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In this study, overloaded elution profiles under ultra-high-pressure liquid chromatographic (UHPLC) conditions and accounting for the severe pressure and temperature gradients generated, are compared with experimental data. The model system consisted of an C18 column packed with 1.7-µm particles (i.e., a UHPLC column) and the solute was 1,3,5-tri‑tert-butylbenzene eluted with a mobile phase composed of 85/15 (v/v) acetonitrile/water. Two thermal modes were considered, and the solute was eluted at the very high inlet pressures necessary to achieve a highly efficient and rapid chromatographic process, as provided by using columns packed with small particles. However, the high inlet pressure and high linear velocity of the mobile phase caused the production of a significant amount of heat, and consequently, the formation of axial and radial temperature gradients. Due to these gradients, the retention and the mobile phase velocity were no longer constant. Thus, simple mathematical models consisting only of the mass balance equations are unsuitable to properly model the elution profiles. Here, the elution concentration profiles were predicted using a combined two-dimensional heat and mass transfer model, also including the calculation of the mobile phase velocity distribution. The isotherm adsorption model was the bi-Langmuir isotherm model with Henry constants that depended on the local temperature and pressure in the column. These adjustments allowed us to precisely account for changes in the shape and retention of the overloaded concentration profiles in the mobile phase. The proposed model provided accurate predictions of the overloaded concentration profiles, demonstrating good agreement with experimental profiles eluted under severe pressure and temperature gradients in the column even in the most extreme cases where the pressure drops reached 846 bar and the temperature gradients equaled 0.15 K mm−1 and 0.95 K mm−1 in the axial and the radial directions, respectiv

In this study, overloaded elution profiles under ultra-high-pressure liquid chromatographic (UHPLC) conditions and accounting for the severe pressure and temperature gradients generated, are compared with experimental data. The model system consisted of an C18 column packed with 1.7-µm particles (i.e., a UHPLC column) and the solute was 1,3,5-tri‑tert-butylbenzene eluted with a mobile phase composed of 85/15 (v/v) acetonitrile/water. Two thermal modes were considered, and the solute was eluted at the very high inlet pressures necessary to achieve a highly efficient and rapid chromatographic process, as provided by using columns packed with small particles. However, the high inlet pressure and high linear velocity of the mobile phase caused the production of a significant amount of heat, and consequently, the formation of axial and radial temperature gradients. Due to these gradients, the retention and the mobile phase velocity were no longer constant. Thus, simple mathematical models consisting only of the mass balance equations are unsuitable to properly model the elution profiles. Here, the elution concentration profiles were predicted using a combined two-dimensional heat and mass transfer model, also including the calculation of the mobile phase velocity distribution. The isotherm adsorption model was the bi-Langmuir isotherm model with Henry constants that depended on the local temperature and pressure in the column. These adjustments allowed us to precisely account for changes in the shape and retention of the overloaded concentration profiles in the mobile phase. The proposed model provided accurate predictions of the overloaded concentration profiles, demonstrating good agreement with experimental profiles eluted under severe pressure and temperature gradients in the column even in the most extreme cases where the pressure drops reached 846 bar and the temperature gradients equaled 0.15 K mm−1 and 0.95 K mm−1 in the axial and the radial directions, respectiv

Hemoglobinopathies constitute one of the most common inherited hematological disorders in the world with an increasing global disease burden each year. One among them is sickle cell disease with diverse genotypes and wide phenotypic heterogenity. Many subgroups exist within the umbrella of sickle cell disease. Hb S/DPunjab, a rare hemoglobinopathy, is one of them, mimics sickle cell disease, and is discussed in the present study. We describe one such unusual clinical case of a young child who presented with intermittent fever and joint problems. The study case was found to have Hb S/DPunjab by high performance liquid chromatography. Clinical and hematological details of this rare condition is only briefly discussed in the literature. Precise diagnosis can be made using high performance liquid chromatography in conjunction with family studies.

This study investigated the influence of pH on the retention of solutes using a mixed-mode column with carboxyl (-COOH) groups acting as weak cation exchanger bonded to the terminal of C18 ligands (C18-WCX column) and a traditional reversed-phase C18 column. First, a model based on electrostatic theory was derived and successfully used to predict the retention of charged solutes (charged, and ionizable) as a function of mobile phase pH on a C18-WCX column. While the Horváth model predicts the pH-dependent retention of ionizable solutes in reversed-phase liquid chromatography (RPLC) solely based on solute ionization, the developed model incorporates the concept of surface potential generated on the surface of the stationary phase and its variation with pH. To comprehensively understand the adsorption process, adsorption isotherms for these solutes were individually acquired on the C18-WCX and reversed-phase C18 columns. The adsorption isotherms followed the Langmuir model for the uncharged solute and the electrostatically modified Langmuir model for charged solutes. The elution profiles for the single components were calculated from these isotherms using the equilibrium dispersion column model and were found to be in close agreement with the experimental elution profiles. To enable modelling of two-component cases involving charged solute(s), a competitive adsorption isotherm model based on electrostatic theory was derived. This model was later successfully used to calculate the elution profiles of two components for scenarios involving (a) a C18 Column: two charged solutes, (b) a C18 Column: one charged and one uncharged solute, and (c) a C18-WCX Column: two charged solutes. The strong alignment between the experimental and calculated elution profiles in all three scenarios validated the developed competitive adsorption model.

This study investigated the influence of pH on the retention of solutes using a mixed-mode column with carboxyl (-COOH) groups acting as weak cation exchanger bonded to the terminal of C18 ligands (C18-WCX column) and a traditional reversed-phase C18 column. First, a model based on electrostatic theory was derived and successfully used to predict the retention of charged solutes (charged, and ionizable) as a function of mobile phase pH on a C18-WCX column. While the Horváth model predicts the pH-dependent retention of ionizable solutes in reversed-phase liquid chromatography (RPLC) solely based on solute ionization, the developed model incorporates the concept of surface potential generated on the surface of the stationary phase and its variation with pH. To comprehensively understand the adsorption process, adsorption isotherms for these solutes were individually acquired on the C18-WCX and reversed-phase C18 columns. The adsorption isotherms followed the Langmuir model for the uncharged solute and the electrostatically modified Langmuir model for charged solutes. The elution profiles for the single components were calculated from these isotherms using the equilibrium dispersion column model and were found to be in close agreement with the experimental elution profiles. To enable modelling of two-component cases involving charged solute(s), a competitive adsorption isotherm model based on electrostatic theory was derived. This model was later successfully used to calculate the elution profiles of two components for scenarios involving (a) a C18 Column: two charged solutes, (b) a C18 Column: one charged and one uncharged solute, and (c) a C18-WCX Column: two charged solutes. The strong alignment between the experimental and calculated elution profiles in all three scenarios validated the developed competitive adsorption model.

In this study, overloaded elution profiles under ultra-high-pressure liquid chromatographic (UHPLC) conditions and accounting for the severe pressure and temperature gradients generated, are compared with experimental data. The model system consisted of an C18 column packed with 1.7-µm particles (i.e., a UHPLC column) and the solute was 1,3,5-tri‑tert-butylbenzene eluted with a mobile phase composed of 85/15 (v/v) acetonitrile/water. Two thermal modes were considered, and the solute was eluted at the very high inlet pressures necessary to achieve a highly efficient and rapid chromatographic process, as provided by using columns packed with small particles. However, the high inlet pressure and high linear velocity of the mobile phase caused the production of a significant amount of heat, and consequently, the formation of axial and radial temperature gradients. Due to these gradients, the retention and the mobile phase velocity were no longer constant. Thus, simple mathematical models consisting only of the mass balance equations are unsuitable to properly model the elution profiles. Here, the elution concentration profiles were predicted using a combined two-dimensional heat and mass transfer model, also including the calculation of the mobile phase velocity distribution. The isotherm adsorption model was the bi-Langmuir isotherm model with Henry constants that depended on the local temperature and pressure in the column. These adjustments allowed us to precisely account for changes in the shape and retention of the overloaded concentration profiles in the mobile phase. The proposed model provided accurate predictions of the overloaded concentration profiles, demonstrating good agreement with experimental profiles eluted under severe pressure and temperature gradients in the column even in the most extreme cases where the pressure drops reached 846 bar and the temperature gradients equaled 0.15 K mm−1 and 0.95 K mm−1 in the axial and the radial directions, respectiv

In this study, overloaded elution profiles under ultra-high-pressure liquid chromatographic (UHPLC) conditions and accounting for the severe pressure and temperature gradients generated, are compared with experimental data. The model system consisted of an C18 column packed with 1.7-µm particles (i.e., a UHPLC column) and the solute was 1,3,5-tri‑tert-butylbenzene eluted with a mobile phase composed of 85/15 (v/v) acetonitrile/water. Two thermal modes were considered, and the solute was eluted at the very high inlet pressures necessary to achieve a highly efficient and rapid chromatographic process, as provided by using columns packed with small particles. However, the high inlet pressure and high linear velocity of the mobile phase caused the production of a significant amount of heat, and consequently, the formation of axial and radial temperature gradients. Due to these gradients, the retention and the mobile phase velocity were no longer constant. Thus, simple mathematical models consisting only of the mass balance equations are unsuitable to properly model the elution profiles. Here, the elution concentration profiles were predicted using a combined two-dimensional heat and mass transfer model, also including the calculation of the mobile phase velocity distribution. The isotherm adsorption model was the bi-Langmuir isotherm model with Henry constants that depended on the local temperature and pressure in the column. These adjustments allowed us to precisely account for changes in the shape and retention of the overloaded concentration profiles in the mobile phase. The proposed model provided accurate predictions of the overloaded concentration profiles, demonstrating good agreement with experimental profiles eluted under severe pressure and temperature gradients in the column even in the most extreme cases where the pressure drops reached 846 bar and the temperature gradients equaled 0.15 K mm−1 and 0.95 K mm−1 in the axial and the radial directions, respectiv

