ABSTRACT
Thirty-eight years after its discovery, the safety of [60]fullerene (C60), the most abundant fullerene with many potential applications, particularly in oxidative stress-related medicine, remains controversial. This is mainly due to the alleged dangers of C60 nanomaterial, which are regularly supported by some publications. While several academic studies have confirmed the safety of C60 in various experimental models, it is well known that C60 aggregates can carry toxic elements. Meanwhile, countless websites offer C60-oily solutions to consumers, without any regulatory consideration. Therefore, an officially certified toxicity study is urgently needed to avoid any public health problems. In this context, we report on the first certified short-term oral toxicity study of soluble C60, designed according to the guidelines of the Organization for Economic Cooperation and Development, with a deviation in the duration (2 weeks instead of 4 weeks) accepted by the U.S. Food and Drug Administration. The results of this study, conducted in an independent accredited European Laboratory, clearly show that C60 in soluble form (0.8 mg/ml of extra virgin olive oil), administered at the highest possible dose of 3.8 mg/kg body weight/day, did not cause any adverse effects in rats after 14 days of daily oral administration. This report should settle the debate on the acute oral toxicity of C60 and pave the way for further preclinical studies. The study is accompanied by a comprehensive report that includes documentation of the raw data.
ABSTRACT
Profiling's of oil yield and fatty acid were monitored during maturation of three different accession of Tunisian apricots (AprB, AprC and AprO) among different days after flowering (DAF) and grown in two different geographical regions of Tunisia. The first results show that a quick distribution started in immature oilseeds apricot and continued until their full maturity. Nine fatty acids were identified in apricot oilseeds such as palmitic, palmitoleic, stearic, oleic, linoleic, linolenic, arachidic, gadoleic and margaric acids. Palmitic, oleic and linoleic acids were determined as major fatty acids in apricot oil varieties. Interestingly, the content of each fatty acid in the three accessions of apricot varied significantly (p < 0.05) during seeds development and especially in wild apricot AprB. PCA analysis in AprB demonstrate that at the time-date of 41 DAF, the production of fatty acids is in its maximum and could have numerous future therapeutics applications.
ABSTRACT
Red blood cells (RBCs) are the subject of clinical attention due to their biological importance. Recently, it has been shown that certain erythrocyte pathologies could be linked to an abnormal lipid composition. In this work, we have developed a simple and fast method using online sample preparation with liquid chromatography coupled to mass spectrometry (SPE-HPLC-MS/MS), to identify a large number of sphingolipids (SL) and phospholipids (PL). The use of online sample preparation considerably reduces analysis times (15 min including extraction and separation of lipids + 2 min for system re-equilibration) and facilitates experimentation while ensuring very good extraction yields. This method was then successfully applied to the quantification of 30 sphingolipids and phospholipids in plasma and erythrocyte extracts from a cohort of individuals with Gaucher disease, treated or not by enzymotherapy. Our results for the study of this disease, led us to establish the lipid profile of the healthy red blood cells, still not very well-known to date. For this, we adopted a semi-targeted approach, based on the use of a triple-quadrupole analyzer and identified more than two hundred different lipid species. These promising results will hopefully enable us to enrich our knowledge of the normal red blood cells lipidome.
