Enhanced formaldehyde oxidation over MnO2 and doped manganese-based catalysts: Experimental and theoretical Insights into mechanism and performance.

Thermal catalytic degradation of formaldehyde (HCHO) over manganese-based catalysts is garnering significant attention. In this study, both theoretical simulations and experimental methods were employed to elucidate the primary reaction pathways of HCHO on the MnO2(110) surface. Specifically, the effects of doping MnO2 with elements such as Fe, Ce, Ni, Co, and Cu on the HCHO oxidation properties were evaluated. Advanced characterization techniques, including X-ray diffraction (XRD), scanning electron microscopy (SEM), Brunauer-Emmett-Teller (BET), and X-ray photoelectron spectroscopy (XPS), were employed to discern the physical properties and chemical states of the active components on the catalyst surface. The comprehensive oxidation pathway of HCHO on the MnO2(110) surface includes O2 adsorption and dissociation, HCHO adsorption and dehydrogenation, CO2 desorption, H2O formation and desorption, oxygen vacancy supplementation, and other elementary reactions. The pivotal rate-determining step was identified as the hydrogen migration process, characterized by an energy barrier of 234.19 kJ mol-1. Notably, HCHOO and *CHOO emerged as crucial intermediates during the reaction. Among the doped catalysts, Fe-doped MnO2 outperformed its counterparts doped with Ce, Ni, Co, and Cu. The optimal degradation rate and selectivity were achieved at a molar ratio of Fe: Mn = 0.1. The superior performance of the Fe-doped MnO2 can be ascribed to its large specific surface area, conducive pore structure for HCHO molecular transport, rich surface-adsorbed oxygen species, and a significant presence of oxygen vacancies.

LC-MS/MS Quantitation of Formaldehyde-Glutathione Conjugates as Biomarkers of Formaldehyde Exposure and Exposure-Induced Antioxidants: A New Look on an Old Topic.

Humans are continuously exposed to formaldehyde via both endogenous and exogenous sources. Prolonged exposure to formaldehyde is associated with many human diseases, such as lung cancer and leukemia. The goal of this study is to develop biomarkers to measure formaldehyde exposure, which could be used to predict the risk of associated diseases. As glutathione (GSH) is well-known for its crucial role in the detoxification of a wide variety of xenobiotics, including formaldehyde, we rigorously quantitated in this study the conjugates formed when formaldehyde reacted with GSH using liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS) coupled with an isotope dilution method. The results showed for the first time that (S)-1-(((R)-2-amino-3-(carboxymethylamino)-3-oxopropylthio)methyl)-5-oxopyrrolidine-2-carboxylic acid (PGF) and thioproline-glycine (SPro-Gly) are major metabolites in both nonenzymatic reactions and formaldehyde-exposed human cells. In particular, over 35% of the formaldehyde from external sources was found to convert to SPro-Gly in the exposed cells. Interestingly, data showed that these exposure-induced adducts exhibited good antioxidative properties, which can protect cells from hydrogen peroxide mediated oxidative insult. It is anticipated that the findings of this study could shed light on developing PGF and SPro-Gly as dietary supplements and on the development of noninvasive methods to assess health risks associated with formaldehyde exposure.

Preparation and Evaluation of '3 Cap' Pulsatile Drug Delivery System of Ramipril.

BACKGROUND: Chronotherapeutics, the drug delivery based on circadian rhythm, is recently gaining much attention worldwide. Various diseases like asthma, hypertension, and arthritis show the circadian variation that demands time scheduled drug release for effective drug action. Therefore, the pulsatile drug delivery system has been designed to confer preprogrammed drug delivery. OBJECTIVE: In the present study, a '3 Cap' pulsatile drug delivery system has been developed, optimized, and characterized in order to achieve the floating and pulsatile release of ramipril. METHODS: An optimal response surface design was employed to investigate the effect of isopropanol: formaldehyde vapors for varying time on drug release from the capsules. '3 Cap' pulsatile drug delivery system was evaluated in terms of floating time, density, the effect of gastric flow rate, and type of dissolution apparatus on drug release. RESULTS: Independent variables exhibited a significant effect on the drug release of the prepared formulations. Results showed that time between the release of fractions of dose increased with an increase in formaldehyde: isopropanol ratio and duration of exposure to formaldehyde vapors with no effect of gastric flow rate. CONCLUSION: The results of the designed system revealed that an optimum exposure of 1:2 of isopropanol: formaldehyde vapors for sixty minutes resulted in the desired release of second pulse of dose after a predetermined lag time of 5 hours as desired. '3Cap' system was successful in achieving floating and pulsed release of hypertensive drug opening a 'new lease of life' to the existing drug molecule.

