Biochemical Profiling Research Articles

Association of C677T Polymorphism of Methylenetetrahydrofolate Reductase with Metabolic Syndrome among Eastern Indian Women: A Case-control Study

Introduction: Polymorphisms of Methylene Tetrahydrofolate Reductase (MTHFR) gene have been associated with hyperhomocysteinaemia, which in turn may lead to hypertension, insulin resistance and abnormality of lipid metabolism. All these abnormalities are also components of Metabolic Syndrome (MetS). Thus, there is possibility of an association between MetS and MTHFR polymorphism. Literature search revealed paucity of data on this association, particularly from India. Aim: To evaluate the association of C677T polymorphism of MTHFR gene with MetS, among women of eastern Indian population. Materials and Methods: A hospital-based case-control study was conducted in the Department of Biochemistry, Calcutta National Medical College, Kolkata, West Bengal, India, between December 2016 to June 2018. Anthropometric and biochemical profiling of all 417 study subjects were done. Genetic workup was done by Polymerase Chain Reaction (PCR), using suitable primer, followed by restriction fragment length polymorphism analysis using Hinf 1 enzyme to identify MTHFR C677T polymorphism. Subjects were divided into two groups according to presence or absence of MetS as per International Diabetic Federation worldwide definition and compared with Mann-Whitney U Test. Logistic regression analysis was also performed. Results: Out of enrolled 417 female subjects, 243 were categorised as MetS group. Mean age (in years), of MetS group (48.70±9.03) was found to be significantly higher (p<0.001) than that of control group (42.83±10.90). A total of 45 (25%) females with MetS exhibited the presence of heterozygous CT genotype; that was significantly higher (p=0.026) than the non MetS group 21 (14.9%). The mutant T allele frequency was also significantly higher among the subjects suffering from MetS; 45 (12.5%)compared to 21 (7.4%) in the control group (p=0.036). Multiple logistic regression analysis revealed the odds of developing MetS among subjects with heterozygous CT genotype was 7.721 times (CI: 2.38-25.05) compared to those with wild genotype. Conclusion: C677T polymorphism of MTHFR gene was associated with the occurrence of MetS among woman.

Biofabrication, biochemical profiling, and in vitro applications of salivary gland decellularized matrices via magnetic bioassembly platforms.

Trending three-dimensional tissue engineering platforms developed via biofabrication and bioprinting of exocrine glands are on the rise due to a commitment to organogenesis principles. Nevertheless, a proper extracellular matrix (ECM) microarchitecture to harbor primary cells is yet to be established towards human salivary gland (SG) organogenesis. By using porcine submandibular gland (SMG) biopsies as a proof-of-concept to mimic the human SG, a new decellularized ECM bioassembly platform was developed herein with varying perfusions of sodium dodecyl sulfate (SDS) to limit denaturing events and ensure proper preservation of the native ECM biochemical niche. Porcine SMG biopsies were perfused with 0.01%, 0.1%, and 1% SDS and bio-assembled magnetically in porous polycarbonate track-etched (PCTE) membrane. Double-stranded DNA (dsDNA), cell removal efficiency, and ECM biochemical contents were analyzed. SDS at 0.1% and 1% efficiently removed dsDNA (< 50ng/mg) and preserved key matrix components (sulfated glycosaminoglycans, collagens, elastin) and the microarchitecture of native SMG ECM. Bio-assembled SMG decellularized ECM (dECM) perfused with 0.1-1% SDS enhanced cell viability, proliferation, expansion confluency rates, and tethering of primary SMG cells during 7 culture days. Perfusion with 1% SDS promoted greater cell proliferation rates while 0.1% SDS supported higher acinar epithelial expression when compared to basement membrane extract and other substrates. Thus, this dECM magnetic bioassembly strategy was effective for decellularization while retaining the original ECM biochemical niche and promoting SMG cell proliferation, expansion, differentiation, and tethering. Altogether, these outcomes pave the way towards the recellularization of this novel SMG dECM in future in vitro and in vivo applications.

Stings on wings: Proteotranscriptomic and biochemical profiling of the lesser banded hornet (Vespa affinis) venom.

