Improving enantiomeric resolutions by avoiding peak distortion effects in on-line coupled liquid chromatography to gas chromatography.

In this work, we study the effect of different variables affecting elution profile distortion on the enantiomeric resolution eventually achievable when working with on-line coupled liquid chromatography to gas chromatography (LC-GC). Specifically, the proposed configuration combines achiral reversed-phase liquid chromatography (RPLC) and chiral gas chromatography (enantio-GC), with heptakis-(2,3,6-tri-O-methyl)-ß-cyclodextrin as enantioselective stationary phase to analyse target fractions transferred (from LC to GC) via the through oven transfer adsorption desorption (TOTAD) interface. The high degree of orthogonality resulting from the combination of two chromatographic columns having very different separation mechanisms (and also requiring mobile phases in distinct physical states), as well as integration of the sample preparation step in the first dimension of the system, significantly contributed to exploit the performance of the proposed two-dimensional approach. Occasional adverse effects, which may result in severe peak distortions during LC-GC analysis and could be explained by flow instabilities due to viscous fingering, are circumvented by using the outstanding capacity of the TOTAD interface for achieving effective elimination of the eluent arriving from the LC preseparation.

Relationship between HLA genetic variations, COVID-19 vaccine antibody response, and risk of breakthrough outcomes.

The rapid global distribution of COVID-19 vaccines, with over a billion doses administered, has been unprecedented. However, in comparison to most identified clinical determinants, the implications of individual genetic factors on antibody responses post-COVID-19 vaccination for breakthrough outcomes remain elusive. Here, we conducted a population-based study including 357,806 vaccinated participants with high-resolution HLA genotyping data, and a subset of 175,000 with antibody serology test results. We confirmed prior findings that single nucleotide polymorphisms associated with antibody response are predominantly located in the Major Histocompatibility Complex region, with the expansive HLA-DQB1*06 gene alleles linked to improved antibody responses. However, our results did not support the claim that this mutation alone can significantly reduce COVID-19 risk in the general population. In addition, we discovered and validated six HLA alleles (A*03:01, C*16:01, DQA1*01:02, DQA1*01:01, DRB3*01:01, and DPB1*10:01) that independently influence antibody responses and demonstrated a combined effect across HLA genes on the risk of breakthrough COVID-19 outcomes. Lastly, we estimated that COVID-19 vaccine-induced antibody positivity provides approximately 20% protection against infection and 50% protection against severity. These findings have immediate implications for functional studies on HLA molecules and can inform future personalised vaccination strategies.

1H-HRMAS NMR study of cold smoked Atlantic salmon (Salmo salar) treated with E-beam.

(1)H High resolution magic angle spinning (HRMAS) NMR spectroscopic data in combination with principal components analysis and analysis of variance were used to differentiate between irradiated and non-irradiated cold-smoked Atlantic salmon (Salmo salar). NMR profiling was obtained, with a spectral acquisition time of less than 8 min, from a small sample size of intact white salmon muscle, by non-destructive analysis that includes a very simple and rapid sample preparation step. Results obtained enable the use of creatine, trimethylamine oxide and the sum of phosphorylcholine and glycerophosphorylcholine as diagnostic compounds to detect irradiation treatment. This study shows the potential of (1)H-HRMAS to be a rapid method for investigating compositional changes due to food processing as well as to confirm the presence or absence of some bioactive compounds in irradiated samples.

A quick method for identifying radiolytic hydrocarbons in low-fat-containing food.

