Search results for: biochemical pathway

Authors: Lorenzo Maiorino, Fabio Fortuna, Giovanni Panebianco, Marco Sanzari, Gabriella Arcese, Valerio Maria Paolozzi

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It is well known that sport is a factor of social cohesion and the breaking down of barriers between people. From this point of view, the aim is to demonstrate how, through the (re)generation of sustainable structures, it is possible to give life to a new social, cultural, and economic pathway, where possible, in peripheral areas with problems of abandonment and degradation. The aim of this paper is, therefore, to study realities such as European programs and funds and to highlight the ways in which planning can be used to respond to critical issues such as urban decay, abandonment, and the mitigation of social differences. For this reason, the analysis will be carried out through the Multiannual Financial Framework (MFF) package, the Next Generation EU, the Recovery and Resilience Facility (RRF), the Cohesion Fund, the European Social Fund, and other managed funds. The procedure will rely on sources and data of unquestionable origin, and the relation to the object of study in question will be highlighted. The project lends itself to be ambitious and exploring a further aspect of the sports theme, which, as we know, is one of the foundations for a healthy society.

Keywords: sport, social inclusion, urban regeneration, sports facilities, European funds

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Authors: Khairi El Battawy, Monika Skalicki

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The present investigation has been done on pure Egyptian Arabian mares that reared in private horse studs. Fifty non-pregnant mares were selected and examined to classify them as either being reproductively healthy or subfertile mares including clinical endometritis, early embryonic death, granulosa cell tumor, repeat breeder (post-breeding endometritis), and anoestrus mares. The purpose of the study was to assess oxidative/antioxidant biochemical metabolites, lipogram, trace elements and reproductive hormones throughout reproductive conditions in mares during regular estrous, anestrum, early pregnancy, granulose cell tumor, ovulation failure, and endometritis. Results showed intensification of the free radical-dependent process in the blood of infertile mare, especially mares with endometritis. Ultrasonography as a diagnostic tool diagnosis of endometritis in mares was an important step as it revealed much information concerning infertility problem.

Keywords: endometritis, ovulation, oxidative, mare

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Authors: Ihtisham Bukhari

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In the current study, we enrolled a 46, XY phenotypically female patient bearing testes in her inguinal canal. DNA sequencing of the AR gene detected a missense mutation C.1715A > G (p. Y572C) in exon 2 which is already known to cause Complete androgen insensitivity syndrome (CAIS). We further studied the effects of this mutation on the testicular histopathology of the patient. No spermatocytes were seen in the surface spreading of testicular tissues while H&E staining showed that seminiferous tubules predominantly have only Sertoli cells. To confirm this meiotic failure is likely due to the current AR mutation we performed mRNA expression of genes associated with AR pathway, expression and location of the associated proteins in testicular tissues. Western blot and real-time PCR data showed that the patient had high levels of expression of AMH, SOX9, and INNB in testis. Tubules were stained with SOX9 and AMH which revealed Sertoli cell maturation arrest. Therefore, we suggest that AR mutation enhances AMH expression which ultimately leads to failure in the maturation of Sertoli cells and failure in spermatogenesis.

Keywords: androgen receptor, spermatogenesis, infertility, Sertoli cell only syndrome

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Authors: Luke Andrighetto, Paul G. Stevenson, Luke C. Henderson, Jim Pearson, Xavier A. Conlan

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As pseudoephedrine, a common ingredient in cold and flu medications is closely monitored and restricted in Australia, alternative methods of accessing it are of interest. The impurities and by-products of every reaction step of pseudoephedrine/ephedrine and methamphetamine synthesis have been mapped in order to develop a chemical fingerprint based on synthetic route. Likewise, seized methamphetamine contains a combination of different cutting agents and starting materials. Therefore, in-silico optimised two-dimensional HPLC with DryLab® and OpenMS® software has been used to efficiently separate complex seizure samples. An excellent match between simulated and real separations was observed. Targeted separation of model compounds was completed with significantly reduced method development time. This study produced a two-dimensional separation regime that offers unprecedented separation power (separation space) while maintaining a rapid analysis time that is faster than those previously reported for gas chromatography, single dimension high performance liquid chromatography or capillary electrophoresis.

Keywords: chemical fingerprint, ephedrine, methamphetamine, two-dimensional HPLC

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Authors: Fazilet Özlem Çekiç, Seyda Yılmaz

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Salinity and drought are important environmental problems in the world and have negative effects on plant metabolism. Gamma-aminobutyric acid (GABA), four-carbon non-protein amino acid, is a significant component of the free amino acid pool. GABA is widely distributed in prokaryotic and eukaryotic organisms. Environmental stress factors increase GABA accumulation in plants. Our aim was to evaluate the effect of gibberellic acid (GA) on GABA metabolism system during drought and salt stress factors in Phaseolus vulgaris L. plants. GABA, Glutamate dehydrogenase (GDH) activity, chlorophyll, and lipid peroxidation (MDA) analyses were determined. According to our results we can suggest that GA play a role in GABA metabolism during salt and drought stresses in bean plants. Also GABA shunt is an important metabolic pathway and key signaling allowing to adapt to drought and salt stresses.

