Next-Generation Phage Display to Identify Peptide Ligands of Deubiquitinases.

Phage display (PD) is a powerful method and has been extensively used to generate monoclonal antibodies and identify epitopes, mimotopes, and protein interactions. More recently, the combination of next-generation sequencing (NGS) with PD (NGPD) has revolutionized the capabilities of the method by creating large data sets of sequences from affinity selection-based approaches (biopanning) otherwise challenging to obtain. NGPD can monitor motif enrichment, allow tracking of the selection process over consecutive rounds, and highlight unspecific binders. To tackle the wealth of data obtained, bioinformatics tools have been developed that allow for identifying specific binding sequences (binders) that can then be validated. Here, we provide a detailed account of the use of NGPD experiments to identify ubiquitin-specific protease peptide ligands.

Obstructive Sleep Apnea and Hypertension: Updates to a Critical Relationship.

PURPOSE OF REVIEW: Obstructive sleep apnea (OSA) is an underdiagnosed illness linked to essential hypertension (HTN), resistant hypertension (r-HTN), and cardiovascular disease (CVD). This review provides updates on the epidemiology, pathophysiology, and treatments of OSA-associated HTN. RECENT FINDINGS: Mild sleep apnea increases the risk for HTN. Eighty-nine percent of young patients aged 18-35 with HTN not attributed to secondary causes have underlying OSA. Home sleep studies are noninferior to formal polysomnography for OSA diagnosis. Nocturnal oxygen desaturation rate is positively correlated with HTN severity. Gut microbiome neo-colonization in response to high-fat diet cravings in patients with OSA alters immune function and worsens HTN. Carbonic anhydrase inhibitors and probiotics show newfound potential for OSA-associated HTN treatment. OSA recognition improves hospital outcomes after a STEMI. Hypoxia-inducible factor (HIF) transcription increases in a dose-dependent manner to hypoxia, and HIFs are strongly linked to cancer growth. OSA and HTN are comorbid conditions with adversely connected pathophysiology including sympathetic hyperactivity, gut dysbiosis, proinflammation, endothelial damage, rostral fluid shifts, pharyngeal collapse, intravascular fluid retention, nocturnal energy expenditure, and metabolic derangements. The dose-response effect of OSA on HTN severity challenges blood pressure (BP) control, so those with refractory HTN should be screened for OSA.

A Simple Whole-Plasmid PCR Method to Construct High-Diversity Synthetic Phage Display Libraries.

Phage display technology utilises peptide and antibody libraries with very high diversities to select ligands with specific binding properties. The production of such libraries can be labour intensive and technically challenging and whilst there are commercial sources of libraries, the exploitation of the resulting binders is constrained by ownership of the libraries. Here, a peptide library of ~ 1 × 109 variants for display on gene VIII was produced alongside three VHH antibody libraries with similar diversity, where 12mer, 16mer or 21mer CDR3s were introduced into the highly stable cAbBCII10 scaffold displayed on gene III. The cloning strategy used a simple whole-plasmid PCR method and type IIS restriction enzyme assembly that facilitate the seamless insertion of diversity into any suitable phage coat protein or antibody scaffold. This method reproducibly produced 1 × 109 variants from just 10 transformations and the four libraries had relatively low bias with 82 to 86% of all sequences present as single copies. The functionality of both peptide and antibody libraries were demonstrated by selection of ligands with specific binding properties by biopanning. The peptide library was used to epitope map a monoclonal antibody. The VHH libraries were pooled and used to select an antibody to recombinant human collagen type 1.

Initial Outcomes Following Introduction of Percutaneous Arteriovenous Fistula Program with Comparison to Historical Surgically Created Fistulas.

