Search results for: Elisa Gonzalez

Authors: Chodidjah, Edi Dharmana, Hardhono, Sarjadi

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Breast cancer treatment options including surgery, radiation therapy, chemotherapy, and immunotherapy have not been effective. Besides, they have side effects. Keladi Tikus (Typhonium flagelliforme) has been shown to improve immune system, suppress tumor growth and induce apoptosis. One of the parameters for immune system, tumor growth and apoptosis is IFNγ (Interferon γ), VEGF (Vascular Endothelial Growth Factor) and Caspase 3 respectively. The aim of this study was to examine the effect of the administration of Keladi Tikus tuber extract at the dose of 200 mg/kgBW, 400 mg/KgBW, and 800 mg/kgBW on the level of IFNγ, VEGF and caspase 3 expression. In this experimental study using post test randomized control group design, 24 CH3 mice with tumor were randomly divided into 4 groups including control group and treated groups: Treated with 0.2 cc extract of Keladi Tikus at the dose of 200 mg/kgBW, 400 mg/kgBW, 800 mg/kgBW, respectively for 30 days. On day 31 the lymphatic tissue was taken and evaluated for its level of IFNγ, using ELISA. The tumor tissue was taken and subjected to immunohistochemistry staining for VEGF and caspase 3 expression evaluation. The data on IFNγ, VEGF and Caspase 3 expression were analyzed using One Way Anova with significant level of 0.05. One Way Anova resulted in p<0.05. LSD test showed that the level of IFNγ and Caspase 3 for control group was different from that of treated groups. There was no significant different between the treated group of 400 mg/KgBW and 800mg/KgBW. VEGF expressions for all the treated groups were significant. In conclusion, the oral administration of ethanolic extract of Keladi Tikus (Typhonium flagelliforme) at the dose of 200mg/kgBW, 400 mg/kgBW,800 mg/kgBW increases IFNγ, Caspase 3 and decreases VEGF expression in C3H mice with adenocarsinoma mamma.

Keywords: Typhonium flagelliforme, IFNγ, caspase 3, VEGF

Procedia PDF Downloads 408

Authors: Garzon V., Bustos R., Salvador J. P., Marco M. P., Pinacho D. G.

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Bacterial resistance to antimicrobial treatment has increased significantly in recent years, making it a public health problem. Large numbers of bacteria are resistant to all or nearly all known antimicrobials, creating the need for the development of new types of antimicrobials or the use of “last line” antimicrobial drug therapies for the treatment of multi-resistant bacteria. Some of the chemical groups of antimicrobials most used for the treatment of infections caused by multiresistant bacteria in the clinic are Glycopeptide (Vancomycin), Polymyxin (Colistin), Lipopeptide (Daptomycin) and Carbapenem (Meropenem). Molecules that require therapeutic drug monitoring (TDM). Due to the above, a methodology based on nanobiotechnology based on an optical and electrochemical biosensor is being developed, which allows the evaluation of the plasmatic levels of some antimicrobials such as glycopeptide, polymyxin, lipopeptide and carbapenem quickly, at a low cost, with a high specificity and sensitivity and that can be implemented in the future in public and private health hospitals. For this, the project was divided into five steps i) Design of specific anti-drug antibodies, produced in rabbits for each of the types of antimicrobials, evaluating the results by means of an immunoassay analysis (ELISA); ii) quantification by means of an electrochemical biosensor that allows quantification with high sensitivity and selectivity of the reference antimicrobials; iii) Comparison of antimicrobial quantification with an optical type biosensor; iv) Validation of the methodologies used with biosensor by means of an immunoassay. Finding as a result that it is possible to quantify antibiotics by means of the optical and electrochemical biosensor at concentrations on average of 1,000ng/mL, the antibodies being sensitive and specific for each of the antibiotic molecules, results that were compared with immunoassays and HPLC chromatography. Thus, contributing to the safe use of these drugs commonly used in clinical practice and new antimicrobial drugs.