In this study, overloaded elution profiles under ultra-high-pressure liquid chromatographic (UHPLC) conditions and accounting for the severe pressure and temperature gradients generated, are compared with experimental data. The model system consisted of an C18 column packed with 1.7-µm particles (i.e., a UHPLC column) and the solute was 1,3,5-tri‑tert-butylbenzene eluted with a mobile phase composed of 85/15 (v/v) acetonitrile/water. Two thermal modes were considered, and the solute was eluted at the very high inlet pressures necessary to achieve a highly efficient and rapid chromatographic process, as provided by using columns packed with small particles. However, the high inlet pressure and high linear velocity of the mobile phase caused the production of a significant amount of heat, and consequently, the formation of axial and radial temperature gradients. Due to these gradients, the retention and the mobile phase velocity were no longer constant. Thus, simple mathematical models consisting only of the mass balance equations are unsuitable to properly model the elution profiles. Here, the elution concentration profiles were predicted using a combined two-dimensional heat and mass transfer model, also including the calculation of the mobile phase velocity distribution. The isotherm adsorption model was the bi-Langmuir isotherm model with Henry constants that depended on the local temperature and pressure in the column. These adjustments allowed us to precisely account for changes in the shape and retention of the overloaded concentration profiles in the mobile phase. The proposed model provided accurate predictions of the overloaded concentration profiles, demonstrating good agreement with experimental profiles eluted under severe pressure and temperature gradients in the column even in the most extreme cases where the pressure drops reached 846 bar and the temperature gradients equaled 0.15 K mm−1 and 0.95 K mm−1 in the axial and the radial directions, respectiv

This study investigated the influence of pH on the retention of solutes using a mixed-mode column with carboxyl (-COOH) groups acting as weak cation exchanger bonded to the terminal of C18 ligands (C18-WCX column) and a traditional reversed-phase C18 column. First, a model based on electrostatic theory was derived and successfully used to predict the retention of charged solutes (charged, and ionizable) as a function of mobile phase pH on a C18-WCX column. While the Horváth model predicts the pH-dependent retention of ionizable solutes in reversed-phase liquid chromatography (RPLC) solely based on solute ionization, the developed model incorporates the concept of surface potential generated on the surface of the stationary phase and its variation with pH. To comprehensively understand the adsorption process, adsorption isotherms for these solutes were individually acquired on the C18-WCX and reversed-phase C18 columns. The adsorption isotherms followed the Langmuir model for the uncharged solute and the electrostatically modified Langmuir model for charged solutes. The elution profiles for the single components were calculated from these isotherms using the equilibrium dispersion column model and were found to be in close agreement with the experimental elution profiles. To enable modelling of two-component cases involving charged solute(s), a competitive adsorption isotherm model based on electrostatic theory was derived. This model was later successfully used to calculate the elution profiles of two components for scenarios involving (a) a C18 Column: two charged solutes, (b) a C18 Column: one charged and one uncharged solute, and (c) a C18-WCX Column: two charged solutes. The strong alignment between the experimental and calculated elution profiles in all three scenarios validated the developed competitive adsorption model.

This study investigated the influence of pH on the retention of solutes using a mixed-mode column with carboxyl (-COOH) groups acting as weak cation exchanger bonded to the terminal of C18 ligands (C18-WCX column) and a traditional reversed-phase C18 column. First, a model based on electrostatic theory was derived and successfully used to predict the retention of charged solutes (charged, and ionizable) as a function of mobile phase pH on a C18-WCX column. While the Horváth model predicts the pH-dependent retention of ionizable solutes in reversed-phase liquid chromatography (RPLC) solely based on solute ionization, the developed model incorporates the concept of surface potential generated on the surface of the stationary phase and its variation with pH. To comprehensively understand the adsorption process, adsorption isotherms for these solutes were individually acquired on the C18-WCX and reversed-phase C18 columns. The adsorption isotherms followed the Langmuir model for the uncharged solute and the electrostatically modified Langmuir model for charged solutes. The elution profiles for the single components were calculated from these isotherms using the equilibrium dispersion column model and were found to be in close agreement with the experimental elution profiles. To enable modelling of two-component cases involving charged solute(s), a competitive adsorption isotherm model based on electrostatic theory was derived. This model was later successfully used to calculate the elution profiles of two components for scenarios involving (a) a C18 Column: two charged solutes, (b) a C18 Column: one charged and one uncharged solute, and (c) a C18-WCX Column: two charged solutes. The strong alignment between the experimental and calculated elution profiles in all three scenarios validated the developed competitive adsorption model.

This study investigated the influence of pH on the retention of solutes using a mixed-mode column with carboxyl (-COOH) groups acting as weak cation exchanger bonded to the terminal of C18 ligands (C18-WCX column) and a traditional reversed-phase C18 column. First, a model based on electrostatic theory was derived and successfully used to predict the retention of charged solutes (charged, and ionizable) as a function of mobile phase pH on a C18-WCX column. While the Horváth model predicts the pH-dependent retention of ionizable solutes in reversed-phase liquid chromatography (RPLC) solely based on solute ionization, the developed model incorporates the concept of surface potential generated on the surface of the stationary phase and its variation with pH. To comprehensively understand the adsorption process, adsorption isotherms for these solutes were individually acquired on the C18-WCX and reversed-phase C18 columns. The adsorption isotherms followed the Langmuir model for the uncharged solute and the electrostatically modified Langmuir model for charged solutes. The elution profiles for the single components were calculated from these isotherms using the equilibrium dispersion column model and were found to be in close agreement with the experimental elution profiles. To enable modelling of two-component cases involving charged solute(s), a competitive adsorption isotherm model based on electrostatic theory was derived. This model was later successfully used to calculate the elution profiles of two components for scenarios involving (a) a C18 Column: two charged solutes, (b) a C18 Column: one charged and one uncharged solute, and (c) a C18-WCX Column: two charged solutes. The strong alignment between the experimental and calculated elution profiles in all three scenarios validated the developed competitive adsorption model.

Tato bakalářská práce se věnuje stanovením biologicky aktivních látek obsažených v merlíku čilském pomocí moderních analytických metod. Nejprve je uveden popis merlíku čilského, chemické složení, zpracování a jeho využití. Dále jsou také uvedeny pozitivní účinky na zdraví a stručně popsány teoretické základy separačních technik v kapalné fázi a metod sledujících antioxidační aktivitu látek. Experimentální část práce je věnována stanovení antioxidační kapacity extraktů připravených z merlíku čilského pomocí spektrofotometrických metod., This bachelor thesis is devoted to the determination of biologically active substances contained in Quinoa using modern analytical methods. Firstly, a description of quinoa, its chemical composition, processing, and uses is given. The health benefits are also presented and the theoretical basis of liquid phase separation techniques and methods monitoring the antioxidant activity of the compounds are briefly described. The experimental part of the work deals with the determination of the antioxidant capacity of extracts prepared from quinoa by spectrophotometric methods., Fakulta chemicko-technologická, Posluchačka seznámila komisi se svojí bakalářskou prací. Dále reagovala na připomínky a zodpověděla otázky členů komise: Zkoušela jste studovanou potravinu zařadit do jídelníčku? Upřesněte účinnost použitých rozpouštědel? Jak se změní antioxidační účinnost po povaření studované potraviny? Vysvětlete použité zkratky. Jak byly stanoveny prezentované chyby?, Dokončená práce s úspěšnou obhajobou

Diabetes mellitus is one of the most common diseases worldwide. The continued prevalence increase, together with the complexity of the disease with severe long-term complications makes efficient diagnostical and monitorial methods very important. The determination of glycated hemoglobin, HbA1c, plays an important role in both, underlying the importance of high-quality analytical instruments. Whenever a new instrument is introduced into routine clinical use an evaluation is needed. This study evaluated the agreement between the new Tosoh G11 HbA1c high performance liquid chromatography system as a replacement for the Bio-Rad D-100 at the Chromatographic Department of the Institute of Laboratory Medicine, Faculty of Medicine, University of Debrecen. In total, 66 samples were analyzed using both analyzers and a Bland-Altman comparison was performed together with a correlation study. Additionally, a linearity check, optimal error and routine error testing was carried out specifically for the Tosoh G11. The results showed high agreement between measurements, r=0.9975 (p<0.001) for HbA1c mmol/mol and r=0.9971 (p<0.001) for HbA1c %. High repeatability and accuracy were observed of the Tosoh G11 with the highest variation coefficient of 1.50 % and linearity r =0.9999. The conducted study supports the replacement of the Bio-Rad D-100 to the Tosoh G11., Diabetes mellitus är en av de vanligaste världsomfattande sjukdomarna. Fortsatt prevalensökning, tillsammans med den komplexa sjukdomsbilden innefattande allvarliga långsiktiga komplikationer gör effektiva diagnostiska och övervakande metoder väldigt viktiga. Bestämning av glykerat hemoglobin, HbA1c, spelar en viktig roll i båda, vilket kräver att analysinstrument av hög kvalitet finns. När ett nytt instrument introduceras inom kliniska rutinanalyser krävs utvärdering. Denna studie utvärderade överrensstämmelsen mellan det nya Tosoh G11 HbA1c högupplösande vätskekromatografisystemet som en ersättning för Bio-Rad D-100 på den kromatografiska enheten av laboratoriemedicin på Debrecens universitet. Totalt analyserades 66 prov med respektive instrument och Bland-Altman jämförelse genomfördes tillsammans med en korrelationsstudie. Dessutom utfördes ett linjäritetstest, optimal- och rutinfeltest specifikt för Tosoh G11. Resultaten visade en hög överrensstämmelse mellan mätningarna, r=9975 (p<0.001) för HbA1c mmol/mol och r =9971 (p<0.001) för HbA1c %. Tosoh G11 visade hög repeterbarhet och noggrannhet, med den högsta variationskoefficient på endast 1.50 % och en linjäritet på r=0.9999. Den genomförda studien stödjer Tosoh G11 som ersättning för Bio-Rad D-100.