Keywords: gibberellic acid, GABA, Phaseolus vulgaris L., salinity, drought

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Authors: Labiba Zerari Bourafa, Djebar Mohamed Reda, Berrebah Houria, Khadri Sihem, Chiheb Linda

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The study of the effect of metal dust (pollutants) was performed on higher plant white beans Phaseolus vulgaris; the experience took place in cellular toxicology laboratory (in vitro culture). The seeds of the bean Phaseolus vulgaris are cultured in a metal contaminated dust medium (a single treatment by different increasing doses), at a rate of 10 seeds per box, for 10 days. The measurement of morpho-metric parameters is performed during the first 96 hours that follow the germination; while the dosage of the proline, the protein content and histological sections are formed on the tenth day (240 h). All morpho-metric and biochemical parameters measured were highly disturbed by metal dust; histological sections confirm this disurbance.

Keywords: conductive fabrics, metal dust, osmoticums, roots, Phaseolus vulgaris

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Authors: Beata Skibska, Andrzej Stanczak, Agnieszka Skibska

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Nanodiamond (ND) is a carbon nanomaterial that has high biocompatibility, and it has a very positive effect on a number of biochemical processes. NDs have great potential in treating multiple inflammation-associated diseases. The purpose of this study was to investigate the anti-inflammatory effect of nanodiamonds and lipoic acid (LA) (as antioxidants) on rats' cardiovascular systems after lipopolysaccharide (LPS) administration. Animal experiments enabled the determination of how nanodiamonds act when applied independently or in combination with lipoic acid. The effect of NDs and LA on C-reactive protein (CRP) levels and heart edema was evaluated. NDs and LA administered after LPS administration attenuated heart edema and significantly decreased the CRP level. The results suggest that NDs and LA play an important role in LPS-induced inflammation in the heart. NDs find new applications in modern biomedical science and biotechnologies.

Keywords: nanodiamonds, lipoic acid, inflammation, cardiovascular system

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Authors: Malik SS, Masood N, Mubarik S, Khadim TM

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Introduction: Xeroderma pigmentosum group G (XPG) gene plays a crucial role in the correction of UV-induced DNA damage through nucleotide excision repair pathway. Single nucleotide polymorphisms in XPG gene have been reported to be associated with different cancers. Current case-control study was designed to evaluate the relationship between one of the most frequently found XPG (rs1047768 T>C) polymorphism and breast cancer risk. Methodology: A total of 200 individuals were screened for this polymorphism including 100 pathologically confirmed breast cancer cases and age-matched 100 controls. Genotyping was carried out using Tetra amplification-refractory mutation system (ARMS) PCR and results were confirmed by gel electrophoresis. Results: Conditional logistic regression analysis showed significant association between TC genotype (OR: 8.9, CI: 2.0 – 38.7) and increased breast cancer risk. Although homozygous CC genotype was more frequent in patients as compared to controls, but it was statistically non-significant (OR: 3.9, CI: 0.4 – 35.7). Conclusion: In conclusion, XPG (rs1047768 T>C) polymorphism may contribute towards increased risk of breast cancer but other polymorphisms may also be evaluated to elucidate their role in breast cancer.

Keywords: XPG, breast cancer, NER, ARMS-PCR

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Authors: Anita Jäger, Andrew Salazar, Jörg von Hagen, Harald Kolmar

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In the treatment of individuals with sensitive and psoriatic skin, several inflammation and itch-related molecular and cellular targets have been identified, but many of these have yet to be characterized. In this study, we present two potential targets in the skin that can be linked to the inflammation and itch cycle. 11ßHSD1 is the enzyme responsible for converting inactive cortisone to active cortisol used to transmit signals downstream. The activation of the receptor NK1R correlates with promoting inflammation and the perception of itch and pain in the skin. In this study, both targets have been investigated based on their involvement in inflammation. The role of both identified targets was characterized based on the secretion of inflammation cytokine- IL6, IL-8, and CCL2, as well as phosphorylation and signaling pathways. It was found that treating skin cells with molecules able to inhibit inflammatory pathways results in the reduction of inflammatory signaling molecules secreted by skin cells and increases their proliferative capacity. Therefore, these molecular targets and their associated pathways show therapeutic potential and can be mitigated via small molecules. This research can be used for further studies in inflammation and itch pathways and can help to treat pathological symptoms.

Keywords: inflammation, itch, signaling pathway, skin

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Authors: Soheil Zorofchian Moghadamtousi, Habsah Abdul Kadir, Mohammadjavad Paydar, Elham Rouhollahi, Hamed Karimian

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The present study was designed to evaluate the molecular mechanisms of Annona muricata leaves ethyl acetate extract (AMEAE) against lung cancer A549 cells. Cell viability analysis revealed the selective cytotoxic effect of AMEAE towards A549 cells. Treatment of A549 cells with AMEAE significantly elevated the reactive oxygen species formation, followed by attenuation of mitochondrial membrane potential via upregulation of Bax and downregulation of Bcl-2, accompanied by cytochrome c release to the cytosol. The released cytochrome c triggered the activation of caspase-9 followed by caspase-3. In addition, AMEAE-induced apoptosis was accompanied by cell cycle arrest at G1 phase. Our data showed for the first time that AMEAE inhibited the proliferation of A549 cells, leading to cell cycle arrest and programmed cell death through activation of the mitochondrial-mediated signaling pathway.