BACKGROUND: Recently, there has been an abundance of encouraging data regarding the creation of percutaneous arteriovenous fistulas. Despite promising data regarding their clinical maturation, a paucity of data exists which provides direct comparison between percutaneously created AVFs (pAVF) and open surgically created AVFs (sAVF). This study has 2 primary objectives: First, to compare clinical outcomes of pAVFs to sAVFs, with emphasis on clinical maturation and frequency of postoperative interventions to facilitate maturation. Second, to contribute toward the evidence-based incorporation of the pAVF procedure into the hemodialysis access algorithm. METHODS: A single-center retrospective review was performed on all consecutive patients undergoing surgically created brachiocephalic arteriovenous fistula (BC-AVF, sAVF group) from January 1, 2018 to December 31, 2018 and Ellipsys-created percutaneous arteriovenous fistula (pAVF group) from January 1, 2019 to December 31, 2019. Comparative analysis between groups was performed. RESULTS: A total of 24 patients underwent Ellipsys-created pAVF with mean age of 56.7 ± 22.6 years (12 males [50%], 12 females [50%]) and 62 patients underwent surgically created BC-AVF with mean age of 62.5 ± 13.2 years (32 males [52%], 30 females [48%]). Both the pAVF and sAVF groups had comparable mean operating times (60 ± 40 vs. 56 ± 25 min, P = 0.67) and frequency of procedural technical success (23 [96%] vs. 62 [100%], P = 0.28), respectively. The pAVF group had a lower clinical maturation rate (12 [52%] vs. 54 [87%], P = 0.003) and a higher primary failure rate (9 [39%] vs. 6 [10%], P = 0.003) when compared to the sAVF group. The pAVF group had an increased overall rate of undergoing a postoperative intervention (18 [78%] vs. 13 [21%], P< 0.001), as well as an increased number of total postoperative interventions (1.1 ± 0.9 vs. 0.3 ± 0.6 interventions, P< 0.001) compared to the sAVF group. Percutaneous transluminal angioplasty of the juxta anastomotic segment was the most prevalent postoperative intervention performed in the pAVF group and occurred at a significantly increased frequency when compared to the sAVF group rate (13 [57%] vs. 5 [8%], P< 0.001). CONCLUSIONS: In our single-center retrospective review, patients undergoing Ellipsys-created pAVF in the first year following introduction of percutaneous endovascular had inferior rates of clinical maturation and underwent more postoperative interventions when compared to historical patients undergoing surgically created BC-AVF. Outcome discrepancies compared to previously reported Ellipsys data demonstrate a need for further studies examining the practical translatability of the pAVF.

Initial 6-month quality review of a percutaneous endovascular arteriovenous fistula program.

BACKGROUND: Percutaneous arteriovenous fistulas have recently proven successful alternatives to surgical arteriovenous fistulas with encouraging initial results. The Ellipsys Endovascular Arteriovenous Fistula System utilizing ultrasound and thermal energy has recently received approval for use in the United States. At the University of New Mexico, we developed an integrated service between Vascular Surgery, Interventional Radiology, and Interventional Nephrology for percutaneous arteriovenous fistulas utilizing Ellipsys. METHODS: We performed a retrospective chart review of the initial 6 months (January 1st 2019 to July 1st 2019) of 18 percutaneous arteriovenous fistula placements to evaluate our initial technical success rate, the number of arteriovenous fistulas meeting maturation characteristics or use in dialysis, and to identify areas for quality improvement. RESULTS: Initial technical success was achieved in 17 out of 18 arteriovenous fistulas (94.4%). Three patients did not report for any follow-up at the end of the initial 6 months. Of the remaining patients, 7 out of 15 were using their arteriovenous fistulas or meeting maturation characteristics at the end of the study (46.7%). Patient loss to follow-up/no-show (16.7%), patient not yet requiring hemodialysis (27.8%), and poor post-surgical maturation and/or need for additional maturation procedures (55.6%) were the predominate reasons for non-use. We identified improved coordination of care, early intervention, and outpatient dialysis center education as the primary areas of focus for quality improvement. CONCLUSION: Initial technical success rate of percutaneous arteriovenous fistulas placement was comparable to published studies. Early and aggressive secondary angiographic interventions of arteriovenous fistulas failing to meet cannulation requirements, improved coordination of post-operative care, and outpatient dialysis center education appear to be the primary targets for quality improvement.

Size-Dependent Lattice Dynamics of Atomically Precise Cadmium Selenide Quantum Dots.

Material properties depend sensitively on the atomic arrangements and atomic bonding, but these are notoriously difficult to measure in nanosized atomic clusters due to the small size of the objects and the challenge of obtaining bulk samples of identical clusters. Here, we have combined the recent ability to make gram quantities of identical semiconductor quantum-dot nanoparticles with the ability to measure lattice dynamics on small sample quantities of hydrogenated materials using high energy resolution inelastic x-ray scattering, to measure the size dependence of the phonon density of states in CdSe quantum dots. The fact that we have atomically precise structural models for these nanoparticles allows the calculation of the phonon density of states using density functional theory, providing both experimental and theoretical confirmations of the important role that the inertia of the surface capping species plays on determining the lattice dynamics.

Bariatric Surgery and Hypertension.

