CALMODULIN6 negatively regulates cold tolerance by attenuating ICE1-dependent stress responses in tomato.

Chilling temperatures induce an increase in cytoplasmic calcium (Ca2+) ions to transmit cold signals, but the precise role of Calmodulins (CaMs), a type of Ca2+ sensor, in plant tolerance to cold stress remains elusive. In this study, we characterized a tomato (Solanum lycopersicum) CaM gene, CALMODULIN6 (CaM6), which responds to cold stimulus. Overexpressing CaM6 increased tomato sensitivity to cold stress whereas silencing CaM6 resulted in a cold-insensitive phenotype. We showed that CaM6 interacts with Inducer of CBF expression 1 (ICE1) in a Ca2+-independent process and ICE1 contributes to cold tolerance in tomato plants. By integrating RNA-sequencing (RNA-seq) and chromatin immunoprecipitation-sequencing (ChIP-seq) assays, we revealed that ICE1 directly altered the expression of 76 downstream cold-responsive (COR) genes that potentially confer cold tolerance to tomato plants. Moreover, the physical interaction of CaM6 with ICE1 attenuated ICE1 transcriptional activity during cold stress. These findings reveal that CaM6 attenuates the cold tolerance of tomato plants by suppressing ICE1-dependent COR gene expression. We propose a CaM6/ICE1 module in which ICE1 is epistatic to CaM6 under cold stress. Our study sheds light on the mechanism of plant response to cold stress and reveals CaM6 is involved in the regulation of ICE1.

The miR164a-NAM3 module confers cold tolerance by inducing ethylene production in tomato.

Because of a high sensitivity to cold, both the yield and quality of tomato (Solanum lycopersicum L.) are severely restricted by cold stress. The NAC transcription factor (TF) family has been characterized as an important player in plant growth, development, and the stress response, but the role of NAC TFs in cold stress and their interaction with other post-transcriptional regulators such as microRNAs in cold tolerance remains elusive. Here, we demonstrated that SlNAM3, the predicted target of Sl-miR164a/b-5p, improved cold tolerance as indicated by a higher maximum quantum efficiency of photosystem II (Fv/Fm), lower relative electrolyte leakage, and less wilting in SlNAM3-overexpression plants compared to wild-type. Further genetic and molecular confirmation revealed that Sl-miR164a/b-5p functioned upstream of SlNAM3 by inhibiting the expression of the latter, thus playing a negative role in cold tolerance. Interestingly, this role is partially mediated by an ethylene-dependent pathway because either Sl-miR164a/b-5p silencing or SlNAM3 overexpression improved cold tolerance in the transgenic lines by promoting ethylene production. Moreover, silencing of the ethylene synthesis genes, SlACS1A, SlACS1B, SlACO1, and SlACO4, resulted in a significant decrease in cold tolerance. Further experiments demonstrated that NAM3 activates SlACS1A, SlACS1B, SlACO1, and SlACO4 transcription by directly binding to their promoters. Taken together, the present study identified the miR164a-NAM3 module conferring cold tolerance in tomato plants via the direct regulation of SlACS1A, SlACS1B, SlACO1, and SlACO4 expression to induce ethylene synthesis.

Protein phosphatase 2A - structure, function and role in neurodevelopmental disorders.

Neurodevelopmental disorders (NDDs), including intellectual disability (ID), autism and schizophrenia, have high socioeconomic impact, yet poorly understood etiologies. A recent surge of large-scale genome or exome sequencing studies has identified a multitude of mostly de novo mutations in subunits of the protein phosphatase 2A (PP2A) holoenzyme that are strongly associated with NDDs. PP2A is responsible for at least 50% of total Ser/Thr dephosphorylation in most cell types and is predominantly found as trimeric holoenzymes composed of catalytic (C), scaffolding (A) and variable regulatory (B) subunits. PP2A can exist in nearly 100 different subunit combinations in mammalian cells, dictating distinct localizations, substrates and regulatory mechanisms. PP2A is well established as a regulator of cell division, growth, and differentiation, and the roles of PP2A in cancer and various neurodegenerative disorders, such as Alzheimer's disease, have been reviewed in detail. This Review summarizes and discusses recent reports on NDDs associated with mutations of PP2A subunits and PP2A-associated proteins. We also discuss the potential impact of these mutations on the structure and function of the PP2A holoenzymes and the etiology of NDDs.

