Analyzing laboratory test utilization trends in belgian primary care: a decade of insights with international perspectives.

BACKGROUND: Clinical laboratory testing, essential for medical diagnostics, represents a significant part of healthcare activity, influencing around 70% of critical clinical decisions. The automation of laboratory equipment has expanded test menus and increased efficiency to meet the growing demands for clinical testing. However, concerns about misutilization remain prevalent. In Belgium, primary care has seen a dramatic increase in lab test usage, but recent utilization data is lacking. METHODS: We conducted a comprehensive retrospective analysis of laboratory test utilization trends within the primary care settings of Belgium over a ten-year period, spanning from 2012 to 2021, incorporating a vast dataset of 189 million test records for almost 1.5 million persons. This was the first study to integrate the metadata from both the INTEGO & THIN databases, which are derived from the two major electronic medical record (EMR) systems used in primary care in Belgium, providing a comprehensive national perspective. This research provides crucial insights into patient-level patterns, test-level utilization, and offers international perspectives through comparative analysis. RESULTS: We found a subtle annual increase in the average number of laboratory tests per patient (ranging from approximately 0.5-1%), indicative of a deceleration in growth in laboratory test ordering when compared to previous decades. We also witnessed stability and consistency of the most frequently ordered laboratory tests across diverse patient populations and healthcare contexts over the years. CONCLUSIONS: These findings emphasize the need for continued efforts to optimize test utilization, focusing not only on tackling overutilization but on enhancing the diagnostic relevance of tests ordered. The frequently ordered tests should be prioritized in these initiatives to ensure their continued effectiveness in patient care. By consolidating extensive datasets, employing rigorous statistical analysis, and incorporating international perspectives, this study provides a solid foundation for evidence-based strategies aimed at refining laboratory test utilization practices. These strategies can potentially improve the quality of healthcare delivery while simultaneously addressing cost-effectiveness concerns in healthcare.

Co/Ni/Cu-NH2BDC MOF@natural Egyptian zeolite ore nanocomposite for calcium ion removal in water softening applications.

Water softening is a treatment process required to remove calcium (Ca(II)) and magnesium (Mg(II)) cations from water streams. Nanocomposites can provide solutions for such multiple challenges and have high performance and low application costs. In this work, a multimetallic cobalt, nickel, and copper 2-aminoterephthalic acid metal-organic framework ((Co/Ni/Cu-NH2BDC) MOF) was synthesized by a simple solvothermal technique. This MOF was supported on an Egyptian natural zeolite ore and was used for the adsorption of Ca(II) ions for water-softening applications. The adsorbent was characterized using Fourier transform infrared (FTIR) spectroscopy, field emission scanning electron microscopy (FESEM), X-ray diffraction (XRD), N2 adsorption-desorption isotherms, and zeta potential measurements. The adsorption isotherm data for the prepared adsorbent toward Ca(II) were best fit using the Redlich-Peterson model and showed a maximum adsorption capacity of 88.1 mg/g. The adsorption kinetics revealed an equilibrium time of 10 min, which was best fit using the Avrami model. The intermolecular interactions of Ca(II) ions with zeolite and MOF were investigated by Monte Carlo simulations, molecular dynamics simulations, and FTIR and XRD analyses. The adsorption sites in the zeolite structure were oxygen atoms, while those in the MOF structure were amine nitrogen atoms. The Ca(II) ions are coordinated with the solvent molecules in both structures. Finally, the in vitro cytotoxicity of this nanocomposite was assessed, revealing viability levels of 74.57 ± 2.1% and 21 ± 2.79% for Vero and African green monkey kidney and human liver (HepG2) cells, respectively. Cytotoxicity assays help assess the environmental impact of these materials, ensuring that they do not harm aquatic organisms or disrupt ecosystems. Thus, this study demonstrated the valorization of MOF/zeolite as a valuable and industry-ready adsorbent that can appropriate Ca(II) contaminants from aqueous streams.

Phytochemical profiling of Livistona carinensis leaf extract via UHPLC-QTOF-MS/MS with assessment of its antiviral mechanisms.