Diabetes mellitus is one of the most common diseases worldwide. The continued prevalence increase, together with the complexity of the disease with severe long-term complications makes efficient diagnostical and monitorial methods very important. The determination of glycated hemoglobin, HbA1c, plays an important role in both, underlying the importance of high-quality analytical instruments. Whenever a new instrument is introduced into routine clinical use an evaluation is needed. This study evaluated the agreement between the new Tosoh G11 HbA1c high performance liquid chromatography system as a replacement for the Bio-Rad D-100 at the Chromatographic Department of the Institute of Laboratory Medicine, Faculty of Medicine, University of Debrecen. In total, 66 samples were analyzed using both analyzers and a Bland-Altman comparison was performed together with a correlation study. Additionally, a linearity check, optimal error and routine error testing was carried out specifically for the Tosoh G11. The results showed high agreement between measurements, r=0.9975 (p<0.001) for HbA1c mmol/mol and r=0.9971 (p<0.001) for HbA1c %. High repeatability and accuracy were observed of the Tosoh G11 with the highest variation coefficient of 1.50 % and linearity r =0.9999. The conducted study supports the replacement of the Bio-Rad D-100 to the Tosoh G11., Diabetes mellitus är en av de vanligaste världsomfattande sjukdomarna. Fortsatt prevalensökning, tillsammans med den komplexa sjukdomsbilden innefattande allvarliga långsiktiga komplikationer gör effektiva diagnostiska och övervakande metoder väldigt viktiga. Bestämning av glykerat hemoglobin, HbA1c, spelar en viktig roll i båda, vilket kräver att analysinstrument av hög kvalitet finns. När ett nytt instrument introduceras inom kliniska rutinanalyser krävs utvärdering. Denna studie utvärderade överrensstämmelsen mellan det nya Tosoh G11 HbA1c högupplösande vätskekromatografisystemet som en ersättning för Bio-Rad D-100 på den kromatografiska enheten av laboratoriemedicin på Debrecens universitet. Totalt analyserades 66 prov med respektive instrument och Bland-Altman jämförelse genomfördes tillsammans med en korrelationsstudie. Dessutom utfördes ett linjäritetstest, optimal- och rutinfeltest specifikt för Tosoh G11. Resultaten visade en hög överrensstämmelse mellan mätningarna, r=9975 (p<0.001) för HbA1c mmol/mol och r =9971 (p<0.001) för HbA1c %. Tosoh G11 visade hög repeterbarhet och noggrannhet, med den högsta variationskoefficient på endast 1.50 % och en linjäritet på r=0.9999. Den genomförda studien stödjer Tosoh G11 som ersättning för Bio-Rad D-100.

Reversed-phase high-performance liquid chromatography (RP-HPLC) is a widely accepted method in drug and toxicological analyses to detect and quantify drugs and metabolites. However, current RP-HPLC methods use mobile phases that contain significant amounts of organic modifiers such as acetonitrile. Although these organic modifiers are effective, they have been linked to negative environmental effects. The focus of this research is to develop a water-rich mobile phase system composed of >90% water and a smaller amount of a nonpolar organic modifier. Long-chain alcohols such as 1-butanol have proved to be suitable organic modifiers for water-rich RP-HPLC analysis. The use of 1-butanol as an organic modifier is studied in the current research to analyze drugs belonging to amphetamine-related drug classes. A series of differing gradients and elution tests were run to observe which method produced the best separation and resolution among the compounds. The preliminary samples include quinine caffeine, vanillin, and acetylsalicylic acid. An isocratic elution using mobile phases containing 0.1% formic acid and butanol in water was able to separate the preliminary sample. The retention times are 1.775, 4.96, 9.769, and 13.373, respectively; additionally, the resolution values are 1.13, 2.53, 2.72, and 1.86. The illicit substances include amphetamine, methamphetamine, and MDEA. An isocratic elution using mobile phases containing a phosphate buffer, pH-adjusted to 7.0 with a 3.5% butanol in water has produced the efficient peak resolution in an illicit drug sample. The retention times are 7.208, 7.932, and 10.053, respectively. A Waters: Atlantis T3 column helped to achieve the separation but could not fully prevent peak tailing. The peak tailing was caused by silanol effects that occur in the stationary phase, but efforts have been made to correct those effects. However, with the minimal peak tailing, the peaks still achieved a resolution of 1.52, 1.74, and 4.50 which mee

The method describes a procedure for determining the individual and total alcoholic compound content of olive oils and olive pomace oils as well as of blends of both. The alcoholic compounds in olive and olive pomace oils comprise aliphatic alcohols, sterols and triterpenic dialcohols.

The flexibility and required softness of polyvinyl chloride (PVC) -one of the most widely used polymeric materials for medical device production- is achieved by the addition of an appropriate amount of plasticizer (up to 40 %) into the polymer matrix. As plasticizers are not chemically bound to PVC they can easily migrate to surrounding media such as saliva, blood, plasma or serum resulting in undesirable toxic effects. Migrations of plasticizers from PVC can be significantly suppressed by embedding plasticized PVC into another -‘less harmful’- polymer, thus preventing its direct contact with the surrounding media. Co-extruded TPU/PVC/TPU catheters were prepared with various thicknesses of thermoplastic polyurethane (TPU) layers to evaluate the retarding effect of the protective TPU layer on releasing bis(2-ethylhexyl) terephthalate plasticizer (the only one approved by European Pharmacopoeia). A substantial difference was found in the migration of plasticizer from non-coated and coated (three-layer) catheters into a saline solution and acetonitrile which was dependent on the time exposed to the concentration of released plasticizer determined simultaneously by reverse-phase liquid chromatographic technique and weight loss analysis. The results proved that the TPU inner and outer layers represent an efficient protective barrier and significantly reduce amounts of released plasticizer. © 2022 Elsevier Ltd

Here, cell wall of a zygomycete fungus, Rhizopus delemar, grown on bread waste was wet spun into monofilaments. Using the whole cell wall material omits the common chitosan isolation and purification steps and leads to higher material utilization. The fungal cell wall contained 36.9% and 19.7% chitosan and chitin, respectively. Solid state NMR of the fungal cell wall material confirmed the presence of chitosan, chitin, and other carbohydrates. Hydrogels were prepared by ultrafine grinding of the cell wall, followed by addition of lactic acid to protonate the amino groups of chitosan, and subsequently wet spun into monofilaments. The monofilament inhibited the growth of Bacillus megaterium (Gram+ bacterium) and Escherichia coli (Gram- bacterium) significantly (92.2% and 99.7% respectively). Cytotoxicity was evaluated using an in vitro assay with human dermal fibroblasts, indicating no toxic inducement from exposure of the monofilaments. The antimicrobial and biocompatible fungal monofilaments, open new avenues for sustainable biomedical textiles from abundant food waste., QC 20230111

Cellulose acetate (CA) is subjected to different aqueous environments (e.g., lake water, seawater, artificial seawater) under controlled laboratory conditions to investigate the degradation potential in natural water systems. The main changes in the CA films are detected during the first months of the 12-month study. Approximately 5% and 10% weight losses are observed during the first month of aging at RT and at 60 °C, respectively. The temperature effect is rather negligible and the weight loss also does not significantly increase on prolonged aging. A quantitative high-performance liquid chromatography analysis shows that this weight loss is mainly caused by deacetylation and depending on the aging conditions, 10–30% of the acetate groups are lost during the first 1–3 months of aging. Fourier transform infrared and nuclear magnetic resonance analyses further support this. It is well established that a high degree of substitution (DS) is the bottleneck for biodegradation of CA and a significantly higher biodegradation rate has been demonstrated for CA materials with degree of substitution, DS<2, compared to those with DS>2. The degree of deacetylation observed here is enough to decrease the DS to below 2, which can therefore have a significant effect on the subsequent biodegradation., QC 20230615

Mitochondria represent an important drug target for many phatology, including neurodegeneration, metabolic disease, heart failure, ischemia-reperfusion injury, and cancer. Mitochondrial dysfunctions are caused by mutation in mitochondrial DNA or in nuclear genes encoding mitochondrial proteins. Cell-penetrating peptides (CPPs) have been employed to overcome biological barriers, target this organelle, and therapeuticaly restore mitochondrial functions. Here, we describe recent methods used to deliver oligonucleotides targeting mitochondrial protein by using mitochondrial penetrating peptides. In particular, we highlight recent advances of formulated peptides/oligonucleotides nanocomplexes as a proof-of-principle for pharmaceutical form of peptide-based therapeutics., QC 20220602

Magenta lilly pilly (Syzygium paniculatum) is an Australian native tree that produces berry fruits that are rich in phytochemicals reportedly beneficial to human health. Here we explored the biological activities of polyphenol-enriched extracts from the magenta lilly pilly fruit, benchmarking it against traditional sources including purple sweet potato and blackberry. We show that the extracts exert potent antioxidant and neuroprotective properties as well as antimicrobial activity against Staphylococcus aureus. The phenolic composition of lilly pilly was investigated using liquid chromatography coupled to mass spectrometry (HPLC-DAD-MS), revealing anthocyanins to be the primary component in high abundance compared to traditional anthocyanin-containing plants. Three anthocyanins from lilly pilly, along with their glycosylation patterns and stability, were characterised. Altogether, our results demonstrate the potential to exploit magenta lilly pilly fruits as a high-yielding source of phenolics with beneficial biological properties of potential interest for multiple downstream applications., QC 20230522