Subject(s)
Gaucher Disease , Tandem Mass Spectrometry , Humans , Chromatography, High Pressure Liquid , Phospholipids , Erythrocytes , SphingolipidsABSTRACT
Essential oils (Eos) have demonstrated antiviral activity, but their toxicity can hinder their use as therapeutic agents. Recently, some essential oil components have been used within safe levels of acceptable daily intake limits without causing toxicity. The "ImmunoDefender," a novel antiviral compound made from a well-known mixture of essential oils, is considered highly effective in treating SARS-CoV-2 infections. The components and doses were chosen based on existing information about their structure and toxicity. Blocking the main protease (Mpro) of SARS-CoV-2 with high affinity and capacity is critical for inhibiting the virus's pathogenesis and transmission. In silico studies were conducted to examine the molecular interactions between the main essential oil components in "ImmunoDefender" and SARS-CoV-2 Mpro. The screening results showed that six key components of ImmunoDefender formed stable complexes with Mpro via its active catalytic site with binding energies ranging from -8.75 to -10.30 kcal/mol, respectively for Cinnamtannin B1, Cinnamtannin B2, Pavetannin C1, Syzyginin B, Procyanidin C1, and Tenuifolin. Furthermore, three essential oil bioactive inhibitors, Cinnamtannin B1, Cinnamtannin B2, and Pavetannin C, had significant ability to bind to the allosteric site of the main protease with binding energies of -11.12, -10.74, and -10.79 kcal/mol; these results suggest that these essential oil bioactive compounds may play a role in preventing the attachment of the translated polyprotein to Mpro, inhibiting the virus's pathogenesis and transmission. These components also had drug-like characteristics similar to approved and effective drugs, suggesting that further pre-clinical and clinical studies are needed to confirm the generated in silico outcomes.
Subject(s)
COVID-19 , Oils, Volatile , Humans , Antiviral Agents/chemistry , Molecular Docking Simulation , Molecular Dynamics Simulation , Oils, Volatile/chemistry , Oils, Volatile/pharmacology , Peptide Hydrolases/metabolism , Protease Inhibitors/chemistry , SARS-CoV-2 , Viral Nonstructural Proteins/metabolismABSTRACT
Voluntary drug intoxication is mainly due to drug overdose or the interaction of several drugs. Coma and its associated complications such as hypoventilation, aspiration pneumopathy, and heart rhythm disorders are the main hallmarks of drug intoxication. Conventional detoxification treatments, including gastric lavage or vomiting, administration of ipecac or activated charcoal (CH), and the use of antidotes, have proven to be inefficient and are generally associated with severe adverse effects. To overcome these limitations, titanate nanotubes (TiNTs) are proposed as an efficient emerging detoxifying agent because of their tubular shape and high adsorption capacity. In the present study, the detoxifying ability of TiNTs was evaluated on paracetamol (PR)-intoxicated rats. Results indicate that the loading ability of PR into TiNTs (70%) was significantly higher than that recorded for CH (38.6%). In simulated intestinal medium, TiNTs showed a controlled drug release of less than 10% after 72 h of incubation. In PR-intoxicated rats, TiNTs treatment resulted in a 64% decrease of PR after 4 h of poisoning versus 40% for CH. Concomitantly, TiNTs efficiently reduced PR absorption by 90% after 24 h of poisoning, attenuated the elevated levels of biochemical markers (i.e., alanine aminotransferase, aspartate aminotransferase, creatinine, and TNF-α) and mitigated oxidative stress by increasing the activity of superoxide dismutase and reducing the oxidized glutathione/total glutathione ratio, suggesting a histoprotective effect of TiNTs against paracetamol-induced toxicity in rats. In addition to their safety and high stability in the entire gastro-intestinal tract, biodistribution analysis revealed that TiNTs exhibited low intestinal absorption owing to their large cluster size of compact aggregate nanomaterials across the intestinal villi hindering the absorption of paracetamol. Collectively, these data provide a new and promising solution for in vivo detoxification. TiNTs are expected to have great potential for the treatment of voluntary and accidental intoxication in emergency care.
ABSTRACT
In humans, tetrahydrobiopterin (H4Bip) is the cofactor of several essential hydroxylation reactions which dysfunction cause very serious diseases at any age. Hence, the determination of pterins in biological media is of outmost importance in the diagnosis and monitoring of H4Bip deficiency. More than half a century after the discovery of the physiological role of H4Bip and the recent advent of gene therapy for dopamine and serotonin disorders linked to H4Bip deficiency, the quantification of quinonoid dihydrobiopterin (qH2Bip), the transient intermediate of H4Bip, has not been considered yet. This is mainly due to its short half-life, which goes from 0.9 to 5 min according to previous studies. Based on our recent disclosure of the specific MS/MS transition of qH2Bip, here, we developed an efficient HPLC-MS/MS method to achieve the separation of qH2Bip from H4Bip and other oxidation products in less than 3.5 min. The application of this method to the investigation of H4Bip autoxidation kinetics clearly shows that qH2Bip's half-life is much longer than previously reported, and mostly longer than that of H4Bip, irrespective of the considered experimental conditions. These findings definitely confirm that an accurate method of H4Bip analysis should include the quantification of qH2Bip.