Among 36 species of the genus Livistona (family Palmae or Arecaceae), L. carinensis is considered the only species native to Africa. Previous studies showed the richness of Livistona fruits in phenolic compounds. The goal of the current study was to investigate the phytochemical composition and assess the antiviral mechanisms of the L. carinensis leaves' ethanolic extract cultivated in Egypt for the first time. The ultra-high performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UHPLC-QTOF-MS/MS) was applied. Moreover, the total crude extract was fractionated using ethyl acetate and n-butanol for phytochemical investigations by various chromatographic and spectroscopic techniques. Besides, the antiviral activity of L. carinensis leaves was assessed using three protocols in vitro using MTT assay compared to acyclovir. UHPLC-QTOF-MS/MS-based analysis resulted in identification of 72 metabolites tentatively. They belonged to diverse phytochemical classes, mainly including flavonoids (29), organic acids (10), and phenolic acids (7). The antiviral activity investigations revealed a direct Adeno virus inactivation mechanism rather than inhibition of virus replication or blocking its attachment to Vero cells. Hence, the plant leaves may be a potential candidate for discovery of novel antiviral drugs owing to the diversity of identified phytochemical classes.

Bioassay-guided isolation of anti-inflammatory and antinociceptive metabolites among three Moroccan Juniperus leaves extract supported with in vitro enzyme inhibitory assays.

ETHNOPHARMACOLOGICAL RELEVANCE: Herbs of the genus Juniperus (family Cupressaceae) have been commonly used in ancestral folk medicine known as "Al'Araar" for treatment of rheumatism, diabetes, inflammation, pain, and fever. Bioassay-guided isolation of bioactives from medicinal plants is recognized as a potential approach for the discovery of novel drug candidates. In particular, non-addictive painkillers are of special interest among herbal phytochemicals. AIM OF THE STUDY: The current study aimed to assess the safety of J. thurifera, J. phoenicea, and J. oxycedrus aqueous extracts in oral treatments; validating the traditionally reported anti-inflammatory and analgesic effects. Further phytochemical investigations, especially for the most bioactive species, may lead to isolation of bioactive metabolites responsible for such bioactivities supported with in vitro enzyme inhibition assays. MATERIALS AND METHODS: Firstly, the acute toxicity study was investigated following the OECD Guidelines. Then, the antinociceptive, and anti-inflammatory bioactivities were evaluated based on chemical and mechanical trauma assays and investigated their underlying mechanisms. The most active J. thurifera n-butanol fraction was subjected to chromatographic studies for isolating the major anti-inflammatory metabolites. Moreover, several enzymatic inhibition assays (e.g., 5-lipoxygenase, protease, elastase, collagenase, and tyrosinase) were assessed for the crude extracts and isolated compounds. RESULTS: The results showed that acute oral administration of the extracts (300-500 mg/kg, p. o.) inhibited both mechanically and chemically triggered inflammatory edema in mice (up to 70% in case of J. thurifera) with a dose-dependent antinociceptive (tail flick) and anti-inflammatory pain (formalin assay) activities. This effect was partially mediated by naloxone inhibition of the opioid receptor (2 mg/kg, i. p.). In addition, 3-methoxy gallic acid (1), quercetin (2), kaempferol (3), and ellagic acid (4) were successfully identified being involved most likely in J. thurifera extract bioactivities. Nevertheless, quercetin was found to be the most potent against 5-LOX, tyrosinase, and protease with IC50 of 1.52 ± 0.01, 192.90 ± 6.20, and 399 ± 9.05 µM, respectively. CONCLUSION: J. thurifera extract with its major metabolites are prospective drug candidates for inflammatory pain supported with inhibition of inflammatory enzymes. Interestingly, antagonism of opioid and non-opioid receptors is potentially involved.

Long-read powered viral metagenomics in the oligotrophic Sargasso Sea.

Dominant microorganisms of the Sargasso Sea are key drivers of the global carbon cycle. However, associated viruses that shape microbial community structure and function are not well characterised. Here, we combined short and long read sequencing to survey Sargasso Sea phage communities in virus- and cellular fractions at viral maximum (80 m) and mesopelagic (200 m) depths. We identified 2,301 Sargasso Sea phage populations from 186 genera. Over half of the phage populations identified here lacked representation in global ocean viral metagenomes, whilst 177 of the 186 identified genera lacked representation in genomic databases of phage isolates. Viral fraction and cell-associated viral communities were decoupled, indicating viral turnover occurred across periods longer than the sampling period of three days. Inclusion of long-read data was critical for capturing the breadth of viral diversity. Phage isolates that infect the dominant bacterial taxa Prochlorococcus and Pelagibacter, usually regarded as cosmopolitan and abundant, were poorly represented.