Obesity continues to increase in prevalence worldwide. Hypertension has long been associated with obesity, and weight loss continues to be a first-line therapy in the treatment of hypertension. Lifestyle modification and pharmacologic therapy, however, often meet with treatment failure. Bariatric surgery continues to be the most successful approach to sustained weight loss. This review focuses on the underlying physiologic mechanisms of obesity-hypertension, and the impact of bariatric surgery on the treatment of hypertension. Current available literature on the physiologic mechanisms of obesity-hypertension, and the major trials, meta-analyses and systematic reviews of the impact of bariatric surgery procedures on hypertension are reviewed. Evidence suggests significant improvement in obesity-hypertension in patients who undergo surgical weight-reduction procedures. Malabsorptive techniques such as the Roux-en-Y gastric bypass or surgical resection techniques such as laparoscopic sleeve gastrectomy appear to offer superior results in regards to hypertension control over restrictive techniques such as Gastric Banding. Though long-term control of hypertension following surgery remains a concern, available follow-up post-operative data of up to 10 years suggests a sustained, if lessened, effect on hypertension control over time.

Tris(2-mercaptoimidazolyl)hydroborato Cadmium Thiolate Complexes, [TmBut]CdSAr: Thiolate Exchange at Cadmium in a Sulfur-Rich Coordination Environment.

A series of cadmium thiolate compounds that feature a sulfur-rich coordination environment, namely [TmBut]CdSAr, have been synthesized by the reactions of [TmBut]CdMe with ArSH (Ar = C6H4-4-F, C6H4-4-But, C6H4-4-OMe, and C6H4-3-OMe). In addition, the pyridine-2-thiolate and pyridine-2-selenolate derivatives, [TmBut]CdSPy and [TmBut]CdSePy have been obtained via the respective reactions of [TmBut]CdMe with pyridine-2-thione and pyridine-2-selone. The molecular structures of [TmBut]CdSAr and [TmBut]CdEPy (E = S or Se) have been determined by X-ray diffraction and demonstrate that, in each case, the [CdS4] motif is distorted tetrahedral and approaches a trigonal monopyramidal geometry in which the thiolate ligand adopts an equatorial position; [TmBut]CdSPy and [TmBut]CdSePy, however, exhibit an additional long-range interaction with the pyridyl nitrogen atoms. The ability of the thiolate ligands to participate in exchange was probed by 1H and 19F nuclear magnetic resonance (NMR) spectroscopic studies of the reactions of [TmBut]CdSC6H4-4-F with ArSH (Ar = C6H4-4-But or C6H4-4-OMe), which demonstrate that (i) exchange is facile and (ii) coordination of thiolate to cadmium is most favored for the p-fluorophenyl derivative. Furthermore, a two-dimensional EXSY experiment involving [TmBut]CdSC6H4-4-F and 4-fluorothiophenol demonstrates that degenerate thiolate ligand exchange is also facile on the NMR time scale.

Transition from Molecular Vibrations to Phonons in Atomically Precise Cadmium Selenide Quantum Dots.

We use micro-Raman spectroscopy to measure the vibrational structure of the atomically precise cadmium selenide quantum dots Cd35Se20X30L30, Cd56Se35X42L42, and Cd84Se56X56L56. These quantum dots have benzoate (X) and n-butylamine (L) ligands and tetrahedral (Td) shape with edges that range from 1.7 to 2.6 nm in length. Investigating this previously unexplored size regime allows us to identify the transition from molecular vibrations to bulk phonons in cadmium selenide quantum dots for the first time. Room-temperature Raman spectra have broad CdSe peaks at 175 and 200 cm-1. Density functional theory calculations assign these peaks to molecular surface and interior vibrational modes, respectively, and show that the interior, surface, and ligand atom motion is strongly coupled. The interior peak intensity increases relative to the surface peak as the cluster size increases due to the relative increase in the polarizability of interior modes with quantum dot size. The Raman spectra do not change with temperature for molecular Cd35Se20X30L30, while the interior peak narrows and shifts to higher energy as temperature decreases for Cd84Se56X56L56, a spectral evolution typical of a phonon. This result shows that the single bulk unit cell contained within Cd84Se56X56L56 is sufficient to apply a phonon confinement model, and that Cd56Se35X42L42, with its 2.1 nm edge length, marks the boundary between molecular vibrations and phonons.

A Trematode Parasite Derived Growth Factor Binds and Exerts Influences on Host Immune Functions via Host Cytokine Receptor Complexes.