Subject(s)
Biopterins , Tandem Mass Spectrometry , Humans , Biopterins/analysis , Biopterins/metabolism , Pterins , KineticsABSTRACT
Inborn errors of monoamine neurotransmitter metabolism are rare genetic diseases classified as catecholamine and serotonin metabolism disorders or neurotransmitter transportopathies. To diagnose these orphan diseases, monoamine metabolites have been identified and validated as cerebrospinal fluid (CSF) biomarkers: 5-hydroxy-tryptophane, 5-hydroxy-indol-acetic acid, 3-ortho-methyl-DOPA, homovanillic acid, and 3-methoxy-4-hydroxyphenylglycol. The present work presents a UHPLC-MS/MS method developed for the quantification of these metabolites in CSF and compares it with a previously described UHPLC with fluorescence detection (UHPLC-FD) method. MS/MS detection was performed in positive electrospray ionization and multiple reaction monitoring mode. The UHPLC-MS/MS and UHPLC-FD methods were validated in terms of accuracy, linearity, precision and matrix effect. The lower limits of quantification (LLOQ) ranged between 0.5 and 10 nm and between 1 and 5 nm for the UHPLC-MS/MS method and the UHPLC-FD one, respectively. We verified the applicability of both methods by analyzing 30 CSF samples. The measured concentrations were comparable with the reference values described in the literature. The two methods allowed pathological samples to be distinguished from healthy ones for clinical diagnosis. UHPLC-MS/MS and UHPLC-FD methods exhibited very close LLOQs. As the UHPLC-MS/MS method is more selective, it allows faster analysis with a run time of 6 min per run vs. 10 min for the UHPLC-FD method.
Subject(s)
Neurotransmitter Agents , Tandem Mass Spectrometry , Tandem Mass Spectrometry/methods , Chromatography, High Pressure Liquid/methods , Limit of Detection , BiomarkersABSTRACT
Despite recent technological developments in analytical chemistry, separation and direct characterization of transient intermediates remain an analytical challenge. Among these, separation and direct characterization of quinonoid dihydrobiopterin (qH2Bip), a transient intermediate of tetrahydrobiopterin (H4Bip)-dependent hydroxylation reactions, essential in living organisms, with important and varied human pathophysiological impacts, are a clear illustration. H4Bip regeneration may be impaired by competitive nonenzymatic autoxidation reactions, such as isomerization of qH2Bip into a more stable 7,8-H2Bip (H2Bip) isomer, and subsequent nonenzymatic oxidation reactions. The quinonoid qH2Bip intermediate thus plays a key role in H4Bip-dependent hydroxylation reactions. However, only a few experimental results have indirectly confirmed this finding while revealing the difficulty of isolating qH2Bip from H4Bip-containing solutions. As a result, no current H4Bip assay method allows this isomer to be quantified even by liquid chromatography-tandem mass spectrometry (MS/MS). Here, we report isolation, structural characterization, and abundance of qH2Bip formed upon H4Bip autoxidation using three methods integrated into MS/MS. First, we characterized the structure of the two observed H2B isomers using IR photodissociation spectroscopy in conjunction with quantum chemical calculations. Then, we used differential ion mobility spectrometry to fully separate all oxidized forms of H4Bip including qH2Bip. These data are consistent and show that qH2Bip can also be unambiguously identified thanks to its specific MS/MS transition. This finding paves the way for the quantification of qH2Bip with MS/MS methods. Most importantly, the half-life value of this intermediate is nearly equivalent to that of H4Bip (tens of minutes), suggesting that an accurate method of H4Bip analysis should include the quantification of qH2Bip.
