Search results for: NOD2 (nucleotide binding oligomerization receptor 2)

Authors: Reza Vakili, Xiaolei Fan, Alex Walton

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The first near ambient pressure (NAP)-XPS study of CO oxidation over Pt nanoparticles (NPs) incorporated into Zr-based UiO (UiO for Universitetet i Oslo) MOFs was carried out. For this purpose, the MOF-based Catalysts were prepared by wetness impregnation (WI-PtNPs@UiO-67) and linker design (LD-PtNPs@UiO-67) methods along with PtNPs@ZrO₂ as the control catalyst. Firstly, the as-synthesized catalysts were reduced in situ prior to the operando XPS analysis. The existence of Pt(II) species was proved in UiO-67 by observing Pt 4f core level peaks at a high binding energy of 72.6 ± 0.1 eV. However, by heating the WI-PtNPs@UiO-67 catalyst in situ to 200 °C under vacuum, the higher BE components disappear, leaving only the metallic Pt 4f doublet, confirming the formation of Pt NPs. The complete reduction of LD-PtNPs@UiO-67 is achieved at 250 °C and 1 mbar H₂. To understand the chemical state of Pt NPs in UiO-67 during catalytic turnover, we analyzed the Pt 4f region using operando NAP-XPS in the temperature-programmed measurements (100-260 °C) with reference to PtNPs@ZrO₂ catalyst. CO conversion during NAP-XPS experiments with the stoichiometric mixture shows that LD-PtNPs@UiO-67 has a better CO turnover frequency (TOF, 0.066 s⁻¹ at 260 °C) than the other two (ca. 0.055 s⁻¹). Pt 4f peaks only show one chemical species present at all temperatures, but the core level BE shifts change as a function of reaction temperature, i.e., Pt 4f peak from 71.8 eV at T < 200 °C to 71.2 eV at T > 200 °C. As this higher BE state of 71.8 eV was not observed after in situ reductions of the catalysts and only once the CO/O₂ mixture was introduced, we attribute it to the surface saturation of Pt NPs with adsorbed CO. In general, the quantitative analysis of Pt 4f data from the operando NAP-XPS experiments shows that the surface chemistry of the Pt active phase in the two PtNPs@UiO-67 catalysts is the same, comparable to that of PtNPs@ZrO₂. The observed difference in the catalytic activity can be attributed to the particle sizes of Pt NPs, as well as the dispersion of active phase in the support, which are different in the three catalysts.

Keywords: CO oxidation, heterogeneous catalysis, MOFs, Metal Organic Frameworks, NAP-XPS, Near Ambient Pressure X-ray Photoelectron Spectroscopy

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Authors: Vikash Yadav, Ashish Arora

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Microorganisms have self-defense mechanisms to protect themselves from toxic environments. Phenolic acid decarboxylase(pad) is responsible for the defense against toxicity caused by phenolic acids, converting them into less toxic vinyl derivatives. The transcription of the pad gene is regulated by a negative transcription factor, phenolic acid decarboxylase regulators (PadR), in a substrate-inducible manner. The PadR family members share the conserved DNA-binding features and interact with the operator DNA using a winged helix-turn-helix (wHTH) motif, which contains a three-helix motif and a β-stranded wing. The members of this family function as transcriptional regulators that are involved in various cellular survival processes, such as toxin production, detoxification, multidrug resistance, antibiotic biosynthesis, and carbon catabolism. Rv1176 of Mycobacterium tuberculosis H37Rv has been assigned to the PadR family protein that remains to be structurally and functionally uncharacterized. To reveal the structural mechanism by which Rv1176 could regulates effector-responsive transcription, several experiments were performed, including Electrophoretic Mobility Shift Assay (EMSA) for DNA protein interaction, differential scanning calorimetry (DSC) and Differential Scanning Fluorimetry (DSF) for temperature and ligand-dependent protein stability, Circular Dichroism (CD) spectroscopy for secondary structure analysis. Further, to evaluate the functional role of Rv1176, the intracellular survival of recombinant M. smegmatis was examined in murine macrophage cell line J774A.1 and different stressed conditions like oxidative, pH, and nutritive stress. All these studies demonstrated that Rv1176 could behave as a transcription regulator and its expression in recombinant M. smegmatis increases intracellular survival.

Keywords: EMSA, Mycobacterium tuberculosis, PadR family protein, transcriptional regulator

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Authors: Murali Aarthy, Sanjeev Kumar Singh

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High risk human papillomaviruses are highly associated with the carcinoma of the cervix and the other genital tumors. Cervical cancer develops through the multistep process in which increasingly severe premalignant dysplastic lesions called cervical intraepithelial neoplastic progress to invasive cancer. The oncoprotein E7 of human papillomavirus expressed in the lower epithelial layers drives the cells into S-phase creating an environment conducive for viral genome replication and cell proliferation. The replication of the virus occurs in the terminally differentiating epithelium and requires the activation of cellular DNA replication proteins. To date, no suitable drug molecule is available to treat HPV infection whereas identification of potential drug targets and development of novel anti-HPV chemotherapies with unique mode of actions are expected. Hence, our present study aimed to identify the potential inhibitors analogous to EGCG, a green tea molecule which is considered to be safe to use for mammalian systems. A 3D similarity search on the natural small molecule library from natural product database using EGCG identified 11 potential hits based on their similarity score. The structure based docking strategies were implemented in the potential hits and the key interacting residues of protein with compounds were identified through simulation studies and binding free energy calculations. The conformational changes between the apoprotein and the complex were analyzed with the simulation and the results demonstrated that the dynamical and structural effects observed in the protein were induced by the compounds and indicated the dominance to the oncoprotein. Overall, our study provides the basis for the structural insights of the identified potential hits and EGCG and hence, the analogous compounds identified can be potent inhibitors against the HPV 16 E7 oncoprotein.

Keywords: EGCG, oncoprotein, molecular dynamics simulation, analogues

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Authors: Magdalena Korytowska, Gunnar Lengstrand, Cecilia Larsson Wexell

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Background: Breast cancer (BC) is the most common cancer among women worldwide, and cases are continuing to increase in Sweden. Bone is the most common metastatic site in breast cancer patients, where > 65-75% of women with advanced breast cancer develop bone metastases during their disease. To prevent the skeletal-related events of metastases (e.g., pathological fractures, bone loss, cancer-induced bone pain, and hypercalcemia bone), two different classes of antiresorptive medications (AR), bisphosphonate and denosumab are typically administered every 3 to 4 weeks. Since 2015, adjuvant bisphosphonate treatment has been used every six months for three to five years in postmenopausal women for the prevention of skeletal metastases and improved survival. Methods: A case-control study was conducted to test the hypotheses that patients treated with high-dose AR are at higher risk of developing MRONJ than breast cancer patients with adjuvant bisphosphonate treatment at a lower dose. Medical and odontological data was collected between 2015-2020. Assessment of oral health and dental care before and during oncological treatment took place at the specialist clinic for Orofacial medicine linked to the specific hospital. Results: In total, 220 patients were included, 101 patients in the high-dose group and 119 patients in the adjuvant BP-treatment group. MRONJ was diagnosed in 13 patients (14%) in the high-dose group. The mandible was affected in most of the cases (84.6%), with a mean duration of high-dose treatment of 19.7 months. In 46.2% of cases, no dental cause of MRONJ could be identified. Overall, estrogen receptor-positive (ER+) BC was the most representative type in 172 patients (78.2%). However, this was 83.9% in the high-dose cases group. The most used drug was denosumab. Twenty-five patients (26.9%) switched their medication from ZOL to denosumab during their oncological treatment. Patients with ER+ breast cancer were reported in 88 patients (87.8%) in the adjuvant group that was treated with ZOL. Conclusions: MRONJ was diagnosed only in the high-dose AR group. Dental assessment and care of patients in the adjuvant group should be considered, with a recommendation to potentially prolong ZOL treatment from 3 to 5 years, with concomitant use of hormonal therapy in patients diagnosed with ER+ breast cancer to prevent bone loss induced by oncological treatment. A new referral for dental assessment is very important in the case of bone metastases when treatment with high dose AR will be required since it is associated with a higher risk of MRONJ.