Chemometric-based drug discovery approaches from natural origins using hyphenated chromatographic techniques.

INTRODUCTION: Isolation and characterization of bioactive components from complex matrices of marine or terrestrial biological origins are the most challenging issues for natural product chemists. Biochemometric is a new potential scope in natural product analytical science, and it is a methodology to find the compound's correlation to their bioactivity with the help of hyphenated chromatographic techniques and chemometric tools. OBJECTIVES: The present review aims to evaluate the application of chemometric tools coupled to chromatographic techniques for drug discovery from natural resources. METHODS: The searching keywords "biochemometric," "chemometric," "chromatography," "natural products bioassay," and "bioassay" were selected to search the published articles between 2010-2023 using different search engines including "Pubmed", "Web of Science," "ScienceDirect," and "Google scholar." RESULTS: An initial stage in natural product analysis is applying the chromatographic hyphenated techniques in conjunction with biochemometric approaches. Among the applied chromatographic techniques, liquid chromatography (LC) techniques, have taken up more than half (53%) and also, mass spectroscopy (MS)-based chromatographic techniques such as LC-MS are the most widely used techniques applied in combination with chemometric methods for natural products bioassay. Considering the complexity of dataset achieved from chromatographic hyphenated techniques, chemometric tools have been increasingly employed for phytochemical studies in the context of determining botanicals geographical origin, quality control, and detection of bioactive compounds. CONCLUSION: Biochemometric application is expected to be further improved with advancing in data acquisition methods, new efficient preprocessing, model validation and variable selection methods which would guarantee that the applied model to have good prediction ability in compound relation to its bioactivity.

Establishing the Reportable Interval for Routine Clinical Laboratory Tests: A Data-Driven Strategy Leveraging Retrospective Electronic Medical Record Data.

BACKGROUND: This paper presents a data-driven strategy for establishing the reportable interval in clinical laboratory testing. The reportable interval defines the range of laboratory result values beyond which reporting should be withheld. The lack of clear guidelines and methodology for determining the reportable interval has led to potential errors in reporting and patient risk. METHODS: To address this gap, the study developed an integrated strategy that combines statistical analysis, expert review, and hypothetical outlier calculations. A large data set from an accredited clinical laboratory was utilized, analyzing over 124 million laboratory test records from 916 distinct tests. The Dixon test was applied to identify outliers and establish the highest and lowest non-outlier result values for each test, which were validated by clinical pathology experts. The methodology also included matching the reportable intervals with relevant Logical Observation Identifiers Names and Codes (LOINC) and Unified Code for Units of Measure (UCUM)-valid units for broader applicability. RESULTS: Upon establishing the reportable interval for 135 routine laboratory tests (493 LOINC codes), we applied these to a primary care laboratory data set of 23 million records, demonstrating their efficacy with over 1% of result records identified as implausible. CONCLUSIONS: We developed and tested a data-driven strategy for establishing reportable intervals utilizing large electronic medical record (EMR) data sets. Implementing the established interval in clinical laboratory settings can improve autoverification systems, enhance data reliability, and reduce errors in patient care. Ongoing refinement and reporting of cases exceeding the reportable limits will contribute to continuous improvement in laboratory result management and patient safety.

Environment-specific virocell metabolic reprogramming.

Viruses impact microbial systems through killing hosts, horizontal gene transfer, and altering cellular metabolism, consequently impacting nutrient cycles. A virus-infected cell, a "virocell," is distinct from its uninfected sister cell as the virus commandeers cellular machinery to produce viruses rather than replicate cells. Problematically, virocell responses to the nutrient-limited conditions that abound in nature are poorly understood. Here we used a systems biology approach to investigate virocell metabolic reprogramming under nutrient limitation. Using transcriptomics, proteomics, lipidomics, and endo- and exo-metabolomics, we assessed how low phosphate (low-P) conditions impacted virocells of a marine Pseudoalteromonas host when independently infected by two unrelated phages (HP1 and HS2). With the combined stresses of infection and nutrient limitation, a set of nested responses were observed. First, low-P imposed common cellular responses on all cells (virocells and uninfected cells), including activating the canonical P-stress response, and decreasing transcription, translation, and extracellular organic matter consumption. Second, low-P imposed infection-specific responses (for both virocells), including enhancing nitrogen assimilation and fatty acid degradation, and decreasing extracellular lipid relative abundance. Third, low-P suggested virocell-specific strategies. Specifically, HS2-virocells regulated gene expression by increasing transcription and ribosomal protein production, whereas HP1-virocells accumulated host proteins, decreased extracellular peptide relative abundance, and invested in broader energy and resource acquisition. These results suggest that although environmental conditions shape metabolism in common ways regardless of infection, virocell-specific strategies exist to support viral replication during nutrient limitation, and a framework now exists for identifying metabolic strategies of nutrient-limited virocells in nature.