Keywords: Annona muricata, lung cancer, apoptosis, mitochondria

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Authors: Esra Yuksel, Dilek Duman, Levent Yeniay, Sezgin Ulukaya

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Sentinel lymph node biopsy (SLNB) is a minimal invasive technique with lower morbidity in axillary staging of breast cancer. Isosulfan blue stain is frequently used in SLNB and regarded as safe. The present case report aimed to report severe decrement in SpO2 following isosulfan blue administration, as well as skin and urine signs and inconsistency with clinical picture in a 67-year-old ,77 kg, ASA II female case that underwent SLNB under general anesthesia. Ten minutes after subcutaneous administration of 10 ml 1% isosulfan blue by the surgeons into the patient, who were hemodynamically stable, SpO2 first reduced to 87% from 99%, and then to 75% in minutes despite 100% oxygen support. Meanwhile, blood pressure and EtCO2 monitoring was unremarkable. After specifying that anesthesia device worked normally, airway pressure did not increase and the endotracheal tube has been placed accurately, the blood sample was taken from the patient for arterial gas analysis. A severe increase was thought in MetHb concentration since SpO2 persisted to be 75% although the concentration of inspired oxygen was 100%, and solution of 2500 mg ascorbic acid in 500 ml 5% Dextrose was given to the patient via intravenous route until the results of arterial blood gas were obtained. However, arterial blood gas results were as follows: pH: 7.54, PaCO2: 23.3 mmHg, PaO2: 281 mmHg, SaO2: %99, and MetHb: %2.7. Biochemical analysis revealed a blood MetHb concentration of 2%.However, since arterial blood gas parameters were good, hemodynamics of the patient was stable and methemoglobin concentration was not so high, the patient was extubated after surgery when she was relaxed, cooperated and had adequate respiration. Despite the absence of respiratory or neurological distress, SpO2 value was increased only up to 85% within 2 hours with 5 L/min oxygen support via face mask in the surgery room as the patient was extubated. At that time, the skin of particularly the upper part of her body has turned into blue, more remarkable on the face. The color of plasma of the blood taken from the patient for biochemical analysis was blue. The color of urine coming throughout the urinary catheter placed in intensive care unit was also blue. Twelve hours after 5 L/min. oxygen inhalation via a mask, the SpO2 reached to 90%. During monitoring in intensive care unit on the postoperative 1st day, facial color and urine color of the patient was still blue, SpO2 was 92%, and arterial blood gas levels were as follows: pH: 7.44, PaO2: 76.1 mmHg, PaCO2: 38.2 mmHg, SaO2: 99%, and MetHb 1%. During monitoring in clinic on the postoperative 2nd day, SpO2 was 95% without oxygen support and her facial and urine color turned into normal. The patient was discharged on the 3rd day without any problem.In conclusion, SLNB is a less invasive alternative to axillary dissection. However, false pulse oximeter reading due to pigment interference is a rare complication of this procedure. Arterial blood gas analysis should be used to confirm any fall in SpO2 reading during monitoring.

Keywords: isosulfan blue, pulse oximetry, SLNB, methemoglobinemia

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Authors: Christy Rosaline, Rathankar Roa, Waheeta Hopper

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Persistence of tuberculosis, emergence of multidrug-resistance and extensively drug-resistant forms of the disease, has increased the interest in developing new antitubercular drugs. Developing inhibitors for 3-deoxy-D-arabino-heptulosonate 7-phosphate synthase from Mycobacterium tuberculosis (MtbDAH7Ps), an enzyme involved in shikimate pathway, gives a selective target for antitubercular agents. MtbDAH7Ps was screened against ZINC database, and shortlisted compounds were subjected to induce fit docking. Prime/Molecular Mechanics Generalized Born Surface Area calculation was used to validate the binding energy of ligand-protein complex. Molecular Dynamics analysis for of the lead compounds–MtbDAH7Ps complexes showed that the backbone of MtbDAH7Ps in their complexes were stable. These results suggest that the shortlisted lead compounds ZINC04097114, ZINC15163225, ZINC16857013, ZINC06275603, and ZINC05331260 could be developed into novel drug leads to inhibit DAH7Ps in Mycobacterium tuberculosis.

Keywords: MtbDAH7Ps, Mycobacterium tuberculosis, HTVS, molecular dynamics

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Authors: Aleksandra Bylinska, Samantha Slight-Webb, Kevin Thomas, Miles Smith, Susan Macwana, Nicolas Dominguez, Eliza Chakravarty, Joan T. Merrill, Judith A. James, Joel M. Guthridge

