Hemoglobin Variants Influence Plasmodium Falciparum Sexual Differentiation

It has long been recognized that individuals who express variations of the hemoglobin-A (HbA) protein experience less severe malaria disease. As malaria remains to be one of the most significant infectious diseases in history, this human adaptation has led to the persistence of HbA variants (HbVARs) in the population. The intricate lifecycle of the parasite which causes the most cases of clinical malaria, Plasmodium falciparum, relies on both asexual and sexual reproductive cycles, with host to vector transmission reliant on sexual stage gametocyte formation. Multiple epidemiological studies have shown that HbVARs may influence gametocyte production during P. falciparum infection, with greater gametocyte numbers reported in individuals with hemoglobin variant containing erythrocytes (Hb VAR-Ery) when compared to hemoglobin A containing erythrocytes (Hb A-Ery). Here we provide experimental support for these studies by showing significantly higher sexual differentiation rates among parasites grown in Hb S containing erythrocytes (Hb S-Ery) obtained from sickle cell patients than those differentiated in Hb A-Ery (p=0.038). Because the digestion of hemoglobin is such an integral part of the intraerythrocytic cycle, we then sought to determine whether there was a difference between the hydrolysis efficiencies of HbA and other hemoglobin variants (HbVAR). By using a prominent recombinant P. falciparum hemoglobinase we found the hydrolysis efficiency of HbA to be significantly (p=0.0058) more efficient after 24 hours compared to a HbVAR sample containing mixed amounts of HbA, HbF, and HbS. To further determine whether there is a link between hemoglobin digestion efficiency and sexual differentiation, we therapeutically inhibited the hemoglobin digestion and hemozoin formation process in a culture of P. falciparum using sub-optimal doses of chloroquine diphosphate. We found a significant difference (p<0.001) among gametocyte conversion rates between treated and non-treated cultures, as well as a moderate negative correlation between hemozoin formation and gametocyte conversion rate (Pearson r=0.72, p=0.008). Gene expression analysis also revealed patterns of expression that were consistent with increased gametocytogenesis. We conclude that hemoglobin type plays a significant role in the process of sexual conversion in P. falciparum. Though further studies should be completed in order to confirm these results, these findings may suggest hemoglobin digestion efficiency as a causative factor for sexual differentiation. As individuals with hemoglobinopathies make up approximately 7% of the global population, and malaria infection rates have been shown to differ depending on these genetic dynamics, these findings may support the creation of targeted initiatives to reduce transmission specifically in areas where there is a high percentage of hemoglobinopathy carriage. Disclosures No relevant conflicts of interest to declare.

Novel AOPs-Based Dual-Environmental Digestion Method for Determination of Total Dissolved Nitrogen in Water

Based on a synergistic digestion method of ultraviolet combined with ozone (UV/O3), this article investigates the reaction characteristics of nitrogen-containing compounds (N-compounds) in water and the influence of ions on digestion efficiency. In this respect, a novel and efficient AOPs-based dual-environmental digestion method for the determination of total dissolved nitrogen (TDN) in waters with complex components is proposed, in the hopes of improving the detection efficiency and accuracy of total nitrogen via online monitoring. The results show that inorganic and organic N-compounds have higher conversion rates in alkaline and acidic conditions, respectively. Meanwhile, the experimental results on the influence of Cl−, CO32−, and HCO3− on the digestion process indicate that Cl− can convert to radical reactive halogen species (RHS) in order to promote digestion efficiency, but CO32− and HCO3− cause a cyclic reaction consuming numerous •OH, weakening the digestion efficiency. Ultimately, to verify the effectiveness of this novel digestion method, total dissolved nitrogen samples containing ammonium chloride, urea, and glycine in different proportions were digested under the optimal conditions: flow rate, 0.6 L/min; reaction temperature, 40 °C; pH in acidic conditions, 2; digestion time in acidic condition, 10 min; pH in alkaline conditions, 11; digestion time in alkaline conditions, 10 min. The conversion rate (CR) of samples varied from 93.23% to 98.64%; the mean CR was greater than 95.30%. This novel and efficient digestion method represents a potential alternative for the digestion of N-compounds in the routine analysis or online monitoring of water quality.

Development of Optimal Digesting Conditions for Microplastic Analysis in Dried Seaweed Gracilaria fisheri

Currently, research on the accumulation of microplastics (MPs) in the marine food web is being highlighted. An accurate and reliable digestion method to extract and isolate MPs from complex food matrices has seldom been validated. This study aimed to compare the efficacy of MP isolation among enzymatic-, oxidative-, and the combination of two digestion methods on red seaweed, Gracilaria fisheri. The dried seaweed sample was digested using three different methods under various conditions using enzymes (cellulase and protease), 30% H2O2, and a combination of enzymes and 30% H2O2. The method possessing the best digestion efficiency and polymer recovery rate of MPs was selected, and its effect on spiked plastic polymer integrity was analyzed by Raman spectroscopy. As a result, the enzymatic method rendered moderate digestion efficiency (59.3–63.7%) and high polymer recovery rate (94.7–98.9%). The oxidative method using 30% H2O2 showed high digestion efficiency (93.0–96.3%) and high polymer recovery rate (>98%). The combination method was the most effective method in terms of digestion efficiency, polymer recovery rate, and expenditure of digestion time. The method also showed no chemical changes in the spiked plastic polymers (PE, PP, PS, PVC, and PET) after the digestion process. All the spiked plastic polymers were identifiable using Raman spectroscopy.