Persistent Shortness of Breath in Post-COVID-19 Patients: Inducible Laryngeal Obstruction Can Be a Cause.

BACKGROUND: Inducible laryngeal obstruction (ILO) is the adduction of the true vocal folds during inspiration or, less frequently, expiration. Its etiology is unknown. PURPOSE: This study aimed to identify ILO as a possible cause of shortness of breath (SOB) in post-COVID-19 patients. METHODS: A retrospective study was conducted on 59 post-COVID-19 adults complaining of SOB. We collected the cases' clinical and chest imaging data, including demographic data, pulmonary function test (PFT), CT chest, and laryngeal endoscopic examination. A visual score was used to assign the severity of the glottic obstruction. The grades of this score ranged from 0 (complete patency) to 3 (almost complete closure). RESULTS: ILO was detected in 8 out of 59 cases (13.5%). Two cases out of eight had glottic closure grade 1, while five cases had glottic closure grade 2, and one had glottic closure grade 3. There was a positive correlation between ILO grades (the severity of glottic closure) and PFT results (the degree of upper airway restriction). PFTs in ILO have a specific pattern. It was typical for the expiratory loop to be normal and the inspiratory loop to be flattened. CONCLUSION: ILO is a possible cause of SOB in post-COVID-19 adult cases. It should be considered during evaluation and management.

Valorization potential of Egyptian mango kernel waste product as analyzed via GC/MS metabolites profiling from different cultivars and geographical origins.

Increasing attention has been given to mango (Mangifera indica) fruits owing to their characteristic taste, and rich nutritional value. Mango kernels are typically discarded as a major waste product in mango industry, though of potential economic value. The present study aims to outline the first comparison of different mango kernel cvs. originated from different localities alongside Egypt, e.g., Sharqia, Suez, Ismailia, and Giza. Gas chromatography-mass spectroscopy (GC-MS) post silylation analysis revealed that sugars were the major class being detected at 3.5-290.9 µg/mg, with some kernels originating from Sharqia province being the richest amongst other cvs. In consistency with sugar results, sugar alcohols predominated in Sharqia cvs. at 1.3-38.1 µg/mg represented by ribitol, iditol, pinitol, and myo-inositol. No major variation was observed in the fatty acids profile either based on cv. type or localities, with butyl caprylate as a major component in most cvs. identified for the first time in mango. Regarding phenolics, Sedeeq cv. represented the highest level at 18.3 µg/mg and showing distinct variation among cvs. posing phenolics as better classification markers than sugars. Multivariate data analyses (MVA) confirmed that the premium cvs "Aweis and Fons" were less enriched in sugars, i.e., fructose, talose, and glucose compared to the other cvs. Moreover, MVA of Zabdeya cv. collected from three localities revealed clear segregation to be chemically distinct. Sharqia originated mango kernels were rich in sugars (e.g., glucose and fructose), whilst sarcosine esters predominated in other origins.

Seasonal variations of the epididymis in donkeys (Equus asinus) with special reference to blood epididymal barrier.