The effect of lignin-alumina hybrid additive on the properties of composition used in abrasive tools.

Lignin-Al2O3 hybrids were tested as effective additives for application in abrasive materials. The main focus was on the reduction of environmental pollution. The emission of volatile compounds, mainly phenol and formaldehyde, was investigated using detailed evolved gas analysis (EGA) performed by means of mass spectroscopy (QMS) in combined differential scanning calorimetry (DSC) and thermogravimetry (TG) analysis. It was established that the addition of lignin-Al2O3 hybrid additives can reduce the emission of phenol and formaldehyde. Crucially, free phenol emission was not detected from the lignin-Al2O3 additives or from lignin itself using the TG-MS method. Moreover, the addition of lignin-type fillers to phenolic composites can lower emissions of the two aforementioned compounds. No emission of other toxic compounds was detected. The mechanical properties of the lignin-alumina hybrids and resin systems were investigated using the three-point flexural test (also as an element of an ageing test), a compressive test, and testing of abrasibility. The results indicate that the lignin and alumina used as a hybrid additive for abrasive materials improve the adhesion between the binder and abrasive grain, and increase the flexibility of the composites, which has a positive impact on the performance of the final products.

High-value utilization of hydroxymethylated lignin in polyurethane adhesives.

Lignin is a good candidate for the polymerization and chemical modification to prepare sustainable chemicals and materials, but a relatively low hydroxyl content becomes an obstacle for the preparation of lignin-based polyurethane (PU) adhesives. In order to improve its reactivity, the acetic acid lignin (AAL) was hydroxymethylated before copolymerized with isocyanate during the preparation of PU adhesives. The hydroxymethylation was carried out in an alkaline formaldehyde solution and it was found that 85 °C is the optimal temperature. On that condition, the free formaldehyde content of the corresponding product HL-6 was as low as 0.32%, while the hydroxymethyl was increased by 189.11% compared with original AAL and reached 2.92 mmol/g. In the polymerization of PU adhesives, the hydroxymethylated lignin with a higher aliphatic hydroxyl content formed a more compact three-dimensional urethane cross-linking network with isocyanate. The mechanical properties and thermal stability of the lignin-based PU adhesive were improved by 15-30 wt% in HL-6, and particularly the tensile strength was increased by 21-41 MPa, which indicated that the hydroxymethylation is an efficient way to enrich the hydroxyl in lignin, and the modified lignin is adequate to partially replace petroleum-based polyols for the preparation of PU adhesives with excellent properties.

Preparation of TTO/UF resin microcapsule via in situ polymerisation and modelling of its slow release.

For slow release of tea tree oil (TTO), TTO were encapsulated by urea-formaldehyde (UF) resin via in situ polymerisation. The effects of curing time and drying condition on particle size, and TTO loading of the TTO/UF microcapsules were investigated. The results indicated that TTO/UF resin microcapsules with curing time of 80 min had narrow size distribution and good wall cover. Drying at ambient was better to maintain the TTO content than drying at oven. The loading of TTO with curing time of 80 min can be up to 45 wt.% of the mass-proportion to the prepared microcapsules, and more than 90 wt.% of the loaded TTO could be sustainably released in about 5 days. Moreover, the release kinetics of TTO/UF microcapsules was well described by Ritger-Peppas model, revealing non-Fickian diffusion. Promisingly, TTO/UF microcapsules with good stability can be used as a slow release vehicle for antibacterial application.