Keywords: antibiotics, electrochemical biosensor, optical biosensor, therapeutic drug monitoring

Procedia PDF Downloads 60

Authors: Rossy G. Olan Villegas, Gerardo Acosta Garcia, Aurea Bernardino Nicanor, Leopoldo Gonzalez Cruz, Humberto Ramirez Medina

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Capsicum annuum is a crop of agricultural and economic importance in Mexico and other countries. The fruit (pepper) contains bioactive components such as carotenoids, phenolic compounds and capsaicinoids that improve health. However, pepper cultivation is affected by biotic and abiotic factors that decrease yield. Some phytohormones like gibberellins and auxins induce the formation and development of fruit in several plants. In this study, we evaluated the effect of the exogenous application of phytohormones like gibberellic acid and indolbutyric acid on fruit development of jalapeno pepper plants, the protein profile of plant tissues, the accumulation of bioactive compounds and antioxidant activity in the pericarp and seeds. For that, plants were sprinkled with these phytohormones. The fruit collection for the control, indolbutyric acid and gibberellic acid treatments was 7 peppers per plant; however, for the treatment that combines indolbutyric acid and gibberellic acid, a fruit with the shortest length (1.52 ± 1.00 cm) and weight (0.41 ± 1.0 g) was collected compared to fruits of plants grown under other treatments. The length (4,179 ± 0,130 cm) and weight of the fruit (8,949 ± 0.583 g) increased in plants treated with indolbutyric acid, but these characteristics decreased with the application of GA3 (length of 3,349 ± 0.127 cm and a weight 4,429 ± 0.144 g). The content of carotenes and phenolic compounds increased in plants treated with GA3 (1,733 ± 0.092 and 1,449 ± 0.009 mg / g, respectively) or indolbutyric acid (1,164 ± 0.042 and 0.970 ± 0.003 mg / g). However, this effect was not observed in plants treated with both phytohormones (0.238 ± 0.021 and 0.218 ± 0.004 mg / g). Capsaicin content was higher in all treatments; but it was more noticeable in plants treated with both phytohormones, the value being 0.913 ± 0.001 mg / g (three times greater in amount). The antioxidant activity was measured by 3 different assays, 2,2-diphenyl-1-picrylhydrazyl (DPPH), antioxidant power of ferric reduction (FRAP) and 2,2'-Azinobis-3-ethyl-benzothiazoline-6-sulfonic acid ( ABTS) to find the minimum inhibitory concentration of the reducing radical (IC50 and EC50). Significant differences were observed from the application of the phytohormone, being the fruits treated with gibberellins, which had a greater accumulation of bioactive compounds. Our results suggest that the application of phytohormones modifies the development of fruit and its content of bioactive compounds.

Keywords: auxins, capsaicinoids, carotenoids, gibberellins

Procedia PDF Downloads 92

Authors: Saad Al-Shibli, Nasser Amjad, Muna Al Kubaisi, Norra Harun, Shaikh Mizan

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Leptin is a multifunctional hormone produced mainly by adipocyte. Leptin and its receptor have long been found associated with breast cancer. The main aim of this study is to investigate the correlation between Leptin/Leptin receptor and the clinicopathological features of breast cancer. Blood samples for ELISA, tissue samples from tumors and adjacent breast tissue were taken from 51 women with breast cancer with a control group of 40 women with a negative mammogram. Leptin and Leptin receptor in the tissues were estimated by immunohistochemistry (IHC). They were localized at the subcellular level by immunocytochemistry using transmission electron microscopy (TEM). Our results showed significant difference in serum leptin level between control and the patient group, but no difference between pre and post-operative serum leptin levels in the patient group. By IHC, we found that the majority of the breast cancer cells studied, stained positively for leptin and leptin receptors with co-expression of leptin and its receptors. No significant correlation was found between leptin/leptin receptors expression with the race, menopausal status, lymph node metastasis, estrogen receptor expression, progesterone receptor expression, HER2 expression and tumor size. Majority of the patients with distant metastasis were associated with high leptin and leptin receptor expression. TEM views both Leptin and Leptin receptor were found highly concentrated within and around the nucleus of the cancer breast cells, indicating nucleus is their principal seat of actions while the adjacent breast epithelial cells showed that leptin gold particles are scattered all over the cell with much less than that of the cancerous cells. However, presence of high concentration of leptin does not necessarily prove its over-expression, because it could be internalized from outside by leptin receptor in the cells. In contrast, leptin receptor is definitely over-expressed in the ductal breast cancer cells. We conclude that reducing leptin levels, blocking its downstream tissue specific signal transduction, and/or blocking the upstream leptin receptor pathway might help in prevention and therapy of breast cancer.