SlMPK1- and SlMPK2-mediated SlBBX17 phosphorylation positively regulates CBF-dependent cold tolerance in tomato.

B-box (BBX) proteins are an important class of zinc finger transcription factors that play a critical role in plant growth and stress response. However, the mechanisms of how BBX proteins participate in the cold response in tomato remain unclear. Here, using approaches of reverse genetics, biochemical and molecular biology we characterized a BBX transcription factor, SlBBX17, which positively regulates cold tolerance in tomato (Solanum lycopersicum). Overexpressing SlBBX17 enhanced C-repeat binding factor (CBF)-dependent cold tolerance in tomato plants, whereas silencing SlBBX17 increased plant susceptibility to cold stress. Crucially, the positive role of SlBBX17 in CBF-dependent cold tolerance was dependent on ELONGATED HYPOCOTYL5 (HY5). SlBBX17 physically interacted with SlHY5 to directly promote the protein stability of SlHY5 and subsequently increased the transcriptional activity of SlHY5 on SlCBF genes under cold stress. Further experiments showed that cold-activated mitogen-activated protein kinases, SlMPK1 and SlMPK2, also physically interact with and phosphorylate SlBBX17 to enhance the interaction between SlBBX17 and SlHY5, leading to enhanced CBF-dependent cold tolerance. Collectively, the study unveiled a mechanistic framework by which SlMPK1/2-SlBBX17-SlHY5 regulated transcription of SlCBFs to enhance cold tolerance, thereby shedding light on the molecular mechanisms of how plants respond to cold stress via multiple transcription factors.

Protein O-glycosylation regulates diverse developmental and defense processes in plants.

Post-translational modifications affect protein functions and play key roles in controlling biological processes. Plants have unique types of O-glycosylation that are different from those of animals and prokaryotes, and they play roles in modulating the functions of secretory proteins and nucleocytoplasmic proteins by regulating transcription and mediating localization and degradation. O-glycosylation is complex because of the dozens of different O-glycan types, the widespread existence of hydroxyproline (Hyp), serine (Ser), and threonine (Thr) residues in proteins attached by O-glycans, and the variable modes of linkages connecting the sugars. O-glycosylation specifically affects development and environmental acclimatization by affecting diverse physiological processes. This review describes recent studies on the detection and functioning of protein O-glycosylation in plants, and provides a framework for the O-glycosylation network that underlies plant development and resistance.

Effect of an low-energy Nd: YAG laser on periodontal ligament stem cell homing through the SDF-1/CXCR4 signaling pathway.

BACKGROUND: The key to the success of endogenous regeneration is to improve the homing rate of stem cells, and low-energy laser is an effective auxiliary means to promote cell migration and proliferation. The purpose of this study was to observe whether low-energy neodymium (Nd: YAG) laser with appropriate parameters can affect the proliferation and migration of periodontal ligament stem cells (PDLSCs) through SDF-1/CXCR4 pathway. METHODS: h PDLSCs were cultured and identified. CCK8 assay was used to detect the proliferation of h PDLSCs after different power (0, 0.25, 0.5, 1, and 1.5 W) Nd: YAG laser (MSP, 10 Hz, 30 s, 300 µ m) irradiation at 2th, 3rd,5th, and 7th days, and the optimal laser irradiation parameters were selected for subsequent experiments. Then, the cells were categorized into five groups: control group (C), SDF-1 group (S), AMD3100 group (A), Nd: YAG laser irradiation group (N), and Nd: YAG laser irradiation + AMD3100 group (N + A). the migration of h PDLSCs was observed using Transwell, and the SDF-1 expression was evaluated using ELISA andRT-PCR. The SPSS Statistics 21.0 software was used for statistical analysis. RESULTS: The fibroblasts cultured were identified as h PDLSCs. Compared with the C, when the power was 1 W, the proliferation rate of h PDLSCs was accelerated (P < 0.05). When the power was 1.5 W, the proliferation rate decreased (P < 0.05). When the power was 0.25 and 0.5 W, no statistically significant difference in the proliferation rate was observed (P > 0.05). The number of cell perforations values as follows: C (956.5 ± 51.74), A (981.5 ± 21.15), S (1253 ± 87.21), N (1336 ± 48.54), and N + A (1044 ± 22.13), that increased significantly in group N (P < 0.05), but decreased in group N + A (P < 0.05). The level of SDF-1 and the expression level of SDF-1 mRNA in groups N and N + A was higher than that in group C (P < 0.05) but lower than that in group A (P < 0.05). CONCLUSIONS: Nd: YAG laser irradiation with appropriate parameters provides a new method for endogenous regeneration of periodontal tissue. SDF-1/CXCR4 signaling pathway may be the mechanism of LLLT promoting periodontal regeneration.