Keywords: antiresorptive therapy, breast cancer, dental care, MRONJ

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Authors: Clotilde Guyon, Nada Jmari, Yen-Chin Li, Jean Denoyel, Noriyuki Fujikado, Christophe Blanchet, David Root, Matthieu Giraud

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Aire induces ectopic expression of a large repertoire of tissue-specific antigen (TSA) genes in thymic medullary epithelial cells (MECs), driving immunological self-tolerance in maturing T cells. Although important mechanisms of Aire-induced transcription have recently been disclosed through the identification and the study of Aire’s partners, the fine transcriptional functions underlied by a number of them and conferred to Aire are still unknown. Alternative cleavage and polyadenylation (APA) is an essential mRNA processing step regulated by the termination complex consisting of 85 proteins, 10 of them have been related to Aire. We evaluated APA in MECs in vivo by microarray analysis with mRNA-spanning probes and RNA deep sequencing. We uncovered the preference of Aire-dependent transcripts for short-3’UTR isoforms and for proximal poly(A) site selection marked by the increased binding of the cleavage factor Cstf-64. RNA interference of the 10 Aire-related proteins revealed that Clp1, a member of the core termination complex, exerts a profound effect on short 3’UTR isoform preference. Clp1 is also significantly upregulated in the MECs compared to 25 mouse tissues in which we found that TSA expression is associated with longer 3’UTR isoforms. Aire-dependent transcripts escape a global 3’UTR lengthening associated with MEC differentiation, thereby potentiating the repressive effect of microRNAs that are globally upregulated in mature MECs. Consistent with these findings, RNA deep sequencing of actinomycinD-treated MECs revealed the increased stability of short 3’UTR Aire-induced transcripts, resulting in TSA transcripts accumulation and contributing for their enrichment in the MECs.

Keywords: Aire, central tolerance, miRNAs, transcription termination

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Authors: Berina Pilipović, Saša Pilipović, Maja Pašić-Kulenović

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Inflammation is the body's response to impaired homeostasis caused by infection, injury or trauma resulting in systemic and local effects. Inflammation causes the body's response to injury and is characterized by a series of events including inflammatory response, response to pain receptors and the recovery process. Inflammation can be acute and chronic. The inflammatory response is described in three different phases. Free radical is an atom or molecule that has the unpaired electron and is therefore generally very reactive chemical species. Biologically important example of reaction with free radicals is called Lipid peroxidation (LP). Lipid peroxidation reactions occur in biological membranes, and if at the outset is not stopped with the action of antioxidants, it will bring damage to the membrane, which results in partial or complete loss of their physiological functions. Calcium antagonists and beta-adrenergic receptor antagonists are known drugs, and for many years and widely used in the treatment of cardiovascular diseases. Some of these compounds also show antioxidant activity. The mechanism of antioxidant activities of calcium antagonists and beta-blockers is unknown, since their structure varies widely. This research investigated the possible local anti-inflammatory activity of ointments containing 1% carvedilol in the white petrolatum USP. Ear inflammation was induced by 3% croton oil acetone solution, in quantity of 10 µl on both mouse ears. Albino Swiss mouse (n = 8) are treated with 2.5 mg/ear ointment, and control group was treated on the same way as previous with hydrocortisone 1% ointment (2.5 mg/ear). The other ear of the same animal was used as control one. Ointments were administered once per day, on the left ear. After treatment, ears were observed for three days. After three days, we measured mass (mg) of 6 mm ear punch of treated and controlled ears. The results of testing anti-inflammatory effects of ointments with carvedilol in the mouse ear model show stronger observed effect than ointment with 1% hydrocortisone in the same basis. Identical results were confirmed by the difference between the mass of 6 mm ears punch. The results were also confirmed by histological examination. Ointments with carvedilol showed significant reduction of the inflammation process caused by croton oil on the mouse inflammation model.

Keywords: antioxidant, carvedilol, inflammation, mouse ear

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Authors: Mohamed A. Etman, Mona G. Abd-Elnasser, Mohamed A. Shams-Eldin, Aly H. Nada

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Orally disintegrating tablets (ODTs) are gaining importance as new drug delivery systems and emerged as one of the popular and widely accepted dosage forms, especially for the pediatric and geriatric patients. Their advantages such as administration without water, anywhere, anytime lead to their suitability to geriatric and pediatric patients. They are also suitable for the mentally ill, the bed-ridden and patients who do not have easy access to water. The benefits, in terms of patient compliance, rapid onset of action, increased bioavailability, and good stability make these tablets popular as a dosage form of choice in the current market. These dosage forms dissolve or disintegrate in the oral cavity within a matter of seconds without the need of water or chewing. Desloratadine is a tricyclic antihistaminic, which has a selective and peripheral H1-antagonist action. It is an antagonist at histamine H1 receptors, and an antagonist at all subtypes of the muscarinic acetylcholine receptor. Desloratadine is the major metabolite of loratadine. Twelve different placebos ODT were prepared (F1-F12) using different functional excipients. They were evaluated for their compressibility, hardness and disintegration time. All formulations were non sticky except four formulations; namely (F8, F9, F10, F11). All formulations were compressible with the exception of (F2). Variable disintegration times were found ranging between 20 and 120 seconds. It was found that (F12) showed the least disintegration time (20 secs) without showing any sticking which could be due to the use of high percentage of superdisintegrants. Desloratadine showed bitter taste when formulated as ODT without any treatment. Therefore, different techniques were tried in order to mask its bitter taste. Using Eudragit EPO resulted in complete masking of the bitter taste of the drug and increased the acceptability to volunteers. The compressible non sticky formulations (F1, F3, F4, F5, F6, F7 and F12) were subjected to further evaluation tests after addition of coated desloratadine, including weight uniformity, wetting time, and friability testing.. Fairly good weight uniformity values were observed in all the tested formulations. F12 exhibiting the shortest wetting time (14.7 seconds) and consequently the lowest (20 seconds) disintegration time. Dissolution profile showed that 100% desloratadine release was attained after only 2.5 minutes from the prepared ODT (F12) with dissolution efficiency of 95%.

Keywords: Desloratadine, orally disintegrating tablets (ODTs), formulations, taste masking

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Authors: K. S. Vimal, D. Bodhini, K. Ramya, N. Lakshmipriya, R. M. Anjana, V. Sudha, J. A. Lovegrove, V. Mohan, V. Radha