The crystal growth of curcumin in pure propan-2-ol and in this solvent containing two structurally related impurities, demethoxycurcumin (DMC) and bisdemethoxycurcumin (BDMC), has been investigated by seeded isothermal desupersaturation experiments at 283, 293 and 308 K. In situ attenuated total reflectance UV-visible spectroscopy and focused beam reflectance measurement were used as process analytical technologies to monitor the solution concentration over time and to track the particle counts. Both impurities are found to slow down the growth of curcumin spherulites. The product particles collected after growth in the presence of the impurities present a rougher and more porous surface appearance in comparison to curcumin crystalline material grown in pure solutions. A detailed analysis of the powder X-ray diffraction patterns along with compositional analysis by high performance liquid chromatography of grown crystals reveals that impurities are not incorporated into the solid phase except at the highest impurity concentrations explored. By molecular modelling it is shown that the influence of impurities on the diffractograms of the material grown at high impurity concentrations is consistent with the formation of a solid solution. By fitting the Birth and Spread theory to the experimental growth data it is found that the interfacial energy for growth in the presence of the impurities is higher than that for growth of curcumin in the absence of impurities. Two scenarios are envisaged explaining the results. The first is that 3-D nucleation occurs on the impurity encumbered CUR crystallite generating new crystalline material on which crystal growth may continue. The second scenario envisages that a molecular cluster from the solution attaches to the impurity-encumbered surface and condenses into a crystalline surface nucleus without perfect lattice matching., QC 20230404

Glycerol-plasticized wheat gluten was explored for producing soft high-density biofoams using dry upscalable extrusion (avoiding purposely added water). The largest pore size was obtained when using the food grade ammonium bicarbonate (ABC) as blowing agent, also resulting in the highest saline liquid uptake. Foams were, however, also obtained without adding a blowing agent, possibly due to a rapid moisture uptake by the dried protein powder when fed to the extruder. ABC's low decomposition temperature enabled extrusion of the material at a temperature as low as 70 °C, well below the protein aggregation temperature. Sodium bicarbonate (SBC), the most common food-grade blowing agent, did not yield the same high foam qualities. SBC's alkalinity, and the need to use a higher processing temperature (120 °C), resulted in high protein cross-linking and aggregation. The results show the potential of an energy-efficient and industrially upscalable low-temperature foam extrusion process for competitive production of sustainable biofoams using inexpensive and readily available protein obtained from industrial biomass (wheat gluten)., QC 20230614

Home Analytical Chemistry for the Study of Paintings and the Detection of Forgeries Chapter Analysis of Natural and Synthetic Organic Lakes and Pigments by Chromatographic and Mass Spectrometric Techniques Francesca Sabatini & Ilaria Degano Chapter First Online: 09 September 2022 390 Accesses Part of the Cultural Heritage Science book series (CUHESC) Abstract This chapter aims to provide an overview of natural and synthetic organic lakes and pigments and of how their analysis by chromatography and mass spectrometry can provide useful information in the examination of works of art. The characteristics of natural organic colourants are reviewed, including information on their provenance and uses in the production of lakes; information on the date of production, availability as paint products and application in artworks of the most important synthetic organic pigments is also provided. This chapter specifically illustrates the state-of-the-art of organic pigments analysis by high performance liquid chromatography with different detectors, and by gas chromatography – mass spectrometry also coupled to analytical pyrolysis. Different mass spectrometry based techniques used both as detector and directly applied on a sample or its extract are also presented. Particular attention is paid to sample preparation; further issues related to difficulties in finding relevant standards or reference materials and understanding photo-degradation processes occurring in degraded organic pigments are also discussed. Several case studies are presented, showing the potentialities of liquid chromatography interfaced with mass spectrometry in determining properties of the samples which can be of assistance for assessing the provenance, dating or even the authenticity of a painting.

Objective. To determine the fatty acid (FA) profile, conjugated linoleic acid (CLA) and isomers in cow's milk produced under organic conditions in southeastern Mexico. Materials and methods. Forty-eight milk samples were collected during one year from three production units (PU 1, 2 and 3) and a collecting tank (CT) in the municipality of Tecpatán, Chiapas (12 months x 4 = 48), following the guidelines established in Mexican regulations. Fat was extracted with detergent solution; AG analysis was performed by gas chromatography with flame ionization detector and ALC and isomers were obtained by high performance liquid chromatography (HPLC) with UV-Vis detector. Results. Chromatographic analyses identified and quantified 23 FA in all milks, from C4 to C20:1. Statistical analysis (ANOVA, Tukey) showed significance (p<0.05) at C4-C10, C16, C18, C18:1n9t, C18:2n6 and C20; in all cases the TC was similar in at least two PU. The mean values over time exhibited homogeneous regularity behavior. CLA and 9 of its isomers were also determined; ANOVA did not show significance. The isomer C18:2 9c-11t had the highest percentage of total CLA with a mean value of 90.06%. Conclusions. Gas and liquid chromatographic analyses allowed characterization of FA, CLA and isomers profiles in organic milk produced in southeastern Mexico, which had not been studied so far., Objetivo. Determinar el perfil de ácidos grasos (AG), ácido linoleico conjugado (ALC) e isómeros en leche de vaca producida en condiciones orgánicas en el sureste de México. Materiales y métodos. Durante un año se colectaron 48 muestras de leche, proveniente de tres unidades de producción (UP 1, 2 y 3) y tanque colector (TC) del municipio de Tecpatán, Chiapas (12 meses x 4 = 48), siguiendo las pautas establecidas en normatividad mexicana. La grasa se extrajo con solución detergente; el análisis de AG se realizó por cromatografía de gases con detector de ionización de flama y el ALC e isómeros se obtuvieron mediante cromatografía líquida de alta resolución (CLAR) con detector UV-Vis. Resultados. Los análisis cromatográficos identificaron y cuantificaron 23 AG en todas las leches, desde el C4 hasta el C20:1. El análisis estadístico (ANOVA, Tukey) arrojó significancia (p<0.05) en C4-C10, C16, C18, C18:1n9t, C18:2n6 y C20; en todos los casos el TC fue similar en al menos dos UP. Los valores medios a través del tiempo exhibieron comportamiento con regularidad homogénea. Asimismo, se determinaron el ALC y 9 de sus isómeros; el ANOVA no arrojó significancia. El isómero C18:2 9c-11t tuvo el mayor porcentaje del ALC total con un valor medio de 90.06%. Conclusiones. Los análisis de cromatografía de gases y de líquidos permitieron caracterizar los perfiles de AG, ALC e isómeros en leche orgánica que se produce en el sureste mexicano, que hasta el momento no se habían estudiado.

The flexibility and required softness of polyvinyl chloride (PVC) -one of the most widely used polymeric materials for medical device production- is achieved by the addition of an appropriate amount of plasticizer (up to 40 %) into the polymer matrix. As plasticizers are not chemically bound to PVC they can easily migrate to surrounding media such as saliva, blood, plasma or serum resulting in undesirable toxic effects. Migrations of plasticizers from PVC can be significantly suppressed by embedding plasticized PVC into another -'less harmful'polymer, thus preventing its direct contact with the surrounding media. Co-extruded TPU/PVC/TPU catheters were prepared with various thicknesses of thermoplastic polyurethane (TPU) layers to evaluate the retarding effect of the protective TPU layer on releasing bis(2-ethylhexyl) terephthalate plasticizer (the only one approved by European Pharmacopoeia). A substantial difference was found in the migration of plasticizer from non-coated and coated (three-layer) catheters into a saline solution and acetonitrile which was dependent on the time exposed to the concentration of released plasticizer determined simultaneously by reverse-phase liquid chromatographic technique and weight loss analysis. The results proved that the TPU inner and outer layers represent an efficient protective barrier and significantly reduce amounts of released plasticizer.

К настоящему времени химическое оружие в Российской Федерации уничтожено. Бывшие предприятия по уничтожению химического оружия подлежат перепрофилированию. Важнейшим этапом этой деятельности является установление безопасности элементов инженерной инфраструктуры объектов для оценки возможности вовлечения их в хозяйственный оборот. Из всех отравляющих веществ, обращавшихся на объектах, наиболее токсичным и персистентным является, так называемый, Ви-газ (вещество VR, О-изобутил-S-[(2-диэтиламино)этил]метилфосфонотиоат). Аттестованные газохроматографические методики, применявшиеся ранее для контроля остаточных количеств этого вещества на поверхностях и в глубинных слоях материалов, основаны на конверсии вещества во фторангидрид на таблетках, импрегнированных фторидом серебра. В настоящее время эти таблетки не производятся, и применение методик невозможно. Применение жидкостной тандемной хроматомасс-спектрометрии обеспечило прямое определение VR на уровне порога токсического действия в материалах различного матричного состава. Наряду с VR в рамках единой методики достигается также ранее методически не обеспеченное определение токсичной кислоты, образующейся при замене в молекуле VR алкильного радикала при кислороде протоном. Предложена методика высокочувствительного определения VR и наиболее токсичного продукта его гидролиза S-[(2-диэтиламино)этил]метилфосфонотиоата в смывах с поверхностей и в глубинных слоях материалов различного состава. Методика основана на внесении в пробу внутреннего стандарта (параоксона), экстракции метанолом, концентрировании экстракта и последующем анализе методом высокоэффективной жидкостной хроматографии с тандемным масс-спектрометрическим детектированием в режиме мониторинга заданных реакций (MRM). Реализация разработанной процедуры позволяет определять аналиты на уровне, соответствующем порогам токсического действия. Особое внимание в методике уделено достоверной идентификации аналитов, что имеет определяющее значение при международных рассл, To date, chemical weapons in the Russian Federation have been completely destroyed. Former chemical weapons destruction facilities (CWDFs) are subject to re-profiling. The most important stage of this activity is to assess the safety of engineering elements of the facilities to find out whether they can be involved in national economy. The most toxic and persistent substance processed at the CWDFs was a V-series agent. The certified gas chromatographic methods previously used for control of residual amounts of this substance on the surfaces and in deep layers of materials were based on its conversion to acid fluoride on silver fluoride pads. The latter are no longer produced, and these methods are impossible to use. The use of liquid tandem chromatography-mass spectrometry (HPLC-MS/MS) provided a direct determination of VR in the different matrices at the level of the toxicity threshold. At the same time HPLC-MS/MS method allows to determine the toxic acid formed by the hydrolysis of the P–O bond in the VR molecule. Previously no methods for the determination of this acid were available. A unified procedure for the sensitive target determination of VR (O-isobutyl S- (2-diethylamino)ethyl methylphosphonothioate) and its most toxic hydrolysis product S-[(2-diethylamino)ethyl] methylphosphonothioic acid in surface wipes and in deep layers of various materials is proposed. The method involves the addition of an internal standard (paraoxon) to the sample, extraction with methanol, concentration of the extract, and analysis by HPLC-MS/MS in the multiple reaction monitoring (MRM) mode. The detection limits of the analytes are close to their toxic threshold levels. Special emphasis in the procedure is paid to the reliability of identification of the analytes, which is of key importance in international investigations under the Chemical Weapons Convention.

