Nonclinical safety and immunogenicity assessment of a combined DTacP vaccine in animal models.

The (diphtheria, tetanus, and pertussis [acellular, component] [DTacP]) vaccine is a combined vaccine designed to prevent three potentially fatal diseases including pertussis, tetanus, and diphtheria in both children and adults. We utilized advanced technology to develop a novel DTacP vaccine that was previously unavailable in China. The nonclinical studies were performed to evaluate the immunogenicity, potential toxicity, and local tolerance of the vaccine in animal models. In the immunogenicity study, three batches of the vaccine were intraperitoneally administered to National Institutes of Health (NIH) mice, resulting in 100% seropositivity for all three batches. Additionally, antibody levels notably increased as the immunization dosage increased. In acute toxicity study, no mortality was observed among the animals during the 14-day observation period, and no abnormalities in clinical signs were reported. Active systemic anaphylaxis assessment in guinea pigs showed no evidence of serious allergic reactions in the vaccine groups. In the repeat-dose toxicity study, where five intramuscular doses were administered every 2 weeks, gross autopsy and histopathological examination revealed no vaccine-related systemic pathological changes in rats, with dose site irritant reactions mostly recovered at the end of recovery period. In conclusion, the vaccine demonstrated good local and systemic tolerance, supporting its clinical development.

{Cu8} Cluster-Sandwiched Polyoxotungstates and Their Polymers: Syntheses, Structures, and Properties.

Four inorganic-organic hybrid octa-Cu cluster sandwiched polyoxotungstates (POTs), [Cu8(H2O)2(en)4(B-α-H2SiW9O34)2] (1), [Cu8(H2O)2(en)4(B-α-H2GeW9O34)2] (2), K2[Cu8(en)4(B-α-HSiW9O34)2]·6H2O (3), and K2[Cu8(en)4(B-α-HGeW9O34)2]·2H2O (4) (en = ethylenediamine), were hydrothermally made and characterized by single-crystal X-ray diffraction, infrared spectra, powder X-ray diffraction, and thermogravimetric analysis, respectively. Structure analysis reveals that the polyoxoanion of 1/2 is a discrete dimer built by two trivalent Keggin [B-α-XW9O34]10- (X = Si/Ge) fragments and one octa-Cu cluster, whereas 3 and 4 display a two-dimensional network built by octa-Cu-sandwiched POT units via substitution of coordinated water on polyanions of 1 and 2 and further expand into a three-dimensional framework via K cation bridges. Ultraviolet-visible diffuse reflectance spectra reveal that 1-4 are potential semiconductor materials. Moreover, its visible light-driven catalytic H2 evolution activity, electrochemical properties, catalysis for oxygenation reactions of thioethers, and magnetic behaviors have been investigated in detail.

Increased Number of Perivascular CD20-Positive B Lymphocytes in the Neuropathology of CLIPPERS: Findings of 6 Patients from Mainland China.

INTRODUCTION: Chronic lymphocytic inflammation with pontine perivascular enhancement responsive to steroids (CLIPPERS) syndrome is a recently described chronic inflammatory disease of the central nervous system. There are few reports of CLIPPERS in the Chinese population to date. We summarized the clinical characteristics of 6 CLIPPERS patients to deepen the understanding of this disease. METHODS: The clinical manifestations and treatment of 6 CLIPPERS patients confirmed by pathology or clinical diagnosis in our hospital were retrospectively analyzed. RESULTS: The common clinical manifestations included ataxia, dysarthria, diplopia, dysphagia, dizziness, cognitive impairment, facial paresthesia, and paralysis. Most of the lesions showed typical symmetric "pepper powder"-like dot and nodular enhancement centered in the pontine and cerebellum except 1 patient with unilateral nodular enhancement. The brain histopathological examination of the 5 biopsied patients indicated that, with the exception of patient 4 with no lymphocyte infiltration, a large amount of perivascular lymphocytic infiltration was found in the other 4 patients, among whom only 1 patient was dominated by CD3+ T cell infiltration and the other 3 patients were dominated by CD20+ B cell infiltration. After treatment with intravenous methylprednisolone, all patients had significant clinical recovery associated with complete or significant MRI recovery, but they were prone to relapse after withdrawal or reduction of the corticosteroid. CONCLUSION: Our reports highlight the importance of neuropathological examinations when encountering atypical imaging manifestations, such as unilateral and large nodular Gd+ lesions, in order to establish a final diagnosis of CLIPPERS. In addition, the lymphocytic infiltration in the lesions of CLIPPERS may be dominated by CD20+ B cells instead of CD3+ T cells.