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Gene-lifestyle interaction studies have been carried out in various populations. However, to date there are no studies in an Asian Indian population. Hence, we examined whether lifestyle factors such as diet and physical activity modify the association between fat mass and obesity–associated (FTO) gene variants and obesity and type 2 diabetes (T2D) in an Asian Indian population. We studied 734 unrelated T2D and 884 normal glucose-tolerant (NGT) participants randomly selected from the Chennai Urban Rural Epidemiology Study (CURES) in Southern India. Obesity was defined according to the World Health Organization Asia Pacific Guidelines (non-obese, BMI < 25 kg/m2; obese, BMI ≥ 25 kg/m2). Six single nucleotide polymorphisms (SNPs) in the FTO gene (rs9940128, rs7193144, rs8050136, rs918031, rs1588413 and rs11076023) identified from recent genome-wide association studies for T2D were genotyped by polymerase chain reaction-restriction fragment length polymorphism and direct sequencing. Dietary assessment was carried out using a validated food frequency questionnaire and physical activity was based upon the self-report. Interaction analyses were performed by including the interaction terms in the model. A joint likelihood ratio test of the main SNP effects and the SNP-diet/physical activity interaction effects was used in the linear regression analyses to maximize statistical power. Statistical analyses were performed using STATA version 13. There was a significant interaction between FTO SNP rs8050136 and carbohydrate energy percentage (Pinteraction=0.04) on obesity, where the ‘A’ allele carriers of the SNP rs8050136 had 2.46 times higher risk of obesity than those with ‘CC’ genotype (P=3.0x10-5) among individuals in the highest tertile of carbohydrate energy percentage. Furthermore, among those who had lower levels of physical activity, the ‘A’ allele carriers of the SNP rs8050136 had 1.89 times higher risk of obesity than those with ‘CC’ genotype (P=4.0x10-5). We also found a borderline interaction between SNP rs11076023 and carbohydrate energy percentage (Pinteraction=0.08) on T2D, where the ‘A’ allele carriers in the highest tertile of carbohydrate energy percentage, had 1.57 times higher risk of T2D than those with ‘TT’ genotype (P=0.002). There was also a significant interaction between SNP rs11076023 and physical activity (Pinteraction=0.03) on T2D. No further significant interactions between SNPs and macronutrient intake or physical activity on obesity and T2D were observed. In conclusion, this is the first study to provide evidence for a gene-diet and gene-physical activity interaction on obesity and T2D in an Asian Indian population. These findings suggest that the association between FTO gene variants and obesity and T2D is influenced by carbohydrate intake and physical activity levels. Greater understanding of how FTO gene influences obesity and T2D through dietary and exercise interventions will advance the development of behavioral intervention and personalised lifestyle strategies predicted to reduce the development of metabolic diseases in ‘A’ allele carriers of both SNPs in this Asian Indian population.

Keywords: dietary intake, FTO, obesity, physical activity, type 2 diabetes, Asian Indian.

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Authors: Uzaira Rafique, Kousar Parveen

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The discharge volume and composition of Textile effluents gains scientific concern due to its hazards and biotoxcity of azo dyes. Azo dyes are non-biodegradable due to its complex molecular structure and recalcitrant nature. Serious attempts have been made to synthesize and develop new materials to combat the environmental problems. The present study is designed for removal of a range of azo dyes (Methyl orange, Congo red and Basic fuchsine) from synthetic aqueous solutions and real textile effluents. For this purpose, Metal (cobalt) doped alumina hybrids are synthesized and applied as adsorbents in the batch experiment. Two different aluminium precursor (aluminium nitrate and spent aluminium foil) and glucose are mixed following sol gel method to get hybrids. The synthesized materials are characterized for surface and bulk properties using FTIR, SEM-EDX and XRD techniques. The characterization of materials under FTIR revealed that –OH (3487-3504 cm-1), C-H (2935-2985 cm-1), Al-O (~ 800 cm-1), Al-O-C (~1380 cm-1), Al-O-Al (659-669 cm-1) groups participates in the binding of dyes onto the surface of hybrids. Amorphous shaped particles and elemental composition of carbon (23%-44%), aluminium (29%-395%), and oxygen (11%-20%) is demonstrated in SEM-EDX micrograph. Time-dependent batch-experiments under identical experimental parameters showed 74% congo red, 68% methyl orange and 85% maximum removal of basic fuchsine onto the surface of cobalt doped alumina hybrids probably through the ion-exchange mechanism. The experimental data when treated with adsorption models is found to have good agreement with pseudo second order kinetic and freundlich isotherm for adsorption process. The present study concludes the successful synthesis of novel and efficient cobalt doped alumina hybrids providing environmental friendly and economical alternative to the commercial adsorbents for the treatment of industrial effluents.

Keywords: alumina hybrid, adsorption, dopant, isotherm, kinetic

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Authors: Tatiana A. Pozdniakova, Maria A. P. Cechinel, Luciana P. Mazur, Rui A. R. Boaventura, Vitor J. P. Vilar.

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Two brown macroalgae, Laminaria hyperborea and Pelvetia canaliculata, were employed as natural cation exchangers in a fixed-bed column for Zn(II) removal from a galvanization wastewater. The column (4.8 cm internal diameter) was packed with 30-59 g of previously hydrated algae up to a bed height of 17-27 cm. The wastewater or eluent was percolated using a peristaltic pump at a flow rate of 10 mL/min. The effluent used in each experiment presented similar characteristics: pH of 6.7, 55 mg/L of chemical oxygen demand and about 300, 44, 186 and 244 mg/L of sodium, calcium, chloride and sulphate ions, respectively. The main difference was nitrate concentration: 20 mg/L for the effluent used with L. hyperborean and 341 mg/L for the effluent used with P. canaliculata. The inlet zinc concentration also differed slightly: 11.2 mg/L for L. hyperborean and 8.9 mg/L for P. canaliculata experiments. The breakthrough time was approximately 22.5 hours for both macroalgae, corresponding to a service capacity of 43 bed volumes. This indicates that 30 g of biomass is able to treat 13.5 L of the galvanization wastewater. The uptake capacities at the saturation point were similar to that obtained in batch studies (unpublished data) for both algae. After column exhaustion, desorption with 0.1 M HNO3 was performed. Desorption using 9 and 8 bed volumes of eluent achieved an efficiency of 100 and 91%, respectively for L. hyperborean and P. canaliculata. After elution with nitric acid, the column was regenerated using different strategies: i) convert all the binding sites in the sodium form, by passing a solution of 0.5 M NaCl, until achieve a final pH of 6.0; ii) passing only tap water in order to increase the solution pH inside the column until pH 3.0, and in this case the second sorption cycle was performed using protonated algae. In the first approach, in order to remove the excess of salt inside the column, distilled water was passed through the column, leading to the algae structure destruction and the column collapsed. Using the second approach, the algae remained intact during three consecutive sorption/desorption cycles without loss of performance.

Keywords: biosorption, zinc, galvanization wastewater, packed-bed column

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Authors: Anamarija Kuhar, Veronika Kloboves Prevodnik, Nataša Nolde, Ulrika Klopčič

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The decision for immunocytochemistry (ICC) is usually made in the basis of the findings in Giemsa- and/or Papanicolaou- smears. More demanding diagnostic cases require preparation of additional cytological preparations. Therefore, it is convenient to suspend cytological samples in a liquid based medium (LBM) that preserve antigen and morphological properties. However, the duration of these properties being preserved in the medium is usually unknown. Eventually, cell morphology becomes impaired and altered, as well as antigen properties may be lost or become diffused. In this study, the influence of cytological sample storage length in in-house liquid based medium on antigen properties and cell morphology is evaluated. The question is how long the cytological samples in this medium can be stored so that the results of immunocytochemical reactions are still reliable and can be safely used in routine cytopathological diagnostics. The stability of 6 ICC markers that are most frequently used in everyday routine work were tested; Cytokeratin AE1/AE3, Calretinin, Epithelial specific antigen Ep-CAM (MOC-31), CD 45, Oestrogen receptor (ER), and Melanoma triple cocktail were tested on methanol fixed cytospins prepared from fresh fine needle aspiration biopsies, effusion samples, and disintegrated lymph nodes suspended in in-house cell medium. Cytospins were prepared on the day of the sampling as well as on the second, fourth, fifth, and eight day after sample collection. Next, they were fixed in methanol and immunocytochemically stained. Finally, the percentage of positive stained cells, reaction intensity, counterstaining, and cell morphology were assessed using two assessment methods: the internal assessment and the UK NEQAS ICC scheme assessment. Results show that the antigen properties for Cytokeratin AE1/AE3, MOC-31, CD 45, ER, and Melanoma triple cocktail were preserved even after 8 days of storage in in-house LBM, while the antigen properties for Calretinin remained unchanged only for 4 days. The key parameters for assessing detection of antigen are the proportion of cells with a positive reaction and intensity of staining. Well preserved cell morphology is highly important for reliable interpretation of ICC reaction. Therefore, it would be valuable to perform a similar analysis for other ICC markers to determine the duration in which the antigen and morphological properties are preserved in LBM.