Keywords: breast cancer, expression, leptin, leptin receptors

Procedia PDF Downloads 115

Authors: María Magdalena Méndez-González, Miguel García Rocha, Carlos Manuel Yermo De la Cruz

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Calcium phosphate (Ca5(PO4)3(X)) is widely used in orthopedic applications and is widely used as powder and granules. However, their presence in bone is in the form of nanometric needles 60 nm in length with a non-stoichiometric phase of apatite contains CO3-2, Na+, OH-, F-, and other ions in a matrix of collagen fibers. The crystal size, morphology control and interaction with cells are essential for the development of nanotechnology. The structural results of calcium phosphate, synthesized by chemical precipitation with crystal size of 22.85 nm are presented in this paper. The calcium phosphate powders were analyzed by X-ray diffraction, energy dispersive spectroscopy (EDS), infrared spectroscopy and FT-IR transmission electron microscopy. Network parameters, atomic positions, the indexing of the planes and the calculation of FWHM (full width at half maximum) were obtained. The crystal size was also calculated using the Scherer equation d (hkl) = cλ/βcosѲ. Where c is a constant related to the shape of the crystal, the wavelength of the radiation used for a copper anode is 1.54060Å, Ѳ is the Bragg diffraction angle, and β is the width average peak height of greater intensity. Diffraction pattern corresponding to the calcium phosphate called hydroxyapatite phase of a hexagonal crystal system was obtained. It belongs to the space group P63m with lattice parameters a = 9.4394 Å and c = 6.8861 Å. The most intense peak is obtained 2Ѳ = 31.55 (FWHM = 0.4798), with a preferred orientation in 121. The intensity difference between the experimental data and the calculated values is attributable to the temperature at which the sintering was performed. The intensity of the highest peak is at angle 2Ѳ = 32.11. The structure of calcium phosphate obtained was a hexagonal configuration. The intensity changes in the peaks of the diffraction pattern, in the lattice parameters at the corners, indicating the possible presence of a dopant. That each calcium atom is surrounded by a tetrahedron of oxygen and hydrogen was observed by infrared spectra. The unit cell pattern corresponds to hydroxyapatite and transmission electron microscopic crystal morphology corresponding to the hexagonal phase with a preferential growth along the c-plane was obtained.

Keywords: structure, nanoparticles, calcium phosphate, metallurgical and materials engineering

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Authors: Enrique González Pendás, Vidal Pérez Hernández, Jorge Ferro Díaz, Nelson Careaga Pendás

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The investigation is carried out on the Protected Area of San Ubaldo, toward the interior of an open pinegrove with palm trees in a dry plainness of quar zitic sands, belonging to the Floristic Managed Reservation San Ubaldo-Sabanalamar, Guane, Pinar del Río, Cuba. This area is characterized by drastic seasonal variations, high temperatures and water evaporation, strong solar radiation, with sandy soils of almost pure quartz, which are very acid and poor in nutrients. The objective of the present work is to determine evidence of facilitation and its relationship with the structure and composition of plant communities in these peculiar ecosystems. For this study six lineal parallel transepts of 100 m are traced, in those, a general recording of the flora is carried out. To establish which plants act as nurses, is taken into account a height over 1 meter, canopy over 1.5 meter and the occurrence of several species under it. Covering was recorded using the line intercept method; the medium values of species richness for the taxa under nurses is compared with those that are located in open spaces among them. Then, it is determined which plants are better recruiter of other species (better nurses). An experiment is made to measure and compare some parameters in pine seedlings under the canopy of the Byrsonima crassifolia (L.) Kunth. and in open spaces, also the number of individuals is counted by species to calculate the frequency and total abundance in the study area. As a result, it is offered an up-to-date floristic list, a phylogenetic tree of the plant community showing a high phylodiversity, it is proven that the medium values of species richness and abundance of species under the nurses, is significantly superior to those occurring in open spaces. Furthermore, by means of phylogenetic trees it is shown that the species which cohabit under the nurses are not phylogenetically related. The former results are cited evidences of facilitation among plants, as well as it is one more time shown the importance of the nurse effect in preserving plant diversity on extreme environments.

Keywords: facilitation, nurse plants, positive interactions, quarzitic sands

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Authors: Aliaa M. Issa, Mahmoud N. ElRouby, Sahar A. S. Ahmad, Mahmoud M. El-Merzabani

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Sider honey is a type of honey produced by bees feeding on the nectar of Sider tree, Ziziphus spina-christi (L) Desf . Honey is an effective agent for preventing, inhibiting and treating the growth of human and animal cancer cell lines in vitro and in vivo. The aim of the present study was to evaluate the impact of different dilutions from crude Sider honey and different duration times of exposure on the growth of six tumor cell lines (human cervical cancer cell line, HeLa; human hepatocellular carcinoma cell line, HepG-2; human larynx carcinoma cell line, Hep-2; brain tumor cell line, U251) as well as one animal cancerous cell line (Ehrlich ascites carcinoma cells line, EAC) and one normal cell line, Homo sapiens, human, (WISH) CCL-25. Different concentrations and treatment durations with Sider honey were tested on the growth of several cancer cell lines types. Histopathological changes in the tumor masses, animal survival, apoptosis and necrosis of the used cancer cell lines (using flow cytometry) were evaluated. Sider honey was administers either to the tumor mass itself by intratumoral injection or via drinking water. One-way ANOVA test was used for the analysis of (the means + standard error) of the optical density obtained from the Elisa reader and flow cytometry. The study revealed that different concentrations of Sider honey affected the growth patterns of all the studied cancer cell lines as well as their histopathological changes, and it depended on the cell line nature and the concentration of honey used. It is obvious that the relative animal survival percentage (bearing Ehrlich ascites carcinoma, EAC cells) was proportionally increased with the increase in the used honey concentrations. The study of apoptosis and necrosis using the flow cytometry technique emphasized the viability results. In conclusion, Sider honey was effective as antitumor agent, in the used concentrations.