Enantiomeric Polyoxometalates Based on Malate Chirality-Inducing Tetra-ZrIV-Substituted Keggin Dimeric Clusters.

On the basis of the synergistic strategy of lacunary polyoxometalate structure-directing function and chiral ligand inducting role, two pairs of enantiomeric polyoxotungstates, (NH4)4(TMA)4[Zr4(µ3-O)2(l-/d-mal)2(B-α-HSiW10O37)2] (TMA = tetramethylammonium, mal = malate (C4H5O5); l-mal for 1a, d-mal for 1b) and (NH4)4(TMA)4[Zr4(µ3-O)2(l-/d-mal)2(B-α-PW10O37)2] (l-mal for 2a, d-mal for 2b), and a mesomeric polyoxotungstate, (NH4)3Na2K5[Zr4(µ3-O)2(l-mal)(d-mal)(B-α-SiW10O37)2] (3), were hydrothermally synthesized. 1a, 2a and 1b, 2b respectively exhibit 1-D 21 right- and left-hand helical chains formed by hydrogen-bonding interactions, and 3 forms a 3-D (3,10)-connected framework by Na+/K+ ions with {418.624.83}{43}2 topology. These homochiral compounds represent the first examples of enantiomerically pure ZrIV-substituted Keggin POMs. In this system, {Zr4(µ3-O)2(l-/d-mal)2} clusters transfer chirality from d- or l-mal to Keggin polyoxotungstate dimeric clusters, which was demonstrated by structural comparison between the homochiral architecture and mesomer as well as circular dichroism spectra of enantiomers. UV-vis diffuse reflectance spectra reveal that 1-3 are potential semiconductor materials. In addition, 1 and 2 exhibit second harmonic generation response with their response intensities of 0.8 times that of KDP.

Transcriptional repressor Blimp1 regulates follicular regulatory T-cell homeostasis and function.

The B-lymphocyte-induced maturation protein 1 (Blimp1) regulates T-cell homeostasis and function. Loss of Blimp1 could double the proportion of follicular regulatory T (Tfr) cells. However, the effects that Blimp1 may have on the function of Tfr cells remain unknown. Here we document the function for Blimp1 in Tfr cells in vitro and in vivo. Data presented in this study demonstrate that Tfr cells indirectly inhibit the activation and differentiation of B cells by negatively regulating follicular helper T cells, so lowering the secretion of antibody. Lack of Blimp1 makes the immune suppression function of Tfr cells impaired in vitro. In the in vivo study, adoptive transfer of Tfr cells could reduce immune responses in germinal centres and relieve the muscle weakness symptoms of mice with experimental autoimmune myasthenia gravis. Blimp1 deficiency resulted in reduced suppressive ability of Tfr cells. This study identifies that Tfr cells are potent suppressors of immunity and are controlled by Blimp1.

Two New Copper Borates with Mesoscale Cubic Supramolecular Cages Assembled from {Cu4 @B20 } Clusters.

Two new copper borates, namely H6 [(µ4 -O)Cu4 @B20 O32 (OH)8 ]⋅25 H2 O (1) and H6 [(µ4 -O)Cu4 @B20 O32 (OH)8 ]⋅34 H2 O⋅8 H3 BO3 (2), with 3D supramolecular framework have been made under solvothermal conditions, which built by novel cubic supramolecular cages with mesoscale cavities via the H-bondings. Interestingly, the cage is assembled by [(µ4 -O)Cu4 @B20 O32 (OH)8 ] ({Cu4 @B20 }) cluster units with different point-group symmetry. Owing to extra H3 BO3 molecules participated in building the supramolecular framework, 2 has a larger cubic cage size and higher non-framework volume, leading to the cage size extended to mesoporous size set as a version of ''1 plus".

CsBxGe6-xO12 (x = 1): A Zeolite Sodalite-Type Borogermanate with a High Ge/B Ratio by Partial Boron Substitution.