The synthesis of purine conjugates with natural amino acids is one of the promising directions in search for novel therapeutic agents, including antimycobacterial agents. The purpose of this study was to synthesize N-(purin-6-yl)dipeptides containing the terminal fragment of (S)-glutamic acid. To obtain the target compounds, two synthetic routes were tested. The first of them is based on coupling of N-(purin-6-yl)-(S)-amino acids to dimethyl (S)-glutamate in the presence of carbodiimide coupling agent followed by the removal of ester groups. However, it turned out that this coupling process was accompanied by racemization of the chiral center of N-(purin-6-yl)-α-amino acids and in all cases led to mixtures of (S,S)- and (R,S)-diastereomers (6:4). Individual (S,S)-diastereomers were obtained using an alternative approach based on the nucleophilic substitution of chlorine in 6-chloropurine or 2-amino-6-chloropurine with corresponding dipeptides as nucleophiles. The enantiomeric purity of the target compounds was confirmed by chiral HPLC. To test the assumption that racemization of the chiral center of N-(purin-6-yl)-α-amino acids occurs with the participation of nitrogen atoms of the imidazole ring via the stage of formation of a chirally labile intermediate, we obtained such structural analogs of N-(purin-6-yl)-(S)-alanine as N-(9-benzylpurin-6-yl)-(S)-alanine and N-(7-deazapurin-6-yl)-(S)-alanine. It was found that coupling of these compounds to dimethyl (S)-glutamate was also accompanied by racemization. This indicates that the imidazole fragment does not play a crucial role in this process. When testing the antimycobacterial activity of some of the obtained compounds, conjugates with moderate activity against the laboratory Mycobacterium tuberculosis H37Rv strain (MIC 3.1–6.25 μg/mL) were identified. © 2021, The Author(s), under exclusive licence to Springer-Verlag GmbH, AT part of Springer Nature.

Рекуррентная аппроксимация абсолютных времен удерживания аналитов (tR) в обращенно-фазовой высокоэффективной жидкостной хроматографии (ОФ ВЭЖХ) при разных содержаниях органического компонента в составе элюента (C) рекомендована как способ выявления обратимого образования гидратов. Критерием их образования являются отклонения зависимостей tR(С + DС) = atR(С) + b (*) от линейности (DС – постоянный «шаг» вариаций концентраций; в нашем случае выбрано DС = 5 % объем.). Однако такие отклонения достаточно невелики и поэтому их измерение предъявляет особые требования к надежности используемого оборудования. Кроме того, их причиной могут быть и другие факторы, например, сложно контролируемое несоответствие реальных расходов элюента их заданным значениям. Сравнение значений tR, полученных для одних и тех же аналитов с применением одной и той же колонки в одинаковых условиях, но на разных приборах, с использованием системы метанол – вода в качестве элюента подтвердило, что один набор значений составляет 76-98 % от величин, измеренных на другом приборе. Следовательно, расходы элюента во втором случае в такой же пропорции меньше, чем в первом. Предложен простой способ выявления подобных отклонений по результатам рекуррентной аппроксимации наборов значений tR = f(C) любых соединений, не образующих гидратов в условиях ОФ ВЭЖХ (в нашем случае выбран хлорбензол). Значения коэффициентов корреляции (R) рекуррентных зависимостей (*) превышающие 0.999 указывают на отсутствие влияния любых искажающих факторов., Parts of experimental results presented in the current paper has been obtained using the equipment of the Resource Centre “Methods of analysis of substances’ properties” of Scientific Park of St. Petersburg State University. The authors are grateful to the staff of this Center for assistance.

Liquid phase microextraction (LPME) into a microfluidic has undergone great advances focused on downscaled and miniaturized devices. In this work, a microfluidic device was developed for the extraction of sulfonamides in order to accelerate the mass transfer and passive diffusion of the analytes from the donor phase to the acceptor phase. The subsequent analysis was carried out by high performance liquid chromatography with UV-DAD (HPLC-DAD). Several parameters affecting the extraction efficiency of the method such as the supported liquid membrane, composition of donor and acceptor phase and flow rate were investigated and optimized. Tributyl phosphate was found to be a good supported liquid membrane which confers not only great affinity for analytes but also long-term stability, allowing more than 20 consecutive extractions without carry over effect. Under optimum conditions, extraction efficiencies were over 96 % for all sulfonamides after 10 minutes extraction and only 10 µL of sample was required. Relative standard deviation was between 3-5 % for all compounds. Method detection limits were 45, 57, 54 and 33 ng mL−1 for sulfadiazine (SDI), sulfamerazine (SMR), sulfamethazine (SMT) and sulfamethoxazole (SMX), respectively. Quantitation limits were 0.15, 0.19, 0.18 and 0.11 µg mL−1 for SDI, SMR, SMT SMX, respectively. The proposed microfluidic device was successfully applied for the determination of sulfonamides in urine samples with extraction efficiencies within the range of 86-106 %. The proposed method improves the procedures proposed to date for the determination of sulfonamides in terms of efficiency, reduction of the sample volume and extraction time.

Background: A research priority in finding a cure for HIV is to establish methods to accurately locate and quantify where and how HIV persists in people living with HIV (PLWH) receiving suppressive antiretroviral therapy (ART). Infusing copper-64 (64Cu) radiolabelled broadly neutralising antibodies targeting HIV envelope (Env) with CT scan and positron emission tomography (PET) identified HIV Env in tissues in SIV infected non-human primates. We aimed to determine if a similar approach was effective in people living with HIV (PLWH). Method(s): Unmodified 3BNC117 was compared with 3BNC117 bound to the chelator MeCOSar and 64Cu (64Cu-3BNC117) in vitro to assess binding and neutralization. In a clinical trial 64Cu-3BNC117 was infused into HIV uninfected (Group 1), HIV infected and viremic (viral load, VL >1000 c/mL; Group 2) and HIV infected aviremic (VL <20 c/mL; Group 3) participants using two dosing strategies: high protein (3mg/kg unlabeled 3BNC117 combined with <5mg 64Cu-3BNC117) and trace (<5mg 64Cu-3BNC117 only). All participants were screened for 3BNC117 sensitivity from virus obtained from viral outgrowth. Magnetic resonance imaging (MRI)/PET and pharmacokinetic assessments (ELISA for serum 3BNC117 concentrations and gamma counting for 64Cu) were performed 1, 24- and 48-hours post dosing. The trial (clincialtrials.gov NCT03063788) primary endpoint was comparison of PET standard uptake values (SUVs) in regions of interest (e.g lymph node groups and gastrointestinal tract). Finding(s): Comparison of unmodified and modified 3BNC117 in vitro demonstrated no difference in HIV binding or neutralisation. 17 individuals were enrolled of which 12 were dosed including Group 1 (n=4, 2 high protein, 2 trace dose), Group 2 (n=6, 2 high protein, 4 trace) and Group 3 (n=2, trace only). HIV+ participants had a mean CD4 of 574 cells/microL and mean age 43 years. There were no drug related adverse effects and no differences in tissue uptake in regions of interest (e.g lymph nod

Background: Uric acid, a metabolic product of purine degradation in humans, is a risk factor for developing gout and type 2 diabetes, and supplementation with quercetin lowers plasma uric acid in mildly hyperuricemic men. Here we examined the mechanism of inhibition of enzymes involved in uric acid metabolism by quercetin, conjugates and microbial catabolites, and measured the effect of lowered circulating uric acid on endothelial cell gene expression. Method(s): Inhibition of adenosine deaminase (ADA), purine nucleoside phosphorylase (PNP) and xanthine oxidoreductase (XOR) activity by quercetin and metabolites was determined by HPLC. Human umbilical vein endothelial cells (HUVECs) were cultured under conditions mimicking blood flow, treated with uric acid (0, 300 or 500 mumol/L), and changes in gene expression measured using transcriptomics and quantitative droplet digital PCR. Result(s): In human plasma, no inhibition of PNP activity was observed, and only quercetin weakly inhibited ADA. XOR was not present at sufficient amount in human plasma to use for testing, but quercetin, quercetin-3'-sulfate and the gut microbial metabolite 3',4'-dihydroxyphenylacetic acid inhibited bovine milk XOR. Several changes were observed in gene expression in HUVECs under flow compared to static conditions, but after uric acid treatment, only very few changes were detected. Conclusion(s): We propose that the main mechanism by which quercetin, as quercetin-3'-sulfate, lowers uric acid in vivo is through inhibition of XOR, and not ADA nor PNP. The pertinent shift in uric acid concentration was not sufficient to produce significant changes in endothelial gene expression in a cell model.Copyright © 2020 Elsevier Inc.