Keywords: antitumor, honey, sider, tumor cell lines

Procedia PDF Downloads 506

Authors: Dwi Arisandi Wijaya, Ernawati Arifin Giri-Rachman, Neni Nurainy

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Typhoid fever disease can be prevented by using a polysaccharide-based vaccine Vi which is a virulence factor of S.typhi. To produce high yield Vi polysaccharide from bacteria, it is important to know the biosynthesis of Vi polysaccharide and the regulators involved. In the In vivo condition, S. typhi faces different osmolarity, and the bacterial two-component system OmpR-EnvZ, regulate by up and down Capsular Vi polysaccharide biosynthesis. A high yielded Vi Polysaccharide strain, S. typhi strain C6524 used to study the effect of media osmolarity on Vi polysaccharide biosynthesis and the osmoregulation pattern of S. typhi strain C6524. The methods were performed by grown S. typhi strain C6524 grown on medium with 50 mM, 100 mM, and 150 mM osmolarity with the batch system. Vi polysaccharide concentration was measured by ELISA method. For further investigation of the osmoregulation pattern of strain C6524, the osmoregulator gene, OmpR, has been isolated and sequenced using the specific primer of the OmpR gene. Nucleotide sequence analysis is done with BLAST and Lallign. Amino Acid sequence analysis is done with Prosite and Multiple Sequence Alignment. The results of cultivation showed the average content of polysaccharide Vi for 50 mM, 100 mM, and 150 mM osmolarities 11.49 μg/mL, 12.06 μg/mL, and 14.53 μg/mL respectively. Analysis using Anova stated that the osmolarity treatment of 150 mM significantly affects Vi content. Analysis of nucleotide sequences shows 100% identity between S. typhi strain C6524 and Ty2. Analysis of amino acid sequences shows that the OmpR response regulator protein of the C6524 strain also has a α4-β5-α5 motif which is important for the regulatory activation system when phosphorylation occurs by domain kinase. This indicates that the regulator osmolarity response of S. typhi strain C6524 has no difference with the response regulator owned by S. typhi strain Ty2. A high Vi response rate in the 150 mM osmolarity treatment requires further research for RcsB-RcsC, another two-component system involved in Vi Biosynthesis.

Keywords: osmoregulator, OmpR, Salmonella, Vi polysaccharide

Procedia PDF Downloads 175

Authors: Abdolamir Allameh, Seyyed Mortaza Haghgoo, Adnan Khosravi, Esmaeil Mortaz, Mihan Pourabdollah-Toutkaboni, Sharareh Seifi

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Non-Small Cell Lung Carcinoma (NSCLC) is associated with a number of gene mutations in epidermal growth factor receptor (EGFR). The prognostic significance of mutations in exons 19 and 21, together with serum levels of EGFR, amphiregulin (AR), and Transforming Growth Factor-alpha (TGF-α) are implicated in diagnosis and treatment. The aim of this study was to examine the relationship of EGFR mutations in selected exons with the expression of relevant ligands in sera samples of NSCLC patients. For this, a group of NSCLC patients (n=98) referred to the hospital for lung surgery with a mean age of 59±10.5 were enrolled (M/F: 75/23). Blood specimen was collected from each patient. Besides, formalin fixed paraffin embedded tissues were processed for DNA extraction. Gene mutations in exons 19 and 21 were detected by direct sequencing, following DNA amplification which was done by PCR (Polymerase Chain Reaction). Also, serum levels of EGFR, AR, and TGF-α were measured by ELISA. The results of our study show that EGFR mutations were present in 37% of Iranian NSCLC patients. The most frequently identified mutations were deletions in exon 19 (72.2%) and substitutions in exon 21 (27.8%). The most frequently identified alteration, which is considered as a rare mutation, was the E872K mutation in exon 21, which was found in 90% (9 out of 10) cases. EGFR mutation detected in exon 21 was significantly (P<0.05) correlated with the levels of its ligands, EGFR and TGF-α in serum samples. Furthermore, it was found that increased serum AR (>3pg/ml) and TGF-α (>10.5 pg/ml) were associated with shorter overall survival (P<0.05). The results clearly showed a close relationship between EGFR mutations and serum EGFR and serum TGF-α. Increased serum EGFR was associated with TGF-α and AR and linked to poor prognosis of NSCLC. These findings are implicated in clinical decision-making related to EGFR-Tyrosine kinase inhibitors (TKIs).