Keywords: cytology samples, cytospins, immunocytochemistry, liquid-based cytology

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Authors: Ahmed Elbeltagy, Francesca Bertolini, Damarius Fleming, Angelica Van Goor, Chris Ashwell, Carl Schmidt, Donald Kugonza, Susan Lamont, Max Rothschild

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The major factor in shaping genomic variation of the African indigenous rural chicken is likely natural selection drives the development genetic footprints in the chicken genomes. To investigate such a hypothesis of a selection footprint, a total of 292 birds were randomly sampled from three indigenous ecotypes from East Africa (Uganda, Rwanda) and North Africa (Egypt) and two registered Egyptian breeds (Fayoumi and Dandarawi), and from the synthetic Kuroiler breed. Samples were genotyped using the Affymetrix 600K Axiom® Array. A total of 526,652 SNPs were utilized in the downstream analysis after quality control measures. The intra-population runs of homozygosity (ROH) that were consensuses in > 50% of individuals of an ecotype or > 75% of a breed were studied. To identify inter-population differentiation due to genetic structure, FST was calculated for North- vs. East- African populations in addition to population-pairwise combinations for overlapping windows (500Kb with an overlap of 250Kb). A total of 28,563 ROH were determined and were classified into three length categories. ROH and Fst detected sweeps were identified on several autosomes. Several genes in these regions are likely to be related to adaptation to local environmental stresses that include high altitude, diseases resistance, poor nutrition, oxidative and heat stresses and were linked to gene ontology terms (GO) related to immune response, oxygen consumption and heme binding, carbohydrate metabolism, oxidation-reduction, and behavior. Results indicated a possible effect of natural selection forces on shaping genomic structure for adaptation to local environmental stresses.

Keywords: African Chicken, runs of homozygosity, FST, selection footprints

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Authors: Daniela Rivera-Tobar Mario Perez-Won, Roberto Lemus-Mondaca, Gipsy Tabilo-Munizaga

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Different dietary trends worldwide seek to consume foods with anti-inflammatory properties, rich in antioxidants, proteins, and unsaturated fatty acids that lead to better metabolic, intestinal, mental, and cardiac health. In this sense, food matrices with high protein content based on macro and microalgae are an excellent alternative to meet the new needs of consumers. An emerging and environmentally friendly technology for producing protein matrices is ultrasonic agglomeration. It consists of the formation of permanent bonds between particles, improving the agglomeration of the matrix compared to conventionally agglomerated products (compression). Among the advantages of this process are the reduction of nutrient loss and the avoidance of binding agents. The objective of this research was to optimize the ultrasonic agglomeration process in matrices composed of Spirulina (Arthrospira platensis) powder and Cochayuyo (Durvillae Antartica) flour, by means of the response variable (Young's modulus) and the independent variables were the process conditions (percentage of ultrasonic amplitude: 70, 80 and 90; ultrasonic agglomeration times and cycles: 20, 25 and 30 seconds, and 3, 4 and 5). It was evaluated using a central composite design and analyzed using response surface methodology. In addition, the effects of agglomeration on thermophysical and microstructural properties were evaluated. It was determined that ultrasonic compression with 80 and 90% amplitude caused conformational changes according to Fourier infrared spectroscopy (FTIR) analysis, the best condition with respect to observed microstructure images (SEM) and differential scanning calorimetry (DSC) analysis, was the condition of 90% amplitude 25 and 30 seconds with 3 and 4 cycles of ultrasound. In conclusion, the agglomerated matrices present good macro and microstructural properties which would allow the design of food systems with better nutritional and functional properties.

Keywords: ultrasonic agglomeration, physical properties of food, protein matrices, macro and microalgae

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Authors: Keith T. S. Tung, H. W. Tsang, Rosa S. Wong, Frederick K. Ho, Patrick Ip

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Objectives: Atopic diseases are increasingly prevalent among children and adolescents, both locally and internationally. One of the possible contributing factors could be the hypercholesterolemia which leads to cholesterol accumulation in macrophages and other immune cells that would eventually promote inflammatory responses, including augmentation of toll-like receptor (TLR). Meanwhile, physical activity is well known for its beneficial effects against the condition of hypercholesterolemia and incidence of atopic diseases. This study, therefore, explored whether atopic diseases were associated with increased cholesterol levels and whether physical activity habit influenced this association. Methods: This is a sub-study derived from the longitudinal cohort study which recruited a group of children at five years of age in Kindergarten 3 (K3) to investigate the long-term impact of family socioeconomic status on child development. In 2018/19, adolescents (average age: 13 years old) were asked to report their physical activity habit and history of any atopic diseases. During health assessment, peripheral blood samples were collected from the adolescents to study their lipid profile [total cholesterol, high-density lipoprotein (HDL)-cholesterol, and low-density lipoprotein (LDL)-cholesterol]. Regression analyses were performed to test the relationships between variables of interest. Results: Among the 315 adolescents, 99 (31.4%) reported to have allergic rhinitis. There were 45 (14.3%) with eczema, 17 (5.4%) with a food allergy, and 12 (3.8%) with asthma. Regression analyses showed that adolescents with a history of any type of atopic diseases had significantly higher total cholesterol (B=13.3, p < 0.01) and LDL cholesterol (B=7.9, p < 0.05) levels. Further subgroup analyses were conducted to examine the effect of physical activity level on the association between atopic diseases and cholesterol levels. We found stronger associations among those who did not meet the World Health Organization recommendation of at least 60 minutes of moderate-to-vigorous activities each day (total cholesterol: B=15.5, p < 0.01; LDL cholesterol: B=10.4, p < 0.05). For those who met this recommendation, the associations between atopic diseases and cholesterol levels became insignificant. Conclusion: Our study results support the current research evidence on the relationship between an elevated level of cholesterol and atopic diseases. More importantly, our results provide preliminary support for the protective effect of regular exercises against elevated cholesterol level due to atopic diseases. The findings highlight the importance of a healthy lifestyle for keeping cholesterol levels in the normal range, which can bring benefits to both physical and mental health.

Keywords: atopic diseases, Chinese adolescents, cholesterol level, physical activity

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Authors: Sayed Mostafa, Siham Mostafa Mohamed Faheid, Ibrahim Rizk Sayed Ahmed, Yasser Fehry Mohamed Kishk, Gamal Hassan Ragab

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Gluten-free foods are still the only treatment for gluten-allergic patients. Consequently, this study is concerned with improving the quality attributes of gluten-free bread using different concentrations (0, 20, 40, 60 and 80ppm) of all maltogenic α-amylase (MA) and xylanase (XY) compared with wheat flour Balady bread and untreated gluten-free Balady bread (GFBB). Pasting properties, falling number, water activity, alkaline water retention capacity (AWRC) and sensory properties (fresh bread, after 24h, after 48h and after 72h) of gluten-free bread were evaluated. Additionally, the effect of merging different concentrations of maltogenic α-amylase and xylanase on stalling behavior (AWRC) and sensory properties of gluten-free Balady bread was investigated. The addition of MA led to a gradually decreased peak viscosity, breakdown, setback and pasting temperature of GFBB with the increasing level of MA. Maltogenic α-amylase and xylanase addition led to a reduction in the FN values compared to the untreated gluten-free sample, noting that the MA-treated samples showed a significant decrease compared to the XY-treated and untreated samples. Wheat flour Balady bread significantly showed a higher value of AWRC compared to untreated gluten-free Balady bread at different storage periods (zero time, after 24h, after 48h and after 72h). MA-treated samples showed higher water binding capacity and water activity (aw)in comparison with XY-treated samples, with significance during all storage periods. Concerning the overall acceptability during the third day, the highest score (4.6) was observed by the GFBB sample containing 40ppm MA, followed by 4.3, which was investigated by the GFBB sample containing 80ppm XY with no significance between them and with significance compared to the other samples.