BACKGROUND: As radiation-induced alterations of the lipid fraction of foods are related to their initial fat content, concentrations of fat degradation products used as irradiation markers are expected to be lower when irradiating low-fat-containing foods. Thus the sensitivity required when applying analytical methods for identifying irradiation markers in foods eventually depends on their respective amounts of fat. The aim of this study was to perform the qualitative analysis of characteristic hydrocarbons resulting from irradiation of samples with a fat content as low as 25 g kg(-1). RESULTS: A rapid extraction using a small amount of ethyl acetate was the unique sample pretreatment required to accomplish the analysis of radiolytic markers by using on-line coupling of reverse phase liquid chromatography and gas chromatography with mass spectrometry detection (RPLC/GC/MS). Efficient elimination of the large volumes (up to 2170 µL) directly transferred from LC to GC was achieved by optimising the operation mode of the through-oven transfer adsorption/desorption system used as interface. CONCLUSION: The reported procedure allowed confirmation, in less than 65 min, of the occurrence of up to five irradiation markers, namely n-pentadecane, 1-hexadecene, 1,7-hexadecadiene, n-heptadecane and 8-heptadecene, in cooked ham irradiated at doses as low as 2 kGy.

Feasibility of family-directed home-based bimanual intensive therapy combined with modified constraint induced movement therapy (h-BITmCI) in very low and low bimanual functional level: A brief report.

OBJECTIVE: To examine the feasibility of a home-based hybrid Bimanual-Intensive-Therapy combined with modified Constraint-Induced-Movement-Therapy (h-BITmCI) in children with spastic unilateral cerebral palsy (SUCP) with low and very low bimanual functional level. METHODS: A single-group of 10 children aged 5-8 years old, performed the hybrid home Bimanual-Intensive-Therapy (BIT, 80 hours) combined with modified Constraint-Induced-Movement-Therapy (mCIMT, 20 hours): h-BITmCI. Thus, Bimanual Functional Performance (BFP), Quality of Life (QoL) and expectations from families were measured through the Assisting Hand Assessment, (AHA), Pediatric Quality of Life Inventory, for Cerebral Palsy, (PedsQLTM v. 3.0, CP) and a specific questionnaire for families for baseline period (week 0), during the treatment phase (week 4 and week 8) and after the intervention (week 10). Repeated measures ANOVA analysis (with post hoc test correction) was used for the BFP and QoL, with a confidence interval (CI) of 95% and with p value <.008 considered statistically significant. RESULTS: Ten children completed the study with an average of 77-hours-BIT and 17-hours-mCIMT. None of the participants dropped out of the study during the follow-up process, and the parents' expectations were fulfilled, indicating high caregiver compliance. During the first 80 hours of BIT, a mean increase of 3.7 AHA units was obtained for the BFP (p = 1.00) and 1.64 points in the QoL (p = 1.00). Clinically relevant changes were observed in the last two weeks (20 hours mCIMT) with a mean increase of 10.6 AHA units in BFP and 6.29 points in QoL (p < .001). CONCLUSIONS: h-BITmCI protocol is feasible to be performed at home with the family's involvement, obtaining the greatest improvements after 100 hours of both therapies. Thus, mCIMT would be a relevant condition to increase the affected upper limb functionality, rather than the dosage used to obtain clinically relevant changes.

Chiral analysis by online coupling of reversed-phase liquid chromatography to gas chromatography and mass spectrometry.

Enantiomeric composition of selected chiral compounds present in complex mixtures is determined by using the online coupling of reversed-phase liquid chromatography (LC) to gas chromatography (GC) and mass spectrometry. Integration of sample preparation into GC analysis, in a completely automated way, is achieved by means of the effective clean-up resulting from both the LC fractionation step and the eluent elimination provided by the through oven transfer adsorption desorption system used for LC-GC interfacing. The possibilities of the technique are illustrated through some examples concerning the stereodifferentiation in essential oils of major and minor chiral compounds via LC-GC transfer of different volume fractions, ranging from 0.5 to 1.9 ml, which show the significance of the window size for the determination of enantiomeric profiles.

Formation, Characterization, and Occurrence of ß-Carboline Alkaloids Derived from α-Dicarbonyl Compounds and l-Tryptophan.