The partial replacement of Ge atoms in tetrahedral positions by a small number of B atoms leads to a new microporous borogermanate, CsBxGe6-xO12 (x = 1), under solvothermal conditions. Its framework shows the highest atomic ratios of Ge/B in reported borogermanates and leads to a new type of zeolite sodalite-type net.

Diverse Ligand-Functionalized Mixed-Valent Hexamanganese Sandwiched Silicotungstates with Single-Molecule Magnet Behavior.

Under hydrothermal conditions, replacement of the water molecules in the [Mn(III) 4 Mn(II) 2 O4 (H2 O)4 ](8+) cluster of mixed-valent Mn6 sandwiched silicotungstate [(B-α-SiW9 O34 )2 Mn(III) 4 Mn(II) 2 O4 (H2 O)4 ](12-) (1 a) with organic N ligands led to the isolation of five organic-inorganic hybrid, Mn6 -substituted polyoxometalates (POMs) 2-6. They were all structurally characterized by IR spectroscopy, elemental analysis, thermogravimetric analysis, diffuse-reflectance spectroscopy, and powder and single-crystal X-ray diffraction. Compounds 2-6 represent the first series of mixed-valent {Mn(III) 4 Mn(II) 2 O4 (H2 O)4-n (L)n } sandwiched POMs covalently functionalized by organic ligands. The preparation of 1-6 not only indicates that the double-cubane {Mn(III) 4 Mn(II) 2 O4 (H2 O)4-n (L)n } clusters are very stable fragments in both conventional aqueous solution and hydrothermal systems and that organic functionalization of the [Mn(III) 4 Mn(II) 2 O4 (H2 O)4 ](8+) cluster by substitution reactions is feasible, but also demonstrates that hydrothermal environments can promote and facilitate the occurrence of this substitution reaction. This work confirms that hydrothermal synthesis is effective for making novel mixed-valent POMs substituted with transition-metal (TM) clusters by combining lacunary Keggin precursors with TM cations and tunable organic ligands. Furthermore, magnetic measurements reveal that 3 and 6 exhibit single-molecule magnet behavior.

Lanthanide Germanate Cluster Organic Frameworks Based on {Ln8Ge12} Clusters: From One-Dimensional Chains to Two-Dimensional Layers and Three-Dimensional Frameworks.

Under hydrothermal conditions, six series of novel lanthanide (Ln) organogermanates (LnGs) [Ln8Ge12(µ3-O)24E12(H2O)16]·14H2O (Ln(3+) = Pr(3+), 1; Nd(3+), 2; Sm(3+), 3; Eu(3+), 4; Gd(3+), 5; one-dimensional (1-D) LnG cluster organic chain (LnGCOC)), {[Nd8Ge12(µ3-O)24E12(H2O)10](µ2-H2O)2[Nd8Ge12(µ3-O)24E12(H2O)16]}·18H2O (6, two-dimensional (2-D) planar LnG cluster organic layer (LnGCOL)), {[Ln2GeE(HO)2O(H2O)(CH3COO)2(CO3)]2[Ln8Ge12E12(µ3-O)24(H2O)10]}·6H2O (Ln(3+) = Pr(3+), 7; Nd(3+), 8; 2-D wave-shaped LnGCOL), [TbGeE(HO)2O(H2O)(pca)]2[Tb8Ge12E12(µ3-O)24(H2O)8]·10H2O (9, three-dimensional (3-D) LnG cluster organic framework (LnGCOF)), {([Nd(pza)2(H2O)2]2[Nd8Ge12E12(µ3-O)24(H2O)12])([Nd(pza)2]2[Nd8Ge12E12(Hpza)2(µ3-O)24(H2O)10])}·4OH·14H2O (10, 3-D LnGCOF), {[Nd8Ge12E12(µ3-O)24(H2O)10][Nd(pca)(pda)(H2O)]2}·12H2O (11, 3-D LnGCOF) and {[Nd8Ge12E12(µ3-O)24(H2O)10][Nd(pza)(pda)(H2O)]2}·12H2O (12, 3-D LnGCOF) (Hpca = 2-picolinic acid, H2pda = 2,6-pyridinedicarboxylic acid, Hpza = 2-pyrazinecarboxylic acid) were prepared by introducing the second auxiliary ligands into the organogermanate-lanthanide-oxide reaction system. The obtainment of these LnGs realized the utilization of the second auxiliary ligands inducing the assembly from 1-D LnGCOCs to 2-D LnGCOLs and 3-D LnGCOFs based on LnG cluster (LnGC) {Ln8Ge12E12(µ3-O)24(H2O)16}({Ln8Ge12}) units and Ln-organic complexes or organic ligand connectors. It should be noted that the well-organized structural constructions of 1-12 can be visualized as the gradual replacement of active water sites located at equatorial and polar positions on the hypothetical [Ln8Ge12(µ3-O)24E12(H2O)18] LnGC core with oxygen or nitrogen atoms from organic ligands. The solid-state luminescent properties of 2, 3, 4, 6, and 8-12 have been investigated at room temperature.