Keywords: celiac disease, gluten-free products, anti-stalling agents, maltogenic α-amylase, xylanase

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Authors: Mohammadjavad Sotoudeheian

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COVID-19, which began in December 2019, uses the angiotensin-converting enzyme 2 (ACE2) receptor to enter and spread through the cells. ACE2 mRNA is present in almost every organ, including nasopharynx, lung, as well as the brain. Ports of entry of SARS-CoV-2 into the central nervous system (CNS) may include arterial circulation, while viremia is remarkable. However, it is imperious to develop neurological symptoms evaluation CSF analysis in patients with COVID-19, but theoretically, ACE2 receptors are expressed in cerebellar cells and may be a target for SARS-CoV-2 infection in the brain. Recent evidence agrees that SARS-CoV-2 can impact the brain through direct and indirect injury. Two biomarkers for CNS injury, glial fibrillary acidic protein (GFAP) and neurofilament light chain (NFL) detected in the plasma of patients with COVID-19. NFL, an axonal protein expressed in neurons, is related to axonal neurodegeneration, and GFAP is over-expressed in CNS inflammation. GFAP cytoplasmic accumulation causes Schwan cells to misfunction, so affects myelin generation, reduces neuroskeletal support over NfLs during CNS inflammation, and leads to axonal degeneration. Interleukin-6 (IL-6), which extensively over-express due to interleukin storm during COVID-19 inflammation, regulates gene expression, as well as GFAP through STAT molecular pathway. IL-6 also impresses the phosphoinositide 3-kinase (PI3K)/STAT/smads pathway. The PI3K/ protein kinase B (Akt) pathway is the main modulator upstream of the mammalian target of rapamycin (mTOR), and alterations in this pathway are common in neurodegenerative diseases. Most neurodegenerative diseases show a disruption of autophagic function and display an abnormal increase in protein aggregation that promotes cellular death. Therefore, induction of autophagy has been recommended as a rational approach to help neurons clear abnormal protein aggregates and survive. The mTOR is a major regulator of the autophagic process and is regulated by cellular stressors. The mTORC1 pathway and mTORC2, as complementary and important elements in mTORC1 signaling, have become relevant in the regulation of the autophagic process and cellular survival through the extracellular signal-regulated kinase (ERK) pathway.

Keywords: mTORC1, COVID-19, PI3K, autophagy, neurodegeneration

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Authors: R. K. Agrawal, Shiv Agrawal

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Fines and micro create fundamental problems of respiration. From mines to mills steel plants generate lot of pollutants. Legislation & Government laws are stricter day by day & each plant has to think of recycling, reuse &reintegration of pollutants generated during the process of steel making. This paper deals with experiments conducted in Bhilai Steel Plant and Real Ispat and Power Limited for reuse, recycle & reintegrate some of the steel making process fines. Iron ore fines with binders have been agglomerated to be used as a part of the charge for small furnaces. This will improve yield at nominal cost. Rolling mill fines have been recycled to increase the yield of sinter making. This will solve the problems of fine disposal. Huge saving on account of recycling will be achieved. Lime fines after briquetting is used along with prime lime. Lime fines have also been used as a binding material during production of fly ash bricks. These fines serve as low-cost binder. Experiments have been conducted along with coke breeze & gas cleaning plant sludge. As a result, the anti-sloping compound has been developed for converter vessels. Dolo char and Char during Sponge Iron production have been successfully used in power generation and brick making. Pellets have been made with ventilation dust & flue dust. These samples have been tried as a coolant in the converter. Pellets have been made with Sinter Plant electrostatic precipitator micro fines with liquid binder. Trials have been conducted to reuse these pellets in sinter making. Coke breeze from coke-ovens fines and mill scale along with binders were agglomerated. This was used in furnace after attaining required screening and reactivity index. These actions will definitely bring social, economic and environment-friendly universe.

Keywords: briquette, dolo char, electrostatic precipitator, pellet, sinter

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Authors: Rowland Brucken

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This paper argues that nation-building theories that prioritize democratic governance best explain the successful post-independence development of Botswana. Three main competing schools of thought exist regarding the sequencing of policies that should occur to re-build weakened or failed states. The first posits that economic development should receive foremost attention, while democratization and a binding sense of nationalism can wait. A second group of experts identified constructing a sense of nationalism among a populace is necessary first, so that the state receives popular legitimacy and obedience that are prerequisites for development. Botswana, though, transitioned into a multi-party democracy and prosperous open economy due to the utilization of traditional democratic structures, enlightened and accountable leadership, and an educated technocratic civil service. With these political foundations already in place when the discovery of diamonds occurred, the resulting revenues were spent wisely on projects that grew the economy, improved basic living standards, and attracted foreign investment. Thus democratization preceded, and therefore provided an accountable basis for, economic development that might otherwise have been squandered by greedy and isolated elites to the detriment of the greater population. Botswana was one of the poorest nations in the world at the time of its independence in 1966, with little infrastructure, a dependence on apartheid South Africa for trade, and a largely subsistence economy. Over the next thirty years, though, its economy grew the fastest of any nation in the world. The transparent and judicious use of diamond returns is only a partial explanation, as the government also pursued economic diversification, mass education, and rural development in response to public needs. As nation-building has become a project undertaken by nations and multilateral agencies such as the United Nations and the North Atlantic Treaty Organization, Botswana may provide best practices that others should follow in attempting to reconstruct economically and politically unstable states.

Keywords: Botswana, democratization, economic development, nation-building

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Authors: Asis Kumar Khan, Rajeev Kumar Goel

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Recent infrastructure growth all around the world lead to increase in demand for concrete day by day. Cement being binding material for concrete the usage of cement also gone up significantly. Cement manufacturing utilizes abundant natural resources and causes environment pollution by releasing a huge quantity of CO₂ into the atmosphere. So, it is high time to look for alternates to reduce the cement consumption in concrete. Geopolymer concrete is one such material which utilizes the industrial waste such as fly ash, ground granulated blast furnace slag and low-cost alkaline liquids such as sodium hydroxide and sodium silicate to produce the concrete. On the other side, river sand is becoming very expensive due to its large-scale depletion at source and the high cost of transportation. In this view, river sand is replaced by crushed sand in this study. In this work, an attempt has been made to understand the durability parameters of geopolymer concrete by partially replacing fly ash with cement. Fly ash is replaced by cement at various levels e.g., from 0 to 50%. Concrete cubes of 100x100x100mm were used for investigating different durability parameters. The various parameters studied includes compressive strength, split tensile strength, drying shrinkage, sodium sulphate attack resistance, sulphuric acid attack resistance and chloride permeability. Highest compressive strength & highest split tensile strength is observed in 30% replacement level. Least drying is observed with 30% replacement level. Very good resistance for sulphuric acid & sodium sulphate is found with 30% replacement. However, it was not possible to find out the chloride permeability due to the high conductivity of geopolymer samples of all replacement levels.

