Rapid changes in Atlantic grey seal milk from birth to weaning – immune factors and indicators of metabolic strain

True seals have the shortest lactation periods of any group of placental mammal. Most are capital breeders that undergo short, intense lactations, during which they fast while transferring substantial proportions of their body reserves to their pups, which they then abruptly wean. Milk was collected from Atlantic grey seals (Halichoerus grypus) periodically from birth until near weaning. Milk protein profiles matured within 24 hours or less, indicating the most rapid transition from colostrum to mature phase lactation yet observed. There was an unexpected persistence of immunoglobulin G almost until weaning, potentially indicating prolonged trans-intestinal transfer of IgG. Among components of innate immune protection were found fucosyllactose and siallylactose that are thought to impede colonisation by pathogens and encourage an appropriate milk-digestive and protective gut microbiome. These oligosaccharides decreased from early lactation to almost undetectable levels by weaning. Taurine levels were initially high, then fell, possibly indicative of taurine dependency in seals, and progressive depletion of maternal reserves. Metabolites that signal changes in the mother’s metabolism of fats, such as nicotinamide and derivatives, rose from virtual absence, and acetylcarnitines fell. It is therefore possible that indicators of maternal metabolic strain exist that signal the imminence of weaning.Publisher PDFPeer reviewe

A method for the analysis of sugars in biological systems using reductive amination in combination with hydrophilic interaction chromatography and high resolution mass spectrometry

Separation of sugar isomers extracted from biological samples is challenging because of their natural occurrence as alpha and beta anomers and, in the case of hexoses, in their pyranose and furanose forms. A reductive amination method was developed for the tagging of sugars with the aim of it becoming part of a metabolomics work flow. The best separation of the common hexoses (glucose, fructose, mannose and galactose) was achieved when 2H5-aniline was used as the tagging reagent in combination with separation on a ZICHILIC column. The method was used to tag a range of sugars including pentoses and uronic acids. The method was simple to perform and was able to improve both the separation of sugars and their response to electrospray ionisation. The method was applied to the profiling of sugars in urine where a number of hexose and pentose isomers could be observed. It was also applied to the quantification of sugars in post-mortem brain samples from three control samples and three samples from individuals who had suffered from bipolar disorder

Effect of Bistorta affinis (D. Don) and Malcolmia cabulica (Boiss) Essential Oils on Human and Foodborne Pathogens, Insects and Termites

This current study investigates the biological efficiency of essential oils extracted from Bistorta affinis and Malcolmia cabulica on human and foodborne pathogens as well as against insects and termites. The oils were obtained by steam distillation using a Clevenger-type system and analyzed for their constituents using GC-MS. Twenty compounds were identified, wherein carvacrol and thymol were the main constituents in both plants. Essential oils exhibited significant activity against all tested foodborne bacteria, fungi, and other pathogens. In addition, essential oils from both plants have shown promising activity against all tested insects, which is a positive sign of being used as an insect repellent. In contrast, no activity was observed against termites. The bioactivities are mainly due to carvacrol and thymol’s levels in the essential oils that known for their effectiveness against foodborne pathogens and pests. The present study constitutes a basis for further investigation and development of bioactive compounds in Bistorta affinis and Malcolmia cabulica

Exogenous γ-aminobutyric acid (GABA) mitigated salinity-induced impairments in mungbean plants by regulating their nitrogen metabolism and antioxidant potential

BackgroundIncreasing soil salinization has a detrimental effect on agricultural productivity.Therefore, strategies are needed to induce salinity-tolerance in crop species for sustainable foodproduction. γ-aminobutyric acid (GABA) plays a key role in regulating plant salinity stresstolerance. However, it remains largely unknown how mungbean plants (Vigna radiata L.) respondto exogenous GABA under salinity stress.MethodsThus, we evaluated the effect of exogenous GABA (1.5 mM) on the growth and physiobiochemicalresponse mechanism of mungbean plants to saline stress (0-, 50-, and 100 mM [NaCland Na2SO4, at a 1:1 molar ratio]).ResultsIncreased saline stress adversely affected mungbean plants' growth and metabolism. Forinstance, leaf-stem-root biomass (34- and 56%, 31- and 53%, and 27- and 56% under 50- and 100mM, respectively]) and chlorophyll concentrations declined. The carotenoid level increased (10%)at 50 mM and remained unaffected at 100 mM. Hydrogen peroxide (H2O2), malondialdehyde(MDA), osmolytes (soluble sugars, soluble proteins, proline), total phenolic content, andenzymatic activities of superoxide dismutase (SOD), ascorbate peroxidase (APX), peroxidase(POD), glutathione reductase (GTR), and polyphenol oxidation (PPO) were significantlyincreased. In leaves, salinity caused a significant increase in Na+ concentration but a decrease inK+ concentration, resulting in a low K+/Na+ concentration (51- and 71% under 50- and 100- mMstress). Additionally, nitrogen concentration and the activities of nitrate reductase (NR) andglutamine synthetase (GS) decreased significantly. The reduction in glutamate synthase (GOGAT)activity was only significant (65%) at 100 mM stress. Exogenous GABA decreased Na+, H2O2,and MDA concentrations but enhanced photosynthetic pigments, K+ and K+/Na+ ratio, Nmetabolism, osmolytes, and enzymatic antioxidant activities, thus reducing salinity-associatedstress damages, resulting in improved growth and biomass.ConclusionExogenous GABA may have improved the salinity tolerance of mungbean plants by maintaining their morpho-physiological responses and reducing the accumulation of harmfulsubstances under salinity. Future molecular studies can contribute to a better understanding of themolecular mechanisms by which GABA regulates mungbean salinity tolerance