Silencing miR-146a influences B cells and ameliorates experimental autoimmune myasthenia gravis.

MicroRNAs have been shown to be important regulators of immune homeostasis as patients with aberrant microRNA expression appeared to be more susceptible to autoimmune diseases. We recently found that miR-146a was up-regulated in activated B cells in response to rat acetylcholine receptor (AChR) α-subunit 97-116 peptide, and this up-regulation was significantly attenuated by AntagomiR-146a. Our data also demonstrated that silencing miR-146a with its inhibitor AntagomiR-146a effectively ameliorated clinical myasthenic symptoms in mice with ongoing experimental autoimmune myasthenia gravis. Furthermore, multiple defects were observed after miR-146a was knocked down in B cells, including decreased anti-R97-116 antibody production and class switching, reduced numbers of plasma cells, memory B cells and B-1 cells, and weakened activation of B cells. Previously, miR-146a has been identified as a nuclear factor-κB-dependent gene and predicted to base pair with the tumour necrosis factor receptor-associated factor 6 (TRAF6) and interleukin-1 receptor-associated kinase 1 (IRAK1) genes to regulate the immune response. However, our study proved that miR-146a inhibition had no effect on the expression of TRAF6 and IRAK1 in B cells. This result suggests that the function of miR-146a in B cells does not involve these two target molecules. We conclude that silencing miR-146a exerts its therapeutic effects by influencing the B-cell functions that contribute to the autoimmune pathogenesis of myasthenia gravis.

A Series of Aluminoborates Templated or Supported by Zinc-Amine Complexes.

A series of open-framework aluminoborates (ABOs), namely, [Zn2 (en)5 ][Al2 B10 O20 ] (1), [Zn(en)(dien)][AlB5 O10 ] (2), [Zn(en)3 ][AlB7 O12 (OH)2 ] (3), [Zn(en)2 ][AlB5 O10 ] (4), K7 {AlO0.5 [BO2 (OH)]Zn@[B12 O21 (OH)3 ]}⋅H2 O (5) (en=ethylenediamine, dien=diethylenetriamine) have been made under mild solvothermal conditions and characterized by single-crystal X-ray diffraction, powder X-ray diffraction, elemental analysis, IR and UV/Vis spectroscopy, thermogravimetric analysis, and nonlinear optical determination. They were classified as two types: Compounds 1-4 contain intersecting channels and exhibit various 4-connected nets built by AlO4 tetrahedra and oxo boron clusters, zinc-amine complexes act as the templates (1-3) or directly bond to the walls of the ABO (4); compound 5 exhibits a double-layer structure made by nanosized [BO2 (OH)]Zn@[B12 O21 (OH)3 ] ({BZn@B12 }) clusters with Al2 O7 dimers, the channels are within the layer. The second harmonic generation (SHG) measurement shows that the SHG responses of 1-3 are about 2.5, 1.6, and 0.5 times that of KH2 PO4 . Compounds 1-2 are type I phase-matchable materials. UV/Vis spectroscopy indicates that compounds 1-5 are wide-band-gap semiconductors.

Unexpected high-temperature stability of ß-Zn4Sb3 opens the door to enhanced thermoelectric performance.

ß-Zn4Sb3 has one of the highest ZT reported for binary compounds, but its practical applications have been hindered by a reported poor stability. Here we report the fabrication of nearly dense single-phase ß-Zn4Sb3 and a study of its thermoelectric transport coefficients across a wide temperature range. Around 425 K we find an abrupt decrease of its thermal conductivity. Past this point, Zn atoms can migrate from crystalline sites to interstitial positions; ß-Zn4Sb3 becomes metastable and gradually decomposes into Zn(hcp) and ZnSb. However, above 565 K it recovers its stability; in fact, the damage caused by decomposition can be repaired completely. This is key to its excellent thermoelectric performance at high temperature: the maximum ZT reaches 1.4. Molecular dynamics simulations are used to shed light on the microscopic behavior of the material.