Subject(s)
Tandem Mass Spectrometry , Biopterins/analogs & derivatives , Chromatography, Liquid , Isomerism , Oxidation-Reduction , Tandem Mass Spectrometry/methodsABSTRACT
To develop novel carbon-based nanocarriers, we proposed grafting on the [60]Fullerene (C60) biologically active molecules. In this process, the formed derivatives described another approach to use photo-cycloaddition reactions for developing the third nanovector generation. As a result, the photoexcitation of C60 and azomethine ylide (AZMYtrp), with visible light, was considered as the most promising pathway to synthesize fulleropyrrolidine (FPL). After complexation with sodium cation (Na+), the error masses of FPL mono-, bis- and tris-adducts were remarkably decreased to -85.93 %, -53.99 % and -99.42 %, respectively. The formed FPL-Na+ complexes presented a significant capacity for trapping OH and OOH free radicals. In fact, their antiradical properties increased when Na+ was bonded with FPL-Na+ mono-adduct carbonyl oxygens. Comparing FPL bis-adducts regioisomers, under three different AZMYtrp forms, the neutral and anionic-neutral forms of FPL cis1 isomer were considered as the most reactive bis-nanocarriers with mole fractions of about 61 % and 46 %, respectively, in contrast to FPL-Na+, when the mixture was dominated by the anionic-neutral form of cis2 isomer with 50.34 %.
Subject(s)
Fullerenes , Carbon , Cycloaddition Reaction , Free Radicals , IsomerismABSTRACT
Gaucher disease (GD) is a genetic disease with mutations in the GBA gene that encodes glucocerebrosidase causing complications such as anaemia and bone disease. GD is characterized by accumulation of the sphingolipids (SL) glucosylceramide (GL1), glucosylsphingosine (Lyso-GL1), sphingosine (Sph) and sphingosine-1-phosphate (S1P). These SL are increased in the plasma of GD patients and the associated complications have been attributed to the accumulation of lipids in macrophages. Our recent findings indicated that red blood cells (RBCs) and erythroid progenitors may play an important role in GD pathophysiology. RBCs abnormalities and dyserythropoiesis have been observed in GD patients. Moreover, we showed higher SL levels in the plasma and in RBCs from untreated GD patients compared with controls. In this study, we quantified SL in 16 untreated GD patients and 15 patients treated with enzyme replacement therapy. Our results showed that the treatment significantly decreases SL levels in the plasma and RBCs. The increased SL content in RBCs correlates with abnormal RBC properties and with markers of disease activity. Because RBCs lack glucocerebrosidase activity, we investigated how lipid overload could occur in these cells. Our results suggested that SL overload in RBCs occurs both during erythropoiesis and during its circulation in the plasma.
Subject(s)
Erythrocytes/metabolism , Gaucher Disease/blood , Glucosylceramidase/genetics , Sphingolipids/blood , Adolescent , Adult , Aged , Aged, 80 and over , Biomarkers/blood , Child , Child, Preschool , Erythropoiesis/genetics , Female , Gaucher Disease/genetics , Gaucher Disease/pathology , Humans , Lysophospholipids/blood , Macrophages/metabolism , Male , Middle Aged , Psychosine/analogs & derivatives , Psychosine/blood , Sphingosine/analogs & derivatives , Sphingosine/blood , Young AdultABSTRACT
Amino acids and related compounds constitute a class of biomarkers which is analyzed for early diagnosis of metabolic diseases (MDs). Protocols based on liquid chromatography hyphenated to tandem mass spectrometry (LC-MS/MS) are routinely used for MD diagnosis. Our ultimate objective is to evaluate the analytical performance of differential mobility spectrometry (DMS) hyphenated to MS/MS, in the perspective of using DMS-MS/MS as an alternative or complementary method for the topics of emergency in metabolic diagnosis and newborn rapid screening. The aim of the present study is to evaluate the robustness of a DMS-MS/MS protocol for the separation, identification, and quantification of amino acids and related compounds. Performance in terms of peak capacity and separation of isobaric and isomeric species is compared to those using drift tube type ion mobility spectrometry instruments. High reproducibility of the measurement of the DMS compensation voltage (CV) of metabolites shows that this CV parameter, or the corresponding electric field, could be used for application in metabolite identification. Multiple measurements show that the CV value of each AA or related compound is stable over a large period of time (6 months). Potential effects of matrix or concentration of the analytes on the DMS identifier are found to be negligible. Quantification of a selected set of metabolites in human plasmas has been carried out. The method linearity, intra-assay and inter-assay precision, detection limit, quantification limit and trueness analysis were assessed as adequate for both physiological and pathological conditions. Concentration levels of metabolites derived with our DMS-MS protocols were found to be in good agreement with those obtained with routine LC-MS/MS protocols used for the diagnosis of MDs at the Hospital Robert Debré (Paris).