Amassing of heavy metals in soils, vegetables and crop plants irrigated with wastewater: Health risk assessment of heavy metals in Dera Ghazi Khan, Punjab, Pakistan

Human health is the main concern related to use of crop products irrigated with contaminated irrigation sources. Present research has been conducted to explore heavy metal status of sewage and industrial wastewater being used up for irrigation purpose in the peri-urban areas of the district Dera Ghazi Khan which has not been explored widely before. The analysis also followed heavy metal detection in the subsequent irrigated soil and vegetables/crop plants in relation to assessment of health risk to the consumer to plan the future monitoring in this area. An unremitting boost of heavy metals into the environment from wastewater irrigation has become a global issue. These heavy metals enter the food chain and pose health assumptions to consumers upon utilization. In the present study, an investigation has been conducted to determine metal concentrations in the wastewater, soil, and different plant species. For wastewater samples, pH, total dissolved solids (TDS), electrical conductivity (EC), and selected heavy metals such as Al, As, Cr, Cu, Fe, Mn, Pb, Zn, and Ni were determined. The mean values of heavy metals in the soil samples were within the WHO/FAO safe limit, while Cr and Pb were the most frequent (100%) among the metals. However, differentiating the sites, the concentration of Cr and Cu, Ni, and Fe were elevated. The metal transfer was highly effective from soil to the growing plants i.e. brinjal, red corn, wheat, tomato, and spinach than other plant species. Among the metals, Cr, Ni, Mn, and Pb in plant samples were exceeding the WHO/FAO safe limit. Health risk index (HRI) have revealed the possible potential risk of heavy metal contaminated plant species in the order of spinach (6.4) > wheat (6.4) > brinjal (5.9) > tomato (4.7) > red corn (4.5) > apple gourd (4.3) > white corn (3.8) > cabbage (3.1) > luffa (2.9). Likewise, HRI of different metals was calculated as Cu (19.6) > Zn (17.9) > Cr (2.95) > Ni (0.85) > Mn (0.48) > Fe (0.15) > Cd (0.11) > Pb (0.05) > As (0.00001). The level of HRI through the use of dietary plants revealed an elevated risk level than the acceptable limit (HRI > 1) for Cu > Zn > Cr in adults. Our findings suggest that there would be a serious health risk to the consumers due to the consumption of these plant species being irrigated with the wastewater. Therefore, a strict regulatory mechanism is proposed for the safety of food plants in the study area including monitoring and recycling of crop plants, and building water treatment plants to remove pollutants and clean wastewater

Potent In Vitro α-Glucosidase and β-Secretase Inhibition of Amyrin-Type Triterpenoid Isolated from Datura metel Linnaeus (Angel’s Trumpet) Fruits

This study deals with α-glucosidase and β-secretase inhibitory screening of extract/fractions and isolated daturaolone (1), namely, 3-oxo-6-β-hydroxy-β-amyrin (daturaolone) from chloroform fraction of Datura metel L. Among entire fractions, the chloroform soluble fraction showed excellent activity against α-glucosidase with % inhibition 90.8 with IC50160.2±1.85 μg and daturaolone (1) with 98.7% inhibition with IC50840.4±1.74 μM, respectively. Similarly, extract and daturaolone (1) also exhibited significant activity against the β-secretase enzyme (BACE1) with % activities 88.27 and 95.19 and with IC50 values 304.21±2.98 μg and 260.70±1.87 μM, respectively, as compared to the standard inhibitor (Ans670, Sta671, Val672)-amyloid-β/A4 precursor protein 770 fragments 662-675) with % activity 94.21 and IC50 value 289.24±1.60 μM. This finding encourages and opens a new window for further detail phytochemical investigation on D. metel in order to isolate novel compounds with promising enzyme inhibitory potential

An exploration of the mechanism of hydrophilic interaction chromatography (HILIC) and the application of HILIC in metabolomic profiling