Distinct animal lineages have convergently recruited venoms as weaponry for prey capture, anti-predator defence, conspecific competition, or a combination thereof. Most studies, however, have been primarily confined to a narrow taxonomic breadth. The venoms of cone snails, snakes, spiders and scorpions remain particularly well-investigated. Much less explored are the venoms of wasps (Order: Hymenoptera) that are infamous for causing excruciating and throbbing pain, justifying their apex position on Schmidt's pain index, including some that are rated four on four. For example, the lesser banded wasp (V. affinis) is clinically important yet has only been the subject of a few studies, despite being commonly found across tropical and subtropical Asia. Stings from these wasps, especially from multiple individuals of a nest, often lead to clinically severe manifestations, including mastocytosis, myasthenia gravis, optic neuropathy, and life-threatening pathologies such as myocardial infarction and organ failure. However, their venom composition and activity remain unexplored in the Indian subcontinent. Here, we report the proteomic composition, transcriptomic profile, and biochemical and pharmacological activities of V. affinis venom from southern India. Our findings suggest that wasp venoms are rich in diverse toxins that facilitate antipredator defence. Biochemical and pharmacological assessments reveal that these toxins can exhibit significantly higher activities than their homologues in medically important snakes. Their ability to exert potent effects on diverse molecular targets makes them a treasure trove for discovering life-saving therapeutics. Fascinatingly, wasp venoms, being evolutionarily ancient, exhibit a greater degree of compositional and sequence conservation across very distant populations/species, which contrasts with the patterns of venom evolution observed in evolutionarily younger lineages, such as advanced snakes and cone snails.

Assessment of biochemical, cooking, sensory and textural properties of the boiled food product of white yam (D. rotundata) genotypes grown at different locations

Specific biochemical properties and textural attributes determine the final quality and acceptability of yam food products. This study assessed the flour and cooking qualities (boiled yam) of sixteen elite white yam genotypes (D. rotundata) grown in three locations. Fresh yam samples were cut into regular-shaped pieces and boiled using the standard procedure. Sub-samples were oven-dried at 65 °C for 72 h and milled to flour. The biochemical profiling for the yam flour showed, on average, 61.35 ± 5.15% starch, 5.35 ± 0.15% sugar, 1.55 ± 0.24% crude fiber, 1.91 ± 0.31% ash, 5.65 ± 0.66% protein, 0.33 ± 0.02% fat and 34.87 ± 1.94% amylose content. The boiled yam's water absorption and cooking time ranged from 0.35 to 5.17% and 7.00–18 min, with an average of 2.74% and 10.64 min, respectively. The hardness of boiled yam from the sensory assay correlated positively with the hardness of instrumental texture analysis (p < 0.001, r = 0.47). In contrast, the hardness of instrumental texture had a significant negative correlation with the chewiness of sensory profile analysis (p < 0.05, r = 0.37). Likewise, water absorption correlated positively and significantly (p < 0.05, r = 0.43) with the chewiness of the sensory analysis. The study shows that the sensory attributes that determine the acceptability of boiled yam could be determined using instrumental measurements to save time and cost.

Characterization of Antagonist Potential of Selected Compost Bacterial Isolates (CBI) against Plant and Human Pathogens

Several fecal and enteric pathogens are present within the primary organic raw materials that are introduced to compost piles. These pathogens may compete with the existing microbiota and limit their efficiency, yielding only partial decomposition of the final compost. These pathogens also affect the process kinetics and persist in the final compost or may even regrow as a result of the declining effect of indigenous antagonistic micro-organisms. In this work, 11 indigenous bacterial isolates were selected from compost piles that were constructed from different percentages of comingled primary organic raw materials. Enzymatic, biochemical, and genetic characterization profiling of these strains was fulfilled. The top hits supplied by GenBank proved the genetic diversity of these strains, which belonged to 6 different families. This diversity, applied also at enzymatic and biochemical levels, showed the different degradation patterns of amino acids, carbohydrates, hormones, and proteins. CBI2 has been shown to be the most active isolate in the degradation of the different types of hormones and proteins from dairy products but lacks the enzymes needed for the degradation of ammonia into nitrogen. The antagonistic potential of recuperated secondary metabolites proved the total inhibition of all strains against Fusarium oxysporum and no growth limitation against Botrytis cinerea. Only the secondary metabolites of CBI1, CBI5, and CBI9 isolates showed inhibitory activity against Salmonella Typhimurium and Escherichia coli, whereas only those of CBI6 and CBI8 inhibited the growth of Salmonella Typhimurium and Listeria monocytogenes accordingly. From that finding, these strains are considered pioneering, with high potential to ensure both the efficient degradation of organic matter and the elimination of existing pathogens when applied to compost piles.