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Background: Systemic Lupus Erythematosus (SLE) is an interferon-related autoimmune disease characterized by B cell dysfunction. One of the main hallmarks is a loss of tolerance to self-antigens leading to increased levels of autoantibodies against nuclear components (ANAs). However, up to 20% of healthy ANA+ individuals will not develop clinical illness. SLE is more prevalent among women and minority populations (African, Asian American and Hispanics). Moreover, African Americans have a stronger interferon (IFN) signature and develop more severe symptoms. The exact mechanisms involved in ethnicity-dependent B cell dysregulation and the progression of autoimmune disease from ANA+ healthy individuals to clinical disease remains unclear. Methods: Peripheral blood mononuclear cells (PBMCs) from African (AA) and European American (EA) ANA- (n=12), ANA+ (n=12) and SLE (n=12) individuals were assessed by multimodal scRNA-Seq/CITE-Seq methods to examine differential gene signatures in specific B cell subsets. Library preparation was done with a 10X Genomics Chromium according to established protocols and sequenced on Illumina NextSeq. The data were further analyzed for distinct cluster identification and differential gene signatures in the Seurat package in R and pathways analysis was performed using Ingenuity Pathways Analysis (IPA). Results: Comparing all subjects, 14 distinct B cell clusters were identified using a community detection algorithm and visualized with Uniform Manifold Approximation Projection (UMAP). The proportion of each of those clusters varied by disease status and ethnicity. Transitional B cells trended higher in ANA+ healthy individuals, especially in AA. Ribonucleoprotein high population (HNRNPH1 elevated, heterogeneous nuclear ribonucleoprotein, RNP-Hi) of proliferating Naïve B cells were more prevalent in SLE patients, specifically in EA. Interferon-induced protein high population (IFIT-Hi) of Naive B cells are increased in EA ANA- individuals. The proportion of memory B cells and plasma cells clusters tend to be expanded in SLE patients. As anticipated, we observed a higher signature of cytokine-related pathways, especially interferon, in SLE individuals. Pathway analysis among AA individuals revealed an NRF2-mediated Oxidative Stress response signature in the transitional B cell cluster, not seen in EA individuals. TNFR1/2 and Sirtuin Signaling pathway genes were higher in AA IFIT-Hi Naive B cells, whereas they were not detected in EA individuals. Interferon signaling was observed in B cells in both ethnicities. Oxidative phosphorylation was found in age-related B cells (ABCs) for both ethnicities, whereas Death Receptor Signaling was found only in EA patients in these cells. Interferon-related transcription factors were elevated in ABCs and IFIT-Hi Naive B cells in SLE subjects of both ethnicities. Conclusions: ANA+ healthy individuals have altered gene expression pathways in B cells that might drive apoptosis and subsequent clinical autoimmune pathogenesis. Increases in certain regulatory pathways may delay progression to SLE. Further, AA individuals have more elevated activation pathways that may make them more susceptible to SLE.

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Authors: Jasminka Mujic, Karin Milde-Langosch, Volkmar Mueller, Mirza Suljagic, Tea Becirevic, Jozo Coric, Daria Ler

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MACC1 (metastasis associated in colon cancer-1) is a prognostic biomarker for tumor progression, metastasis, and survival of a variety of solid cancers. MACC1 also causes tumor growth in xenograft models and acts as a master regulator of the HGF/MET signaling pathway. In breast cancer, the expression of MACC1 determined by immunohistochemistry was significantly associated with positive lymph node status and advanced clinical stage. The aim of the present study was to further investigate the prognostic or predictive value of MACC1 expression in breast cancer using western blot analysis and immunohistochemistry. The results of our study have shown that high MACC1 expression in breast cancer is associated with shorter disease-free survival, especially in node-negative tumors. The MACC1 might be a suitable biomarker to select patients with a higher probability of recurrence which might benefit from adjuvant chemotherapy. Our results support a biologic role and potentially open the perspective for the use of MACC1 as predictive biomarker for treatment decision in breast cancer patients.

Keywords: breast cancer, biomarker, HGF/MET, MACC1

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Authors: Mudasir Ahmad Syed, Raashid Ahmd Wani, Mashooq Ahmad Dar, Uneeb Urwat, Riaz Ahmad Shah, Nazir Ahmad Ganai

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Salmonella enterica serovar Typhimurium (Salmonella Typhimurium) is a primary avian pathogen responsible for severe intestinal pathology in younger chickens and economic losses. However, the Salmonella Typhimurium is also able to cause infection in humans, described by typhoid fever and acute gastro-intestinal disease. A study was conducted at days to investigate pathological, histopathological, haemato-biochemical, immunological and expression kinetics of NRAMP (natural resistance associated macrophage protein) gene family (NRAMP1 and NRAMP2) in broiler chickens following experimental infection of Salmonella Typhimurium at 0,1,3,5,7,9,11,13 and 15 days respectively. Infection was developed in birds through oral route at 2×108 CFU/ml. Clinical symptoms appeared 4 days post infection (dpi) and after one-week birds showed progressive weakness, anorexia, diarrhea and lowering of head. On postmortem examination, liver showed congestion, hemorrhage and necrotic foci on surface, while as spleen, lungs and intestines revealed congestion and hemorrhages. Histopathological alterations were principally observed in liver in second week post infection. Changes in liver comprised of congestion, areas of necrosis, reticular endothelial hyperplasia in association with mononuclear cell and heterophilic infiltration. Hematological studies confirm a significant decrease (P<0.05) in RBC count, Hb concentration and PCV. White blood cell count showed significant increase throughout the experimental study. An increase in heterophils was found up to 7dpi and a decreased pattern was observed afterwards. Initial lymphopenia followed by lymphocytosis was found in infected chicks. Biochemical studies showed a significant increase in glucose, AST and ALT concentration and a significant decrease (P<0.05) in total protein and albumin level in the infected group. Immunological studies showed higher titers of IgG in infected group as compared to control group. The real time gene expression of NRAMPI and NRAMP2 genes increased significantly (P<0.05) in infected group as compared to controls. The peak expression of NRAMP1 gene was seen in liver, spleen and caecum of infected birds at 3dpi, 5dpi and 7dpi respectively, while as peak expression of NRAMP2 gene in liver, spleen and caecum of infected chicken was seen at 9dpi, 5dpi and 9dpi respectively. This study has role in diagnostics and prognostics in the poultry industry for the detection of salmonella infections at early stages of poultry development.