ß-Carbolines (ßCs) are naturally occurring bioactive alkaloids, whereas α-dicarbonyl compounds are reactive substances generated in foods and in vivo. In this work, l-tryptophan reacted with α-dicarbonyl compounds affording new ß-carbolines. Glyoxal afforded 1-hydroxymethyl-ß-carboline (HME-ßC) and its 3-carboxylic acid, and methylglyoxal afforded 1-(1-hydroxyethyl)-ß-carboline (HET-ßC) and its 3-carboxylic acid. 3-Deoxyglucosone afforded 1-(1,3,4,5-tetrahydroxypent-1-yl)-ß-carboline isomers (1a/b), 1-(1,4,5-trihydroxypent-1-yl)-ß-carboline (2), and 1-(1,5-dihydroxypent-3-en-1-yl)-ß-carboline (3). The formation of these ßCs increased under acidic conditions and with increasing temperature. A mechanism is proposed explaining the conversion of a carbonyl into a hydroxy group based on tautomerism and cyclization to the dihydro-ßC-3-COOH intermediates, which were isolated and gave the ßCs. These α-dicarbonyl-derived ßCs occurred in model reactions of l-tryptophan with fructose or glucose incubated under heating and can be considered as advanced glycation end products (AGEs). They were also present in foods and formed during heating processes. HET-ßC appeared in processed foods, reaching up to 309 ng/g, with the highest amount found in dried tomato, fried onion, toasted bread, and Manuka honey. HME-ßC was only detected in some foods with lower amounts than HET-ßC. HET-ßC appeared in foods as a racemic mixture of enantiomers suggesting the same mechanism of formation as the synthetized product. α-Dicarbonyl-derived ßCs (HET-ßC, HME-ßC, and 1a/b-3) occur in foods and food processing and, therefore, they are ingested during diet.

Arsenate and arsenite differential toxicity in Tetrahymena thermophila.

A comparative analysis of toxicities of both arsenic forms (arsenite and arsenate) in the model eukaryotic microorganism Tetrahymena thermophila (ciliate protozoa) has shown the presence of various detoxification mechanisms and cellular effects comparable to those of animal cells under arsenic stress. In the wild type strain SB1969 arsenate is almost 2.5 times more toxic than arsenite. According to the concentration addition model used in binary metallic mixtures their toxicities show an additive effect. Using fluorescent assays and flow cytometry, it has been detected that As(V) generates elevated levels of ROS/RNS compared to As(III). Both produce the same levels of superoxide anion, but As(V) also causes greater increases in hydrogen peroxide and peroxynitrite. The mitochondrial membrane potential is affected by both As(V) and As(III), and electron microscopy has also revealed that mitochondria are the main target of both arsenic ionic forms. Fusion/fission and swelling mitochondrial and mitophagy, together with macroautophagy, vacuolization and mucocyst extruction are mainly associated to As(V) toxicity, while As(III) induces an extensive lipid metabolism dysfunction (adipotropic effect). Quantitative RT-PCR analysis of some genes encoding antioxidant proteins or enzymes has shown that glutathione and thioredoxin metabolisms are involved in the response to arsenic stress. Likewise, the function of metallothioneins seems to be crucial in arsenic detoxification processes, after using both metallothionein knockout and knockdown strains and cells overexpressing metallothionein genes from this ciliate. The analysis of the differential toxicity of As(III) and As(V) shown in this study provides cytological and molecular tools to be used as biomarkers for each of the two arsenic ionic forms.

Alternatives to continuous positive airway pressure treatment in sleep apnoeas and hypopnoeas syndrome related to myofunctional and postural reeducation therapy: protocol for a systematic review.