Biochemical metabolomic profiling of the Crown-of-Thorns Starfish (Acanthaster): New insight into its biology for improved pest management

The Crown-of-Thorns Starfish (COTS), Acanthaster species, is a voracious coral predator that destroys coral reefs when in outbreak status. The baseline metabolite and lipid biomolecules of 10 COTS tissues, including eggs from gravid females, were investigated in this study to provide insight into their biology and identify avenues for control. Targeted and untargeted metabolite- and lipidomics-based mass spectrometry approaches were used to obtain tissue-specific metabolite and lipid profiles. Across all COTS tissues, 410 metabolites and 367 lipids were identified. Most abundant were amino acids and peptides (18.7%) and wax esters (17%). There were 262 metabolites and 192 lipids identified in COTS eggs. Wax esters were more abundant in the eggs (30%) followed by triacylglycerols (TG), amino acids, and peptides. The diversity of asterosaponins in eggs (34) was higher than in tissues (2). Several asterosaponins known to modulate sperm acrosome reaction were putatively identified, including glycoside B, asterosaponin-4 (Co-Aris III), and regularoside B (asterosaponin A). The saponins saponin A, thornasteroside A, hillaside B, and non-saponins dictyol J and axinellamine B which have been shown to possess defensive properties, were found in abundance in gonads, skin, and radial nerve tissues. Inosine and 2-hexyldecanoic acid are the most abundant metabolites in tissues and eggs. As a secondary metabolite of purine degradation, inosine plays an important role in purine biosynthesis, while 2-hexyldecanoic acid is known to suppress side-chain crystallization during the synthesis of amphiphilic macromolecules (i.e., phospholipids). These significant spatial changes in metabolite, lipid, and asterosaponin profiles enabled unique insights into key biological tissue-specific processes that could be manipulated to better control COTS populations. Our findings highlight COTS as a novel source of molecules with therapeutic and cosmetic properties (ceramides, sphingolipids, carnosine, and inosine). These outcomes will be highly relevant for the development of strategies for COTS management including chemotaxis-based biocontrol and exploitation of COTS carcasses for the extraction of commercial molecules.

Identification of Subproteomic Markers for Skeletal Muscle Profiling.

The biochemical and cell biological profiling of contractile fiber types and subcellular structures plays a central role in basic and applied myology. Mass spectrometry-based proteomics presents an ideal approach for the systematic identification of proteomic and subproteomic markers. These representative components of fast versus slow muscle fibers and their subcellular fractions are highly useful for in-depth surveys of skeletal muscle adaptations to physiological challenges, as well as the improvement of diagnostic, prognostic, and therapy-monitoring methodologies in muscle pathology. This chapter outlines the identification of subproteomic markers for skeletal muscle profiling based on bottom-up and top-down approaches, including fluorescence two-dimensional difference gel electrophoresis (2D-DIGE).

Genomic analysis and biochemical profiling of an unaxenic strain of Synechococcus sp. isolated from the Peruvian Amazon Basin region.

