix-bulge motifs play the main role in the recognition and excision of introns, while the eukaryal enzymes use a measuring mechanism to determine the positions of the universally positioned splice sites relative to the conserved domain of pre-tRNA. Two crystallographic structures of tRNA intron-splicing endonuclease from Thermoplasma acidophilum DSM 1728 (EndA(Ta)) have been solved to 2.5-A and 2.7-A resolution by molecular replacement, using the 2.7-A resolution data as the initial model and the single-wavelength anomalous-dispersion phasing method using selenomethionine as anomalous signals, respectively. The models show that EndA(Ta) is a homodimer and that it has overall folding similar to that of other archaeal tRNA endonucleases. From structural and mutational analyses of H236A, Y229F, and K265I in vitro, we have demonstrated that they play critical roles in recognizing the splice site and in cleaving the pre-tRNA substrate.

Rheumatoid arthritis (RA) is a chronic inflammatory disease that is characterized by hyperplasia of the synovial fibroblasts, which is partly the result of decreased apoptosis. This study investigated the mechanisms through which curcumin, a polyphenolic compound from the rhizome of Curcuma longa, exerts its anti-proliferative action in the synovial fibroblasts obtained from patients with RA. Exposure of the synovial fibroblasts to curcumin resulted in growth inhibition and the induction of apoptosis, as measured by MTT assay, fluorescent microscopy and Annexin-V-based assay. RT-PCR and immunoblotting showed that treating the cells with curcumin resulted in the down-regulation of anti-apoptotic Bcl-2 and the X-linked inhibitor of the apoptosis protein as well as the up-regulation of pro-apoptotic Bax expression in a concentration-dependent manner. Curcumin-induced apoptosis was also associated with the proteolytic activation of caspase-3 and caspase-9, and the concomitant degradation of poly(ADP-ribose) polymerase protein. Furthermore, curcumin decreased the expression levels of the cyclooxygenase (COX)-2 mRNA and protein without causing significant changes in the COX-1 levels, which was correlated with the inhibition of prostaglandin E(2) synthesis. These results show that curcumin might help identify a new therapeutic pathway against hyperplasia of the synovial fibroblasts in RA.

Two isolates of Tricholoma matsutake T-008 and T-034, preserved in Entomopathogenic Fungal Culture Collection (EFCC) of Korea, were used in the present study. The isolates had 100% Bootstrap homology with Tricholoma matsutake U62964 and T. matsutake AB188557 and AF309538 preserved in Gene Bank of NCBI. Mycelial growth of T. matsutake was highest in TMM and MYA at 25℃. The highest dry wt. of mycelium was obtained after 65 days of culture, when 6 mycelial discs were inoculated in 100 ml of broth in 250 ml shaking flask. Mycelial mats were observed in clumped condition at the inoculation sites of pine forest after two weeks of inoculation. After 5 months of inoculation, mycelia mats were observed growing inside soil and walls of a few inoculation sites, while mycelial mats growth up to 5~8 cm were observed in the roots of pine tree after 6 months. The survival rate of the inoculum was about 40% of the total inoculation sites. The survival rate was found below 20% when the mycelium was inoculated in the summer. The reasons for low survival rates of the mycelium were mainly due to dry season and the soil-borne small animals such as earthworm and mole. After one year of inoculation, no external difference was observed between the artificially inoculated mycelia and the naturally existing mycelia of T. matsutake. The present study showed that fruiting bodies of T. matsutake could be produced by artificial inoculation under the appropriate environmental conditions.

In vitro fruiting bodies were produced from ten different isolates of Cordyceps militaris EFCC C-5736, EFCC C-5941, EFCC C-5976, EFCC C-6040, EFCC C-6849, EFCC C-7268, EFCC C-7342, EFCC C-7992, EFCC C-8027 and EFCC C-8549. Single ascospores were isolated from in vitro grown fruiting bodies and used for fruiting body production in brown rice medium by both intra-strain crossing and out-crossing. Length and dry wt. of stromata grown in vitro were measured. Strains producing highest dry wt. of stromata were selected. Both intra-strain crossings and inter-strain crossings of single ascospore strains were found to produce profuse fruiting bodies of C. militaris.

Dideoxypetrosynol A, a polyacetylene from the marine sponge Petrosia sp., is known to exhibit significant selective cytotoxic activity against a small panel of human tumor cell lines, the mechanisms of which however, are poorly understood. The aim of the present study was to further elucidate the possible mechanisms by which dideoxypetrosynol A exerts its anti-proliferative action in cultured human monocytic leukemia U937 cells. We observed that the proliferation-inhibitory effect of dideoxypetrosynol A was due to the induction of G1 arrest in the cell cycle, the effects of which were associated with up-regulation of cyclin D1 and down-regulation of cyclin E, in a concentration-dependent manner without any change in cyclin-dependent-kinases (Cdks) expression. Dideoxypetrosynol A markedly induced the levels of Cdk inhibitor p16/INK4a expression. Furthermore, down-regulation of phosphorylation of retinoblastoma protein (pRB) by this compound was associated with enhanced binding of pRB and transcription factor E2F-1. Overall, our results demonstrate a combined mechanism involving the inhibition of pRB phosphorylation and induction of p16 as targets for dideoxypetrosynol A, may explain some of its anti-cancer effects.