INTRODUCTION: The main treatment for sleep apnoeas and hypopnoeas syndrome (SAHS) is continuous positive airway pressure (CPAP). However, patients sometimes do not adhere to the treatment protocol. Supplementary and complementary therapies have appeared as alternatives. Some of the therapies which are especially important are those related to myofunctional (MFT) and postural therapy (PT), as all of them are non-invasive, and their application is simpler than that of CPAP. We aim to present a protocol for a systematic review and meta-analysis for investigating new SAHS treatments, including the protocols and frequency of use and the effects they have on patient signs, symptoms and quality of life. METHODS AND ANALYSIS: The literature search will be conducted using the Cochrane, Web of Science, Medline (via PubMed) and Scopus databases, from January 2020 to December 2020. All types of studies written in English and Spanish that investigate the use of alternative SAHS treatments related to MFT, or more importantly, the combination of MFT and PRT, will be selected. To evaluate their quality, the Critical Appraisal Checklist for Analytical Cross-Sectional will be applied. The primary factor valued in the studies will be the inclusion of MFT and PT reeducation in the treatment. Subgroup analyses will be carried out evaluating the specific type of treatments chosen and the improvements or deteriorations in the level of health and quality of life in the patients. Finally, several patient-related outcomes, namely sleep quality, quality of life and sleep Apnoeas and Hypoapnoeas Index, will be examined. ETHICS AND DISSEMINATION: In this case, ethical approval is not necessary. The data used in the review will be exclusively obtained from published studies, implying there are no privacy concerns. The information obtained will be relevant to understand if the new treatments applied in SAHS are effective, and if postural and MFT therapy used together can be considered an appropriate approach to treat this disease.The results will be published in a peer-reviewed journal.

1H-HRMAS NMR study of smoked Atlantic salmon (Salmo salar).

High-resolution magic angle spinning (HRMAS) NMR spectroscopic data of smoked Atlantic salmon (Salmo salar) were fully assigned by combination of one- and two-dimensional-HRMAS experiments. Complete representative spectra, obtained after few minutes of analysis time, revealed a large number of minor and major compounds in the sample. The methodology is limited by the low sensitivity of NMR, and therefore HRMAS only enables the determination of the most relevant components. These were fatty acids (FAs), carbohydrates, nucleoside derivatives, osmolytes, amino acids, dipeptides and organic acids. For the first time, spectra were resolved sufficiently to allow semiquantitative determination in intact muscle of the highly polyunsaturated FA 22:6 omega-3. Additionally, the feasibility of (1)H-HRMAS NMR metabolite profiling was tested to identify some bioactive compounds during storage. This profiling was carried out by the non-destructive and direct analysis (i.e. without requiring sample preparation and multiple step procedures) of intact salmon muscle. The proposed procedure can be applied to a large number of samples with high throughput due to the short time of analysis and quick evaluation of the data.

Use of absorbent materials in on-line coupled reversed-phase liquid chromatography-gas chromatography via the through oven transfer adsorption desorption interface.

The use of absorbents as retaining materials in the through oven transfer adsorption desorption interface (TOTAD) of an on-line coupled reversed-phase liquid chromatography-gas chromatography system (RPLC-GC) is proposed for the first time. A comparative study of an adsorbent (Tenax TA) and two absorbents, namely polydimethylsiloxane and poly(50% phenyl/50% methylsiloxane) is performed to establish the best experimental conditions for the automated and simultaneous determination of 15 organophosphorus and organochlorine pesticide residues in olive oil. The proposed method provides satisfactory repeatability (RSDs lower, in general, than 8.5%) and sensitivity (limits of detection ranging from 0.6 to 81.9 microg/L) for the investigated compounds.

Absorbents as packing materials in on-line coupling of reversed phase liquid chromatography and gas chromatography via a programmed temperature vaporizer.

A method based on the use of absorbents as packing materials in the interface of the direct coupling between reversed phase liquid chromatography and gas chromatography (RPLC-GC) is proposed. To that end, a comparative study on different adsorbents and absorbents was carried out. Specifically, Tenax TA and Gaschrom were used as adsorbents while polydimethylsiloxane and poly(50% phenyl:50% methylsiloxane) were the absorbents tested. Some experimental variables involved in the solvent elimination were separately optimised for adsorbent and absorbent materials. Relative standard deviations (RSD) lower than 10% were achieved in all cases but the use of absorbents showed interesting advantages with respect to adsorbents, namely a simpler performance of the experimental work, which facilitates the sample preparation step and the subsequent gas chromatographic analysis to be performed.

Polydimethylsiloxane as a packing material in a programmed temperature vaporizer to introduce large-volume samples in capillary gas chromatography.