Cyanobacteria are diverse photosynthetic microorganisms able to produce a myriad of bioactive chemicals. To make possible the rational exploitation of these microorganisms, it is fundamental to know their metabolic capabilities and to have genomic resources. In this context, the main objective of this research was to determine the genome features and the biochemical profile of Synechococcus sp. UCP002. The cyanobacterium was isolated from the Peruvian Amazon Basin region and cultured in BG-11 medium. Growth parameters, genome features, and the biochemical profile of the cyanobacterium were determined using standardized methods. Synechococcus sp. UCP002 had a specific growth rate of 0.086 ± 0.008μ and a doubling time of 8.08 ± 0.78h. The complete genome of Synechococcus sp. UCP002 had a size of ∼3.53Mb with a high coverage (∼200x), and its quality parameters were acceptable (completeness = 99.29%, complete and single-copy genes = 97.5%, and contamination = 0.35%). Additionally, the cyanobacterium had six plasmids ranging from 24 to 200kbp. The annotated genome revealed ∼3,422 genes, ∼ 3,374 protein-coding genes (with ∼41.31% hypothetical protein-coding genes), two CRISPR Cas systems, and 61 non-coding RNAs. Both the genome and plasmids had the genes for prokaryotic defense systems. Additionally, the genome had genes coding the transcription factors of the metalloregulator ArsR/SmtB family, involved in sensing heavy metal pollution. The biochemical profile showed primary nutrients, essential amino acids, some essential fatty acids, pigments (e.g., all-trans-β-carotene, chlorophyll a, and phycocyanin), and phenolic compounds. In conclusion, Synechococcus sp. UCP002 shows biotechnological potential to produce human and animal nutrients and raw materials for biofuels and could be a new source of genes for synthetic biological applications.

In-situ multi-mode extraction (iMME) sampler for a wide-scope analysis of chemical and biological targets in water in urbanized and remote (off-the-grid) locations

Chemical pollution (including chemicals of emerging concern – CECs) continues to gain increasing attention as a global threat to human health and the environment, with numerous reports on the adverse and sometimes devastating effects upon ecosystems the presence of these chemicals can have. Whilst many studies have investigated presence of CECs in aquatic environments, these studies have been often focused on higher income countries, leaving significant knowledge gaps for many low-middle income countries. This study proposes a new integrated powerless, in-situ multi-mode extraction (iMME) sampler for the analysis of chemicals (105 CECs) and biological (5 genes) markers in water in contrasting settings: an urbanized Avon River in the UK and remote Olifants River in Kruger National Park in South Africa. The overarching goal was to develop a sampling device that maintains integrity of a diverse range of analytes via analyte immobilization using polymeric and glass fibre materials, without access to power supply or cold chain (continuous chilled storage) for sample transportation. Chemical analysis was achieved using ultra-performance liquid chromatography coupled with tandem mass spectrometry. Several mobile CECs showed low stability in river water, at room temperature and typical 24 h sampling/transport time. It is therefore recommended that, in the absence of cooling, environmental water samples are spiked with internal standards on site, immediately after collection and analyte immobilization option is considered, in order to allow fully quantitative analysis. iMME has proven effective in immobilization, concentration and increased stability of CECs at room temperature (and at least 7 days storage) allowing for sample collection at remote locations. The results from the River Avon and Olifants River sampling indicate that the pristine environment of Olifants catchment is largely unaffected by CECs common in the urbanized River Avon in the UK with a few exceptions: lifestyle chemicals (e.g., caffeine, nicotine and their metabolites), paracetamol and UV filters due to tourism and carbamazepine due to its persistent nature. iMME equipped with an additional gene extraction capability provides an exciting new opportunity of comprehensive biochemical profiling of aqueous samples with one powerless in-situ device. Further work is required to provide full integration of the device and comprehensive assessment of performance in both chemical and biological targets.

Case report on clinical effect of Khadir compound drug (Kitibha har kashay)inPsoriasis and its cell line study

Psoriasis is a common skin disease which is inflammatory, proliferative, autoimmune, and chronic with relapsing nature. It resembles with Kitibha Kustha roga ofAyurveda which is described asrelapsing disease with dry, rough, discolored skin lesions. This is a case report of psoriasis patientwho was treated conservatively with Kitibha har kashay for 3 months. The assessment of clinical improvement was made, along with the biochemical profiling and photographic analysis. Patient was also instructed to follow the do’s and don’ts of the disease for diet and lifestyle also called pathya and apthya. Parameters that were used to assess the clinical improvement were PASI score, IGA scale and grading of Ayurvedicclinical symptoms of Kitibha.The patient was given the medication prepared from the AyurvedicPharmacy, Faculty of Ayurveda, BHU in ghan form. The crude and prepared drug was standardized from Institute itself and the data of the patient was maintained by regular follow-ups. The cell line study of water extract of Khadir compound was also done to evaluate the proliferative or antiproliferative action by MTT assay. The clinical result have shown that this drug posses a very effective antipsoriatic action that has significantly reduced PASI score to nil and marked improvement in IGA scale, clinical symptoms andphotographic improvement was found. The cell line study by MTT assay has shown the antiproliferative action of Daruharidra (Berberis aristata) that was one of the constituent of Kitibha har kashay.Further study of this drug on psoriasis specific cytokines is needed to be evaluated by biochemical analysis to evaluate its role in breaking the pathology of psoriasis.