Keywords: biochemistry, histopathology, NRAMP, poultry, real time expression, Salmonella Typhimurium

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Authors: Tatiane Benvenuti, Fernando Hamerski, Alexandre Giacobbo, Andrea M. Bernardes, Marco A. S. Rodrigues

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Population growth in urban areas has caused damages to the environment, a consequence of the uncontrolled dumping of domestic and industrial wastewater. The capacity of some plants to purify domestic and agricultural wastewater has been demonstrated by several studies. Since natural wetlands have the ability to transform, retain and remove nutrients, constructed wetlands have been used for wastewater treatment. They are widely recognized as an economical, efficient and environmentally acceptable means of treating many different types of wastewater. T. domingensis Pers. species have shown a good performance and low deployment cost to extract, detoxify and sequester pollutants. Constructed Floating Wetlands (CFWs) consist of emergent vegetation established upon a buoyant structure, floating on surface waters. The upper parts of the vegetation grow and remain primarily above the water level, while the roots extend down in the water column, developing an extensive under water-level root system. Thus, the vegetation grows hydroponically, performing direct nutrient uptake from the water column. Biofilm is attached on the roots and rhizomes, and as physical and biochemical processes take place, the system functions as a natural filter. The aim of this study is to diagnose the application of macrophytes in artificial floating in the treatment of domestic sewage in south Brazil. The T. domingensis Pers. plants were placed in a flotation system (polymer structure), in full scale, in a sewage treatment plant. The sewage feed rate was 67.4 m³.d⁻¹ ± 8.0, and the hydraulic retention time was 11.5 d ± 1.3. This CFW treat the sewage generated by 600 inhabitants, which corresponds to 12% of the population served by this municipal treatment plant. During 12 months, samples were collected every two weeks, in order to evaluate parameters as chemical oxygen demand (COD), biochemical oxygen demand in 5 days (BOD5), total Kjeldahl nitrogen (TKN), total phosphorus, total solids, and metals. The average removal of organic matter was around 55% for both COD and BOD5. For nutrients, TKN was reduced in 45.9% what was similar to the total phosphorus removal, while for total solids the reduction was 33%. For metals, aluminum, copper, and cadmium, besides in low concentrations, presented the highest percentage reduction, 82.7, 74.4 and 68.8% respectively. Chromium, iron, and manganese removal achieved values around 40-55%. The use of T. domingensis Pers. in artificial floating for sewage treatment is an effective and innovative alternative in Brazilian sewage treatment systems. The evaluation of additional parameters in the treatment system may give useful information in order to improve the removal efficiency and increase the quality of the water bodies.

Keywords: constructed wetland, floating system, sewage treatment, Typha domingensis Pers.

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Authors: Mohammad Reza Hossein Zadeh

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Plants have different layers of defense responses against biotic and abiotic stresses. One of the well-defined defense mechanism in plants is systemic acquired resistance (SAR) against a broad-range of pathogens. Salicylic acid (SA) plays a crucial role in regulation of the SAR pathway. It has been proved that Chemically SA-like compounds can mimic the SA signaling role. Imidocloprid is an insecticide being used to control whiteflies on crop plants. In order to study the possible role of Imidocloprid as an elicitor of SAR in plants, experiments were conducted in a completely randomized design frame with three treatments and duplicates on the detached leaves and whole Nicotiana tabacum var. Samsun NN. plants inoculated with Tobacco mild green mosaic virus (TMGMV). Compared with the effect of other SAR-inducers such as SA, Imidoclorid conferred a robust SAR induction in the infected plants. The results suggested that Imidocloprid even more powerful than SA can be considered as strong SAR inducer in the infected plants with viruses, which develop the local lesion symptoms.

Keywords: imidocloprid, Nicotiana tabacum var. Samsun NN, SAR, tobacco mild green, mosaic virus

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Authors: Nouf A. Aldarmahi, Nesrin I. Tarbiah, Nuha A. Alkhattabi, Huda F. Alshaibi

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Nigella sativa which is known as dark cumin is a well-known example for a widely applicable herbal medicine. Nigella sativa can be effective in a variety of diseases such as hypertension, diabetes, bronchitis, gastrointestinal upset, and cancer. The anticancer effect of Nigella sativa appeared to be mediated by immune-modulatory effect through stimulating human natural killer (NK) cells. This is a type of lymphocytes which is part of the innate immunity, also known as the first line of defense in the body against pathogens. This study investigated the effect of thymoquinone as a major component of Nigella sativa on the molecular cytotoxic pathway of NK cell and the role of thymoquinone therapeutic effect on NK cells. NK cells were cultured with breast tumor cells in different ways and cultured media was collected and the concentration of perforin, granzyme B and interferon-α were measured by ELISA. The cytotoxic effect of NK cells on breast tumor cells was enhanced in the presence of thymoquinone, with increased activity of perforin in NK cells. This improved anticancer effect of thymoquinone on breast cancer cells.

Keywords: breast cancer, cancer cells, natural killer cells, thymoquinone

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Authors: Elisha Didam Markus, Thapelo Maqame

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This paper focuses on studying the effects of magnetism on water and their impact to plant growth. Magnetic fields are known to induce higher rate of biochemical reaction and therefore can be used for growth related reactions in plants. For the purpose of this study, two 2 litres bottles were taken, one with two opposite poles magnets (500 mT) one on top and one at the bottom of the bottle. Another bottle was not altered in any way (used as control). Each bottle contained tap water stored up for 24 hours. Plants planted into different pots were watered using water from these bottles. Four pots with soil and manure equally mixed were used and equal volume of radish berry seeds were planted. Two pots were watered with magnetised water and the other two with normal tap water. The developments of plants were monitored in terms of their lengths for a period of 21 days. After 21 days, the lengths of plants watered with magnetised water were found to be 5.6% longer than those watered with tap water.