Subject(s)
Amino Acids , Tandem Mass Spectrometry , Chromatography, Liquid , Humans , Infant, Newborn , Reproducibility of Results , Spectrum AnalysisABSTRACT
The aim of the present research is to investigate the effect of three harvest date on the composition of apricot seed. Indeed, triacylglycerols (TAGs) content and composition were studied in developing Tunisian apricot varieties bitter (Bargoug), semi-sweet (Oud Rhayem) and sweet (Chechi Bazza) cultivars at intervals of early (14 DAP), mid phase (28 DAP) and full phase (55 DAP) of oil accumulation by UHPLC-ESI-MS method. Eleven molecular species of triacylglycerols were detected and identified as LLL, LLO, LLP, LOO, LLS/LOP, LPP, OOO, LOS, OOP, POP and OOS. At 14 DAP, LLO was the major TAGs molecular species with 35.4-52.6% (maximum reached in semi-sweet apricot). Others major TAGs were founded at lower content as LOO (17.5-40.3%) and OOO (5.7-12.7%). However, among maturity, three distinct profiles of TAGs molecular species were observed: bitter apricot was significantly richer in OOO molecular species than cultivars ones. However, semi-sweet and sweet cultivars were richer in LLO and LOO molecular species at different time-dates. These latter may provide a schedule for harvesting Tunisian apricot seeds with high quality of oil content.
Subject(s)
Chromatography, High Pressure Liquid/methods , Prunus armeniaca/chemistry , Prunus armeniaca/growth & development , Seeds/chemistry , Seeds/growth & development , Spectrometry, Mass, Electrospray Ionization/methods , Tandem Mass Spectrometry/methods , Triglycerides/chemistry , Triglycerides/isolation & purificationABSTRACT
Here we describe a new HPLC-MS/MS method using a mixed mode stationary phase and a binary gradient of elution for the rapid separation and quantification of AAs in human plasma without derivatization or ion pairing reagent addition. The sample preparation procedure consists in a single dilution step after protein precipitation with sulfosalicylic acid. The proposed method allows for the unambiguous identification and analysis of 52 AAs and related compounds including the separation of isomers and isobars in an 18 min chromatographic run including the conditioning and the equilibration times. AAs were detected by selective reaction monitoring. Internal calibration was used for the quantification of 37 AAs, including 25 using the corresponding isotopically labeled internal standards. External calibration (no internal standard) was used for five additional analytes. Qualitative detection was achieved for the remaining compounds. Validation studies evaluated accuracy, linearity, within- and between-run precision, lower limits of detection and quantification for 37 amino acids present in commonly used quality control samples. For within-run precision CVs averaged 3.8 % (nâ¯=â¯30) for all compounds. For between-run precision, CVs averaged 8.6 % for all compounds (nâ¯=â¯20). Correlation with the common standard ion-exchange chromatography with post-column derivatization method was also performed for 32 plasma samples. While the proposed method is at least 50 times more sensitive, the data showed good correlation with slopes equal or higher than 0.9 and correlation coefficients mostly higher than 0.90. The method was successfully applied for analysis of plasma samples for detection of inherited disorders of amino acid metabolism.