An Innovative Digestion Method: Ultrasound-Assisted Electrochemical Oxidation for the Onsite Extraction of Heavy Metal Elements in Dairy Farm Slurry

Dairy farm slurry is an important biomass resource that can be used as a fertilizer and in energy utilization and chemical production. This study aimed to establish an innovative ultrasound-assisted electrochemical oxidation (UAEO) digestion method for the rapid and onsite analysis of the heavy metal (HM) contamination level of dairy slurry. The effects of UAEO operating parameters on digestion efficiency were tested based on Cu and Zn concentrations in a dairy slurry sample. The results showed that Cu and Zn digestion efficiency was (96.8 ± 2.6) and (98.5 ± 2.9)%, respectively, with the optimal UAEO operating parameters (digestion time: 45 min; ultrasonic power: 400 W; NaCl concentration: 10 g/L). The digestion recovery rate experiments were then operated with spiked samples to verify the digestion effect on broad-spectrum HMs. When the digestion time reached 45 min, all digestion recovery rates exceeded 90%. Meanwhile, free chlorine concentration, particle size distribution, and micromorphology were investigated to demonstrate the digestion mechanism. It was found that 414 mg/L free chorine had theoretically enough oxidative ability, and the ultrasound intervention could deal with the blocky undissolved particles attributed to its crushing capacity. The results of particle size distribution showed that the total volume and bulky particle proportion had an obvious decline. The micromorphology demonstrated that the ultrasound intervention fragmented the bulky particles, and electrochemical oxidation made irregular blocky structures form arc edge and cellular structures. The aforementioned results indicated that UAEO was a novel and efficient method. It was fast and convenient. Additionally, it ensured digestion efficiency and thus had a good application prospect.

N-Glycomics of Cerebrospinal Fluid: Method Comparison

Cerebrospinal fluid (CSF) contains valuable biological and neurological information. However, its glycomics analysis is hampered due to the low amount of protein in the biofluid, as has been demonstrated by other glycomics studies using a substantial amount of CSF. In this work, we investigated different N-glycan sample preparation approaches to develop a more sensitive method. These methods, one with an increased amount of buffer solution during the N-glycan release step with a lower amount of sample volume and the other with Filter-Aided N-Glycan Separation (FANGS), were compared with recent work to demonstrate their effectiveness. It was demonstrated that an increased amount of buffer solution showed higher intensity in comparison to the previously published method and FANGS. This suggested that digestion efficiency during the N-glycan release step was not in an optimal condition from the previously published method, and that there is a substantial loss of sample with FANGS when preparing N-glycans from CSF.

Development of an Eco-friendly Sample Preparation protocol for metals determination in food samples: Oxygen Pressurized Single Reaction Chamber using Diluted Nitric Acid

Many efforts have been recently made to improve the digestion efficiency by using powerful equipment or by using auxiliary reagent. In this work, an alternative method is reported, which explores...

CHEMOMETRIC STRATEGY FOR OPTIMIZATION OF AN ACID DECOMPOSITION METHOD TO DETERMINE THE MINERAL COMPOSITION IN ALMOND PULP (Terminalia catappa Linn.)

In this work, multivariate optimization techniques were applied to develop an acid digestion procedure in digester block using “cold finger” as reflux system to determine minerals in almond pulp samples by inductively coupled plasma optical emission spectrometry. Two-level full factorial design and Box-Behnken design were applied to evaluate and optimize the factors involved in the acid digestion process. In both experimental designs, Function D and Function MR multiple responses were used to establish the method condition for all analytes and with greater digestion efficiency. Two apple and spinach leaves certified reference materials were analyzed to confirm the proposed method accuracy. The digestion efficiency was evaluated by residual carbon content which showed 1.32-1.77% range. The concentration values found for each element in almond pulp collected in Salvador, Bahia, Brazil in mg 100 g-1 were: 3.08 (Ca), 0.209 (Cu), 0.407 (Fe), 356 (K), 21.5 (Mg), 0.096 (Mn), 34.4 (P) and 0.289 (Zn). The developed method was simple and efficient for almond pulp mineral composition evaluation. This unconventional fruit has nutritional relevance with future application for new food recipes and pharmaceutical products formulation.