The epididymis, a key component of the male reproductive system, controls spermatozoa's maturation, fertility, and storage. The objective of this study is to evaluate the histological, ultrastructural, and immunohistochemical variations in the epididymis of donkeys that occur throughout the year. During the breeding season (spring) and nonbreeding seasons (summer, autumn, and winter), 20 epididymis were collected from adult, clinically healthy donkeys. Compared to non-breeding seasons, the epididymal duct displayed a more active lining epithelium and more sperm in the lumen during the breeding season. The epithelial height is the lowest and the lumen is the widest during the breeding season. Furthermore, the epididymal epithelium in the tail region exhibits undulations with polyps-like projections. The epididymal epithelium is composed mainly of the principal, basal, and dark cells. Tight junction between adjacent principal cells is more obvious in the breeding season as compared to the non-breeding seasons. However, intraepithelial lymphocytes, phagocytic, and other immune cells are more frequent in non-breeding seasons. ß-catenin, which is a component of the adherent junctions between adjacent PCs, exhibits more immunoreactivity during the spring. On the other hand, iNOS, an indicator of oxidative stress, reacts positively during the summer. Additionally, during non-breeding seasons, autophagy was detected within the epididymal epithelium which may be linked to stress adaptation. In conclusion, our findings suggest that the histological and ultrastructural characteristics of the epididymal epithelium are more active during spring compared to other seasons of the year. RESEARCH HIGHLIGHTS: The study aimed to evaluate the histological, ultrastructural, and immunohistochemical variations in the blood epididymal barrier (BEB) and epididymal epithelium of donkeys that occur throughout the year. In comparison to non-breeding seasons, the epididymal duct displayed a more active lining epithelium and more sperm in the lumen during the breeding season. The epithelial height is the lowest and the lumen is the widest during the breeding season. The epididymal epithelium in the tail region exhibits undulations with polyps-like projections that increase the surface area. ß-catenin, which is a component of the adherent junctions between adjacent PCs, exhibits more immunoreactivity during the spring. On the other hand, iNOS, an indicator of oxidative stress, reacts positively during the summer.

Nanotechnology in leukemia: diagnosis, efficient-targeted drug delivery, and clinical trials.

Leukemia is a group of malignant disorders which affect the blood and blood-forming tissues in the bone marrow, lymphatic system, and spleen. Many types of leukemia exist; thus, their diagnosis and treatment are somewhat complicated. The use of conventional strategies for treatment such as chemotherapy and radiotherapy may develop many side effects and toxicity. Hence, modern research is concerned with the development of specific nano-formulations for targeted delivery of anti-leukemic drugs avoiding toxic effects on normal cells. Nanostructures can be applied not only in treatment but also in diagnosis. In this article, types of leukemia, its causes, diagnosis as well as conventional treatment of leukemia shall be reviewed. Then, the use of nanoparticles in diagnosis of leukemia and synthesis of nanocarriers for efficient delivery of anti-leukemia drugs being investigated in in vivo and clinical studies. Therefore, it may contribute to the discovery of novel and emerging nanoparticles for targeted treatment of leukemia with less side effects and toxicities.

Aroma-based discrimination of Egyptian versus Indian guava fruits and in response to probiotics as analyzed via SPME/GC-MS and chemometric tools.

Guava tree (Psidium guajava L., Myrtaceae) is an economic grown worldwide, particularly in tropical and subtropical regions. Guavas encompass numerous cultivars (cvs.) that were discriminated in previous studies based on leaf morphological features and profile of volatile organic compounds (VOCs). Nevertheless, fruit VOCs have also shown outstanding potential for discrimination of other plant taxa, which has not been utilized in guava. Hence, the current study investigates the various guava cvs. harvested from India and Egypt. A total of 5 samples were analyzed by solid phase microextraction coupled to gas chromatography/mass spectrometry. Results led to the detection of 42 VOCs belonging to aldehydes, alcohols, esters, ketones, aliphatic and aromatic hydrocarbons, in addition to monoterpene and sesquiterpene hydrocarbons. Butylated hydroxytoluene and ß-caryophyllene were predominant reaching 77% and 41% in Egyptian and Indian guava, respectively. The impact of probiotic fermentation, i.e., Lactobacillus acidophilus and L. plantarum on aroma profile was not significantly different (p > 0.05). Multivariate data analyses were further applied for samples classification and markers determination, including principal component analysis (PCA) and orthogonal partial least squares discriminant analysis (OPLS-DA). PCA score plot showed clear segregation of Egyptian from Indian specimens, whereas OPLS-DA revealed that ß-caryophyllene was associated with white fruit versus 3-butenyl isothiocyanate and muurolol in red fruit type in the case of Indian guava. The richness of Egyptian guava in butylated hydroxytoluene in addition to the presence of vitamin C may potentiate its antioxidant activity, to be followed in subsequent studies regarding its health effects.