Subject(s)
Amino Acids/blood , Chromatography, Liquid/methods , Tandem Mass Spectrometry/methods , Amino Acids/chemistry , Calibration , Chromatography, High Pressure Liquid/methods , Female , Humans , Infant, Newborn , Reference Standards , Reproducibility of ResultsABSTRACT
Gaucher disease (GD) is a recessively inherited lysosomal storage disorder in which sphingolipids accumulates in the macrophages that transform into Gaucher cells. A growing body of evidence indicates that red blood cells (RBCs) represent important actors in GD pathophysiology. We previously demonstrated that altered RBC properties including increased Lyso-GL1 levels, dyserythropoiesis, and iron metabolism defect in GD patients contribute to anemia and hyperferritinemia. Since RBC defects also correlated well with markers of GD severity and were normalized under enzyme replacement therapy (ERT), the identification of molecules that are deregulated in GD RBCs represents an important issue in the search of pertinent markers of the disease. Here, we found a decreased expression of the GPI-anchored cell surface protein Semaphorin 7A (Sema7A) in RBCs from untreated GD (GD UT) patients, in parallel with increased levels of the soluble form in the plasma. Sema7A plays a role in neural guidance, atherosclerosis, and inflammatory diseases and represents a promigratory cue in physiological and pathological conditions. We showed that the decreased expression of Sema7A in RBCs correlated with their abnormal properties and with markers of GD activity. Interestingly, ERT restored the level of Sema7A to normal values both in RBCs and in plasma from GD patients. We then proposed that SemaA7A represents a simple and pertinent marker of inflammation in GD. Finally, because Sema7A is known to regulate the activity of immune cells, the increased level of soluble Sema7A in GD patients could propagate inflammation in several tissues.
Subject(s)
Gaucher Disease/drug therapy , Semaphorins/therapeutic use , Case-Control Studies , Female , Humans , Male , Prospective Studies , Semaphorins/pharmacologyABSTRACT
Since the early nineties countless publications have reported promising medicinal applications for [60]fullerene (C60) related to its unparalleled affinity towards free radicals. Yet, until now no officially approved C60-based drug has reached the market, notably because of the alleged dangers of C60. Nevertheless, since the publication of the effects of C60 on the lifespan of rodents, a myriad of companies started selling C60 worldwide for human consumption without any approved clinical trial. Nowadays, several independent teams have confirmed the safety of pure C60 while demonstrating that previously observed toxicity was due to impurities present in the used samples. However, a purity criterion for C60 samples is still lacking and there are no regulatory recommendations on this subject. In order to avoid a public health issue and for regulatory considerations, a quality-testing strategy is urgently needed. Here we have evaluated several analytical tools to verify the purity of commercially available C60 samples. Our data clearly show that differential scanning calorimetry is the best candidate to establish a purity criterion based on the sc-fcc transition of a C60 sample (Tonset ≥ 258 K, ∆sc-fccH ≥ 8 J g-1).
ABSTRACT
In the present work, a high-performance liquid chromatographic method coupled with mass spectrometry (HILIC-HPLC /ESI-MS) was used for the characterization and the quantification of glycerophospholipids (GPLs) classes and their molecular species in three genetically different Tunisian apricot cultivars (bitter, sweet and semi-sweet apricots). The application of the proposed method to the analysis of apricot oil allowed to separate and identify 74 molecular species of GPLs. Phosphatidylcholine (PC) class was found to be the most abundant GLPs in the three seed oils (38.6-62.4%) especially in bitter apricot, followed by phosphatidylinositol (PI) and phosphatidylethanolamine (PE) classes with values of 8.3-38.9% and 1.7-25.4% respectively. Phosphatidic acid (PA), phosphatidylglycerol (PG) and lysophosphatidylcholine (LPC) compounds were minor ones with maximums of 11.3%, 9.8% and 9.2% respectively. The results we obtained for the three Tunisian apricot seed varieties clearly indicate that the phospholipids of Tunisian apricot are of great interest. In fact, the high content of phosphatidylcholine (PC) determines it as a suitable and valuable source for obtaining corresponding phospholipids concentrates.