Liquid phase microextraction (LPME) into a microfluidic has undergone great advances focused on downscaled and miniaturized devices. In this work, a microfluidic device was developed for the extraction of sulfonamides in order to accelerate the mass transfer and passive diffusion of the analytes from the donor phase to the acceptor phase. The subsequent analysis was carried out by high performance liquid chromatography with UV-DAD (HPLC-DAD). Several parameters affecting the extraction efficiency of the method such as the supported liquid membrane, composition of donor and acceptor phase and flow rate were investigated and optimized. Tributyl phosphate was found to be a good supported liquid membrane which confers not only great affinity for analytes but also long-term stability, allowing more than 20 consecutive extractions without carry over effect. Under optimum conditions, extraction efficiencies were over 96 % for all sulfonamides after 10 minutes extraction and only 10 µL of sample was required. Relative standard deviation was between 3-5 % for all compounds. Method detection limits were 45, 57, 54 and 33 ng mL−1 for sulfadiazine (SDI), sulfamerazine (SMR), sulfamethazine (SMT) and sulfamethoxazole (SMX), respectively. Quantitation limits were 0.15, 0.19, 0.18 and 0.11 µg mL−1 for SDI, SMR, SMT SMX, respectively. The proposed microfluidic device was successfully applied for the determination of sulfonamides in urine samples with extraction efficiencies within the range of 86-106 %. The proposed method improves the procedures proposed to date for the determination of sulfonamides in terms of efficiency, reduction of the sample volume and extraction time.

Background: A research priority in finding a cure for HIV is to establish methods to accurately locate and quantify where and how HIV persists in people living with HIV (PLWH) receiving suppressive antiretroviral therapy (ART). Infusing copper-64 (64Cu) radiolabelled broadly neutralising antibodies targeting HIV envelope (Env) with CT scan and positron emission tomography (PET) identified HIV Env in tissues in SIV infected non-human primates. We aimed to determine if a similar approach was effective in people living with HIV (PLWH). Method(s): Unmodified 3BNC117 was compared with 3BNC117 bound to the chelator MeCOSar and 64Cu (64Cu-3BNC117) in vitro to assess binding and neutralization. In a clinical trial 64Cu-3BNC117 was infused into HIV uninfected (Group 1), HIV infected and viremic (viral load, VL >1000 c/mL; Group 2) and HIV infected aviremic (VL <20 c/mL; Group 3) participants using two dosing strategies: high protein (3mg/kg unlabeled 3BNC117 combined with <5mg 64Cu-3BNC117) and trace (<5mg 64Cu-3BNC117 only). All participants were screened for 3BNC117 sensitivity from virus obtained from viral outgrowth. Magnetic resonance imaging (MRI)/PET and pharmacokinetic assessments (ELISA for serum 3BNC117 concentrations and gamma counting for 64Cu) were performed 1, 24- and 48-hours post dosing. The trial (clincialtrials.gov NCT03063788) primary endpoint was comparison of PET standard uptake values (SUVs) in regions of interest (e.g lymph node groups and gastrointestinal tract). Finding(s): Comparison of unmodified and modified 3BNC117 in vitro demonstrated no difference in HIV binding or neutralisation. 17 individuals were enrolled of which 12 were dosed including Group 1 (n=4, 2 high protein, 2 trace dose), Group 2 (n=6, 2 high protein, 4 trace) and Group 3 (n=2, trace only). HIV+ participants had a mean CD4 of 574 cells/microL and mean age 43 years. There were no drug related adverse effects and no differences in tissue uptake in regions of interest (e.g lymph nod

Background: Uric acid, a metabolic product of purine degradation in humans, is a risk factor for developing gout and type 2 diabetes, and supplementation with quercetin lowers plasma uric acid in mildly hyperuricemic men. Here we examined the mechanism of inhibition of enzymes involved in uric acid metabolism by quercetin, conjugates and microbial catabolites, and measured the effect of lowered circulating uric acid on endothelial cell gene expression. Method(s): Inhibition of adenosine deaminase (ADA), purine nucleoside phosphorylase (PNP) and xanthine oxidoreductase (XOR) activity by quercetin and metabolites was determined by HPLC. Human umbilical vein endothelial cells (HUVECs) were cultured under conditions mimicking blood flow, treated with uric acid (0, 300 or 500 mumol/L), and changes in gene expression measured using transcriptomics and quantitative droplet digital PCR. Result(s): In human plasma, no inhibition of PNP activity was observed, and only quercetin weakly inhibited ADA. XOR was not present at sufficient amount in human plasma to use for testing, but quercetin, quercetin-3'-sulfate and the gut microbial metabolite 3',4'-dihydroxyphenylacetic acid inhibited bovine milk XOR. Several changes were observed in gene expression in HUVECs under flow compared to static conditions, but after uric acid treatment, only very few changes were detected. Conclusion(s): We propose that the main mechanism by which quercetin, as quercetin-3'-sulfate, lowers uric acid in vivo is through inhibition of XOR, and not ADA nor PNP. The pertinent shift in uric acid concentration was not sufficient to produce significant changes in endothelial gene expression in a cell model.Copyright © 2020 Elsevier Inc.

Research background. Apple juice is one of the most popular and liked beverages worldwide. Due to the increased health consciousness among consumers, beetroot and chokeberry juices have also rising consumption trends. Despite representing a consider-able percentage of the processed fruit and rich source of bioactive compounds, fruit pom-ace, remaining after juice production, has still been underutilised. Here, the possibility of using apple, beetroot and chokeberry pomace in liqueur formulations is investigated. Experimental approach. Apple and chokeberry liqueurs were produced from apple and chokeberry pomace extracts, respectively. Apple/chokeberry and apple/beetroot liqueurs were obtained by combining apple pomace with chokeberry and beetroot pomace extracts in ratios 50:50 and 70:30, respectively. The sensory quality and acceptability of freshly prepared liqueurs were evaluated by experts and consumers. Sugars and phenolics were identified and quantified by high-performance anion-exchange chromatography with pulsed-amperometric detection (HPAEC-PAD) and high-performance liquid chromatog-raphy–diode array detection–tandem mass spectrometry (HPLC–DAD–MS/MS), respec-tively. Storability was preliminarily evaluated based on monitoring of total phenolic con-centration, antioxidant activity and colour each month during 6 months of storage at 4 and 22 °C. Results and conclusions. The expert and the consumer testing indicated that apple and chokeberry pomace could be used as raw materials without any flavour corrections while apple/beetroot pomace liqueur would require modification. High total phenolic content and antioxidant activity were found in all freshly prepared liqueurs, with chokeberry liqueur being by far superior. Among identified phenolics, ellagic acid and phlorizin were quantified as the most prominent, except in chokeberry liqueur, where phlorizin was not quantified. Despite the decrease in total phenolic concentration and antioxidant activity after 6 mont

A high-performance liquid chromatographic analytical method has been developed for the determination of chlorine and nitrophenols at trace level (µg/L) in water, using a reversed-phase elution gradient and an ultraviolet (UV) detector. A Solid Phase Extraction (SPE) method was employed, which involved the design of a two-dimensional pre-column system coupled with high-performance liquid chromatography (HPLC) in order to preconcentrate, purify and isolate the solutes in environmental aqueous matrices. These phenolic compounds are considered priority pollutants by the United States Environmental Protection Agency (USEPA); the compounds are 4,6-dinitro-2-methyl phenol, 2,4-dimethylphenol, 4-chloro-3-methyl phenol, 2,4-dichlorophenol, 2,4,6-trichlorophenol, and pentachlorophenol. The developed method is simple, fast, accurate, and precise. Recoveries greater than 90 % were obtained for the phenols 4,6-dinitro-2-methyl phenol, 2,4-dichlorophenol and 2,4,6-trichlorophenol, approximately 80 % for pentachlorophenol and 52 % for 2,4-dimethylphenol. In addition, an acceptable precision (CV ˂ 5 %) was obtained for all solutes at these concentration levels., Se ha desarrollado un método analítico por cromatografía de líquidos de alta eficiencia para la determinación de cloro y nitrofenoles al nivel de trazas (µg/L) en agua, empleando un gradiente de elución de fase reversa y un detector ultravioleta (UV). Se empleó un método de Extracción en Fase Sólida (EFS), que implicó el diseño de un sistema bidimensional de pre-columnas acoplado con la cromatografía de líquidos de alta eficiencia (CLAE) con el fin de efectuar la preconcentración, purificación y aislamiento de los solutos en matrices acuosas ambientales. Estos compuestos fenólicos son considerados contaminantes prioritarios por la Agencia de Protección Ambiental de los Estados Unidos (USEPA); los compuestos son: 4,6-dinitro-2-metilfenol, 2,4-dimetilfenol, 4-cloro-3-metilfenol, 2,4-diclorofenol, 2,4,6-triclorofenol y pentaclorofenol. El método desarrollado es simple, rápido, exacto y preciso. Se obtuvieron recuperaciones mayores del 90 % para los fenoles 4,6-dinitro-2-metilfenol, 2,4-diclorofenol y 2,4,6-triclorofenol, aproximadamente 80 % para el pentaclorofenol y 52 % para el 2,4-dimetilfenol. Además, se obtuvo una precisión (CV ˂ 5 %) aceptable para todos los solutos a estos niveles de concentración.

Gene-environment interaction (GxE) determines the vulnerability of an individual to a spectrum of stress-related neuropsychiatric disorders. Increased impulsivity, excessive aggression, and other behavioural characteristics are associated with variants within the tryptophan hydroxylase-2 (Tph2) gene, a key enzyme in brain serotonin synthesis. This phenotype is recapitulated in naïve mice with complete, but not with partial Tph2 inactivation. Tph2 haploinsufficiency in animals reflects allelic variation of Tph2 facilitating the elucidation of respective GxE mechanisms. Recently, we showed excessive aggression and altered serotonin brain metabolism in heterozygous Tph2-deficient male mice (Tph2+/−) after predator stress exposure. Here, we sought to extend these studies by investigating aggressive and anxiety-like behaviours, sociability, and the brain metabolism of dopamine and noradrenaline. Separately, Tph2+/− mice were examined for exploration activity in a novel environment and for the potentiation of helplessness in the modified swim test (ModFST). Predation stress procedure increased measures of aggression, dominancy, and suppressed sociability in Tph2+/− mice, which was the opposite of that observed in control mice. Anxiety-like behaviour was unaltered in the mutants and elevated in controls. Tph2+/− mice exposed to environmental novelty or to the ModFST exhibited increased novelty exploration and no increase in floating behaviour compared to controls, which is suggestive of resilience to stress and despair. High-performance liquid chromatography (HPLC) revealed significant genotype-dependent differences in the metabolism of dopamine, and norepinephrine within the brain tissue. In conclusion, environmentally challenged Tph2+/− mice exhibit behaviours that resemble the behaviour of non-stressed null mutants, which reveals how GxE interaction studies can unmask latent genetically determined predispositions. © 2020 The Authors.