Keywords: crushed sand, compressive strength, drying shrinkage, geopolymer concrete, split tensile strength, sodium sulphate attack resistance, sulphuric acid attack resistance

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Authors: Samia Elfekih, Wee Tek Tay, Karl Gordon, Paul De Barro

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Invasive species represent an increasing threat to food biosecurity, causing significant economic losses in agricultural systems. An example is the sweet potato whitefly, Bemisia tabaci, which is a complex of morphologically indistinguishable species causing average annual global damage estimated at US$2.4 billion. The Bemisia complex represents an interesting model for evolutionary studies because of their extensive distribution and potential for invasiveness and population expansion. Within this complex, two species, Middle East-Asia Minor 1 (MEAM1) and Mediterranean (MED) have invaded well beyond their home ranges whereas others, such as Indian Ocean (IO) and Australia (AUS), have not. In order to understand why some Bemisia species have become invasive, genome-wide sequence scans were used to estimate population dynamics over time and relate these to climate. The Bayesian Skyline Plot (BSP) method as implemented in BEAST was used to infer the historical effective population size. In order to overcome sampling bias, the populations were combined based on geographical origin. The datasets used for this particular analysis are genome-wide SNPs (single nucleotide polymorphisms) called separately in each of the following groups: Sub-Saharan Africa (Burkina Faso), Europe (Spain, France, Greece and Croatia), USA (Arizona), Mediterranean-Middle East (Israel, Italy), Middle East-Central Asia (Turkmenistan, Iran) and Reunion Island. The non-invasive ‘AUS’ species endemic to Australia was used as an outgroup. The main findings of this study show that the BSP for the Sub-Saharan African MED population is different from that observed in MED populations from the Mediterranean Basin, suggesting evolution under a different set of environmental conditions. For MED, the effective size of the African (Burkina Faso) population showed a rapid expansion ≈250,000-310,000 years ago (YA), preceded by a period of slower growth. The European MED populations (i.e., Spain, France, Croatia, and Greece) showed a single burst of expansion at ≈160,000-200,000 YA. The MEAM1 populations from Israel and Italy and the ones from Iran and Turkmenistan are similar as they both show the earlier expansion at ≈250,000-300,000 YA. The single IO population lacked the latter expansion but had the earlier one. This pattern is shared with the Sub-Saharan African (Burkina Faso) MED, suggesting IO also faced a similar history of environmental change, which seems plausible given their relatively close geographical distributions. In conclusion, populations within the invasive species MED and MEAM1 exhibited signatures of population expansion lacking in non-invasive species (IO and AUS) during the Pleistocene, a geological epoch marked by repeated climatic oscillations with cycles of glacial and interglacial periods. These expansions strongly suggested the potential of some Bemisia species’ genomes to affect their adaptability and invasiveness.

Keywords: whitefly, RADseq, invasive species, SNP, climate change

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Authors: Ugo Rovigatti

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Neuroblastoma (NBL) has epitomized for at least 40 years our understanding of cancer cellular and molecular biology and its potential applications to novel therapeutic strategies. This includes the discovery of the very first oncogene aberrations and tumorigenesis suppression by differentiation in the 80s; the potential role of suppressor genes in the 90s; the relevance of immunotherapy in the millennium first, and the discovery of additional mutations by NGS technology in the millennium second decade. Similar discoveries were achieved in the majority of human cancers, and similar therapeutic interventions were obtained subsequently to NBL discoveries. Unfortunately, targeted therapies suggested by specific mutations (such as MYCN amplification –MNA- present in ¼ or 1/5 of cases) have not elicited therapeutic successes in aggressive NBL, where the prognosis is still dismal. The reasons appear to be linked to Tumor Heterogeneity, which is particularly evident in NBL but also a clear hallmark of aggressive human cancers generally. The new avenue of cancer immunotherapy (CIT) provided new hopes for cancer patients, but we still ignore the cellular or molecular targets. CIT is emblematic of high-risk disease (HR-NBL) since the mentioned GD2 passive immunotherapy is still providing better survival. We recently critically reviewed and evaluated the literature depicting the genomic landscapes of HR-NBL, coming to the qualified conclusion that among hundreds of affected genes, potential targets, or chromosomal sites, none correlated with anti-GD2 sensitivity. A better explanation is provided by the Micro-Foci inducing Virus (MFV) model, which predicts that neuroblasts infection with the MFV, an RNA virus isolated from a cancer-cluster (space-time association) of HR-NBL cases, elicits the appearance of MNA and additional genomic aberrations with mechanisms resembling chromothripsis. Neuroblasts infected with low titers of MFV amplified MYCN up to 100 folds and became highly transformed and malignant, thus causing neuroblastoma in young rat pups of strains SD and Fisher-344 and larger tumor masses in nu/nu mice. An association was discovered with GD2 since this glycosphingolipid is also the receptor for the family of MFV virus (dsRNA viruses). It is concluded that a dsRNA virus, MFV, appears to provide better explicatory mechanisms for the genesis of i) specific genomic aberrations such as MNA; ii) extensive tumor heterogeneity and chromothripsis; iii) the effects of passive immunotherapy with anti-GD2 monoclonals and that this and similar models should be further investigated in both pediatric and adult cancers.

Keywords: neuroblastoma, MYCN, amplification, viruses, GD2

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Authors: Prince Vivek, Vijay K. Bharti, Manishi Mukesh, Ankita Sharma, Om Prakash Chaurasia, Bhuvnesh Kumar

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High performance of endurance in equid requires adaptive changes involving physio-biochemical, and molecular responses in an attempt to regain homeostasis. We hypothesized that the identification of the suitable reference genes might be considered for assessing of endurance related traits in pony at high altitude and may ensure for individuals struggling to potent endurance trait in ponies at high altitude. A total of 12 mares of ponies, Zanskar breed, were divided into three groups, group-A (without load), group-B, (60 Kg) and group-C (80 Kg) on backpack loads were subjected to a load carry protocol, on a steep climb of 4 km uphill, and of gravel, uneven rocky surface track at an altitude of 3292 m to 3500 m (endpoint). Blood was collected before and immediately after the load carry on sodium heparin anticoagulant, and the peripheral blood mononuclear cell was separated for total RNA isolation and thereafter cDNA synthesis. Real time-PCR reactions were carried out to evaluate the mRNAs expression profile of a panel of putative internal control genes (ICGs), related to different functional classes, namely glyceraldehyde 3-phosphate dehydrogenase (GAPDH), β₂ microglobulin (β₂M), β-actin (ACTB), ribosomal protein 18 (RS18), hypoxanthine-guanine phosophoribosyltransferase (HPRT), ubiquitin B (UBB), ribosomal protein L32 (RPL32), transferrin receptor protein (TFRC), succinate dehydrogenase complex subunit A (SDHA) for normalizing the real-time quantitative polymerase chain reaction (qPCR) data of native pony’s. Three different algorithms, geNorm, NormFinder, and BestKeeper software, were used to evaluate the stability of reference genes. The result showed that GAPDH was best stable gene and stability value for the best combination of two genes was observed TFRC and β₂M. In conclusion, the geometric mean of GAPDH, TFRC and β₂M might be used for accurate normalization of transcriptional data for assessing endurance related traits in Zanskar ponies during load carrying.

Keywords: endurance exercise, ubiquitin B (UBB), β₂ microglobulin (β₂M), high altitude, Zanskar ponies, reference gene

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Authors: Lea Boidin, Martha Baydoun, Bertrand Leroux, Olivier Morales, Samir Acherar, Celine Frochot, Nadira Delhem

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Ovarian cancer (OC) is one of the most defying diseases in gynecologic oncology. Even though surgery remains crucial in the therapy of patients with primary ovarian cancer, recurrent recidivism calls for the development of new therapy protocols to propose for patients dealing with this cancer. FRα is described as a tumor‐associated antigen in OC, where FRα expression is usually linked with more poorly differentiated, aggressive tumors. The Photodynamic treatment (PDT) available data have shown improvements in the uptake of small tumors and in the induction of a proper anti-tumoral immune response. In order to target specifically peritoneal metastatis, which overexpress FRα, a new-patented PS coupled with folic acid has been developed in our team. Herein we propose PDT using this new patented PS for PDT applied in an in vivo mice model. The efficacy of the treatment was evaluated in mice without and with PBMC reconstitution. Mice were divided into four groups: Non-Treated, PS, Light Only, and PDT Treated and subjected to illumination by laser set at 668nm with a duration of illumination of 45 minutes (or 1 min of illumination followed by 2 minutes of pause repeated 45 times). When mice were not reconstituted and after fractionized PDT protocol, a significant decrease in the tumor volume was noticed. An induction in the anti-tumoral cytokine IFNγ chaperoned this decrease while a subsequent inhibition in the cytokine TGFβ. Even more crucial, when mice were reconstituted and upon PDT, the fold of tumor decrease was even higher. An immune response was activated decoded with an increase in NK, CD3 +, LT helper and Cytotoxic T cells. Thereafter, an increase in the expression of the cytokines IFNγ and TNFα were noticed while an inhibition in TGFβ, IL8 and IL10 accompanied this immune response activation. Therefore, our work has shown for the first time that a fractionized PDT protocol using a folate-targeted PDT is effective for treatment of ovarian cancer. The interest in using PDT in this case, goes beyond the local induction of tumor apoptosis only, but can promote subsequent anti-tumor response. Most of the therapies currently used to treat ovarian cancer, have an uncooperative outcomes on the host immune response. The readiness of a tumor adjuvant treatment like PDT adequate in eliminating the tumor and in concert stimulating anti-tumor immunity would be weighty.