Keywords: magnetised water, radish berry, growth percentage, magnetic fields

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Authors: Adenike Adesanya, Nonthaphat Wong, Xiang-Yun Lan, Shea Ping Yip, Chien-Ling Huang

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Background: The most challenging mutation of the oncokinase BCR-ABL protein T315I, which is commonly known as the “gatekeeper” mutation and is notorious for its strong resistance to almost all tyrosine kinase inhibitors (TKIs), especially imatinib. Therefore, this study aims to identify T315I-dependent downstream microRNA (miRNA) pathways associated with drug resistance in chronic myeloid leukemia (CML) for prognostic and therapeutic purposes. Methods: T315I-carrying K562 cell clones (K562-T315I) were generated by the CRISPR-Cas9 system. Imatinib-treated K562-T315I cells were subjected to small RNA library preparation and next-generation sequencing. Putative lncRNA-miRNA-mRNA networks were analyzed with (i) DESeq2 to extract differentially expressed miRNAs, using Padj value of 0.05 as cut-off, (ii) STarMir to obtain potential miRNA response element (MRE) binding sites of selected miRNAs on lncRNA H19, (iii) miRDB, miRTarbase, and TargetScan to predict mRNA targets of selected miRNAs, (iv) IntaRNA to obtain putative interactions between H19 and the predicted mRNAs, (v) Cytoscape to visualize putative networks, and (vi) several pathway analysis platforms – Enrichr, PANTHER and ShinyGO for pathway enrichment analysis. Moreover, mitochondria isolation and transcript quantification were adopted to determine the new mechanism involved in T315I-mediated resistance of CML treatment. Results: Verification of the CRISPR-mediated mutagenesis with digital droplet PCR detected the mutation abundance of ≥80%. Further validation showed the viability of ≥90% by cell viability assay, and intense phosphorylated CRKL protein band being detected with no observable change for BCR-ABL and c-ABL protein expressions by Western blot. As reported by several investigations into hematological malignancies, we determined a 7-fold increase of H19 expression in K562-T315I cells. After imatinib treatment, a 9-fold increment was observed. DESeq2 revealed 171 miRNAs were differentially expressed K562-T315I, 112 out of these miRNAs were identified to have MRE binding regions on H19, and 26 out of the 112 miRNAs were significantly downregulated. Adopting the seed-sequence analysis of these identified miRNAs, we obtained 167 mRNAs. 6 hub miRNAs (hsa-let-7b-5p, hsa-let-7e-5p, hsa-miR-125a-5p, hsa-miR-129-5p, and hsa-miR-372-3p) and 25 predicted genes were identified after constructing hub miRNA-target gene network. These targets demonstrated putative interactions with H19 lncRNA and were mostly enriched in pathways related to cell proliferation, senescence, gene silencing, and pluripotency of stem cells. Further experimental findings have also shown the up-regulation of mitochondrial transcript and lncRNA MALAT1 contributing to the lncRNA-miRNA-mRNA networks induced by BCR-ABL T315I mutation. Conclusions: Our results have indicated that lncRNA-miRNA regulators play a crucial role not only in leukemogenesis but also in drug resistance, considering the significant dysregulation and interactions in the K562-T315I cell model generated by CRISPR-Cas9. In silico analysis has further shown that lncRNAs H19 and MALAT1 bear several complementary miRNA sites. This implies that they could serve as a sponge, hence sequestering the activity of the target miRNAs.

Keywords: chronic myeloid leukemia, imatinib resistance, lncRNA-miRNA-mRNA, T315I mutation

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Authors: Kun-Ting Song, Christian Schott, Peter Schneider, Sebastian Watzele, Regina Kluge, Elena Gubanova, Aliaksandr S. Bandarenka

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The combination of electrochemical impedance spectroscopy (EIS) and the hydrodynamic technique like rotation disc electrode (RDE) provides a critical method for quantitively investigating mechanisms of hydrogen evolution reaction (HER) in acidic and alkaline media. Pt5Gd represented higher HER activities than polycrystalline Pt (Pt(pc)) by means of the surface strain effects. The model of the equivalent electric circuit to fit the impedance data under the RDE configurations is developed. To investigate the relative reaction contribution, the ratio of the charge transfer reactions of the Volmer-Heyrovsky and Volmer-Tafel pathways on Pt and Pt5Gd electrodes is determined. The ratio remains comparably similar in acidic media, but it changes in alkaline media with Volmer–Heyrovsky pathway dominating. This combined approach of EIS and RDE can help to study the electrolyte effects and other essential reactions for electrocatalysis in future work.

Keywords: hydrogen evolution reaction, electrochemical impedance spectroscopy, hydrodynamic methods, electrocatalysis, electrochemical interface

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Authors: Sami El Khatib, Agnès Leroux, Jean-Louis Merlin, François Guillemin, Marie-Ange D’Hallewin