Keywords: lung cancer, Iranian patients, epidermal growth factor, mutation, prognosis

Procedia PDF Downloads 61

Authors: M. Belén Prendes-Gero, Celestino González-Nicieza, M. Inmaculada Alvarez-Fernández

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Roof collapse is one of the problems with a higher frequency in most of the mines of all countries, even now. There are many reasons that may cause the roof to collapse, namely the mine stress activities in the mining process, the lack of vigilance and carelessness or the complexity of the geological structure and irregular operations. This work is the result of the analysis of one accident produced in the “Mary” coal exploitation located in northern Spain. In this accident, the roof of a crossroad of excavated galleries to exploit the “Morena” Layer, 700 m deep, collapsed. In the paper, the work done by the forensic team to determine the causes of the incident, its conclusions and recommendations are collected. Initially, the available documentation (geology, geotechnics, mining, etc.) and accident area were reviewed. After that, laboratory and on-site tests were carried out to characterize the behaviour of the rock materials and the support used (metal frames and shotcrete). With this information, different hypotheses of failure were simulated to find the one that best fits reality. For this work, the software of finite differences in three dimensions, FLAC 3D, was employed. The results of the study confirmed that the detachment was originated as a consequence of one sliding in the layer wall, due to the large roof span present in the place of the accident, and probably triggered as a consequence of the existence of a protection pillar insufficient. The results allowed to establish some corrective measures avoiding future risks. For example, the dimensions of the protection zones that must be remained unexploited and their interaction with the crossing areas between galleries, or the use of more adequate supports for these conditions, in which the significant deformations may discourage the use of rigid supports such as shotcrete. At last, a grid of seismic control was proposed as a predictive system. Its efficiency was tested along the investigation period employing three control equipment that detected new incidents (although smaller) in other similar areas of the mine. These new incidents show that the use of explosives produces vibrations which are a new risk factor to analyse in a next future.

Keywords: forensic analysis, hypothesis modelling, roof failure, seismic monitoring

Procedia PDF Downloads 99

Authors: Mario J. Meco-Gutierrez, Francisco Perez-Hidalgo, Juan R. Heredia-Larrubia, Antonio Ruiz-Gonzalez, Francisco Vargas-Merino

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More and more applications power inverters connected to transformers, for example, the connection facilities to the power grid renewable generation. It is well known that the quality of signal power inverters it is not a pure sine. The harmonic content produced negative effects, one of which is the heating of electrical machines and therefore, affects the life of the machines. The decrease of life of transformers can be calculated by Arrhenius or Montsinger equation. Analyzing this expression any (long-term) decrease of a transformer temperature for 6º C - 7º C means doubles its life-expectancy. Methodologies: This work presents the technique of pulse width modulation (PWM) with an injection of harmonic and triangular frequency carrier modulated in frequency. This technique is used to improve the quality of the output voltage signal of the power inverters controlled PWM. The proposed technique increases in the fundamental term and a significant reduction in low order harmonics with the same commutations per time that control sine PWM. To achieve this, the modulating wave is compared to a triangular carrier with variable frequency over the period of the modulator. Therefore, it is, advantageous for the modulating signal to have a large amount of sinusoidal “information” in the areas of greater sampling. A triangular signal with a frequency that varies over the modulator’s period is used as a carrier, for obtaining more samples in the area with the greatest slope. A power inverter controlled by PWM proposed technique is connected to a transformer. Results: In order to verify the derived thermal parameters under different operation conditions, another ambient and loading scenario is involved for a further verification, which was sampled from the same power transformer. Temperatures of different parts of the transformer will be exposed for each PWM control technique analyzed. An assessment of the temperature be done with different techniques PWM control and hence the life of the transformer is calculated for each technique. Conclusion: This paper analyzes such as transformer heating produced by this technique and compared with other forms of PWM control. In it can be seen as a reduction the harmonic content produces less heat transformer and therefore, an increase in the life of the transformer.