Keywords: folate receptor, ovarian cancer, photodynamic therapy, humanized mice model

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Authors: Manali Jain, Neeta Singh, Raunaq Fatima, Soniya Nityanand, Mandakini Pradhan, Chandra Prakash Chaturvedi

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Isolated Congenital Heart Defect (ICHD) is the major cause of neonatal death worldwide among all forms of CHDs. A significant proportion of fetuses with ICHD die in the neonatal period if no treatment is provided. Recently, stem cell therapies have emerged as a potential approach to ameliorate ICHD in children. ICHD is characterized by cardiac structural abnormalities during embryogenesis due to alterations in the cardiomyogenic properties of a pool of cardiac progenitors/ stem cells associated with fetal heart development. The stem cells present in the amniotic fluid (AF) are of fetal origin and may reflect the physiological and pathological changes in the fetus during embryogenesis. Therefore, in the present study, the cardiomyogenic potential of AF-MSCs derived from fetuses with ICHD (ICHD AF-MSCs) has been evaluated and compared with that of AF-MSCs of structurally normal fetuses (normal AF-MSCs). Normal and ICHD AF-MSC were analyzed for the expression of cardiac progenitor markers viz., stage-specific embryonic antigen-1 (SSEA-1), vascular endothelial growth factor 2 (VEGFR-2) and platelet-derived growth factor receptor-alpha (PDGFR-α) by flow cytometry. The immunophenotypic characterization revealed that ICHD AF-MSCs have significantly lower expression of cardiac progenitor markers VEGFR-2 (0.14% ± 0.6 vs.48.80% ± 0.9; p <0.01), SSEA-1 (70.86% ± 2.4 vs. 88.36% ±2.7; p <0.01), and PDGFR-α (3.92% ± 1.8 vs. 47.59% ± 3.09; p <0.01) in comparison to normal AF-MSCs. Upon induction with 5’-azacytidine for 21 days, ICHD AF-MSCs showed a significantly down-regulated expression of cardiac transcription factors such as GATA-4 (0.4 ± 0.1 vs. 6.8 ± 1.2; p<0.01), ISL-1 (2.3± 0.6 vs. 14.3 ± 1.12; p<0.01), NK-x 2-5 (1.1 ± 0.3 vs. 14.1 ±2.8; p<0.01), TBX-5 (0.4 ± 0.07 vs. 4.4 ± 0.3; p<0.001), and TBX-18 (1.3 ± 0.2 vs. 4.19 ± 0.3; p<0.01) when compared with the normal AF-MSCs. Furthermore, immunocytochemical staining revealed that both types of AF-MSCs could differentiate into cardiovascular lineages and express cardiomyogenic, endothelial, and smooth muscle actin markers, viz., cardiac troponin (cTNT), CD31, and alpha-smooth muscle actin (α-SMA). However, normal AF-MSCs showed an enhanced expression of cTNT (p<0.001), CD31 (p<0.01), and α-SMA (p<0.05), compared to ICHD AF-MSCs. Overall, these results suggest that the ICHD-AF-MSCs have a defective cardiomyogenic differentiation potential and that the defects in these stem cells may have a role in the pathogenesis of ICHD.

Keywords: amniotic fluid, cardiomyogenic potential, isolated congenital heart defect, mesenchymal stem cells

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Authors: Vanessa Morgado, Ricardo Bettencourt da Silva, Carla Palma

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The environmental pollution by microplastics is well recognized. Microplastics were already detected in various matrices from distinct environmental compartments worldwide, some from remote areas. Various methodologies and techniques have been used to determine microplastic in such matrices, for instance, sediment samples from the ocean bottom. In order to determine microplastics in a sediment matrix, the sample is typically sieved through a 5 mm mesh, digested to remove the organic matter, and density separated to isolate microplastics from the denser part of the sediment. The physical analysis of microplastic consists of visual analysis under a stereomicroscope to determine particle size, colour, and shape. The chemical analysis is performed by an infrared spectrometer coupled to a microscope (micro-FTIR), allowing to the identification of the chemical composition of microplastic, i.e., the type of polymer. Creating legislation and policies to control and manage (micro)plastic pollution is essential to protect the environment, namely the coastal areas. The regulation is defined from the known relevance and trends of the pollution type. This work discusses the assessment of contamination trends of a 700 km² oceanic area affected by contamination heterogeneity, sampling representativeness, and the uncertainty of the analysis of collected samples. The methodology developed consists of objectively identifying meaningful variations of microplastic contamination by the Monte Carlo simulation of all uncertainty sources. This work allowed us to unequivocally conclude that the contamination level of the studied area did not vary significantly between two consecutive years (2018 and 2019) and that PET microplastics are the major type of polymer. The comparison of contamination levels was performed for a 99% confidence level. The developed know-how is crucial for the objective and binding determination of microplastic contamination in relevant environmental compartments.

Keywords: measurement uncertainty, micro-ATR-FTIR, microplastics, ocean contamination, sampling uncertainty

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Authors: Pooja Kumari, Anandkumar Tengli

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Aurora kinase enzyme, a protein on overexpression, leads to metastasis and is extremely important for women’s health in terms of prevention or treatment. While creating a targeted technique, the aim of the work is to design purine molecules that inhibit in aurora kinase enzyme and helps to suppress breast cancer. Purine molecules attached to an amino acid in DNA block protein synthesis or halt the replication and metastasis caused by the aurora kinase enzyme. Various protein related to the overexpression of aurora protein was docked with purine molecule using Biovia Drug Discovery, the perpetual software. Various parameters like X-ray crystallographic structure, presence of ligand, Ramachandran plot, resolution, etc., were taken into consideration for selecting the target protein. A higher negative binding scored molecule has been taken for simulation studies. According to the available research and computational analyses, purine compounds may be powerful enough to demonstrate a greater affinity for the aurora target. Despite being clinically effective now, purines were originally meant to fight breast cancer by inhibiting the aurora kinase enzyme. In in-silico studies, it is observed that purine compounds have a moderate to high potency compared to other molecules, and our research into the literature revealed that purine molecules have a lower risk of side effects. The research involves the design, synthesis, and identification of active purine molecules against breast cancer. Purines are structurally similar to the normal metabolites of adenine and guanine; hence interfere/compete with protein synthesis and suppress the abnormal proliferation of cells/tissues. As a result, purine target metastasis cells and stop the growth of kinase; purine derivatives bind with DNA and aurora protein which may stop the growth of protein or inhibits replication and stop metastasis of overexpressed aurora kinase enzyme.