HERV-M (human endogenous retrovirus M), related to the super family of HERV-K, has a methionine (M) tRNA primer-binding site, and is located within the periphilin gene on human chromosome 12q12. HERV-M has been integrated into the periphilin gene as the truncated form, 5'LTR-gag-pol-3'LTR. Polymerase chain reaction (PCR) and reverse transcription-polymerase chain reaction (RT-PCR) approaches were conducted to investigate its evolutionary origins. Interestingly, the insertion of retroelements in a common ancestor genome can make different transcript variants in different species. In the case of the periphilin gene, human (10 variants) and mouse (2 variants) lineages show different transcript variants. Insertion of HERV-M (variant 1-3) could affect the protein-coding region. Also, Alusq/x (variant 4-9) and L1ME4a (mammalian-wide subfamilies of LINE-1) (variant 10) in humans and SINE (short interspersed repetitive element) and RLTR15 (the mouse putative long terminal repeat) (variant 2) in mice could be driving forces in transcript diversification of the periphilin gene during mammalian evolution. The HERV-M derived transcripts (variant 1-3) were expressed in different human tissues, whereas they were not detected in crab-eating monkey and squirrel monkey tissues by RT-PCR amplification. Taken together, HERV-M seems to have been integrated into our common ancestor genome after the divergence of simians and prosimians, and then was actively expressed during hominoid evolution.

The DNA topoisomerase inhibitor beta-lapachone is a quinone obtained from the bark of the lapacho tree (Tabebuia avellanedae) in South America. It has been reported to possess a wide range of pharmacological properties, and is a promising cancer chemopreventive agent. In this study, the effects of beta-lapachone on the growth of the human hepatoma cell line HepG2 were investigated. The results showed that beta-lapachone inhibits the viability of HepG2 by inducing apoptosis, as evidenced by the formation of apoptotic bodies and DNA fragmentation. Reverse transcription-polymerase chain reaction and immunoblotting results indicated that treatments of cells with beta-lapachone resulted in down-regulation of anti-apoptotic Bcl-2 and Bcl-X(L) and up-regulation of pro-apoptotic Bax expression. beta-Lapachone-induced apoptosis was associated with a proteolytic activation of caspase-3 and -9 and degradation of poly(ADP-ribose) polymerase protein. However, beta-lapachone treatment did not affect the inhibitor of apoptosis proteins family and the Fas/FasL system. Taken together, our study indicated that beta-lapachone may have potential as a chemopreventive agent for liver cancer.

Cordyceps militaris specimens were continuously collected by Entomopathogenic Fungal Culture Collection (EFCC), Kangwon National University from different mountains, national parks and recreation parks of Korea from 1986 to 2002, mainly from late May to October of each year. Dry specimens of C. militaris along with their isolates have been preserved in EFCC. Fruiting of C. militaris was induced from single ascospore isolates as well as their combinations in brown rice medium. Fruiting experiments showed that combinations of single ascospore isolates produced fertile fruiting bodies, but single isolates could not produce any fruiting bodies. It was shown that two isolates of the opposite mating types were required to produce fertile stromata. However, combinations of the same mating type isolates produced no fruiting body, showing that C. militaris is a bipolar, heterothallic fungus.

The tRNA-specific adenosine deaminase from the pathogenic bacteria Streptococcus pyogenes (spTAD) has been overexpressed in Escherichia coli and crystallized in the presence of Zn2+ ion at 295 K using ammonium sulfate as a precipitant. Flash-cooled crystals of spTAD diffracted to 2.0 A using 30%(v/v) glycerol as a cryoprotectant. X-ray diffraction data have been collected to 2.0 A using synchrotron radiation. The crystal belongs to the tetragonal space group P4(2)2(1)2, with unit-cell parameters a = b = 81.042, c = 81.270 A. The asymmetric unit contains one subunit of spTAD, with a corresponding crystal volume per protein weight (VM) of 3.3 A3 Da(-1) and a solvent content of 62.7%.

Wikyungtang, an oriental herbal formulation, has been known to exert anti-inflammatory and anti-tumoral activity. However, its molecular mechanism of action is not understood. The purpose of the present study was to examine the effect of the water extract of Wikyungtang (WKT) on the growth of A549 human lung cancer cells. Treatment with WKT resulted in a dose-dependent growth inhibition coupled with the characteristic morphological features of apoptosis. Apoptosis-inducing concentrations of WKT induced caspase-3 and caspase-9 activation accompanied by proteolytic degradation of poly(ADP-ribose)-polymerase and phospholipase C-gamma1. In addition, WKT-induced apoptosis in A549 cells was associated with a decreased expression of the anti-apototic Bcl-XL expression. WKT treatment also inhibited the expression of cyclooxygenase (COX)-2 and the accumulation of prostaglandin E2 without significant changes in the levels of COX-1. Taken together, these findings provide important new insights into the possible molecular mechanisms of the anti-cancer activity of WKT.

Resveratrol, a polyphenolic phytoalexin found in grapes, may have the potential for prevention and therapy for human cancer. We report here that resveratrol inhibits the growth of human lung carcinoma A549 cells and provides molecular understanding of this effect. Resveratrol treatment of A549 cells resulted in a concentration-dependent induction of S phase arrest in cell cycle progression. This anti-proliferative effect of resveratrol was associated with a marked inhibition of the phosphorylation of the retinoblastoma protein (pRB) and concomitant induction of cyclin-dependent kinase (Cdk) inhibitor p21WAF1/CIP, which appears to be transcriptionally upregulated and is p53- dependent. In addition, resveratrol treatment resulted in induction of apoptosis as determined by fluorescence microscopy and flow cytometric analysis. These effects were found to correlate with an activation of caspase-3 and a shift in Bax/Bcl-xL ratio more towards apoptosis. Resveratrol treatment also inhibited the transcriptional activity of nuclear transcription factor kappaB (NF-kappaB). Taken together, these findings suggest that resveratrol has strong potential for development as an agent for prevention against human lung cancer.