Subject(s)
Glycerophospholipids/genetics , Glycerophospholipids/isolation & purification , Prunus armeniaca/chemistry , Prunus armeniaca/genetics , Seeds/chemistry , Chromatography, High Pressure Liquid/methods , Glycerophospholipids/analysis , Spectrometry, Mass, Electrospray Ionization/methodsABSTRACT
OBJECTIVE: To describe the phenotype and the response to folinic acid supplementation of cerebral folate deficiency (CFD) in adults, a disorder diagnosed on low 5-methyltetrahydro-folate (5MTHF) in cerebrospinal fluid (CSF), which can correspond to a inherited disorder of folate metabolism (IDFM) or to a metabolic consequence of various neurological diseases. METHODS: We conducted a retrospective study on 224 adult patients with neurological symptoms who had a 5MTHF CSF dosage, collecting their neurologic and neuroimaging data. RESULTS: 69 patients had CFD (CSF 5MTHF levelâ¯<â¯41â¯nmol/L), 25 of them had severe CFD (sCFD; ≤25â¯nmol/L) with adult onset neurological symptoms in 41%. 56% of sCFD patients had an underlying identified neurologic disorder, mainly mitochondrial diseases, hepatic encephalopathy and primary brain calcifications (no identified IDFM), the others were classified as undiagnosed. sCFD patients presented most frequently pyramidal syndrome (75%), movement disorders (56%), cerebellar syndrome (50%) and intellectual disability (46%). MRI findings mostly showed white matter abnormalities (WMA; 32%) and calcifications (12%), and were normal in 23%. The clinico-radiological phenotype of sCFD patients was not clearly different from non CFD patients in terms of manifestations frequency. However, their neurological picture was more complex with a higher number of combined neurological symptoms (4.7±1.6 vs 3.4±1.7, pâ¯=â¯.01). In Magnetic Resonance Spectroscopy (MRS), Choline/Creatine (Cho/Cr) ratio was lower in sCFD patients (nâ¯=â¯7) compared to non-CFD patients (nâ¯=â¯73) (pâ¯=â¯.005), with good sensitivity (71%) and excellent specificity (92%). Among twenty-one CFD patients treated with folinic acid, nine had a sustained improvement, all with sCFD but one (50% of sCFD patients improved). In two undiagnosed patients with extremely low 5MTHF CSF values, MRI WMA and low Cho/Cr ratios, folinic acid treatment leaded to a dramatic clinical and radiological improvement. CONCLUSION: CSF 5MTHF dosage should be considered in patients with mitochondrial diseases, primary brain calcifications and unexplained complex neurological disorders especially if associated with WMA, since folinic acid supplementation in patients with sCFD is frequently efficient.
Subject(s)
Cerebellar Diseases/complications , Folic Acid Deficiency/complications , Folic Acid Deficiency/genetics , Mutation/genetics , Proteins/genetics , Adolescent , Adult , Aged , Calcinosis/diagnostic imaging , Calcinosis/etiology , Calcinosis/genetics , Cerebellar Diseases/cerebrospinal fluid , Cerebellar Diseases/diagnostic imaging , Cerebellar Diseases/genetics , Child , Child, Preschool , Female , Folic Acid/cerebrospinal fluid , Folic Acid Deficiency/cerebrospinal fluid , Folic Acid Deficiency/diagnostic imaging , Follow-Up Studies , Humans , Infant , Infant, Newborn , Intellectual Disability , Magnetic Resonance Imaging , Male , Middle Aged , Mitochondrial Diseases/diagnostic imaging , Mitochondrial Diseases/etiology , Retrospective Studies , Tetrahydrofolates/cerebrospinal fluid , White Matter/diagnostic imaging , White Matter/pathology , Young AdultABSTRACT
Since the early nineties, countless publications have been devoted to the study of possible uses of [60] fullerene (C60) and its derivatives in the fields of materials and nano-biomedical sciences. However, in spite of the importance of conformers notably from the pharmacological point of view, the cis/trans isomerization of C60 mono-adducts has been rarely seldom investigated. Here we present the results of DFT calculations of the structural, vibrational and NMR properties of both cis and trans isomers of fulleropyrrolidine mono-adduct obtained by photo-addition of glycine methyl ester to C60. Taken together, our results have shown that the cis isomer is more stable than the trans one. For the cis conformation, the simulated vibrational spectrum shows a more intense peak at 1298 cm-1. While 13C spectra revealed no significant differences between the two isomers as compared to experimental results, the calculated 1H chemical shifts show a significant difference between the two conformers in both the gas phase and in solution. The trans isomer presents a proton at 5.86 ppm, which is more deshielded than the proton of the cis conformer (5.24 ppm).