The trematode Fasciola hepatica is responsible for chronic zoonotic infection globally. Despite causing a potent T-helper 2 response, it is believed that potent immunomodulation is responsible for rendering this host reactive non-protective host response thereby allowing the parasite to remain long-lived. We have previously identified a growth factor, FhTLM, belonging to the TGF superfamily can have developmental effects on the parasite. Herein we demonstrate that FhTLM can exert influence over host immune functions in a host receptor specific fashion. FhTLM can bind to receptor members of the Transforming Growth Factor (TGF) superfamily, with a greater affinity for TGF-ß RII. Upon ligation FhTLM initiates the Smad2/3 pathway resulting in phenotypic changes in both fibroblasts and macrophages. The formation of fibroblast CFUs is reduced when cells are cultured with FhTLM, as a result of TGF-ß RI kinase activity. In parallel the wound closure response of fibroblasts is also delayed in the presence of FhTLM. When stimulated with FhTLM blood monocyte derived macrophages adopt an alternative or regulatory phenotype. They express high levels interleukin (IL)-10 and arginase-1 while displaying low levels of IL-12 and nitric oxide. Moreover they also undergo significant upregulation of the inhibitory receptor PD-L1 and the mannose receptor. Use of RNAi demonstrates that this effect is dependent on TGF-ß RII and mRNA knock-down leads to a loss of IL-10 and PD-L1. Finally, we demonstrate that FhTLM aids newly excysted juveniles (NEJs) in their evasion of antibody-dependent cell cytotoxicity (ADCC) by reducing the NO response of macrophages-again dependent on TGF-ß RI kinase. FhTLM displays restricted expression to the F. hepatica gut resident NEJ stages. The altered fibroblast responses would suggest a role for dampened tissue repair responses in facilitating parasite migration. Furthermore, the adoption of a regulatory macrophage phenotype would allow for a reduced effector response targeting juvenile parasites which we demonstrate extends to an abrogation of the ADCC response. Thus suggesting that FhTLM is a stage specific evasion molecule that utilises host cytokine receptors. These findings are the first to clearly demonstrate the interaction of a helminth cytokine with a host receptor complex resulting in immune modifications that facilitate the non-protective chronic immune response which is characteristic of F. hepatica infection.

Exchange of Alkyl and Tris(2-mercapto-1-t-butylimidazolyl)hydroborato Ligands Between Zinc, Cadmium and Mercury.

The tris(2-mercaptoimidazolyl)hydroborato ligand, [TmBut ], has been used to investigate the exchange of alkyl and sulfur donor ligands between the Group 12 metals, Zn, Cd and Hg. For example, [TmBut ]2Zn reacts with Me2Zn to yield [TmBut ]ZnMe, while [TmBut ]CdMe is obtained readily upon reaction of [TmBut ]2Cd with Me2Cd. Ligand exchange is also observed between different metal centers. For example, [TmBut ]CdMe reacts with Me2Zn to afford [TmBut ]ZnMe and Me2Cd. Likewise, [TmBut ]HgMe reacts with Me2Zn to afford [TmBut ]ZnMe and Me2Hg. However, whereas the [TmBut ] ligand transfers from mercury to zinc in the methyl system, [TmBut ]HgMe/Me2Zn, transfer of the [TmBut ] ligand from zinc to mercury is observed upon treatment of [TmBut ]2Zn with HgI2 to afford [TmBut ]HgI and [TmBut ]ZnI. These observations demonstrate that the phenomenological preference for the [TmBut ] ligand to bind one metal rather than another is strongly influenced by the nature of the co-ligands.

Synthesis and structures of cadmium carboxylate and thiocarboxylate compounds with a sulfur-rich coordination environment: carboxylate exchange kinetics involving tris(2-mercapto-1-t-butylimidazolyl)hydroborato cadmium complexes, [Tm(Bu(t))]Cd(O2CR).