Transcriptional and biochemical profiling of defense enzymes in Citrus sinensis during salicylic acid and cinnamon mediated suppression of green and blue mold.

Green and blue mold of citrus are threatening diseases that continuously inflict economic post-harvest loss. The suppressive effect of salicylic (SA) and Cinnamomum verum (CV) on green and blue mold of sweet oranges was investigated in this study. Among five tested plant extracts methanolic extract of Cinnamon caused the highest colony growth inhibition of P. digitatum and P. italicum in an in vitro antifungal assay.The methanolic extract of Cinnamon in combination with SA showed the lowest disease incidence and severity of green and blue mold on citrus fruit without affecting the fruit quality. Transcriptional profiling of defense enzymes revealed that the polyphenol oxidase (PPO), phenylalanine ammonia-lyase (PAL), and peroxidase (POD) genes were upregulated in fruit treated with CV, SA, and their combination compared to the control. The treatment SA+CV caused the highest upsurge in PPO, POD, and PAL gene expression than the control. Furthermore, the biochemical quantification of PPO, POD and PAL also revealed a similar pattern of activity. The present findings unravel the fact that the escalation in the activity of tested defense enzymes is possibly associated with the reduced incidence of blue and green molds. In conclusion, the study unveils the promising suppressive potential of SA+CV against green and blue mold by regulating the expression of PPO, POD, and PAL genes. Therefore, these treatments can find a role as safer alternatives to chemicals in the management of post-harvest green and blue mold.

Comparative Biochemical Profiling of Aluminum Chloride and Sodium Azide Induced Neuroinflammation and Cardiometabolic Disturbance

Comorbidities in human beings signify the numerous risk factors that increase the incidences of neuro- and cardio-metabolic disorders. Experimental models depicting comorbidities are important to explore the molecular pathophysiology that can help suggest appropriate treatment strategies. Tissue-accumulating potential and pathological effects of aluminium chloride (AlCl3)and sodium azide (NaN3) are well recognized. Hence, in the current work, we have for the first time aimed to investigate the unexplored potential of graded dose effects of AlCl3 and NaN3 in inducing early inflammation and cardiometabolic toxicity via comparative biochemical analysis of AlCl3 and/or NaN3. Rats were allocated into seven groups (n = 6). Group 1 was normal control. Remaining groups were given graded doses of AlCl3 and/or NaN3, as LD-AlCl3 (AlCl3 40 mg), MD-AlCl3 (AlCl3 45 mg), and HD-AlCl3 (AlCl3 50 mg) representing low dose, medium dose, and high dose of AlCl3, respectively, and the remaining as LD-NaN3 (NaN3 13 mg), MD-NaN3 (NaN3 15 mg), and HD-NaN3 (NaN3 17 mg) representing low dose, medium dose, and high dose of NaN3, respectively. Serum levels of glucose, insulin, lipid profile, inflammatory mediators like IL-6 and oxidative stress marker, and malondialdehyde (MDA) were analyzed. Likewise, subacute toxicity parameters were analyzed. Immunohistochemistry (IHC) and histopathology (H&E/Masson's trichrome staining) of brain, heart, and pancreatic tissues were done. ECG pattern of all groups was observed. HD-AlCl3 was associated with elevated levels of inflammatory biomarkers, MDA, and glycemic and lipid profiles, whereas it decreased the insulin levels. HD-NaN3 also showed the similar effects of aggravated inflammatory biomarkers, impaired glycemic and lipid profiles, but depicted the maximum mortality rate as compared to HD-AlCl3. IHC showed prominent amyloid plaques and neurofibrillary tangle formation with MD-AlCl3 and HD-AlCl3 as compared to NaN3-treated groups. Likewise, in brain tissues, vacuolation of white matter, vascular congestion, and hemorrhage were seen in HD-AlCl3 treated group, while HD-NaN3 induced death in animals. AlCl3 exposure resulted in an inverted QRS complex, while exposure to NaN3 showed ST depression but with increased mortality. AlCl3 has better controlled results as compared to NaN3 for induction of comorbid experimental animal model depicting early neuroinflammation and cardiometabolic disruption. These determined efforts facilitate the researchers for the development of clinically effective treatment strategies using such experimental models.