Enantioselective Total Synthesis of (-)-Cephalotanin B.

Cephalotaxus diterpenoids are attractive natural products with intriguing molecular frameworks and promising biological features. As a structurally unusual member, (-)-cephalotanin B possesses an extraordinarily congested heptacyclic skeleton, three lactone units, and nine consecutive stereocenters. Herein, we report an enantioselective total synthesis of (-)-cephalotanin B based on a divergent asymmetric Michael addition reaction, a novel Pauson-Khand/deacyloxylation process discovered in the development of a second-generation stereoselective Pauson-Khand reaction protocol, and an epoxide-opening/elimination/dual-lactonization cascade to construct the challenging propeller-shaped A-B-C ring system as key transformations.

The X-linked intellectual disability gene product and E3 ubiquitin ligase KLHL15 degrades doublecortin proteins to constrain neuronal dendritogenesis.

Proper brain development and function requires finely controlled mechanisms for protein turnover, and disruption of genes involved in proteostasis is a common cause of neurodevelopmental disorders. Kelch-like 15 (KLHL15) is a substrate adaptor for cullin3-containing E3 ubiquitin ligases, and KLHL15 gene mutations were recently described as a cause of severe X-linked intellectual disability. Here, we used a bioinformatics approach to identify a family of neuronal microtubule-associated proteins as KLHL15 substrates, which are themselves critical for early brain development. We biochemically validated doublecortin (DCX), also an X-linked disease protein, and doublecortin-like kinase 1 and 2 as bona fide KLHL15 interactors and mapped KLHL15 interaction regions to their tandem DCX domains. Shared with two previously identified KLHL15 substrates, a FRY tripeptide at the C-terminal edge of the second DCX domain is necessary for KLHL15-mediated ubiquitination of DCX and doublecortin-like kinase 1 and 2 and subsequent proteasomal degradation. Conversely, silencing endogenous KLHL15 markedly stabilizes these DCX domain-containing proteins and prolongs their half-life. Functionally, overexpression of KLHL15 in the presence of WT DCX reduces dendritic complexity of cultured hippocampal neurons, whereas neurons expressing FRY-mutant DCX are resistant to KLHL15. Collectively, our findings highlight the critical importance of the E3 ubiquitin ligase adaptor KLHL15 in proteostasis of neuronal microtubule-associated proteins and identify a regulatory network important for development of the mammalian nervous system.

Tomato SlMYB15 transcription factor targeted by sly-miR156e-3p positively regulates ABA-mediated cold tolerance.