Keywords: heating, power-inverter, PWM, transformer

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Authors: Yasser Y. Lenis, Amy Jo Montgomery, Diego F. Carrillo-Gonzalez

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Introduction: The use of didactic instruments in different learning environments plays a pivotal role in enhancing the level of knowledge in veterinary science students. The direct instruction of basic animal reproduction concepts in students enrolled in veterinary medicine programs allows them to elucidate the biological and molecular mechanisms that perpetuate the animal species in an ecosystem. Therefore, universities must implement didactic strategies that facilitate the teaching and learning processes for students and, in turn, enrich learning environments. Objective: to evaluate the effect of the use of a didactic textbook on the level of theoretical knowledge in embryo-maternal recognition for veterinary medicine students. Methods: the participants (n=24) were divided into two experimental groups: control (Ctrl) and treatment (Treat). Both groups received 4 hours of theoretical training regarding the basic concepts in bovine embryo-maternal recognition. However, the Treat group was also exposed to a guided lecture and the activity play-to-learn from a cow reproduction didactic textbook. A pre-test and a post-test were applied to assess the prior and subsequent knowledge in the participants. Descriptive statistics were applied to identify the success rates for each of the tests. Afterwards, a repeated measures model was applied where the effect of the intervention was considered. Results: no significant difference (p>0,05) was observed in the number of right answers for groups Ctrl (54,2%±12,7) and Treat (40,8%±16,8) in the pre-test. There was no difference (p>0,05) compering the number of right answers in Ctrl pre-test (54,2%±12,7) and post-test (60,8±18,8). However, the Treat group showed a significant (p>0,05) difference in the number of right answers when comparing pre-test (40,8%±16,8) and post-test (71,7%±14,7). Finally, after the theoretical training and the didactic activity in the Treat group, an increase of 10.9% (p<0,05) in the number of right answers was found when compared with the Ctrl group. Conclusion: the use of didactic tools that include guided lectures and activities like play-to-learn from a didactic textbook enhances the level of knowledge in an animal reproduction course for veterinary medicine students.

Keywords: animal reproduction, pedagogic, level of knowledge, learning environment

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Authors: Aisha Khodijah Kholib Jati, Suharni Mohamad, Azlan Husin, Wan Suriana Wan Ab Rahman

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Introduction: Toxoplasmosis is caused by an obligate intracellular parasite, Toxoplasma gondii (T. gondii). It is commonly asymptomatic in normal individual, but it can be fatal to immunocompromised patients as it can lead to severe complications such as encephalitis, chorioetinitis and myocarditis. Objective: The aim of the study was to determine the seroprevalence of toxoplasmosis and its association with socio-demographic and behavioral characteristics among hemato-oncology patients in Hospital USM. Methods: In this cross-sectional study, 56 hemato-oncology patients were screened for immunoglobulin M (IgM) antibodies, immunoglobulin G (IgG) antibodies, and IgG avidity of T. gondii by using ELISA Kit (BioRad, USA). For anti-T. gondii IgG antibody, titer ≥ 9 IU/ml was considered as recent infection, while for IgM, ratio ≥ 1.00 was considered as reactive for the anti-T. gondii IgM antibodies. Low avidity index is considered as recent infection within 20 weeks while high avidity considered as past infection. T. gondii exposure, socio-demographic and behavioral characteristics was assessed by a questionnaire and interview. Results: A total of 28 (50.0%) hemato-oncology patients were seropositive for T. gondii antibodies. Out of that total, 27 (48.21%) patients were IgG+/IgM- and one patient (1.79%) was IgG+/IgM+ with high avidity index. Univariate analysis showed that age, gender, ethnicity, marital status, educational level, employment status, stem cell transplant, blood transfusion, close contact with cats, water supply, and consumption of undercooked meat were not significantly associated with Toxoplasma seropositivity rate. Discussion: The seropositivity rate of IgG anti-T. gondii was high among hemato-oncology patients in Hospital USM. With impaired immune system, these patients might have a severe consequence if the infection reactivated. Therefore, screening for anti-T. gondii may be considered in the future. Moreover, health programme towards healthy food and good hygiene practice need to be implemented.