Biochemical profiling, pharmacological management and clinical outcomes in type 2 diabetes in Danish primary care from 2001 to 2015

AimsPrimary care plays an integral role in the management of type 2 diabetes (T2D). We investigated in a large group of individuals in this setting the biochemical profiles, pharmacological management and clinical outcomes as well as their changes over time. MethodsThis is a register-based study including relevant laboratory test results requested between 2000 and 2015 by general practitioners in the greater Copenhagen area. We identified 72,044 individuals with T2D on whom data concerning prescription medicine and clinical outcomes were obtained from national registries. ResultsThe number of individuals with T2D greatly increased from 2001 to 2015. Hemoglobin A1c, estimated glomerular filtration rate and urine albumin creatinine ratio did not change, but cholestrol levels improved. The proportion redeeming anti-diabetics remained around 80%, with an increase for metformin. The use of cardiovascular drugs increased. All-cause and especially cardiovascular mortality decreased over the period. Hospital admissions for non-fatal cardiovascular events dropped. ConclusionThe number of individuals with T2D in primary care increased dramatically whereas pharmacological management, control of risk factors and clinical outcomes seem to have improved. Nevertheless, a conspicuous minority did not receive diabetes-related medication.

Biochemical profiling of metabolomics in heavy metal-intoxicated impaired metabolism and its amelioration using plant-based bioactive compound

Exposure to Pb is widely spreading and has far-reaching negative effects on living systems. This study aimed to investigate the toxic effects of Pb, through biochemical profiling and the ameliorative effects of quercetin against Pb-toxicity. Twenty-five male Wistar albino mice were divided into the following five groups. The CON-group received normal saline; the Pb-group received PbAc; the Pb + Q-CRN group received lead acetate followed by quercetin; the Q-CRN group received quercetin; and the CRN group received corn oil. After 4weeks, the mice were euthanized. It was speculated that Pb significantly increased the levels of serine, threonine, and asparagine and decreased the levels of valine, lysine, and glutamic acid in the plasma of Pb-group, thus impairing amino acid metabolism. However, in the Pb + Q-CRN group, the level of these six amino acids was restored significantly due to the ameliorative effect of quercetin. The presence of lipid metabolites (L-carnitine, sphinganine, phytosphingosine, and lysophosphatidylcholine) in mice serum was confirmed by ESI/MS. The GPx, SOD, GSH, and CAT levels were significantly decreased, and the MDA level was significantly increased, thus confirming the oxidative stress and lipid peroxidation in the Pb group. The antioxidant effect of quercetin was elucidated in the Pb + Q-CRN group. Expression of CPT-I, CPT-II, LCAT, CROT, CACT, and MTR genes was significantly upregulated in the liver of Pb goup mice. Hence, the findings of this study proved that Pb exposure induced oxidative stress, upregulated gene expression, and impaired the lipid and amino acid metabolism in mice.

Biochemical and Molecular Profiling of Wild Edible Mushrooms from Huila, Angola.