Discovery of Kynurenines Containing Oligopeptides as Potent Opioid Receptor Agonists.

Kynurenine (kyn) and kynurenic acid (kyna) are well-defined metabolites of tryptophan catabolism collectively known as "kynurenines", which exert regulatory functions in host-microbiome signaling, immune cell response, and neuronal excitability. Kynurenine containing peptides endowed with opioid receptor activity have been isolated from natural organisms; thus, in this work, novel opioid peptide analogs incorporating L-kynurenine (L-kyn) and kynurenic acid (kyna) in place of native amino acids have been designed and synthesized with the aim to investigate the biological effect of these modifications. The kyna-containing peptide (KA1) binds selectively the m-opioid receptor with a Ki = 1.08 ± 0.26 (selectivity ratio m/d/k = 1:514:10000), while the L-kyn-containing peptide (K6) shows a mixed binding affinity for m, d, and k-opioid receptors, with efficacy and potency (Emax = 209.7 + 3.4%; LogEC50 = -5.984 + 0.054) higher than those of the reference compound DAMGO. This novel oligopeptide exhibits a strong antinociceptive effect after i.c.v. and s.c. administrations in in vivo tests, according to good stability in human plasma (t1/2 = 47 min).

Computational tomography and CFD simulation of a biofilter treating a toluene, formaldehyde and benzo[α]pyrene vapor mixture.

In this work, a 3D computational tomography (CT) of the packing material of a laboratory column biofilter is used to model airflow containing three contaminants. The degradation equations for toluene, formaldehyde and benzo[α]pyrene (BaP), were one-way coupled to the CFD model. Physical validation of the model was attained by comparing pressure drops with experimental measurement, while experimental elimination capacities for the pollutants were used to validate the biodegradation kinetics. The validated model was used to assess the existence of channeling and to predict the impact of the three-dimensional porous geometry on the mass transfer of the contaminants in the gas phase. Our results indicate that a physically meaningful simulation can be obtained using the techniques and approach presented in this work, without the need of performing experiments to obtain macroscopic parameters such as gas-phase axial and radial dispersion coefficients and porosities.

DFT and MD Studies of Formaldehyde-Derived DNA Adducts: Molecular-Level Insights into the Differential Mispairing Potentials of the Adenine, Cytosine, and Guanine Lesions.

4-(Methylnitrosamino)-1-(3-pyridyl)-1-butanone is a potent nicotine-based carcinogen that generates many DNA lesions, including the HOCH2-C, HOCH2-G, and HOCH2-A hydroxymethyl adducts. Despite all lesions containing an altered exocyclic amino group, which allows the hydroxymethyl group to be directed away from the Watson-Crick binding face, only the most persistent adenine adduct is mutagenic. As a first step toward understanding this differential mutagenicity, density functional theory (DFT) and molecular dynamics (MD) simulations were used to gain atomic-level structural details of these DNA damage products. DFT calculations reveal that all three lesions exhibit conformational diversity. However, regardless of the hydroxymethyl-nucleobase orientation, both DFT and MD simulations highlight that HOCH2-C and HOCH2-G form pairs with the canonical complementary base (G and C, respectively) that are structural and energetically preferred over mispairs. In contrast, depending on the hydroxymethyl-nucleobase orientation, the Watson-Crick HOCH2-A:T pair can become significantly destabilized relative to undamaged A:T. As a result, HOCH2-A mispairs with G, C, and A are energetically accessible and maintain key geometrical features of canonical DNA. Overall, our data directly correlate with the reported differential mutagenicity of the hydroxylmethyl lesions and will encourage future studies to further uncover the cellular impact of the most persistent adenine lesion.