Cold is a common abiotic stress that seriously affects plant growth and development. MYB transcription factors are regulatory molecules that play important roles in various biological processes. We have previously demonstrated that SlMYB15 positively regulates cold tolerance in tomato. However, the underlying mechanism of SlMYB15-induced cold tolerance remains largely unexplored. Here, cold-induced SlMYB15 was found to be targeted by Solanum lycopersicum (sly)-miR156e-3p, which was decreased by cold stimulus in tomato. Tomato plants overexpressing sly-MIR156e-3p displayed significant enhancement in susceptibility to cold stress, while silencing of sly-miR156e-3p by an artificial microRNA interference strategy caused tomato plants to be more tolerant to cold. Moreover, both overexpression of SlMYB15 and silencing of sly-miR156e-3p increased the accumulation of ABA. SlMYB15 directly binds to the promoter regions of ABA biosynthesis and signalling genes, SlNCED1 and SlABF4, resulting in enhanced cold tolerance. Further experiments showed that SlMYB15 and sly-miR156e-3p also coordinated the cold tolerance of tomato via the reactive oxygen species (ROS) signalling pathway, as reflected by the increased expression of SlRBOH1, enhanced H2O2 and O2•-accumulation, and amplified activity of antioxidant enzymes in SlMYB15-overexpressing and sly-miR156e-3p-silenced plants. Taken together, our results demonstrate that SlMYB15 targeted by sly-miR156e-3p confers higher survivability to cold stress via ABA and ROS signals. This study provides valuable information for breeding improved crop cultivars better equipped with cold tolerance.

Effect of Twinlight Laser on the Attachment of Human Gingival Fibroblasts to the Root Surface In Vitro.

BACKGROUND This study aimed to compare the effectiveness of subgingival scaling and root planing with the Twinlight laser, Er: YAG laser, and hand instrumentation on the removal of endotoxin and attachment of human gingival fibroblasts (HGFs) to cementum surfaces in vitro. MATERIAL AND METHODS Single-rooted teeth extracted for periodontal disease were collected and divided into 3 groups: group A, root planing with Gracey curet no. 5/6; group B, irradiation with Er: YAG laser; group C, irradiation with Er: YAG laser and Nd: YAG laser. Endotoxins were determined by the limulus amebocyte lysate test. Cell attachment and proliferation of HGFs on root specimens were evaluated by cell counting kit-8 assay. The root surface and cell morphology were observed by scanning electron microscope. RESULTS A flat root surface with scratches was found in group A, Group B had a homogeneous rough morphology without carbonization, and group C had a non-homogeneous rough morphology with ablation. The endotoxin concentration was highest in group A (P<0.05) and lowest in group C (P>0.05). HGFs cultured in group B showed significantly increased adhesion and proliferation compared with groups A and C (P<0.05). HGFs in group B were well attached, covered densely by pseudopodia. HGFs in group A were round with poor extension and short pseudopodia, while the cells in the group C were in narrow, triangular, or polygonal shapes. CONCLUSIONS Twinlight laser-assisted periodontal treatment effectively improved the biocompatibility of root surface and promoted the attachment and proliferation of fibroblasts by removing calculus and reducing the concentration of endotoxins.

Effect of Er:YAG laser pretreatment on glass-ceramic surface in vitro.

This study investigated the feasibility of using an Er:YAG laser to pretreat glass-ceramic surface and evaluate the effect of the treatment on the bonding strength and marginal adaptation between glass-ceramic and dentin. Glass-ceramic samples (CEREC Blocs) and third molars were cut into 6 mm × 6 mm × 2 mm plates. Thirty ceramic plates were randomly divided into 5 groups: group A (control), group B (pretreated with 9.6% hydrofluoric acid [HF]), group C (pretreated with the Er:YAG laser at 300 mJ and 15 Hz), group D (pretreated with the Er:YAG laser at 400 mJ and 15 Hz), and group E (pretreated with the Er:YAG laser at 500 mJ and 15 Hz). The surface morphologies of the samples in each group were studied under a scanning electron microscope, and the sample displaying optimal etching parameters was selected for subsequent experiments. Based on the surface treatments, 30 ceramic and dentin plates were randomly allocated into 3 groups: the control, laser, and acid-etching groups. After bonding a ceramic plate to a dentin plate, the microleakage and bonding strength were measured, and the pretreatment effects of the Er:YAG laser and 9.6% HF were compared. Group E exhibited an etching effect that was more pronounced and uniform than that in groups C and D. Microleakage and bonding strength analyses revealed that the laser and acid-etching groups differed significantly from the control group in dye penetration depth and shear strength (P < 0.05), although the laser and acid-etching groups did not differ from each other. Both 9.6% hydrofluoric acid and Er:YAG laser pretreatments can coarsen glass-ceramic surfaces, improve the marginal adaptation and bonding strength between the glass-ceramic and dentin, and decrease microleakage of the materials. The two treatments showed no apparent differences in pretreatment outcomes.