A high-performance liquid chromatographic analytical method has been developed for the determination of chlorine and nitrophenols at trace level (µg/L) in water, using a reversed-phase elution gradient and an ultraviolet (UV) detector. A Solid Phase Extraction (SPE) method was employed, which involved the design of a two-dimensional pre-column system coupled with high-performance liquid chromatography (HPLC) in order to preconcentrate, purify and isolate the solutes in environmental aqueous matrices. These phenolic compounds are considered priority pollutants by the United States Environmental Protection Agency (USEPA); the compounds are 4,6-dinitro-2-methyl phenol, 2,4-dimethylphenol, 4-chloro-3-methyl phenol, 2,4-dichlorophenol, 2,4,6-trichlorophenol, and pentachlorophenol. The developed method is simple, fast, accurate, and precise. Recoveries greater than 90 % were obtained for the phenols 4,6-dinitro-2-methyl phenol, 2,4-dichlorophenol and 2,4,6-trichlorophenol, approximately 80 % for pentachlorophenol and 52 % for 2,4-dimethylphenol. In addition, an acceptable precision (CV ˂ 5 %) was obtained for all solutes at these concentration levels., Se ha desarrollado un método analítico por cromatografía de líquidos de alta eficiencia para la determinación de cloro y nitrofenoles al nivel de trazas (µg/L) en agua, empleando un gradiente de elución de fase reversa y un detector ultravioleta (UV). Se empleó un método de Extracción en Fase Sólida (EFS), que implicó el diseño de un sistema bidimensional de pre-columnas acoplado con la cromatografía de líquidos de alta eficiencia (CLAE) con el fin de efectuar la preconcentración, purificación y aislamiento de los solutos en matrices acuosas ambientales. Estos compuestos fenólicos son considerados contaminantes prioritarios por la Agencia de Protección Ambiental de los Estados Unidos (USEPA); los compuestos son: 4,6-dinitro-2-metilfenol, 2,4-dimetilfenol, 4-cloro-3-metilfenol, 2,4-diclorofenol, 2,4,6-triclorofenol y pentaclorofenol. El método desarrollado es simple, rápido, exacto y preciso. Se obtuvieron recuperaciones mayores del 90 % para los fenoles 4,6-dinitro-2-metilfenol, 2,4-diclorofenol y 2,4,6-triclorofenol, aproximadamente 80 % para el pentaclorofenol y 52 % para el 2,4-dimetilfenol. Además, se obtuvo una precisión (CV ˂ 5 %) aceptable para todos los solutos a estos niveles de concentración.

Liquid phase microextraction (LPME) into a microfluidic has undergone great advances focused on downscaled and miniaturized devices. In this work, a microfluidic device was developed for the extraction of sulfonamides in order to accelerate the mass transfer and passive diffusion of the analytes from the donor phase to the acceptor phase. The subsequent analysis was carried out by high performance liquid chromatography with UV-DAD (HPLC-DAD). Several parameters affecting the extraction efficiency of the method such as the supported liquid membrane, composition of donor and acceptor phase and flow rate were investigated and optimized. Tributyl phosphate was found to be a good supported liquid membrane which confers not only great affinity for analytes but also long-term stability, allowing more than 20 consecutive extractions without carry over effect. Under optimum conditions, extraction efficiencies were over 96 % for all sulfonamides after 10 minutes extraction and only 10 µL of sample was required. Relative standard deviation was between 3-5 % for all compounds. Method detection limits were 45, 57, 54 and 33 ng mL−1 for sulfadiazine (SDI), sulfamerazine (SMR), sulfamethazine (SMT) and sulfamethoxazole (SMX), respectively. Quantitation limits were 0.15, 0.19, 0.18 and 0.11 µg mL−1 for SDI, SMR, SMT SMX, respectively. The proposed microfluidic device was successfully applied for the determination of sulfonamides in urine samples with extraction efficiencies within the range of 86-106 %. The proposed method improves the procedures proposed to date for the determination of sulfonamides in terms of efficiency, reduction of the sample volume and extraction time.

Given an increasing focus on environmental sustainability, microbial oils have been suggested as an alternative to petroleum-based products. However, microbial oil production relies on the use of costly sugar-based feedstocks. Substrate limitation, elevated costs, and risk of contamination have sparked the search for alternatives to sugar-based platforms. Volatile fatty acids are generated during anaerobic digestion of organic waste and are considered a promising substrate for microbial oil production. In the present study, two freshwater and one marine microalga along with two thraustochytrids were evaluated for their potential to produce lipids when cultivated on volatile fatty acids generated from food waste via anaerobic digestion using a membrane bioreactor. Freshwater microalgae Auxenochlorella protothecoides and Chlorella sorokiniana synthesized lipids rich in palmitic acid (C16:0), stearic acid (C18:0), oleic acid (C18:1), and linoleic acid (C18:2). This composition corresponds to that of soybean and jatropha oils, which are used as biodiesel feedstock. Production of added-value polyunsaturated fatty acids (PUFA) mainly omega-3 fatty acids was examined in three different marine strains: Aurantiochytrium sp. T66, Schizochytrium limacinum SR21, and Crypthecodinium cohnii. Only Aurantiochytrium sp. T66 seemed promising, generating 43.19% docosahexaenoic acid (DHA) and 13.56% docosapentaenoic acid (DPA) in total lipids. In summary, we show that A. protothecoides, C. sorokiniana, and Aurantiochytrium sp. T66 can be used for microbial oil production from food waste material.

The edible filamentous fungi are hot candidate for future supply of functional food and feed with e.g. protein, essential amino acids, and compounds with immunostimulant activity. L-carnitine that plays a crucial role in energy metabolism represents a functional compound normally produced by Zygomycetes filamentous fungus Rhizopus oligosporus in solid-state fermentation. The present study provides the first insights on production of L-carnitine-enriched edible fungal biomass through submerged cultivation of several Ascomycetes and Zygomycetes including Aspergillus oryzae, Neurospora intermedia, Rhizopus oryzae, and Rhizopus oligosporus. A. oryzae with 3 mg L-carnitine yield per gram of fungal biomass, indicates great potential on production of this bioactive compound which is remarkably higher than the other tested fungi in this work and also previous studies. In addition to fungal strain, other factors such as cultivation time and presence of yeast extract were found to play a role. Further studies on submerged growth optimization of A. oryzae in both high-quality recipes and in medium based on low-value substrates are proposed in order to clarify its potential for production of L-carnitine-enriched fungal biomass.

Rapid industrialization has provided comforts to mankind but has also impacted the environment harmfully. There has been severe increase in the pollution due to several industries, in particular due to dye industry, which generate huge quantities of wastewater containing hazardous chemicals. Although tremendous developments have taken place for the treatment and management of such wastewater through chemical or biological processes, there is an emerging shift in the approach, with focus shifting on resource recovery from such wastewater and also their management in sustainable manner. This review article aims to present and discuss the most advanced and state-of-art technical and scientific developments about the treatment of dye industry wastewater, which include advanced oxidation process, membrane filtration technique, microbial technologies, bio-electrochemical degradation, photocatalytic degradation, etc. Among these technologies, microbial degradation seems highly promising for resource recovery and sustainability and has been discussed in detail as a promising approach. This paper also covers the challenges and future perspectives in this field.