The viability of using polydimethylsiloxane (PDMS) as a retaining material inside a programmed temperature vaporizer injector for the introduction of large-volume samples in gas chromatography is assessed. To that end, materials made up of Volaspher A-2 and coated with different percentages of PDMS (5%, 15%, and 50%) are considered. In addition, adsorbent (Tenax TA) and absorbent (PDMS) materials are comparatively studied in terms of their retention capacity. A relative standard deviation lower than 5.0% is obtained from the injection of PDMS, whereas values up to 49% are provided by Tenax TA. Significantly higher amounts of different volatile compounds are retained by PDMS in comparison with Tenax TA. In conclusion, the use of PDMS as a packing material seems to be viable for large-volume sampling and particularly recommendable for minor compounds occurring in complex matrices.

Nature and origin of the violet stains on the walls of a Roman tomb.

The Circular Mausoleum tomb (Roman Necropolis of Carmona, Spain) dates back from the first century AD and is characterized by a dense microbial (phototrophic) colonization on the walls and ceiling. However, some walls exhibited an important number of violet stains of unknown origin. The microbial communities of these violet stains are mainly composed of cyanobacteria, streptomycetes and fungi. A strain of Streptomyces parvus, isolated from the walls, produces a violet pigment in culture media. High performance liquid chromatography-mass spectrometry of the culture extracts obtained from this Streptomyces revealed the presence of a few granaticins, pigments with a benzoisochromanequinone structure. When metabolically active in the tomb, S. parvus synthesizes the pigments that diffuse into the mortar. During rain and/or wetting periods, the pigments are solubilized by alkaline waters and elute from the starting position to the surrounding mortar, enlarging the pigmented area and thus contributing to this exceptional biodeterioration phenomenon.

Activation of peroxisome proliferator-activated receptor-alpha and -gamma in auricular tissue from heart failure patients.

OBJECTIVE: Peroxisome proliferator-activated receptors (PPARs), key transcriptional regulators of lipid and energy metabolism in cardiomyocytes, have recently been proposed to modulate cardiovascular pathophysiological responses in experimental models. However, there is little information about the functional activity of PPARs in human heart failure. AIMS: To investigate PPAR-alpha and -gamma expression and activity, and the association with ET-1 production and fibrosis, in cardiac biopsies from patients with end-stage heart failure due to ischemic cardiomyopathy (ICM) in comparison and from non-failing donor hearts. All samples were obtained during cardiac transplantation. METHODS AND RESULTS: Morphological analysis (by Masson trichrome and image analysis) did not detect fibrosis in the left atrium from non-failing donors (NFLA) or from ICM patients (FLA). However, left ventricles from failing hearts (FLV) contained a greater number of fibrotic areas (NFLA: 3.21+/-1.15, FLA: 1.63+/-0.83, FLV: 14.5+/-3.45%; n = 9, P<0.05). By RT-PCR, preproET-1 expression was similar in the non-failing and failing atrium but was significantly higher in the ventricles from failing hearts (NFLA: 1.00+/-0.06, FLA: 1.08+/-0.11, FLV: 1.74+/-0.19; n = 9, P<0.05). PPAR-alpha and PPAP-gamma mRNA (by RT-PCR) and protein (by Western blot) levels were higher in the ventricles from failing hearts compared with the atrium from failing and non-failing hearts. Electrophoretic mobility shift assays showed that PPAR-alpha and PPAP-gamma were not activated in the ventricles (NFLA: 1.00+/-0.11, FLA: 1.89+/-0.24, FLV: 0.95+/-0.07; n = 9, P<0.05). CONCLUSIONS: These data suggest that PPAR-alpha and PPAP-gamma are selectively activated in the atria from ICM patients and might be functionally important in the maintenance of atrial morphology.

Long-term organ protection by doxazosin and/or quinapril as antihypertensive therapy.