Robust superhydrophobicity: mechanisms and strategies.

Superhydrophobic surfaces hold great prospects for extremely diverse applications owing to their water repellence property. The essential feature of superhydrophobicity is micro-/nano-scopic roughness to reserve a large portion of air under a liquid drop. However, the vulnerability of the delicate surface textures significantly impedes the practical applications of superhydrophobic surfaces. Robust superhydrophobicity is a must to meet the rigorous industrial requirements and standards for commercial products. In recent years, major advancements have been made in elucidating the mechanisms of wetting transitions, design strategies and fabrication techniques of superhydrophobicity. This review will first introduce the mechanisms of wetting transitions, including the thermodynamic stability of the Cassie state and its breakdown conditions. Then we highlight the development, current status and future prospects of robust superhydrophobicity, including characterization, design strategies and fabrication techniques. In particular, design strategies, which are classified into passive resistance and active regeneration for the first time, are proposed and discussed extensively.

High Nitric Oxide Concentration Inhibits Photosynthetic Pigment Biosynthesis by Promoting the Degradation of Transcription Factor HY5 in Tomato.

Photosynthetic pigments in higher plants, including chlorophyll and carotenoid, are crucial for photosynthesis and photoprotection. Previous studies have shown that nitric oxide (NO) plays a dual role in plant photosynthesis. However, how pigment biosynthesis is suppressed by NO remains unclear. In this study, we generated NO-accumulated gsnor mutants, applied exogenous NO donors, and used a series of methods, including reverse transcription quantitative PCR, immunoblotting, chromatin immunoprecipitation, electrophoretic mobility shift, dual-luciferase, and NO content assays, to explore the regulation of photosynthetic pigment biosynthesis by NO in tomato. We established that both endogenous and exogenous NO inhibited pigment accumulation and photosynthetic capacities. High levels of NO stimulated the degradation of LONG HYPOCOTYL 5 (HY5) protein and further inactivated the transcription of genes encoding protochlorophyllide oxidoreductase C (PORC) and phytoene synthase 2 (PSY2)-two enzymes that catalyze the rate-limiting steps in chlorophyll and carotenoid biosynthesis. Our findings provide a new insight into the mechanism of NO signaling in modulating HY5-mediated photosynthetic pigment biosynthesis at the transcriptional level in tomato plants.

The effects of Twinlight laser treatment on the titanium surface proliferation and osteogenic differentiation of mesenchymal stem cells.

BACKGROUND: The study aimed to observe the effects of a Twinlight laser on the titanium surface proliferation of inflammatory Mesenchymal stem cells (MSCs), inflammatory cytokine expression, and osteogenic differentiation. METHODS: The MSCs were collected from bone tissue of healthy individuals.The cellular inflammatory model was established with 1 µg/mL lipopolysaccharide (LPS).Under the cellular inflammatory model,divided into five groups: the normal control group (C); the inflammatory control group (L); Er:YAG laser group (L + E); Nd:YAG laser group (L + N); Er:YAG laser and Nd:YAG laser group (L + E + N). The treated cells were inoculated onto titanium disks.The normal and inflammatory MSCs on the surface of titanium surface were examined by CCK-8, scanning election microscopy (SEM), quantitative real-time polymerase chain reaction (qRT­PCR) and other methods for their proliferation, growth pattern, expression of inflammatory factors Interleukin-6 (IL-6), Interleukin-8 (IL-8) and osteogenic genes Runx2 (Runt-related transcription factor 2) and alkaline phosphatase (ALP), providing the theoretical basis and experimental data for the Twinlight laser-assisted treatment of peri-implantitis. Statistical analyses were performed using a Student's t test with SPSS 17.0 software. RESULTS: Through observation using SEM, the cell densities of the L + E + N, L + E, and L + N groups were similar, but cell bodies in the L + E + N group were fuller and each had more than two pseudopodia. The expression level of IL-6 mRNA in the L, L + N, L + E, and L + E + N groups was higher than in group C (P < 0.05), and the expression level of IL-8 mRNA in the L + E + N group was significantly lower than in group L (P < 0.0001). On day 7, the expression level of ALP mRNA in the L, L + N, L + E, and L + E + N groups was lower than in group C (P < 0.05). On day 14, there was no significant difference in the expression level of ALP mRNA among the L + N, L + E + N, and C groups (P > 0.05). On day 7, the expression level of RUNX2 mRNA in the L + E + N group was higher than in group L (P < 0.001). On day 14, the expression level of RUNX2 mRNA in the L + E + N group was higher than in group L (P < 0.01). CONCLUSION: Twinlight laser treatment promoted cell proliferation, inhibited the expression of inflammatory cytokines, and effectively enhanced the osteogenic differentiation of cells on a titanium surface.