A 3D aluminoborate open framework interpenetrated by 2D zinc-amine coordination-polymer networks in its 11-ring channels.

A new inorganic-organic hybrid solid, [Zn(dap)2 ][AlB5 O10 ], combining the structural features of 3D open-framework inorganic solids and 2D metal-organic coordination polymers has been synthesized under solvothermal conditions. The compound displays extensive luminescence and moderate second-harmonic-generation efficiency.

Marcasite/pyrite nanocomposites confined in N,S-doped carbon nanoboxes for boosted alkali metal ion storage.

FeS2 is a promising electrode material for alkali metal ion storage due to its high theoretical capacity. However, it still faces critical issues such as suboptimal rate and cycling performances owing to sluggish charge transport and significant volume variations. Herein, we constructed FeS2 (m-FeS2) and pyrite FeS2 (p-FeS2) nanocomposites embedded in N,S-doped carbon nanoboxes (m/p-FeS2@NSCN) to conquer such challenges. The microstructure design of nanoboxes effectively alleviates the stress caused by the volume expansion of FeS2 during lithiation processes, thereby improving the cycling stability of the FeS2 electrode. The marcasite/pyrite compositing design further increases the electronic conductivity of FeS2 and optimizes ion migration. As expected, the target m/p-FeS2@NSCN exhibits improved rate capability (595.5 mA h g-1 at 5.0 A g-1) and robust cycling stability (500 cycles without significant capacity decay at 0.1 A g-1) in lithium-ion batteries. Furthermore, m/p-FeS2@NSCN also shows excellent battery performances and potential application prospects in the field of sodium-ion batteries. It achieves a capacity of 355 mA h g-1 at 10.0 A g-1 and sustains 800 cycles without noticeable capacity decay at 0.5 A g-1. This work offers valuable guidance for rationally designing high-performance energy storage materials for alkali metal ion storage.

Three new mixed-alkali- and alkaline-earth-metal borates: from 1D chain to 2D layer to 3D framework.

Three new mixed-metal borates, K4Ba2[B14O20(OH)10]·3H2O (1), LiSr2[B10O16(OH)3] (2), and LiBa[B9O15] (3), have been made under hydro(solvo)thermal conditions and characterized by means of IR, UV-vis-near-IR, thermogravimetric analysis, powder X-ray diffraction, and single-crystal X-ray diffraction, respectively. 1 is a 1D chain constructed from B14O21(OH)10(10-) cluster units, 2 is of a 2D layer with nine-membered-ring windows built up of B10O19(OH)3(9-) cluster units, while 3 exhibits a 3D framework with 12-membered-ring channels composed of B3O7 cluster units.

Liposome and QS-21 Combined Adjuvant Induces theHumoral and Cellular Responses of Acellular Pertussis Vaccine in a Mice Model.

The resurgence of pertussis in vaccinated communities may be related to the reduced long-term immunity induced by acellular pertussis vaccines. Therefore, developing improved pertussis vaccine candidates that could induce strong Th1 or Th17 cellular immunity is an urgent need. The use of new adjuvants may well meet this requirement. In this research, we developed a novel adjuvant candidate by combining liposome and QS-21 adjuvant. Adjuvant activity, protective efficacy, the level of neutralizing antibody against PT, and the resident memory T (TRM) cells in lung tissue after vaccination were studied. We then performed B. pertussis respiratory challenge in mice after they received vaccination with traditional aluminum hydroxide and the novel adjuvant combination. Results showed that the liposome + QS-21 adjuvant group had a rapid antibody and higher antibody (PT, FHA, Fim) level, induced anti-PT neutralizing antibody and recruited more IL-17A-secreting CD4+ TRM cells along with IL-17A-secreting CD8+ TRM cells in mice, which provided robust protection against B. pertussis infection. These results provide a key basis for liposome + QS-21 adjuvant as a promising adjuvant candidate for developing an acellular pertussis vaccine that elicits protective immunity against pertussis.

Safety and immunogenicity of a combined DTacP-sIPV-Hib vaccine in animal models.