Choklad erhålls från kakaobönan som är frukten på kakaoträdet (Theobroma cacao). För att skapa choklad bearbetas kakaobönan vilket påverkar halten av bioaktiva ämnen i produkten. Choklad innehåller flera typer av bioaktiva ämnen där kakaoflavanolerna epikatechin och katechin är de mest framträdande. De hör till klassen flavanoider som i tidigare forskning har kopplats till hälsosamma effekter som sänkt blodtryck, minskad risk för hjärt- och kärlsjukdomar och ökad antioxidantkapacitet. I förordningen nr 432/2012 över godkända hälsopåståenden för livsmedel som EU-kommissionen fastställt framgår att kakaoflavanoler kan bidra till en normal blodcirkulation genom att blodkärlen hålls elastiska. Påståendet får användas för mörk choklad men konsumenten måste upplysas om att ett dagligt intag av 200 mg kakaoflavanoler krävs för att uppnå den gynnsamma effekten. I aktuell studie har kakaobönor från Madagaskar och 70% mörk choklad producerad på kakaobönor från Peru, Mexiko, Filippinerna och Ecuador analyserats i samarbete med Johannas Choklad i Kalmar. Syftet med studien var att undersöka hur halten flavanoider och främst kakaoflavanolerna epikatechin och katechin påverkas vid hantverksmässig chokladtillverkning. Resultatet skulle kunna ge information åt Johannas Choklad vilka bearbetningssteg i processen från kakaoböna till färdig chokladkaka som påverkar halterna mest. Det skulle även kunna ge indikationer på om någon av deras chokladprodukter uppfyller kraven för att erhålla EU:s hälsopåstående angående mörk choklad. Ett antal analyser genomfördes genom att bioaktiva ämnen extraherades med metanol och den slutgiltiga supernatanten indunstades och återstoden löstes i MQ-vatten (v/w =10). Lösningen filtrerades och analyserades därefter med RP-HPLC med UV-detektor (Reversed phase - High Performance Liquid Chromatography). Resultatet från analyserna gav indikationer på att variationen av rostningsgrad (110-135 °C) och processningsförfarandet vid chokladtillverkningen inte påve, Chocolate is obtained from the cocoa bean which is the fruit of the cocoa tree (Theobroma cacao). To create chocolate, the cocoa bean is processed, which affects the content of bioactive substances in the product. Chocolate contains several types of bioactive substances, of which the cocoa flavanols epicatechin and catechin are the most prominent. They belong to the class of flavonoids which in previous research have been linked to health effects such as lowered blood pressure, reduced risk of cardiovascular disease, and increased antioxidant capacity. Regulation No. 432/2012 on approved health claims for food, established by the European Commission, states that cocoa flavanols can contribute to the normal blood circulation by keeping the blood vessels elastic. The claim may be used for dark chocolate, but the consumer must be informed that a daily intake of 200 mg of cocoa flavanols is required to achieve the beneficial effect. In the current study, cocoa beans from Madagascar and 70% dark chocolate produced on cocoa beans from Peru, Mexico, the Philippines, and Ecuador have been analyzed in collaboration with Johanna's Chocolate in Kalmar. The purpose of the study was to investigate how the content of flavonoids and mainly the cocoa flavanols epicatechin and catechin are affected by artisanal chocolate production. The result could provide information to Johanna's Chocolate which processing steps in the process from cocoa bean to finished chocolate cake that affects the levels the most. It could also provide indications as to whether any of their chocolate products meet the requirements for obtaining the EU dark chocolate health claim. A number of assays were performed by extracting bioactive substances with methanol and evaporating the final supernatant and dissolving the residue in MQ water (v/w = 10). The solution was filtered and then analyzed by RP-HPLC with a UV detector (Reversed phase - High Performance Liquid Chromatography). The results of the analyzes ga

Given an increasing focus on environmental sustainability, microbial oils have been suggested as an alternative to petroleum-based products. However, microbial oil production relies on the use of costly sugar-based feedstocks. Substrate limitation, elevated costs, and risk of contamination have sparked the search for alternatives to sugar-based platforms. Volatile fatty acids are generated during anaerobic digestion of organic waste and are considered a promising substrate for microbial oil production. In the present study, two freshwater and one marine microalga along with two thraustochytrids were evaluated for their potential to produce lipids when cultivated on volatile fatty acids generated from food waste via anaerobic digestion using a membrane bioreactor. Freshwater microalgae Auxenochlorella protothecoides and Chlorella sorokiniana synthesized lipids rich in palmitic acid (C16:0), stearic acid (C18:0), oleic acid (C18:1), and linoleic acid (C18:2). This composition corresponds to that of soybean and jatropha oils, which are used as biodiesel feedstock. Production of added-value polyunsaturated fatty acids (PUFA) mainly omega-3 fatty acids was examined in three different marine strains: Aurantiochytrium sp. T66, Schizochytrium limacinum SR21, and Crypthecodinium cohnii. Only Aurantiochytrium sp. T66 seemed promising, generating 43.19% docosahexaenoic acid (DHA) and 13.56% docosapentaenoic acid (DPA) in total lipids. In summary, we show that A. protothecoides, C. sorokiniana, and Aurantiochytrium sp. T66 can be used for microbial oil production from food waste material.

Rapid industrialization has provided comforts to mankind but has also impacted the environment harmfully. There has been severe increase in the pollution due to several industries, in particular due to dye industry, which generate huge quantities of wastewater containing hazardous chemicals. Although tremendous developments have taken place for the treatment and management of such wastewater through chemical or biological processes, there is an emerging shift in the approach, with focus shifting on resource recovery from such wastewater and also their management in sustainable manner. This review article aims to present and discuss the most advanced and state-of-art technical and scientific developments about the treatment of dye industry wastewater, which include advanced oxidation process, membrane filtration technique, microbial technologies, bio-electrochemical degradation, photocatalytic degradation, etc. Among these technologies, microbial degradation seems highly promising for resource recovery and sustainability and has been discussed in detail as a promising approach. This paper also covers the challenges and future perspectives in this field.

The edible filamentous fungi are hot candidate for future supply of functional food and feed with e.g. protein, essential amino acids, and compounds with immunostimulant activity. L-carnitine that plays a crucial role in energy metabolism represents a functional compound normally produced by Zygomycetes filamentous fungus Rhizopus oligosporus in solid-state fermentation. The present study provides the first insights on production of L-carnitine-enriched edible fungal biomass through submerged cultivation of several Ascomycetes and Zygomycetes including Aspergillus oryzae, Neurospora intermedia, Rhizopus oryzae, and Rhizopus oligosporus. A. oryzae with 3 mg L-carnitine yield per gram of fungal biomass, indicates great potential on production of this bioactive compound which is remarkably higher than the other tested fungi in this work and also previous studies. In addition to fungal strain, other factors such as cultivation time and presence of yeast extract were found to play a role. Further studies on submerged growth optimization of A. oryzae in both high-quality recipes and in medium based on low-value substrates are proposed in order to clarify its potential for production of L-carnitine-enriched fungal biomass.

The present study investigated the changes in total and individual glucosinolates (GSLs) in roots and leaves of different clubroot-resistant and -susceptible oilseed rape cultivars following artificial inoculation with Plasmodiophora brassicae isolates with different virulence. The results showed significant differences in clubroot incidence and severity as well as in the amount of total and individual glucosinolates between oilseed rape cultivars in response to virulence of the pathogen. Single among with total aliphatic and total indolic glucosinolate contents were significantly lower in leaves of susceptible cultivars compared to resistant ones due to the infection. Similarly, single and total aliphatic as well as indolic glucosinolate contents in roots were lower in susceptible cultivars compared to resistant cultivars analyzed. The different isolates of P. brassicae seem to differ in their ability to reduce gluconasturtiin contents in the host. The more aggressive isolate P1 (+) might be able to suppress gluconasturtiin synthesis of the host in a more pronounced manner compared to the isolate P1. A possible interaction of breakdown products of glucobrassicin with the auxin receptor transport inhibitor response 1 (TIR1) is hypothesized and its possible effects on auxin signaling in roots and leaves of resistant and susceptible cultivars is discussed. A potential interplay between aliphatic and indolic glucosinolates that might be involved in water homeostasis in resistant cultivars is explained. © 2021 by the authors. Licensee MDPI, Basel, Switzerland.

Cryoturbated peat circles (pH 4) in the Eastern European Tundra harbor up to 2 mM pore water nitrate and emit the greenhouse gas N2O like heavily fertilized agricultural soils in temperate regions. The main process yielding N2O under oxygen limited conditions is denitrification, which is the sequential reduction of nitrate/nitrite to N2O and/or N2. N2O reduction to N2 is impaired by pH < 6 in classical model denitrifiers and many environments. Key microbes of peat circles are important but largely unknown catalysts for C- and N-cycling associated N2O fluxes. Thus, we hypothesized that the peat circle community includes hitherto unknown taxa and is essentially unable to efficiently perform complete denitrification, i.e., reduce N2O, due to a low in situ pH. 16S rRNA analysis indicated a diverse active community primarily composed of the bacterial class-level taxa Alphaproteobacteria, Acidimicrobiia, Acidobacteria, Verrucomicrobiae, and Bacteroidia, as well as archaeal Nitrososphaeria. Euryarchaeota were not detected. 13C2- and 12C2-acetate supplemented anoxic microcosms with endogenous nitrate and acetylene at an in situ near pH of 4 were used to assess acetate dependent carbon flow, denitrification and N2O production. Initial nitrate and acetate were consumed within 6 and 11 days, respectively, and primarily converted to CO2 and N2, suggesting complete acetate fueled denitrification at acidic pH. Stable isotope probing coupled to 16S rRNA analysis via Illumina MiSeq amplicon sequencing identified acetate consuming key players of the family Burkholderiaceae during complete denitrification correlating with Rhodanobacter spp. The archaeal community consisted primarily of ammonia-oxidizing Archaea of Nitrososphaeraceae, and was stable during the incubation. The collective data indicate that peat circles (i) host acid-tolerant denitrifiers capable of complete denitrification at pH 4–5.5, (ii) other parameters like carbon availability rather than pH are possible reason

Background and aim: Hemoglobin (Hb) variants can clinically range from being completely asymptomatic to frequent requirement of transfusions. Some individuals may become aware of a variant only when a complete blood count (CBC), hemoglobin analysis or genetic testing is done for a different reason. These individuals are typically heterozygous for an autosomal recessive variant. A study was conducted to find out the different types of hemoglobinopathies in adults presenting to a tertiary care hospital for routine health check and its geographic distribution. Objective: 1) To find out the severity of anemia associated with different hemoglobinopathies. 2) To find the relevance of glycated hemoglobin (HbA1c) in the presence of various hemoglobinopathies. Methods: A retrospective cross-sectional observational study was done in 111 consecutive patients who were found to have hemoglobinopathies during routine health checks in a tertiary care hospital in South India from 2013 to 2021. Results: One hundred eleven patients were found to have abnormality in Hb electrophoresis and there was a male predominance (69 patients, 62.2%). Majority of patients with beta-thalassemia (28 patients, 63.6%) were from West Bengal. Both HbE trait (17 patients, 65.4%) and homozygous HbE (23 patients, 62.2%) were from Assam. There was statistically significant distribution (p value 0.0001). HbA1c detected Hb variant in those with HbE disease. Conclusion: Hemoglobinopathies constitute a huge hereditary burden and a serious healthcare concern in India. Hence, it is the need of the hour to pick up such asymptomatic cases and provide appropriate premarital and prenatal counseling. Also, it is essential to devise strategies other than routine HbA1c testing to guide blood sugar control.