Metabolism of natural and synthetic bioactive compounds in Cunninghamella fungi and their applications in drug discovery.

Investigation of xenobiotic metabolism is a key step for drug discovery. Since the in vivo investigations may be associated with harmful effects attributed to production of toxic metabolites, it is deemed necessary to predict their structure especially at the preliminary clinical studies. Furthermore, the application of microorganisms that are capable of metabolizing drugs mimic human metabolism and consequently may predict possible metabolites. The genus Cunninghamella has been proven to be a potential candidate, which mimics xenobiotic metabolism occurring inside the human body, including phase I and II metabolic reactions. Moreover, biotransformation with Cunninghamella showed chemical diversity, where a lot of products were detected in relation to the initial substrates after being modified by oxidation, hydroxylation, and conjugation reactions. Some of these products are more bioactive than the parent compounds. The current review presents a comprehensive literature overview regarding the Cunninghamella organisms as biocatalysts, which simulate mammalian metabolism of natural secondary and synthetic compounds.

Evening primrose oil: a comprehensive review of its bioactives, extraction, analysis, oil quality, therapeutic merits, and safety.

Oil crops have become increasingly farmed worldwide because of their numerous functions in foods and health. In particular, oil derived from the seeds of evening primrose (Oenothera biennis) (EPO) comprises essential fatty acids of the omega-6 (ω-6) series. It is well recognized to promote immune cells with a healthy balance and management of female ailments. The nutrients of interest in this oil are linoleic acid (LA, 70-74%) and γ-linolenic acid (GLA, 8-10%), which are polyunsaturated fatty acids (PUFA) that account for EPO's popularity as a dietary supplement. Various other chemicals in EPO function together to supply the body with PUFA, elevate normal ω-6 essential fatty acid levels, and support general health and well-being. The inclusive EPO biochemical analysis further succeeded in identifying several other components, i.e., triterpenes, phenolic acids, tocopherols, and phytosterols of potential health benefits. This comprehensive review capitalizes on EPO, the superior product of O. biennis, highlighting the interrelationship between various methods of cultivation, extraction, holistic chemical composition, sensory characters, and medicinal value. Besides the literature review, this study restates the numerous health advantages of primrose oil and possible drug-EPO interactions since a wide spectrum of drugs are administered concomitantly with EPO. Modern techniques to evaluate EPO chemical composition are addressed with emphasis on the missing gaps and future perspectives to ensure best oil quality and nutraceutical benefits.

Efficient dye removal from industrial wastewater using sustainable activated carbon and its polyamide nanocomposite derived from agricultural and industrial wastes in column systems.

Sugar beet crown (SBC) waste was employed to produce sustainable activated carbon (AC) by a thermo-chemical activation procedure using a fixed ratio of H3PO4/SBC (1 : 1 w/w ratio) at 550 °C/2 h. An activated carbon/polyamide nano-composite (AC/PA) was also prepared through the polymerization of the fabricated AC (90%) with polyamide (PA, 10%) synthetic textile waste using a proper dissolving agent at a specified w/w ratio with the employed polymer (formic acid/PA = 82/18%). Both AC and its derivative AC/PA were employed in the remediation of dyes from industrial wastewater in column systems, and their efficiencies were compared at various applied experimental conditions. The adsorption of the industrial dye waste (IDW) was a pH-, flow rate-, and bed thickness-controlled process by the regarded adsorbents. Kinetic studies confirmed the suitability of the Thomas equation over the Yoon and Nelson model in predicting the dynamic adsorption process of IDW by AC and AC/PA as was assured by the close agreement among the calculated and experimental uptake capacities of both adsorbents at the same applied flow rates, suggesting the chemisorption nature of IDW adsorption. Additionally, electrostatic attraction was the leading mechanism of IDW adsorption by AC and AC/PA composite with some advantages of the former over the latter.

Ocean-wide comparisons of mesopelagic planktonic community structures.