The DTacP-sIPV-Hib combination vaccine can replace the single-component acellular pertussis, diphtheria, tetanus, polio, and Haemophilus influenzae type B vaccines. In this study, we evaluated the safety and immunogenicity of a newly developed DTacP-sIPV-Hib combination vaccine in animal models. We used 40 mice and 46 cynomolgus monkeys to evaluate acute and long-term toxicity. Thirty-six guinea pigs were used for sensitization assessment. For immunogenicity assessment, 50 NIH mice and 50 rats were equally randomized to receive 3 doses of 3 different batches of the tested vaccine at an interval of 21 d, or physiological saline solution (0.5 mL). Orbital blood was collected at an interval of 21 d post inoculation to detect related antibody titers or neutralizing antibody titers against poliovirus. Gross autopsy and histopathological examination revealed no abnormal toxicity or irritation in mice and cynomolgus monkeys. Sensitization assessment in guinea pigs indicated the lack of evident allergic symptoms in the high- and low-dose vaccine groups within 30 min after repeated stimulation. The DTacP-sIPV-Hib combination vaccine induced significant immune responses in mice, rats, and cynomolgus monkeys, with 100% seroconversion rates after 3 doses. The DTacP-sIPV-Hib combination vaccine is safe and immunogenic in animal models. Three doses of the vaccine elicited satisfactory antibody responses in mice, rats, and cynomolgus monkeys.

Effectiveness, immunogenicity, and safety of influenza vaccines with MF59 adjuvant in healthy people of different age groups: A systematic review and meta-analysis.

BACKGROUND: Influenza is a severe disease burden among all age groups. This study aimed to review the efficacy of inactivated influenza vaccines with MF59 adjuvant and non-adjuvanted inactivated influenza vaccines among all age groups against specific influenza vaccine strains. METHODS: Literature search of PubMed, Embase, Medline, OVID, and Cochrane Library Trials (CENTRAL) was implemented up to March 1, 2019. Homogeneity qualified studies were included forData were extracted such as study country location, demographic characteristics, and measure outcomes, and were analyzed by a random effect model and sensitivity analyses to identify heterogeneity. Risk of bias was evaluated using the Cochrane Risk of Bias Tool. RESULTS: We retrieved 1,021 publications and selected 31 studies for full review, including 17 trials for meta-analysis and 6 trials for qualitative synthesis. MF59-adjuvanted influenza vaccines demonstrated better immunogenicity against specific vaccine virus strains compared to non-adjuvanted influenza vaccine both in healthy adult group (RR = 2.10; 95% CI: 1.28-3.44) and the healthy aged (RR = 1.26; 95% CI: 1.10-1.44). CONCLUSION: The quality of evidence is moderate to high for seroconversion and seroprotection rates of influenza vaccine. MF59-adjuvanted influenza vaccines are superior to non-adjuvanted influenza vaccines to enhance immune responses of vaccination in healthy adults and older adults, and could be considered for routine use especially the monovalent prepandemic influenza vaccines.

Development and verification of chemical chromogenic method for determination of residual cetyltrimethyl ammonium bromide content in ACYW135 meningococcal polysaccharide stock solution / 中国生物制品学杂志

@#Objective To develop and verify a chemical choromogenic method for the determination of residual cetyltrimethylammonium bromide(CTAB)content in ACYW135 meningococcal polysaccharide stock solution. Methods A chemical chromogenic method was developed for the determination of residual CTAB content by using titan yellow as chromogenic reagent,and verified for the linear range,intermediate precision and accuracy. ACYW135 meningococcal polysaccharide stock solution were determined for residual CTAB content by the developed method. Results The CTAB reference at concentrations of 4. 0～10. 0 μg/mL showed good linear relationship to A_(500),with a R~2 value of more than 0. 990. The recovery rates of CTAB standard at concentrations of 5. 0,7. 0,and 9. 0 μg/mL were all within 95% ～ 110% in six repeated tests,with the CV values of determination results of less than 10%. All the residual CTAB contents in four batches of meningococcal polysaccharide stock solutions were less than 8. 0 μg/mL. Conclusion The chemical chromogenic method showed good linearity,intermediate precision and accuracy,which might be used for the determination of residual CTAB content in ACYW135 meningococcal polysaccharide stock solution.