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Photodynamic therapy (PDT) is a treatment modality based on the cytotoxic effect occurring on the target tissues by interaction of a photosensitizer with light in the presence of oxygen. One of the major advances in PDT can be attributed to the use of topical aminolevulinic (ALA) to induce Protoporphyrin IX (PpIX) for the treatment of early stage cancers as well as diagnosis. ALA is a precursor of the heme synthesis pathway. Locally delivered to the target tissue ALA overcomes the negative feedback exerted by heme and promotes the transient formation of PpIX in situ to reach critical effective levels in cells and tissue. Whereas early steps of the heme pathway occur in the cytosol, PpIX synthesis is shown to be held in the mitochondrial membranes and PpIX fluorescence is expected to accumulate in close vicinity of the initial building site and to progressively diffuse to the neighboring cytoplasmic compartment or other lipophylic organelles. PpIX is known to be highly reactive and will be degraded when irradiated with light. PpIX photobleaching is believed to be governed by a singlet oxygen mediated mechanism in the presence of oxidized amino acids and proteins. PpIX photobleaching and subsequent spectral phototransformation were described widely in tumor cells incubated in vitro with ALA solution, or ex vivo in human and porcine mucosa superfused with hexylaminolevulinate (hALA). PpIX photobleaching was also studied in vivo, using animal models such as normal or tumor mice skin and orthotopic rat bladder model. Hexyl aminolevulinate a more potent lipophilic derivative of ALA was proposed as an adjunct to standard cystoscopy in the fluorescence diagnosis of bladder cancer and other malignancies. We have previously reported the effectiveness of hALA mediated PDT of rat bladder cancer. Although normal and tumor bladder epithelium exhibit similar fluorescence intensities after intravesical instillation of two hALA concentrations (8 and 16 mM), the therapeutic response at 8mM and 20J/cm2 was completely different from the one observed at 16mM irradiated with the same light dose. Where the tumor is destroyed, leaving the underlying submucosa and muscle intact after an 8 mM instillation, 16mM sensitization and subsequent illumination results in the complete destruction of the underlying bladder wall but leaves the tumor undamaged. The object of the current study is to try to unravel the underlying mechanism for this apparent contradiction. PpIX extraction showed identical amounts of photosensitizer in tumor bearing bladders at both concentrations. Photobleaching experiments revealed mono-exponential decay curves in both situations but with a two times faster decay constant in case of 16mM bladders. Fluorescence microscopy shows an identical fluorescence pattern for normal bladders at both concentrations and tumor bladders at 8mM with bright spots. Tumor bladders at 16 mM exhibit a more diffuse cytoplasmic fluorescence distribution. The different response to PDT with regard to the initial pro-drug concentration can thus be attributed to the different cellular localization.

Keywords: bladder cancer, hexyl-aminolevulinate, photobleaching, confocal fluorescence microscopy

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Authors: Zhipeng Chen, Lingfeng Wang, Shuang Qiu, Shijian Ge

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Macroalgae are larger in size compared with microalgae; hence, they imposed lower separation and drying costs. To explore the potential for enhancing cultivation conditions in macroalgae Chaetomorpha linum (C. linum)-based bioreactor for nutrient recovery from municipal wastewaters and examine the biochemical composition of the macroalgae for the potential downstream production of biofuels, screening experiments were performed. This study suggested that C. linum grew well on primary (PW), secondary (SW), and centrate wastewater (CW). A step feeding approach was shown to significantly enhance biomass productivity when grown on 10% CW; meanwhile, nitrogen and phosphorus removal efficiencies increased to 86.8 ± 1.1% and 92.6 ± 0.2%, respectively. The CO₂-supplemented SW cultures were 1.20 times more productive than the corresponding controls without CO₂ supplementation. These findings demonstrate that C. linum could represent a promising and efficient wastewater treatment alternative which could also provide a feedstock for downstream processing to biofuels.

Keywords: biofuel production, macroalgae, nutrient removal, wastewater

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Authors: Michael Miller, G. David Gearhart

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The role of the American college presidency continues to evolve, increasingly incorporating varied elements of responsibility of institutional management. Once primarily focused on the academic operation of an institution, the role has changed to incorporate all of the business-related and public agency operations of an institution. This means that the modern college president must be capable of persuasively talking to legislators and donors, as well as students taking classes and the faculty who teach them. In addition to these dimensions, the contemporary college president must also be an expert on state and federal compliance issues, and must have the talent to steer marketing and public relations activities in a persuasive manner. This paper will report the findings of a spring 2020 survey of nearly 300 college presidents in the United States concerning their perceptions about the changing nature of the college presidency, with special consideration given to media relations, fundraising, business development, internationalization, and facility management.Survey results will be analyzed based on institutional type, gender, length of tenure in the presidential position, and career pathway to the presidency.

Keywords: college president, leadership, education management, neo-liberal higher education

Procedia PDF Downloads 119

Authors: Wael El-Deeb

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The present study aimed to investigate the diagnostic importance of acute phase proteins in urinary tract infection (UTI) in cattle. We describe the clinical, bacteriological and biochemical findings in 99 lactating cows. Blood and urine samples from diseased (n=84) and control healthy cows (n=15) were submitted to laboratory investigations. The urine analysis revealed hematuria and pyuria in UTI group. The isolated bacteria were E.coli (43/84) Corynebacterium spp, (31/84), Proteus spp. (6/84) and Streptococcus spp (4/84). The concentrations of Haptoglobin (Hp), serum amyloid A (SAA), α1-Acid glycoprotein (AGP), fibrinogen (Fb), total protein, albumen, and globulin were higher in cows with UTI when compared to healthy ones. Fifty-one of 84 cows with UTI were successfully treated. The levels of Hp, SAA, AGP, total protein, and globulin were associated with the odds of treatment failure. Conclusively, acute phase proteins could be used as diagnostic and prognostic biomarkers in cows with UTI.