The influence of storage method on the transparency of pig crystalline lens / A influência do método de armazenamento na transparência do cristalino de porco

ABSTRACT Purpose: The porcine eye is frequently used as a research model. This paper analyzes the effect of different storage methods on the transparency of pig crystalline lens. Methods: A spectral transmission curve (from 220 to 780 nm) for the crystalline lens was determined experimentally after storage in different conditions: saline solution, formalin, castor oil, and freezing at -80°C. The total transmission in the visible spectrum, which was used as an index of transparency, was calculated from these curves. For comparative purposes, fresh lenses were evaluated and used as controls. Results: Storing the porcine crystalline lens in saline solution or castor oil resulted in a transparency loss of approximately 10% after 24 h and storage in formalin resulted in a loss of nearly 30%. Storage by freezing at -80°C for 4 weeks maintained the transparency of the crystalline lens; the spectral transmission measured immediately after defrosting at room temperature coincided exactly with that of the freshly extracted lens. Conclusions: The transparency of porcine crystalline lens is affected by the storage method. The visible spectrum is the most affected, evidenced by the effect on the transparency and consequently the amount of light transmitted. The results show that freezing at -80°C maintains the transpa rency of the crystalline lens for at least 4 weeks.

RESUMO Objetivos: Olho de porco é frequentemente usa do como modelos de pesquisa. Este estudo analisa o efeito de di ferentes métodos de armazenamento na preservação da transparência do cristalino de porco. Métodos: Uma curva de transmissão espectral (de 220 até 780 nm) para o cristalino foi experimentalmente determinada após armazenamento em diferentes condições: solução salina, formol, óleo de mamona e congelamento a -80°C. Transmissão total do espectro visível, que foi usada como um índice de transparência foi calculada a partir dessas curvas. Para fins comparativos, lentes frescas foram avaliadas e usadas como controles. Resultados: O armazenamento do cristalino suíno em solução salina ou óleo de mamona resultou uma perda de transparência de aproximadamente 10% após 24 h e o armazenamento em formol resultou uma perda de quase 30%. O armazenamento por congelamento a -80°C durante 4 semanas manteve a transparência do cristalino; a transmissão espectral medida imediatamente após o descongelamen to à temperatura ambiente coincidiu exatamente com a da lente extraída recentemente. Conclusão: A transparência do cristalino suíno é afetada pelo método de armazenamento. O espectro visível é o mais afetado, evidenciado pelo efeito sobre a transparência e consequentemente a quantidade de luz transmitida. Os resultados mostram que o congelamento a -80°C mantém a transparência do cristalino suíno por pelo menos 4 semanas.

The influence of storage method on the transparency of pig crystalline lens.

PURPOSE: The porcine eye is frequently used as a research model. This paper analyzes the effect of different storage methods on the transparency of pig crystalline lens. METHODS: A spectral transmission curve (from 220 to 780 nm) for the crystalline lens was determined experimentally after storage in different conditions: saline solution, formalin, castor oil, and freezing at -80°C. The total transmission in the visible spectrum, which was used as an index of transparency, was calculated from these curves. For comparative purposes, fresh lenses were evaluated and used as controls. RESULTS: Storing the porcine crystalline lens in saline solution or castor oil resulted in a transparency loss of approximately 10% after 24 h and storage in formalin resulted in a loss of nearly 30%. Storage by freezing at -80°C for 4 weeks maintained the transparency of the crystalline lens; the spectral transmission measured immediately after defrosting at room temperature coincided exactly with that of the freshly extracted lens. CONCLUSIONS: The transparency of porcine crystalline lens is affected by the storage method. The visible spectrum is the most affected, evidenced by the effect on the transparency and consequently the amount of light transmitted. The results show that freezing at -80°C maintains the transpa rency of the crystalline lens for at least 4 weeks.

Exome Sequencing of Fresh-frozen or Formalin-fixed Paraffin-embedded B6C3F1/N Mouse Hepatocellular Carcinomas Arising Either Spontaneously or due to Chronic Chemical Exposure.