A series of cadmium carboxylate compounds in a sulfur-rich environment provided by the tris(2-tert-butylmercaptoimidazolyl)hydroborato ligand, namely, [Tm(Bu(t))]CdO2CR, has been synthesized via the reactions of the cadmium methyl derivative [Tm(Bu(t))]CdMe with RCO2H. Such compounds mimic aspects of cadmium-substituted zinc enzymes and also the surface atoms of cadmium chalcogenide crystals, and have therefore been employed to model relevant ligand exchange processes. Significantly, both (1)H and (19)F NMR spectroscopy demonstrate that the exchange of carboxylate groups between [Tm(Bu(t))]Cd(κ(2)-O2CR) and the carboxylic acid RCO2H is facile on the NMR time scale, even at low temperature. Analysis of the rate of exchange as a function of concentration of RCO2H indicates that reaction occurs via an associative rather than dissociative pathway. In addition to carboxylate compounds, the thiocarboxylate derivative [Tm(Bu(t))]Cd[κ(1)-SC(O)Ph] has also been synthesized via the reaction of [Tm(Bu(t))]CdMe with thiobenzoic acid. The molecular structure of [Tm(Bu(t))]Cd[κ(1)-SC(O)Ph] has been determined by X-ray diffraction, and an interesting feature is that, in contrast to the carboxylate derivatives [Tm(Bu(t))]Cd(κ(2)-O2CR), the thiocarboxylate ligand binds in a κ(1) manner via only the sulfur atom.

Electrical transport and grain growth in solution-cast, chloride-terminated cadmium selenide nanocrystal thin films.

We report the evolution of electrical transport and grain size during the sintering of thin films spin-cast from soluble phosphine and amine-bound, chloride-terminated cadmium selenide nanocrystals. Sintering of the nanocrystals occurs in three distinct stages as the annealing temperature is increased: (1) reversible desorption of the organic ligands (≤150 °C), (2) irreversible particle fusion (200-300 °C), and (3) ripening of the grains to >5 nm domains (>200 °C). Grain growth occurs at 200 °C in films with 8 atom % Cl(-), while films with 3 atom % Cl(-) resist growth until 300 °C. Fused nanocrystalline thin films (grain size = 4.5-5.5 nm) on thermally grown silicon dioxide gate dielectrics produce field-effect transistors with electron mobilities as high as 25 cm(2)/(Vs) and on/off ratios of 10(5) with less than 0.5 V hysteresis in threshold voltage without the addition of indium.

The importance of nanocrystal precursor conversion kinetics: mechanism of the reaction between cadmium carboxylate and cadmium bis(diphenyldithiophosphinate).

We describe the synthesis of cadmium bis(diphenyldithiophosphinate) (Cd(S(2)PPh(2))(2)) from secondary phosphine sulfides and its conversion to cadmium sulfide nanocrystals. Heating Cd(S(2)PPh(2))(2) and cadmium tetradecanoate (≥4 equiv) to 240 °C results in complete conversion of Cd(S(2)PPh(2))(2) to cadmium sulfide nanocrystals with tetradecanoate surface termination. The nanocrystals have a narrow size distribution (d = 3.8-4.1 nm, σ < 10%) that is evident from the line width of the lowest energy absorption feature (λ = 412-422 nm, fwhm = 0.17 eV) and display bright photoluminescence (PLQY(band edge+trap) = 36%). Interestingly, the final diameter is insensitive to the reaction conditions, including the total concentration of precursors and initial cadmium to sulfur ratio. Monitoring the reaction with (31)P NMR, UV-visible, and infrared absorption spectroscopies shows that the production of cadmium diphenylphosphinate (Cd(O(2)PPh(2))(2)) and tetradecanoic anhydride co-products is coupled with the formation of cadmium sulfide. From these measurements we propose a balanced chemical equation for the conversion reaction and use it to optimize a synthesis that affords CdS nanocrystals in quantitative yield. In light of these results we discuss the importance of well-defined precursor reactivity to reproducible conversion kinetics and the synthesis of nanocrystals with unambiguous chemical composition.

Precursor conversion kinetics and the nucleation of cadmium selenide nanocrystals.

The kinetics of cadmium selenide (CdSe) nanocrystal formation was studied using UV-visible absorption spectroscopy integrated with an automated, high-throughput synthesis platform. Reaction of anhydrous cadmium octadecylphosphonate (Cd-ODPA) with alkylphosphine selenides (1, tri-n-octylphosphine selenide; 2, di-n-butylphenylphosphine selenide; 3, n-butyldiphenylphosphine selenide) in recrystallized tri-n-octylphosphine oxide was monitored by following the absorbance of CdSe at λ = 350 nm, where the extinction coefficient is independent of size, and the disappearance of the selenium precursor using {(1)H}(31)P NMR spectroscopy. Our results indicate that precursor conversion limits the rate of nanocrystal nucleation and growth. The initial precursor conversion rate (Q(o)) depends linearly on [1] (Q(o)(1) = 3.0-36 µM/s) and decreases as the number of aryl groups bound to phosphorus increases (1 > 2 > 3). Changes to Q(o) influence the final number of nanocrystals and thus control particle size. Using similar methods, we show that changing [ODPA] has a negligible influence on precursor reactivity while increasing the growth rate of nuclei, thereby decreasing the final number of nanocrystals. These results are interpreted in light of a mechanism where the precursors react in an irreversible step that supplies the reaction medium with a solute form of the semiconductor.