BACKGROUND: Even with optimal blood pressure control, organ protection may also depend on the selected therapeutic regime. Angiotensin-converting enzyme inhibitors have been shown to provide excellent organ protection in hypertension, and may show dose-dependent protective effects. Adrenergic alpha blockers have been associated with an increased rate of heart failure in the Antihypertensive and Lipid-Lowering Treatment to Prevent Heart Attack Trial (ALLHAT) and Vasodilator-Heart Failure Trial (V-HeFT). This has been related to a proapoptotic effect of this drug in cardiomyocytes. Our purpose is to compare the heart and renal protection of a high quinapril dose, with a combined low quinapril dose plus doxazosin, in an animal model of chronic hypertension. METHODS: Uninephrectomized spontaneously hypertensive 12-week-old rats were treated for 36 weeks with either quinapril or a combination of doxazosin plus a low quinapril dose. Tight blood pressure control was achieved with both treatments. Renal and cardiac protection was assessed by different parameters, and cardiac apoptosis was evaluated by active caspase-3, apoptotic protein and heat shock protein levels. Untreated hypertensive and normotensive rats were included as controls. RESULTS: Both treatments showed significant heart and renal protection compared with untreated animals. Both therapeutic regimes showed similar protection in renal and cardiac pathology, coronary media fibrosis, myocardial apoptosis and cardiac index. Proteinuria and left ventricular hypertrophy regression were significantly lower in the quinapril group compared with the combined treatment group. CONCLUSIONS: Blood pressure control with a high quinapril dose provided higher organ protection than a combined therapy with a lower quinapril dose. This effect was not due to a deleterious effect of doxazosin.

Use of the enantiomeric composition for the assessment of the authenticity of fruit beverages.

Enantiomeric compositions of chiral terpenes in commercial fruit beverages were examined by solid phase microextraction-gas chromatography (SPME-GC). Optimization of the method was accomplished on the basis of some parameters involved in the extraction, such as heating temperature and extraction time, that provided the highest peak areas, 60 degrees C and 2 min being the optimal values. With the proposed method relative standard deviation (RSD) values from three replicates ranging from 2 to 12% were obtained. The enantiomeric distribution of some terpenes remained constant, whereas other terpenes (linalool, terpinen-4-ol, and alpha-terpineol) exhibited a considerable variation among samples. This can be indicative of the eventual addition of aromas to some fruit beverages.

Supercritical fluid extraction of all-trans-lycopene from tomato.

A procedure is proposed for the supercritical fluid extraction of all-trans-lycopene from tomato using carbon dioxide at 40 degrees C without modifier. The present method minimizes the risk of degradation via isomerization and oxidation of health-promoting ingredients, such as lycopene. The effect of different experimental variables on the solvating power of the supercritical fluid was evaluated in terms of both the selectivity achievable in the process and the yield of the extraction of all-trans-lycopene. Satisfactory separations of the all-trans-lycopene isomers from the cis counterparts were achieved using a C(30) column. The obtained extract contained 88% all-trans-lycopene and 12% cis-lycopene.

Stereodifferentiation of chiral compounds using reversed-phase liquid chromatography coupled with capillary gas chromatography.

A method is described for the enantiomeric quantitation of some chiral compounds via online coupling of reversed-phase liquid chromatography-gas chromatography. The evaluation of some variables affecting the experimentation (i.e., the packing material used in the interface, volume of the transferred fraction, desorption time, initial temperature of the interface, and purge time) makes it possible to optimize the recoveries obtained for some chiral terpenes and lactones using a capillary column of beta-cyclodextrin dissolved in OV-1701. The proposed method allows the enantiomeric analysis of aqueous matrices obtaining relative standard deviations lower than 9% and detection limits ranging from 0.26-0.93 ppm for the investigated compounds.

A comparative study of the ability of different techniques to extract menthol from Mentha piperita.

Supercritical fluid extraction, direct thermal desorption, hydroalcoholic extraction, and atomization are used to extract menthol from leaf plants of Mentha piperita. The investigated methods are comparatively evaluated on the basis of their reliability to determine the enantiomeric distribution of menthol. The enantioselectivity required for the gas chromatographic analysis is achieved using Octakis (2,6-di-O-n-pentyl-3-O-butyryl)-gamma-cyclodextrin as the stationary phase. From the obtained results, it is established that there is a significant effect of the combination of pressure and temperature to achieve the effective isolation and fractionation of the less and most volatile compounds using supercritical fluids.