Hepatocellular carcinoma (HCC) is the third leading cause of cancer-related death worldwide; however, the mutational properties of HCC-associated carcinogens remain largely uncharacterized. We hypothesized that mechanisms underlying chemical-induced HCC can be characterized by evaluating the mutational spectra of these tumors. To test this hypothesis, we performed exome sequencing of B6C3F1/N HCCs that arose either spontaneously in vehicle controls ( n = 3) or due to chronic exposure to gingko biloba extract (GBE; n = 4) or methyleugenol (MEG; n = 3). Most archived tumor samples are available as formalin-fixed paraffin-embedded (FFPE) blocks, rather than fresh-frozen (FF) samples; hence, exome sequencing from paired FF and FFPE samples was compared. FF and FFPE samples showed 63% to 70% mutation concordance. Multiple known (e.g., Ctnnb1T41A, BrafV637E) and novel (e.g., Erbb4C559S, Card10A700V, and Klf11P358L) mutations in cancer-related genes were identified. The overall mutational burden was greater for MEG than for GBE or spontaneous HCC samples. To characterize the mutagenic mechanisms, we analyzed the mutational spectra in the HCCs according to their trinucleotide motifs. The MEG tumors clustered closest to Catalogue of Somatic Mutations in Cancer signatures 4 and 24, which are, respectively, associated with benzo(a)pyrene- and aflatoxin-induced HCCs in humans. These results establish a novel approach for classifying liver carcinogens and understanding the mechanisms of hepatocellular carcinogenesis.

Allergoids for Allergy Treatment.

BACKGROUND: Chemically modified allergen extracts, known as allergoids, are commonly used for treating allergic patients. In general terms, the concept of allergoids implies allergen extracts with a reduction of their allergenicity maintaining their immunogenicity. Different methods to obtain allergoids have been developed in the past years, opening attractive lines of research. OBJECTIVE: To review the different approaches to allergoid development as well as their characterization, mechanism of action and efficacy and safety issues. METHODS: A revision and analysis of the different types of allergoids has been performed, with special attention to patents submitted and granted in the last years. Additionally, updated information about the mechanism of action and clinical evidence and safety of allergoids has been discussed. RESULTS: Principally, allergoids are obtained by the polymerization of native allergen extracts with aldehydes, including formaldehyde or glutaraldehyde. However, recent patents and publications about different chemical modifications have been presented, as well as about the use of new adjuvants with allergoids. Regarding the characterization, allergoids require more sophisticated analytical methods than native extracts, as a consequence of their properties and characteristics. CONCLUSION: In the last years, the partial understanding of the mechanism of action and the generation of clinical evidence of different types of allergoids, linked to their excellent safety profile and their convenience for a quick build up phase, have made of allergoids an excellent product for allergy treatment.

Mammals divert endogenous genotoxic formaldehyde into one-carbon metabolism.

The folate-driven one-carbon (1C) cycle is a fundamental metabolic hub in cells that enables the synthesis of nucleotides and amino acids and epigenetic modifications. This cycle might also release formaldehyde, a potent protein and DNA crosslinking agent that organisms produce in substantial quantities. Here we show that supplementation with tetrahydrofolate, the essential cofactor of this cycle, and other oxidation-prone folate derivatives kills human, mouse and chicken cells that cannot detoxify formaldehyde or that lack DNA crosslink repair. Notably, formaldehyde is generated from oxidative decomposition of the folate backbone. Furthermore, we find that formaldehyde detoxification in human cells generates formate, and thereby promotes nucleotide synthesis. This supply of 1C units is sufficient to sustain the growth of cells that are unable to use serine, which is the predominant source of 1C units. These findings identify an unexpected source of formaldehyde and, more generally, indicate that the detoxification of this ubiquitous endogenous genotoxin creates a benign 1C unit that can sustain essential metabolism.

Reductive amination derivatization for the quantification of garlic components by isotope dilution analysis.