Light-induced HY5 Functions as a Systemic Signal to Coordinate the Photoprotective Response to Light Fluctuation.

Optimizing the photoprotection of different leaves as a whole is important for plants to adapt to fluctuations in ambient light conditions. However, the molecular basis of this leaf-to-leaf communication is poorly understood. Here, we used a range of techniques, including grafting, chlorophyll fluorescence, revers transcription quantitative PCR, immunoblotting, chromatin immunoprecipitation, and electrophoretic mobility shift assays, to explore the complexities of leaf-to-leaf light signal transmission and activation of the photoprotective response to light fluctuation in tomato (Solanum lycopersicum). We established that light perception in the top leaves attenuated the photoinhibition of both PSII and PSI by triggering photoprotection pathways in the bottom leaves. Local light promoted the accumulation and movement of LONG HYPOCOTYL5 from the sunlit local leaves to the systemic leaves, priming the photoprotective response of the latter to light fluctuation. By directly activating the transcription of PROTON GRADIENT REGULATION5 and VIOLAXANTHIN DE-EPOXIDASE, LONG HYPOCOTYL5 induced cyclic electron flow, the xanthophyll cycle, and energy-dependent quenching. Our findings reveal a systemic signaling pathway and provide insight into an elaborate regulatory network, demonstrating a pre-emptive advantage in terms of the activation of photoprotection and, hence, the ability to survive in a fluctuating light environment.

The discovery of quinoline derivatives, as NF-κB inducing kinase (NIK) inhibitors with anti-inflammatory effects in vitro, low toxicities against T cell growth.

NIK is a critical regulatory protein of the non-classical NF-kB pathway, and its dysregulated activation has been proved to be one of the pathogenic factors in a variety of autoimmune diseases and inflammatory diseases. Nevertheless, its corresponding development of inhibitors faces many obstacles, including the lack of structure types of known inhibitors, immature activity evaluation methods of compounds in vitro. In this study, a series of quinoline derivatives were obtained through rational design and chemical synthesis. Among them, the representative compounds 17c and 24c have excellent inhibitory activities on LPS-induced macrophage (J774) nitric oxide release and anti-Con A-stimulated primary T cell proliferation. This evaluation method has good universality and overcomes the obstacles mentioned above, which are faced by the current inhibitor research to a certain extent. Besides, the compound's toxicity against the growth of T cells under non-stress conditions was evaluated, for the first time, as an indicator for the investigation to avoid potential safety risks. Pharmacokinetic properties evaluation of the less toxic compound 24c confirmed its good metabolic behavior (especially oral properties, F% = 21.7%), and subsequent development value.

A robust and economical pulse-chase protocol to measure the turnover of HaloTag fusion proteins.