The harvesting, processing, and sale of wild edible mushrooms (WEM) is a relevant economic activity in Angola and a good example of the use of non-wood forest products for food. Although there is deep traditional knowledge about the general properties of WEMs, a huge gap remains in detailed scientific knowledge. Thus, this study aimed to investigate the socio-economic importance of the species sold at local markets in Huila, Angola, from their molecular identification to the assessment of their nutritional, chemical, and bioactive profiles. From the eight WEM morphotypes studied, five were identified based on phenotypical and molecular approaches (four Russula spp., and Amanita loosei). The studied mushrooms proved to be a rich source of carbohydrates, proteins, and ashes, also presenting low amounts of fat. Chemical analyses further revealed mannitol as the main free sugar in all samples, and organic acids, namely, oxalic, quinic, malic, citric, and fumaric acids in low amounts. Additionally, the α-tocopherol isoform and monounsaturated fatty acids were predominant. Regarding phenolic acids, protocatechuic, p-hydroxybenzoic, p-coumaric, and cinnamic acids were detected in all mushroom hydroethanolic extracts, being responsible for their antioxidant, antibacterial, and antifungal activities. Our investigation contributes to the identification and knowledge of WEMs as important complementary food sources in Angola, some of which were reported for the first time, promoting their utilization as a basis of nutritional and functional ingredients, as being able to be part of a balanced diet and to be used in new bio-based formulations.

Genome analysis and phenotypic characterization of Halomonas hibernica isolated from a traditional food process with novel quorum quenching and catalase activities

Traditional food processes can utilize bacteria to promote positive organoleptic qualities and increase shelf life. Wiltshire curing has a vital bacterial component that has not been fully investigated from a microbial perspective. During the investigation of a Wiltshire brine, a culturable novel bacterium of the genus Halomonas was identified by 16S rRNA gene (MN822133) sequencing and analysis. The isolate was confirmed as representing a novel species (Halomonas hibernica B1.N12) using a housekeeping (HK) gene phylogenetic tree reconstruction with the selected genes 16S rRNA, 23S rRNA, atpA, gyrB, rpoD and secA. The genome of the new isolate was sequenced and annotated and comparative genome analysis was conducted. Functional analysis revealed that the isolate has a unique phenotypic signature including high salt tolerance, a wide temperature growth range and substrate metabolism. Phenotypic and biochemical profiling demonstrated that H. hibernica B1.N12 possesses strong catalase activity which is an important feature for an industrial food processing bacterium, as it can promote an increased product shelf life and improve organoleptic qualities. Moreover, H. hibernica exhibits biocontrol properties based on its quorum quenching capabilities. Our work on this novel isolate advances knowledge on potential mechanistic interplays operating in complex microbial communities that mediate traditional food processes.

Antifungal Activity and Biochemical Profiling of Exudates from Germinating Maize Nostrano di Storo Local Variety.

Plant pathogens are responsible for important damages to valuable crops causing important economic losses. Agrobiodiversity protection is crucial for the valorization of local varieties that could possess higher resistance to biotic and abiotic stress. At the beginning of germination, seeds are susceptible to pathogens attacks, thus they can release endogenous antimicrobial compounds of different natures in the spermosphere, to contrast proliferation of microorganisms. The work aimed at characterizing the maize of local variety Nostrano di Storo seed exudates secreted during the first phases of germination, to identify compounds active in the defense towards pathogens. Storo seed exudates were proven to inhibit F. verticilloides germination. In order to investigate the cause of the described effect, compositional profiling of the exudates was performed through NMR, lipidomic, and proteomic analyses. This study suggests an important role of microbial endophytic communities in the protection of the seed during the early phases of the germination process and their interplay with fatty acids released by the seeds, rather than a specific antifungal compound. The valorization of agronomically acceptable maize lines with pre-harvest enhanced resistances to pathogens contamination could lead, in the near future, to commercially available varieties potentially requiring more limited chemical protective treatments.

Comparative kinase and cancer cell panel profiling of kinase inhibitors approved for clinical use from 2018 to 2020.