For decades, marine plankton have been investigated for their capacity to modulate biogeochemical cycles and provide fishery resources. Between the sunlit (epipelagic) layer and the deep dark waters, lies a vast and heterogeneous part of the ocean: the mesopelagic zone. How plankton composition is shaped by environment has been well-explored in the epipelagic but much less in the mesopelagic ocean. Here, we conducted comparative analyses of trans-kingdom community assemblages thriving in the mesopelagic oxygen minimum zone (OMZ), mesopelagic oxic, and their epipelagic counterparts. We identified nine distinct types of intermediate water masses that correlate with variation in mesopelagic community composition. Furthermore, oxygen, NO3- and particle flux together appeared as the main drivers governing these communities. Novel taxonomic signatures emerged from OMZ while a global co-occurrence network analysis showed that about 70% of the abundance of mesopelagic plankton groups is organized into three community modules. One module gathers prokaryotes, pico-eukaryotes and Nucleo-Cytoplasmic Large DNA Viruses (NCLDV) from oxic regions, and the two other modules are enriched in OMZ prokaryotes and OMZ pico-eukaryotes, respectively. We hypothesize that OMZ conditions led to a diversification of ecological niches, and thus communities, due to selective pressure from limited resources. Our study further clarifies the interplay between environmental factors in the mesopelagic oxic and OMZ, and the compositional features of communities.

A spectroscopic response factor-based toluidine blue assay towards a universal assay protocol for sulfated polysaccharides: Application to fucoidan content in crude extract.

Sulfated polysaccharides (SPS) have attracted a lot of interest because of their diverse pharmacological functions. Numerous scientific studies have shown that SPS exhibit better biological activity than those that are not sulfated, such as immunomodulatory, anti-viral, and antioxidant activities. A crucial step to a better understanding of the mechanism of action and health effects is the production of high purity SPS. This calls for the development of selective assay techniques that can identify SPS preferentially without being influenced by other substances or the co-extracted polysaccharides. A universal modified toluidine blue (TB) assay was developed in this study to detect SPS. The assay procedures were conducted using different SPS standards including fucoidans from different biogenic sources, in addition to heparin and dextran sulfate. Spectroscopic response factor was calculated for each SPS which showed very good correlation (R2 = 0.998) with the corresponding sulfation degree. The proposed method was applied for determination of SPS content of crude fucoidan product using five different SPS standards. The method was cross validated by conducting ANOVA test to the obtained % recovery revealing that there is no significant difference between the results obtained by identical reference standard and four nonidentical natural SPS standards. This is the first report of a selective universal assay of SPS that enables the selective determination of SPS using a nonidentical reference standard.

Methyl jasmonate elicitation effect on the metabolic profile of cambial meristematic cells culture derived from sweet basil (Ocimum basilicum L.) in relation to antioxidant activity: Untargeted metabolomics study in a time-based approach.

The undifferentiated cambial meristematic cell (CMC) has been recognized as a value-added production platform for plant natural products in comparison to the dedifferentiated plant cell line (DDC). In a time-based approach at 0, 24, 48, and 72 h, the present study aimed at investigating the phytochemical metabolome of methyl jasmonate (MeJA)-elicited CMC cultures derived from sweet basil (Ocimum basilicum L.), including primary and secondary metabolites analyzed using GC/TOF-MS post-silylation and RP-UPLC-C18-FT-MS/MS, respectively, as well as the analysis of aroma composition using headspace SPME-GC-MS. The results revealed a stress response in primary metabolism manifested by an increase in amino and organic acids reaching their maximum levels after 48 (1.3-fold) and 72 (1.7-fold) h, respectively. In addition, phenolic acids (e.g., sagerinic acid, rosmarinic acid, and 3-O-methylrosmarinic acid) followed by flavonoid aglycones (e.g., salvigenin and 5,6,4'-trihydroxy-7,3'-dimethoxyflavone) were the most abundant with prominent increases at 48 (1.2-fold) and 72 (2.1-fold) h, respectively. The aroma was intensified by the elicitation along the time, especially after 48 and 72 h. Furthermore, multivariate data analyses, including principal component analysis (PCA) and orthogonal partial least squares discriminant analysis (OPLS-DA) confirmed elicitation effect, especially post 48 and 72 h. The study further assessed the effect of MeJA elicitation on the antioxidant and polyphenolic content. The cultures at 48 h demonstrated a significant (p < 0.05) antioxidant activity concurrently with correlation with total polyphenolic content using Pearson's correlation. Our study provides new insights to the elicitation impact on primary and secondary metabolism, in addition to aroma profile, to orchestrate the stress response and in relation to antioxidant effect.