The self-labeling protein HaloTag has been used extensively to determine the localization and turnover of proteins of interest at the single-cell level. To this end, halogen-substituted alkanes attached to diverse fluorophores are commercially available that allow specific, irreversible labeling of HaloTag fusion proteins; however, measurement of protein of interest half-life by pulse-chase of HaloTag ligands is not widely employed because residual unbound ligand continues to label newly synthesized HaloTag fusions even after extensive washing. Excess unlabeled HaloTag ligand can be used as a blocker of undesired labeling, but this is not economical. In this study, we screened several inexpensive, low-molecular-weight haloalkanes as blocking agents in pulse-chase labeling experiments with the cell-permeable tetramethylrhodamine HaloTag ligand. We identified 7-bromoheptanol as a high-affinity, low-toxicity HaloTag-blocking agent that permits protein turnover measurements at both the cell population (by blotting) and single-cell (by imaging) levels. We show that the HaloTag pulse-chase approach is a nontoxic alternative to inhibition of protein synthesis with cycloheximide and extend protein turnover assays to long-lived proteins.

Calculation of hot spots for protein-protein interaction in p53/PMI-MDM2/MDMX complexes.

The recently developed MM/GBSA_IE method is applied to computing hot and warm spots in p53/PMI-MDM2/MDMX protein-protein interaction systems. Comparison of the calculated hot (>2 kcal/mol) and warm spots (>1 kcal/mol) in P53 and PMI proteins interacting with MDM2 and MDMX shows a good quantitative agreement with the available experimental data. Further, our calculation predicted hot spots in MDM2 and MDMX proteins in their interactions with P53 and PMI and they help elucidate the interaction mechanism underlying this important PPI system. In agreement with the experimental result, the present calculation shows that PMI has more hot and warm spots and binds stronger to MDM2/MDMX. The analysis of these hot and warm spots helps elucidate the fundamental difference in binding between P53 and PMI to the MDM2/MDMX systems. Specifically, for p53/PMI-MDM2 systems, p53 and PMI use essentially the same residues (L54, I61, Y67, Q72, V93, H96, and I99) of MDM2 for binding. However, PMI enhanced interactions with residues L54, Y67, and Q72 of MDM2. For the p53/PMI-MDMX system, p53 and PMI use similar residues (M53, I60, Y66, Q71, V92, and Y99) of MDMX for binding. However, PMI exploited three extra residues (M61, K93, and L98) of MDMX for enhanced binding. In addition, PMI enhanced interaction with four residues (M53, Y66, Q71, and Y99) of MDMX. These results gave quantitative explanation on why the binding affinities of PMI-MDM2/MDMX interactions are stronger than that of p53-MDM2/MDMX although their binding modes are similar. © 2018 Wiley Periodicals, Inc.

The efficacy of different sources of mesenchymal stem cells for the treatment of knee osteoarthritis.

Osteoarthritis (OA) is a common cause of chronic pain and disability. Regenerative therapies using mesenchymal stem cells (MSCs) provide an option for OA treatment as it could potentially regenerate the damaged cartilage. Bone marrow, adipose tissue and synovium are common MSC sources. The aim is to compare the therapeutic effect of MSCs from bone marrow, adipose tissue and synovium; combining its differentiation potential and accessibility, to decide the optimal source of MSCs for the treatment of knee OA. A comparison of preclinical and clinical studies using MSCs has been made with regard to treatment outcomes, isolation procedure and differentiation potential. All types of MSCs are effective at improving the clinical and structural condition of OA patients, but the longevity of the treatment, i.e. an effect that is maintained for at least 2 years, cannot be guaranteed. This review highlighted great variations in selection criteria and culture expansion conditions of MSCs between the literature and clinical trials. It also emphasised a substantial diversity and lack of consistency in the assessment mythology of clinical outcome after completion of MSC therapies procedures. A more cohesive methodology is required to evaluate the outcome of MSC treatments using quantitative and standardised frameworks in order to be able to directly compare results. Larger population of patients are recommended to assess the quality of MSC when designing studies and clinical trials to reaffirm the efficacy of MSC treatment prior to and within the clinical trials and follow up studies.

Correction to: The efficacy of different sources of mesenchymal stem cells for the treatment of knee osteoarthritis.

There is an error in the Original Publication of this paper. The author names were incorrectly presented.