Keywords: aurora kinases, in silico studies, medicinal chemistry, combination therapies, chronic cancer, clinical translation

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Authors: Fernanda Bravo Cornejo, Camilo Cerda Sarabia, Belén Díaz Díaz, Diego Santibañez Oyarce, Esteban Gómez Terán, Hugo Osses Prado, Raúl Caulier-Cisterna, Jorge Vergara-Quezada, Ana Moya-Beltrán

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Streptococcus pyogenes is a gram-positive bacteria involved in a wide range of diseases and is a major-human-specific bacterial pathogen. In Chile, this year the 'Ministerio de Salud' declared an alert due to the increase in strains throughout the year. This increase can be attributed to the multitude of factors including antimicrobial resistance (AMR) and Virulence Factors (VF). Understanding these VF and AMR is crucial for developing effective strategies and improving public health responses. Moreover, experimental identification and characterization of these pathogenic mechanisms are labor-intensive and time-consuming. Therefore, new computational methods are required to provide robust techniques for accelerating this identification. Advances in Machine Learning (ML) algorithms represent the opportunity to refine and accelerate the discovery of VF associated with Streptococcus pyogenes. In this work, we evaluate the accuracy of various machine learning models in predicting the virulence factors and antimicrobial resistance of Streptococcus pyogenes, with the objective of providing new methods for identifying the pathogenic mechanisms of this organism.Our comprehensive approach involved the download of 32,798 genbank files of S. pyogenes from NCBI dataset, coupled with the incorporation of data from Virulence Factor Database (VFDB) and Antibiotic Resistance Database (CARD) which contains sequences of AMR gene sequence and resistance profiles. These datasets provided labeled examples of both virulent and non-virulent genes, enabling a robust foundation for feature extraction and model training. We employed preprocessing, characterization and feature extraction techniques on primary nucleotide/amino acid sequences and selected the optimal more for model training. The feature set was constructed using sequence-based descriptors (e.g., k-mers and One-hot encoding), and functional annotations based on database prediction. The ML models compared are logistic regression, decision trees, support vector machines, neural networks among others. The results of this work show some differences in accuracy between the algorithms, these differences allow us to identify different aspects that represent unique opportunities for a more precise and efficient characterization and identification of VF and AMR. This comparative analysis underscores the value of integrating machine learning techniques in predicting S. pyogenes virulence and AMR, offering potential pathways for more effective diagnostic and therapeutic strategies. Future work will focus on incorporating additional omics data, such as transcriptomics, and exploring advanced deep learning models to further enhance predictive capabilities.

Keywords: antibiotic resistance, streptococcus pyogenes, virulence factors., machine learning

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Authors: Adel Dalilottojari, Bahman Delalat, Frances J. Harding, Michaelia P. Cockshell, Claudine S. Bonder, Nicolas H. Voelcker

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Endothelial Progenitor Cell (EPC) based therapies continue to be of interest to treat ischemic events based on their proven role to promote blood vessel formation and thus tissue re-vascularisation. Current strategies for the production of clinical-grade EPCs requires the in vitro isolation of EPCs from peripheral blood followed by cell expansion to provide sufficient quantities EPCs for cell therapy. This study aims to examine the use of different biomolecules to significantly improve the current strategy of EPC capture and expansion on collagen type I (Col I). In this study, four different biomolecules were immobilised on a surface and then investigated for their capacity to support EPC capture and proliferation. First, a cell microarray platform was fabricated by coating a glass surface with epoxy functional allyl glycidyl ether plasma polymer (AGEpp) to mediate biomolecule binding. The four candidate biomolecules tested were Col I, collagen type II (Col II), collagen type IV (Col IV) and vascular endothelial growth factor A (VEGF-A), which were arrayed on the epoxy-functionalised surface using a non-contact printer. The surrounding area between the printed biomolecules was passivated with polyethylene glycol-bisamine (A-PEG) to prevent non-specific cell attachment. EPCs were seeded onto the microarray platform and cell numbers quantified after 1 h (to determine capture) and 72 h (to determine proliferation). All of the extracellular matrix (ECM) biomolecules printed demonstrated an ability to capture EPCs within 1 h of cell seeding with Col II exhibiting the highest level of attachment when compared to the other biomolecules. Interestingly, Col IV exhibited the highest increase in EPC expansion after 72 h when compared to Col I, Col II and VEGF-A. These results provide information for significant improvement in the capture and expansion of human EPC for further application.

Keywords: biomolecules, cell microarray platform, cell therapy, endothelial progenitor cells, high throughput screening

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Authors: Nicholas C. Rose, Christopher D. Spicer

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The regeneration of damaged or diseased tissues would provide an invaluable therapeutic tool in biological research and medicine. Cells must be provided with a number of different biochemical signals in order to form mature tissue through complex signaling networks that are difficult to recreate in synthetic materials. The ability to attach and detach bioactive proteins from material in an iterative and dynamic manner would therefore present a powerful way to mimic natural biochemical signaling cascades for tissue growth. We propose to reversibly attach these bioactive proteins using ortho-boronoimine (oBI) linkages and related derivatives formed by the reaction of an ortho-boronobenzaldehyde with a nucleophilic amine derivative. To enable the use of oBIs for biomaterial modification, we have studied binding and cleavage processes with precise detail in the context of small molecule models. A panel of oBI complexes has been synthesized and screened using a novel Förster resonance energy transfer (FRET) assay, using a cyanine dye FRET pair (Cy3 and Cy5), to identify the most reactive boron-aldehyde/amine nucleophile pairs. Upon conjugation of the dyes, FRET occurs under Cy3 excitation and the resultant ratio of Cy3:Cy5 emission directly correlates to conversion. Reaction kinetics and equilibria can be accurately quantified for reactive pairs, with dissociation constants of oBI derivatives in water (KD) found to span 9-orders of magnitude (10⁻²-10⁻¹¹ M). These studies have provided us with a better understanding of oBI linkages that we hope to exploit to reversibly attach bioconjugates to materials. The long-term aim of the project is to develop a modular biomaterial platform that can be used to help combat chronic diseases such as osteoarthritis, heart disease, and chronic wounds by providing cells with potent biological stimuli for tissue engineering.

Keywords: dynamic, bioconjugation, bornoimine, rapid, physiological

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Authors: Kevin A. Droguett, Edwin G. Pérez, Denis Fuentealba, Margarita E. Aliaga, Angélica M. Fierro

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Supramolecular chemistry is a field of growing interest. Moreover, studying the formation of host-guest complexes between macrocycles and dyes is highly attractive due to their potential applications. Examples of the above are drug delivery, catalytic process, and sensing, among others. There are different dyes of interest in the literature; one example is the quinolinone derivatives. Those molecules have good optical properties and chemical and thermal stability, making them suitable for developing fluorescent probes. Secondly, several macrocycles can be seen in the literature. One example is the cucurbiturils. This water-soluble macromolecule family has a hydrophobic cavity and two identical carbonyl portals. Additionally, the thermodynamic analysis of those supramolecular systems could help understand the affinity between the host and guest, their interaction, and the main stabilization energy of the complex. In this work, two 7-(diethylamino) quinoline-2 (1H)-one derivative (QD1-2) and their interaction with cucurbit[7]uril (CB[7]) were studied from an experimental and in-silico point of view. For the experimental section, the complexes showed a 1:1 stoichiometry by HRMS-ESI and isothermal titration calorimetry (ITC). The inclusion of the derivatives on the macrocycle lends to an upward shift in the fluorescence intensity, and the pKa value of QD1-2 exhibits almost no variation after the formation of the complex. The thermodynamics of the inclusion complexes was investigated using ITC; the results demonstrate a non-classical hydrophobic effect with a minimum contribution from the entropy term and a constant binding on the order of 106 for both ligands. Additionally, dynamic molecular studies were carried out during 300 ns in an explicit solvent at NTP conditions. Our finding shows that the complex remains stable during the simulation (RMSD ~1 Å), and hydrogen bonds contribute to the stabilization of the systems. Finally, thermodynamic parameters from MMPBSA calculations were obtained to generate new computational insights to compare with experimental results.

Keywords: host-guest complexes, molecular dynamics, quinolin-2(1H)-one derivatives dyes, thermodynamics

Procedia PDF Downloads 92