In this work, we synthesized internal standards for four garlic organosulfur compounds (OSCs) by reductive amination with 13C, D2-formaldehyde, and developed an isotope dilution analysis method to quantitate these organosulfur components in garlic samples. Internal standards were synthesized for internal absolute quantification of S-allylcysteine (SAC), S-allylcysteine sulfoxide (alliin), S-methylcysteine (SMC), and S-ethylcysteine (SEC). We used a multiple reaction monitoring (MRM) to detect 13C, D2-formaldehyde-modified OSCs by ultrahigh-performance liquid phase chromatography coupled with tandem mass spectrometry (UHPLC-MS/MS) and obtained MS spectra showing different ratios of 13C, D2-formaldehyde-modified and H2-formaldehyde-modified compounds. The resulting labeled and unlabeled OSCs were exhibited correlation coefficient (R2) ranged from 0.9989 to 0.9994, respectively. The average recoveries for four OSCs at three concentration levels ranged from 89% to 105%. By 13C, D2-formaldehyde and sodium cyanoborohydride, the reductive amination-based method can be utilized to generate novel internal standard for isotope dilution and to extend the quantitative application.

Highly stable microwave susceptible agents via encapsulation of Ti-mineral superfine powders in urea-formaldehyde resin microcapsules for tumor hyperthermia therapy.

In this study, Ti-mineral superfine powders (Ti-MSP) encapsulated in urea-formaldehyde resin microcapsules (Ti-MSP@UF-MC) were successfully prepared via a one-step microemulsion method for the first time. Because of the strong confinement effects, the Ti-MSP@UF-MC possessed perfect microwave heating effects. The temperature was 9.3 °C higher than that of the saline solution, superior to UF-MC (no significant microwave heating effect, 0 °C) and Ti-MSP (5.1 °C). The Ti-MSP@UF-MC showed low toxicity and good biocompatibility via a series of studies, including a hemolysis study and the MTT assay in vitro and in vivo. When the concentration was below 1000 µg mL(-1), the hemolysis rate was lower than 5% (hemolysis study). When the concentration was below 400 µg mL(-1), the cell activity was higher than 80% (MTT assay). Moreover, the Ti-MSP@UF-MC exhibited an ideal CT imaging effect in vivo owing to the large molecular weight of Ti-MSP. The Ti-MSP@UF-MC showed a favorable microwave therapy effect in vivo. Using mice bearing H22 tumor cells as an animal model, the tumor suppression rate could reach 100%.

Guiding Principles of Hydrogenase Catalysis Instigated and Clarified by Protein Film Electrochemistry.

Protein film electrochemistry (PFE) is providing cutting-edge insight into the chemical principles underpinning biological hydrogen. Attached to an electrode, many enzymes exhibit "reversible" electrocatalytic behavior, meaning that a catalyzed redox reaction appears reversible or quasi-reversible when viewed by cyclic voltammetry. This efficiency is most relevant for enzymes that are inspiring advances in renewable energy, such as hydrogen-activating and CO2-reducing enzymes. Exploiting the rich repertoire of available instrumental methods, PFE experiments yield both a general snapshot and fine detail, all from tiny samples of enzyme. The dynamic electrochemical investigations blaze new trails and add exquisite detail to the information gained from structural and spectroscopic studies. This Account describes recent investigations of hydrogenases carried out in Oxford, including ideas initiated with PFE and followed through with complementary techniques, all contributing to an eventual complete picture of fast and efficient H2 activation without Pt. By immobilization of an enzyme on an electrode, catalytic electron flow and the chemistry controlling it can be addressed at the touch of a button. The buried nature of the active site means that structures that have been determined by crystallography or spectroscopy are likely to be protected, retained, and fully relevant in a PFE experiment. An electrocatalysis model formulated for the PFE of immobilized enzymes predicts interesting behavior and gives insight into why some hydrogenases are H2 producers and others are H2 oxidizers. Immobilization also allows for easy addition and removal of inhibitors along with precise potential control, one interesting outcome being that formaldehyde forms a reversible complex with reduced [FeFe]-hydrogenases, thereby providing insight into the order of electron and proton transfers. Experiments on O2-tolerant [NiFe]-hydrogenases show that O2 behaves like a reversible inhibitor: it is also a substrate, and implicit in the description of some hydrogenases as "H2/O2 oxidoreductases" is the hypothesis that fast and efficient multielectron transfer is a key to O2 tolerance because it promotes complete reduction of O2 to harmless water. Not only is a novel [4Fe-3S] cluster (able to transfer two electrons consecutively) an important component, but connections to additional electron sources (other Fe-S clusters, an electrode, another quaternary structure unit, or the physiological membrane itself) ensure that H2 oxidation can be sustained in the presence of O2, as demonstrated with enzyme fuel cells able to operate on a H2/air mixture. Manipulating the H-H bond in the active site is the simplest proton-coupled electron-transfer reaction to be catalyzed by an enzyme. Unlike small molecular catalysts or the surfaces of materials, metalloenzymes are far better suited to engineering the all-important outer-coordination shell. Hence, recent successful site-directed mutagenesis of the conserved outer-shell "canopy" residues in a [NiFe]-hydrogenase opens up new opportunities for understanding the mechanism of H2 activation beyond the role of the inner coordination shell.