Keywords: immunocompromised, seroprevalence, socio-demographic, toxoplasmosis

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Authors: Pudji Astuti, C. P. C. Putro, C. M. Airin, L. Sjahfirdi, S. Widiyanto, H. Maheshwari

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Stress of slaughter animals starting long before until at the time of process of slaughtering which cause misery and decrease of meat quality. Meanwhile, determination of animal stress using hormonal such as cortisol is expensive and less practical so that portable stress indicator for cows based on Fourier Transform Infrared Spectroscopy (FTIR) must be provided. The aims of this research are to find out the comparison process of slaughter between Rope Casting Local (RCL) and Restraining Box Method (RBM) by measuring of cortisol and wavelength in FTIR methods. Thirty two of male Ongole crossbred cattle were used in this experiment. Blood sampling was taken from jugular vein when they were rested and repeated when slaughtered. All of blood samples were centrifuged at 3000 rpm for 20 minutes to get serum, and then divided into two parts for cortisol assayed using ELISA and for measuring the wavelength using FTIR. The serum then measured at the wavelength between 4000-400 cm-1 using MB3000 FTIR. Band data absorption in wavelength of FTIR is analyzed descriptively by using FTIR Horizon MBTM. For RCL, average of serum cortisol when the animals rested were 11.47 ± 4.88 ng/mL, when the time of slaughter were 23.27 ± 7.84 ng/mL. For RBM, level of cortisol when rested animals were 13.67 ± 3.41 ng/mL and 53.47 ± 20.25 ng/mL during the slaughter. Based on student t-Test, there were significantly different between RBM and RCL methods when beef cattle were slaughtered (P < 0.05), but no significantly different when animals were rested (P > 0.05). Result of FTIR with the various of wavelength such as methyl group (=CH3) 2986cm-1, methylene (=CH2) 2827 cm-1, hydroxyl (-OH) 3371 cm-1, carbonyl (ketones) (C=O) 1636 cm-1, carboxyl (COO-1) 1408 cm-1, glucosa 1057 cm-1, urea 1011 cm-1have been obtained. It can be concluded that the RCL slaughtered method is better than the RBM method based on the increase of cortisol as an indicator of stress in beef cattle (P<0.05). FTIR is really possible to be used as stub of stress tool due to differentiate of resting and slaughter condition by recognizing the increase of absorption and the separation of component group at the wavelength.

Keywords: cows, cortisol, FTIR, RBM, RCL, stress indicator

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Authors: Vera Sovkova, Andrea Mickova, Matej Buzgo, Karolina Vocetkova, Eva Filova, Evzen Amler

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PCL (poly-ε-caprolactone) nanofibrous scaffolds with adhered liposomes were prepared and tested as a possible drug delivery system for various synthetic growth factors. TGFβ, bFGF, and IGF-I have been shown to increase hMSC (human mesenchymal stem cells) proliferation and to induce hMSC differentiation. Functionalized PCL nanofibers were prepared with synthetic growth factors encapsulated in liposomes adhered to them in three different concentrations. Other samples contained PCL nanofibers with adhered, free synthetic growth factors. The synthetic growth factors free medium served as a control. The interaction of liposomes with the PCL nanofibers was visualized by SEM, and the release kinetics were determined by ELISA testing. The potential of liposomes, immobilized on the biodegradable scaffolds, as a delivery system for synthetic growth factors, and as a suitable system for MSCs adhesion, proliferation and differentiation in vitro was evaluated by MTS assay, dsDNA amount determination, confocal microscopy, flow cytometry and real-time PCR. The results showed that the growth factors adhered to the PCL nanofibers stimulated cell proliferation mainly up to day 11 and that subsequently their effect was lower. By contrast, the release of the lowest concentration of growth factors from liposomes resulted in gradual proliferation of MSCs throughout the experiment. Moreover, liposomes, as well as free growth factors, stimulated type II collagen production, which was confirmed by immunohistochemical staining using monoclonal antibody against type II collagen. The results of this study indicate that growth factors enriched liposomes adhered to surface of PCL nanofibers could be useful as a drug delivery instrument for application in short timescales, be combined with nanofiber scaffolds to promote local and persistent delivery while mimicking the local microenvironment. This work was supported by project LO1508 from the Ministry of Education, Youth and Sports of the Czech Republic

Keywords: drug delivery, growth factors, hMSC, liposomes, nanofibres

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Authors: Lívia Pimentel Sant'ana Dourado, Raquel Das Neves Almeida, Luís Henrique Costa Corrêa Neto, Nayara Soares, Kelly Grace Magalhães