Subject(s)
Fullerenes/chemistry , Pyrrolidines/chemistry , Carbon-13 Magnetic Resonance Spectroscopy , Density Functional Theory , Isomerism , Models, Molecular , Molecular Conformation , Proton Magnetic Resonance Spectroscopy , VibrationABSTRACT
In this paper, we perform the synthesization of carbon nanoparticles for active principle vectorization, with the suggestion of a reaction mechanism of tryptophan methyl ester addition on [60]fullerene. Firstly, we studied the effect of tryptophan form on its addition reaction on [60]fullerene. So, in order to determine the preferred environment that makes this reaction the most favorable, we considered all tryptophan possible forms in our investigation: the molecular, the zwitterionic, and the dibasic forms. Secondly, we investigate the proposed reaction mechanism of tryptophan methyl ester addition on [60]fullerene using theoretical thermodynamic calculation. Our hypothesis suggests the formation of azomethine ylide molecule in a first step followed by its addition on [60]fullerene in the second step by the photo-addition reaction involving the oxygen in its singlet state. The stability of each reactive intermediate involved in this mechanism is verified thermodynamically. The 12 most stable conformations of azomethine ylide were observed through potential energy surface analysis. They were obtained by a relaxed scan of the four dihedral angles. The calculations were conducted on the optimized geometry of fulleropyrrolidine mono-adduct and the bulk values of its thermodynamic constants were also determined. Infrared spectra observed in 100-4000 cm-1 region confirmed our hypothesis suggesting the first step of azomethine ylide formation followed by the second step of azomethine ylide addition on [60]fullerene by ν(Caliphatic-C-N), ν(Caromatic-C-N) and δ(N-H) coupled with ν(C-N) absorption bond. Graphical abstract Optimized geometry of the Fulleropyrrolidine monoaduct molecule.
ABSTRACT
Due to their central role in biochemical processes, fast separation and identification of amino acids (AA) is of importance in many areas of the biomedical field including the diagnosis and monitoring of inborn errors of metabolism and biomarker discovery. Due to the large number of AA together with their isomers and isobars, common methods of AA analysis are tedious and time-consuming because they include a chromatographic separation step requiring pre- or post-column derivatization. Here, we propose a rapid method of separation and identification of sarcosine, a biomarker candidate of prostate cancer, from isomers using differential ion mobility spectrometry (DIMS) interfaced with a tandem mass spectrometer (MS/MS) instrument. Baseline separation of protonated sarcosine from α- and ß-alanine isomers can be easily achieved. Identification of DIMS peak is performed using an isomer-specific activation mode where DIMS- and mass-selected ions are irradiated at selected wavenumbers allowing for the specific fragmentation via an infrared multiple photon dissociation (IRMPD) process. Two orthogonal methods to MS/MS are thus added, where the MS/MS(IRMPD) is nothing but an isomer-specific multiple reaction monitoring (MRM) method. The identification relies on the comparison of DIMS-MS/MS(IRMPD) chromatograms recorded at different wavenumbers. Based on the comparison of IR spectra of the three isomers, it is shown that specific depletion of the two protonated α- and ß-alanine can be achieved, thus allowing for clear identification of the sarcosine peak. It is also demonstrated that DIMS-MS/MS(IRMPD) spectra in the carboxylic C=O stretching region allow for the resolution of overlapping DIMS peaks. Graphical Abstract á .