A novel reductive amination method with isotopic formaldehydes for the preparation of internal standard and standards for determining organosulfur compounds in garlic.

Garlic (Allium sativum) is a long-cultivated plant that is widely utilized in cooking and has been employed as a medicine for over 4000 years. In this study, we fabricated standards and internal standards (ISs) for absolute quantification via reductive amination with isotopic formaldehydes. Garlic has four abundant organosulfur compounds (OSCs): S-allylcysteine, S-allylcysteinine sulfoxide, S-methylcysteine, and S-ethylcysteine are abundant in garlic. OSCs with primary amine groups were reacted with isotopic formaldehydes to synthesize ISs and standards. Cooked and uncooked garlic samples were compared, and we utilized tandem mass spectrometry equipped with a selective reaction monitoring technique to absolutely quantify the four organosulfur compounds.

Targeted DNA and RNA sequencing of fine-needle biopsy FFPE specimens in patients with unresectable hepatocellular carcinoma treated with sorafenib.

The multi-kinase inhibitor sorafenib is now used as standard therapy for advanced hepatocellular carcinoma (HCC). Predictive biomarkers of response to sorafenib are thus necessary. The purpose of this study was to assess the feasibility of using targeted DNA and RNA sequencing to elucidate candidate biomarkers of sorafenib response using fine-needle biopsy, formalin-fixed paraffin-embedded (FFPE) specimens in patients with HCC. Targeted DNA and RNA deep sequencing were feasible for the evaluation of fine-needle biopsy FFPE specimens obtained from 46 patients with HCC treated with sorafenib. Frequent mutations of suppressor genes, such as CTNNB1 (34.8%) and TP53 (26.1%), were detected in the HCC tumors. After excluding these suppressor genes, the average numbers of detected oncogene mutations differed significantly between the non-PD and PD groups (P = 0.0446). This result suggests that the oncogene mutational burden in the tumor might be associated with the clinical response to sorafenib. We have identified candidate gene expression (TGFa, PECAM1, and NRG1) in tumor for the prediction of sorafenib response and PFS by RNA sequencing. Our findings provide new insights into biomarkers for sorafenib therapy and allow us to discuss future therapeutic strategies.

Computational protein design enables a novel one-carbon assimilation pathway.

We describe a computationally designed enzyme, formolase (FLS), which catalyzes the carboligation of three one-carbon formaldehyde molecules into one three-carbon dihydroxyacetone molecule. The existence of FLS enables the design of a new carbon fixation pathway, the formolase pathway, consisting of a small number of thermodynamically favorable chemical transformations that convert formate into a three-carbon sugar in central metabolism. The formolase pathway is predicted to use carbon more efficiently and with less backward flux than any naturally occurring one-carbon assimilation pathway. When supplemented with enzymes carrying out the other steps in the pathway, FLS converts formate into dihydroxyacetone phosphate and other central metabolites in vitro. These results demonstrate how modern protein engineering and design tools can facilitate the construction of a completely new biosynthetic pathway.