During the last two decades, kinase inhibitors have become the major drug class for targeted cancer therapy. Although the number of approved kinase inhibitors increases rapidly, comprehensive in vitro profiling and comparison of inhibitor activities is often lacking in the public domain. Here we report the extensive profiling and comparison of 21 kinase inhibitors approved by the FDA for oncology indications since June 2018 and 13 previously approved comparators on panels of 255 biochemical kinase assays and 134 cancer cell line viability assays. Comparison of the cellular inhibition profiles of the EGFR inhibitors gefitinib, dacomitinib, and osimertinib identified the uncommon EGFR p.G719S mutation as a common response marker for EGFR inhibitors. Additionally, the FGFR inhibitors erdafitinib, infigratinib, and pemigatinib potently inhibited the viability of cell lines which harbored oncogenic alterations in FGFR1-3, irrespective of the specific clinical indications of the FGFR inhibitors. These results underscore the utility of in vitro kinase inhibitor profiling in cells for identifying new potential stratification markers for patient selection. Furthermore, comparison of the in vitro inhibition profiles of the RET inhibitors pralsetinib and selpercatinib revealed they had very similar biochemical and cellular selectivity. As an exception, an NTRK3 fusion-positive cell line was potently inhibited by pralsetinib but not by selpercatinib, which could be explained by the targeting of TRK kinases in biochemical assays by pralsetinib but not selpercatinib. This illustrates that unexpected differences in cellular activities between inhibitors that act through the same primary target can be explained by subtle differences in biochemical targeting. Lastly, FLT3-mutant cell lines were responsive to both FLT3 inhibitors gilteritinib and midostaurin, and the PI3K inhibitor duvelisib. Biochemical profiling revealed that the FLT3 and PI3K inhibitors targeted distinct kinases, indicating that unique dependencies can be identified by combined biochemical and cellular profiling of kinase inhibitors. This study provides the first large scale kinase assay or cell panel profiling study for newly approved kinase inhibitors, and shows that comprehensive in vitro profiling of kinase inhibitors can provide rationales for therapy selection and indication expansion of approved kinase inhibitors.

EP07.01-021 Clinical and Biochemical Profiling of Pleural Mesothelioma Patients Treated With Immunotherapy

Pleural mesothelioma (PM) is a rare and very deadly disease for which there is the necessity to predict treatment’s response and to estimate the overall prognosis. For decades, the only approved treatment for PM has been platinum-based chemotherapy with pemetrexed. Recently the combination of ipilimumab plus nivolumab (I-N) and nivolumab monotherapy (N) have demonstrated a survival benefit in these patients (pts). Nevertheless, any predictive factors of response to immune checkpoint inhibitors (IO) have been identified yet.

Evaluation of serum lipid profiles, uric acid, and high sensitivity C-reactive protein levels between pregnancy-induced hypertension and normotensive pregnant women attending Ambo University Referral Hospital, Ambo, Ethiopia, 2020: A case-control study.

Background and AimsPregnancy‐induced hypertension is one of the top three ranked diseases during pregnancy that cause maternal, fetal, and neonatal morbidity and mortality worldwide. To provide adequate information to clinicians and researchers who are striving for potential interventions, biochemical profiling of such patients is required.MethodsA hospital‐based case‐control study design was conducted from August 2020 to May 2021 to evaluate serum lipid profile, uric acid, and high sensitivity C‐reactive protein (hs‐CRP) among women with pregnancy‐induced hypertension compared to normotensive pregnant women. Data were entered and analyzed using SPSS version 25. Independent t‐test and χ 2 were used to compare the relationship of variables between the two groups. A p‐value less than 0.05 was used to test statistical significance.ResultsThe result of this study showed that while the levels (mean ± SD) of serum total cholesterol (TC), triglyceride (TG), low‐density lipoprotein‐cholesterol (LDL‐C), TC/high‐density lipoprotein‐cholesterol (HDL‐C), TG/HDL‐C, LDL‐C/HDL‐C were significantly elevated, HDL‐C was decreased among women with pregnancy‐induced hypertension than normotensive pregnant women (p < 0.0001). The levels (mean ± SD) of uric acid and hs‐CRP were significantly higher among women with pregnancy‐induced hypertension compared to normotensive pregnant women (p < 0.0001).ConclusionThis study indicated that pregnancy‐induced hypertension women have lipid abnormalities, increased systemic inflammatory markers, and hyperuricemia compared to normotensive pregnant women. Thus, women with PIH showing high dyslipidemia, hyperuricemia, and inflammation are likely to develop hypertension. Therefore, evaluation of these potential biomarkers during early antenatal care services may help seek interventions in PIH.