Reaction chemistry and ligand exchange at cadmium-selenide nanocrystal surfaces.

The surface chemistry of cadmium selenide nanocrystals, prepared from tri-n-octylphosphine selenide and cadmium octadecylphosphonate in tri-n-octylphosphine oxide, was studied with 1H and {1H}31P NMR spectroscopy as well as ESI-MS and XPS. The identity of the surface ligands was inferred from reaction of nanocrystals with Me3Si-X (X = -S-SiMe3, -Se-SiMe3, -Cl and -S-(CH2CH2O)4OCH3)) and unambiguous assignment of the organic byproducts, O,O'-bis(trimethylsilyl)octadecylphosphonic acid ester and O,O'-bis(trimethylsilyl)ocatdecylphosphonic acid anhydride ester. Nanocrystals isolated from these reactions have undergone exchange of the octadecylphosphonate ligands for -X as was shown by 1H NMR (X = -S-(CH2CH2O)4OCH3) and XPS (X = -Cl). Addition of free thiols to as prepared nanocrystals results in binding of the thiol to the particle surface and quenching of the nanocrystal fluorescence. Isolation of the thiol-ligated nanocrystals shows this chemisorption proceeds without displacement of the octadecylphosphonate ligands, suggesting the presence of unoccupied Lewis-acidic sites on the particle surface. In the presence of added triethylamine, however, the octadecylphosphonate ligands are readily displaced from the particle surface as was shown with 1H and {1H}31P NMR. These results, in conjunction with previous literature reports, indicate that as-prepared nanocrystal surfaces are terminated by X-type binding of octadecylphosphonate moieties to a layer of excess cadmium ions.

Kinetics and mechanism of methane, methanol, and dimethyl ether C-H activation with electrophilic platinum complexes.

The relative rates of C-H activation of methane, methanol, and dimethyl ether by [(N-N)PtMe(TFE-d(3))](+) ((N-N) = ArN=C(Me)-C(Me)=NAr; Ar = 3,5-di-tert-butylphenyl, TFE-d(3) = CF(3)CD(2)OD) (2(TFE)) were determined. Methane activation kinetics were conducted by reacting 2(TFE)-(13)C with 300-1000 psi of methane in single-crystal sapphire NMR tubes; clean second-order behavior was obtained (k = 1.6 +/- 0.4 x 10(-3) M(-1) s(-1) at 330 K; k = 2.7 +/- 0.2 x 10(-4) M(-1) s(-1) at 313 K). Addition of methanol to solutions of 2(TFE) rapidly establishes equilibrium between methanol (2(MeOD)) and trifluoroethanol (2(TFE)) adducts, with methanol binding preferentially (K(eq) = 0.0042 +/- 0.0006). C-H activation gives [(N-N)Pt(CH(2)OD)(MeOD)](+) (4), which is unstable and reacts with [(RO)B(C(6)F(5))(3)](-) to generate a pentafluorophenyl platinum complex. Analysis of kinetics data for reaction of 2 with methanol yields k = 2.0 +/- 0.2 x 10(-3) M(-1) s(-1) at 330 K, with a small kinetic isotope effect (k(H)/k(D) = 1.4 +/- 0.1). Reaction of dimethyl ether with 2(TFE) proceeds similarly (K(eq) = 0.023 +/- 0.002, 313 K; k = 5.5 +/- 0.5 x 10(-4) M(-1) s(-1), k(H)/k(D) = 1.5 +/- 0.1); the product obtained is a novel bis(alkylidene)-bridged platinum dimer, [(diimine)Pt(mu-CH(2))(mu-(CH(OCH(3)))Pt(diimine)](2+) (5). Displacement of TFE by a C-H bond appears to be the rate-determining step for all three substrates; comparison of the second-order rate constants (k((methane))/k((methanol)) = 1/1.3, 330 K; k((methane))/k((dimethy)(l e)(ther)) = 1/2.0, 313 K) shows that this step is relatively unselective for the C-H bonds of methane, methanol, or dimethyl ether. This low selectivity agrees with previous estimates for oxidations with aqueous tetrachloroplatinate(II)/hexachloroplatinate(IV), suggesting a similar rate-determining step for those reactions.