The aim of this study was to investigate hydrophilic interaction chromatography (HILIC) and its application to metabolic profiling. The effect of different mobile phase modifiers on the retention behaviour of acidic and neutral test probes was studied on silica gel based columns which confirmed the hypothesis that HILIC is due to a combination of hydrophilic interactions and electrostatic interactions. Among the acidic probes tested, the more negatively charged maleic acid was not strongly retained until the strength of the ammonium ion in the mobile phase was high enough to shield the acid from repulsion by charged silanol groups on the silica gel surface. The neutral test probes were increasingly retained with increasing ionic strength of the mobile phase suggesting that the thickness of the water layer on the surface of the silica gel increased with the strength of the ionic modifier. The utility of three silica hydride columns, Cogent silica C, Cogent Phenyl Hydride and Cogent UDC cholesterol, for separating mixture of metabolites was successfully assessed with an alternating gradient from high organic to high aqueous and then from high aqueous to high organic, thus permitting two orthogonal selectivities to be obtained on the same column. A method for the reductive amination was developed for the analysis of sugars and sugar phosphates. This method was very successful with deuterated aniline as a tag which led to the separation of the three common hexoses glucose, galactose and mannose. The method was applied to the profiling of sugars in milk and in brain tissue. An extensive study of seal milk in comparison with other mammalian milks was also conducted which found that unlike the milks of terrestrial mammals, seal milk had very little lactose. Thus seal pups appear to rely on fat metabolism to produce energy. A notable feature of the milk was that it contained high levels of nicotinamide, an essential precursor for the production of the cofactor NAD+ which is required for the β-oxidation of fats.The aim of this study was to investigate hydrophilic interaction chromatography (HILIC) and its application to metabolic profiling. The effect of different mobile phase modifiers on the retention behaviour of acidic and neutral test probes was studied on silica gel based columns which confirmed the hypothesis that HILIC is due to a combination of hydrophilic interactions and electrostatic interactions. Among the acidic probes tested, the more negatively charged maleic acid was not strongly retained until the strength of the ammonium ion in the mobile phase was high enough to shield the acid from repulsion by charged silanol groups on the silica gel surface. The neutral test probes were increasingly retained with increasing ionic strength of the mobile phase suggesting that the thickness of the water layer on the surface of the silica gel increased with the strength of the ionic modifier. The utility of three silica hydride columns, Cogent silica C, Cogent Phenyl Hydride and Cogent UDC cholesterol, for separating mixture of metabolites was successfully assessed with an alternating gradient from high organic to high aqueous and then from high aqueous to high organic, thus permitting two orthogonal selectivities to be obtained on the same column. A method for the reductive amination was developed for the analysis of sugars and sugar phosphates. This method was very successful with deuterated aniline as a tag which led to the separation of the three common hexoses glucose, galactose and mannose. The method was applied to the profiling of sugars in milk and in brain tissue. An extensive study of seal milk in comparison with other mammalian milks was also conducted which found that unlike the milks of terrestrial mammals, seal milk had very little lactose. Thus seal pups appear to rely on fat metabolism to produce energy. A notable feature of the milk was that it contained high levels of nicotinamide, an essential precursor for the production of the cofactor NAD+ which is required for the β-oxidation of fats

A comparison of the chromatographic properties of silica gel and silicon hydride modified silica gels

The retention properties of a silica gel column and a type C silica (silicon hydride) column for bases, sugars and polar acids were compared in hydrophilic interaction chromatography (HILIC) mode with formic acid or ammonium acetate as aqueous phase modifiers. The type C silica column was much more retentive for a series of model bases than the silica gel column and, surprisingly, retention of bases increased on the type C silica column when, the higher pH, ammonium acetate containing mobile phase was used. The retention of sugars was greater on the type C silica column than on the silica gel column and also increased on the type C silica column with increased pH suggesting either a silanophilic mechanism of retention or some unknown mechanism. Three type C silica based columns, type C silica, cogent diamond hydride and a β-pinene modified column, which it was hoped might exert some additional stereochemical discrimination, were tested for metabolomic profiling of urine. In general the unmodified type C silica column gave the strongest retention of the many polar metabolites in urine and could provide a useful complement to established HILIC methods for metabolomic profiling

Development and Validation of a Versatile UPLC-PDA Method for Simultaneous Determination of Paracetamol, Tizanidine, Aceclofenac, and Nimesulide in Their New Combinations

A simple, rapid, and validated UPLC method was developed for the simultaneous quantitation of paracetamol (PAR), tizanidine (TIZ), aceclofenac (ACF), and nimesulide (NIM) either in pure forms or in their different tablet dosage forms. Chromatographic separation was attained on an ACQUITY UPLC™ BEH C18 column (100 mm × 2.1 mm, 1.7 μm) with a mobile phase consisting of 20 mM phosphate buffer (pH 7.0) : acetonitrile in the proportion (60 : 40 v/v) isocratically pumped at a flow rate of 1.25 mL·min−1, and detection was monitored at 305 nm. All analytes were separated simultaneously at a retention time (tr) of 1.42, 2.31, 3.63, and 5.62 min for PAR, TIZ, ACF, and NIM, respectively, with a total run time less than 6.0 min. The proposed method was validated according to ICH guidelines with respect to accuracy, precision, linearity, limit of detection, limit of quantitation, and robustness. Linearity was obtained over a concentration range of 81.25–487.5, 0.5–3.5, 25–150, and 25–150 µg·mL−1 for PAR, TIZ, ACF, and NIM, respectively. The development method can be successfully employed in QC laboratories for the routine analysis of the investigated drugs in their new combination