Keywords: cows, urinary, infections, haptoglobin, serum Amyloid A

Procedia PDF Downloads 724

Authors: Anwar Ali Abdulla, Thekra Abdulaali Abed Al-Chaabawi, Anwar Kadhim Al-Saffar, Hussein Kadhim Al-Saffar

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Lactic acid bacteria are very significant to human health due to the production of some antimicrobial substances and ability to inhibit pathogenic bacteria. Furthermore, the bacteria are also used as starter culture in the production of various foods. The present study was focused on isolation and characterization of Lactobacillus acidophilus from yogurt and to demonstrate some of probiotic properties of these isolates. All isolates were phenotypically characterized including studying, biochemical, effect of sodium chloride and pH during growth, carbohydrates test and characterizing the antimicrobial activity of Lactobacillus acidophilus against pathogens. The present study demonstrates that Lactobacillus acidophilus produced a bacteriocin- like inhibitory substance with a broad spectrum of antimicrobial activity directed against pathogenic indicator organism suggesting its protective value against enteric pathogens.

Keywords: lactobacillus acidophilus, bacteriocin, antimicrobial activity, probiotic

Procedia PDF Downloads 539

Authors: Li Yuchen, Wu Qingxin, Jin Yuxing, Yang Qian

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Interleukin-11 (IL-11), a well-known anti-inflammatory factor, helps to protect against intestinal epithelium damage caused by physical or chemical factors. However, little is known about the role of IL-11 during viral infection. Herein, high mRNA and protein levels of IL-11 were found in epithelial cells and jejunum of piglets during porcine epidemic diarrhea virus (PEDV) infection, and IL-11 expression was positively correlated with the level of viral infection. Pretreatment with recombinant porcine IL-11 (pIL-11) suppressed PEDV replication in Vero E6 cells, while IL-11 knockdown promoted viral infection. Furthermore, pIL-11 inhibited viral infection by preventing PEDV-mediated apoptosis of cells through activating the IL-11/STAT3 signal pathway. Conversely, application of a STAT3 phosphorylation inhibitor significantly antagonized the anti-apoptosis function of pIL-11 and counteracted its inhibition of PEDV. Our data suggested that that IL-11 is a novel PEDV-inducible cytokine, and its production enhances the anti-apoptosis ability of epithelial cells against PEDV infection. The potential uses of IL-11 as a novel therapeutic against devastating viral diarrhea in piglets deserves more attention and study.

Keywords: Interleukin-11, Porcine epidemic diarrhea virus, STAT3, anti-apoptosis

Procedia PDF Downloads 137

Authors: Hafiz Usama Imad, Muhammad Akram Akhund, Tauha Hussain Ali, Ali Raza Khoso, Fida Hussain Siddiqui

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Construction industry plays a significant role in fulfilling the major requirements of the human being. It is one of the major constituents of every developed country. Although the construction industry of both the developing and developed countries encompasses a major part of the economy, and millions of rupees are utilized every year on various kinds of construction projects. But, this industry is facing numerous hurdles in terms of its budget and timely completion. Construction projects generally consist of several phases like planning, designing, execution, and finishing. This research study aims to determine the significant factors of time overrun in pre-construction planning (PCP) phase of construction projects in Pakistan. Questionnaires were distributed by various means and responses of respondents were compiled and collected data were then analyzed through a statistical technique using SPSS version 24. Major causes of time overrun in pre-construction planning phase; which is an extremely important phase of construction projects, were revealed. The research conclusion will provide a pathway for stakeholders to pay attention to the mentioned causes to overcome the major issue of time overrun.

Keywords: construction industry, Pakistan, pre-construction planning phase, time overrun

Procedia PDF Downloads 255

Authors: Sourita Ghosh, Falguni Pati, Suhanya Duraiswamy

Abstract:

Spheroid, a simple three-dimensional cellular aggregate, allows us to simulate the in-vivo complexity of cellular signaling and interactions in greater detail than traditional 2D cell culture. It can be used as an in-vitro model for drug toxicity testing, tumor modeling and many other such applications specifically for cancer. Our work is focused on the development of an affordable, user-friendly, robust, reproducible, high throughput microfluidic device for water in oil droplet production, which can, in turn, be used for spheroids manufacturing. Here, we have investigated the droplet breakup between two non-Newtonian fluids, viz. silicone oil and decellularized liver matrix, which acts as our extra cellular matrix (ECM) for spheroids formation. We performed some biochemical assays to characterize the liver ECM, as well as rheological studies on our two fluids and observed a critical dependence of capillary number (Ca) on droplet breakup and homogeneous drop formation

Keywords: chip less, droplets, extracellular matrix, liver spheroid

Procedia PDF Downloads 89

Authors: Gulsen Tozsin, Fatma Bakir, Cemil Acar, Ercument Koc

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Kurtbogazi Dam has gained special meaning for Ankara, Turkey for the last decade due to the rapid depletion of nearby resources of drinking water. In this study, the results of the analyses of Kurtbogazi Dam outlet water and the rivers flowing into the Kurtbogazi Dam were discussed for the period of last five years between 2008 and 2012. The quality of these surface water resources were evaluated in terms of pH, temperature, biochemical oxygen demand (BOD5), nitrate, phosphate and chlorine. They were classified according to the Council Directive (75/440/EEC). Moreover, the properties of these surface waters were assessed to determine the quality of water for drinking and irrigation purposes using Piper, US Salinity Laboratory and Wilcox diagrams. The results revealed that the quality of all the investigated water sources are generally at satisfactory level as surface water except for Pazar Stream in terms of ortho-phosphate and BOD5 concentration for 2008.

Keywords: Kurtbogazi dam, water quality assessment, Ankara water, water supply

Procedia PDF Downloads 377