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Introduction: Obesity and high-fat diet intake have a crucial impact on immune cells and inflammatory profile, highlighting an emerging realization that obesity is an inflammatory disease. In the present work, we aimed to characterize the role of caspase-1/11 and NLRP3 inflammasome in the establishment of mice obesity and modulation of inflammatory lipid metabolism induced by high fat diet intake. Methods and results: Wild type, caspase-1/11 and NLRP3 knockout mice were fed with standard fat diet (SFD) or high fat diet (HFD) for 90 days. The weight of animals was measured weekly to monitor the weight gain. After 90 days, the blood, peritoneal lavage cells, heart and liver were collected from mice studied here. Cytokines were measured in serum by ELISA and analyzed in spectrophotometry. Lipid antigen presentation molecule CD1d expression, reactive oxygen species (ROS) generation and lipid droplets biogenesis were analyzed in cells from mice peritoneal cavity by flow cytometry. Liver histopathology was performed for morphological evaluation of the organ. The absence of caspase-1/11, but not NLRP3, in mice fed with HFD favored the mice weight gain, increased liver size, induced development of hepatic steatosis and IL-12 secretion in mice compared to mice fed with SFD. In addition, caspase-1/11 knockout mice fed with HFD presented an increased CD1d molecule expression, as well as higher levels of lipid droplets biogenesis and ROS generation compared to wild type mice also fed with HFD. Conclusion: Our data suggest that caspase-1/11 knockout mice have greater susceptibility to obesity as well as increased activation of lipid metabolism and inflammatory markers.

Keywords: caspase 1, caspase 11, inflamassome, obesity, lipids

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Authors: Elisa Maître

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The redeployment of tram networks in French cities has considerably modified public spaces and the way citizens use them. Above and beyond the image that trams have of contributing to the sustainable urban development, the question of safety for users in these spaces has not been studied much. This study is based on an analysis of use of public spaces laid out for trams, from the standpoint of legibility and safety concerns. The study also examines to what extent the complexity of the design process, with many interactions between numerous and varied players in this process has a role in the genesis of these problems. This work is mainly based on the analysis of links between the uses of these re-designed public spaces (through observations, interviews of users and accident studies) and the analysis of the design conditions and processes of the projects studied (mainly based on interviews with the actors of these projects). Practical analyses were based three points of view: that of the planner, that of the user (based on observations and interviews) and that of the road safety expert. The cities of Montpellier, Marseille and Nice are the three fields of study on which the demonstration of this thesis is based. On part, the results of this study allow showing that the insertion of tram poses some problems complication of public areas of French cities. These complications related to the restructuring of public spaces for the tram, create difficulties of use and safety concerns. On the other hand, interviews depth analyses, fully transcribed, have led us to develop particular dysfunction scenarios in the design process. These elements lead to question the way the legibility and safety of these new forms of public spaces are taken into account. Then, an in-depth analysis of the design processes of public spaces with trams systems would also be a way of better understanding the choices made, the compromises accepted, and the conflicts and constraints at work, weighing on the layout of these spaces. The results presented concerning the impact that spaces laid out for trams have on the difficulty of use, suggest different possibilities for improving the way in which safety for all users is taken into account in designing public spaces.

Keywords: public spaces, road layout, users, design process of urban projects

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Authors: Ambrish Tiwari, Sudhasini Panda, Archana Singh, Kalpana Luthra, S. K. Sharma

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Introduction: On the basis of available literature we know that various host factors play a role in outcome of Tuberculosis (TB) infection by modulating innate immunity. One such factor is Inducible Nitric Oxide Synthase enzyme (iNOS) which help in the production of Nitric Oxide (NO), an antimicrobial agent. Expression of iNOS is in control of various host factors in which Vitamin D along with its nuclear receptor Vitamin D receptor (VDR) is one of them. Vitamin D along with its receptor also produces cathelicidin (antimicrobicidal agent). With this background, we attempted to investigate the levels of Vitamin D and NO along with their associated molecules in tuberculosis patients and household contacts as compared to healthy controls and assess the implication of these findings in susceptibility to tuberculosis (TB). Study subjects and methods: 100 active TB patients, 75 household contacts, and 70 healthy controls were taken. VDR and iNOS mRNA levels were studied using real-time PCR. Serum VDR, cathelicidin, iNOS levels were measured using ELISA. Serum Vitamin D levels were measured in serum samples using chemiluminescence based immunoassay. NO was measured using colorimetry based kit. Results: VDR and iNOS mRNA levels were found to be lower in active TB group compared to household contacts and healthy controls (P=0.0001 and 0.005 respectively). The serum levels of Vitamin D were also found to be lower in active TB group as compared to healthy controls (P =0.001). Levels of cathelicidin and NO was higher in patient group as compared to other groups (p=0.01 and 0.5 respectively). However, the expression of VDR and iNOS and levels of vitamin D was significantly (P < 0.05) higher in household contacts compared to both active TB and healthy control groups. Inference: Higher levels of Vitamin D along with VDR and iNOS expression in household contacts as compared to patients suggest that vitamin D might have a protective role against TB which prevents activation of the disease. From our data, we can conclude that decreased vitamin D levels could be implicated in disease progression and we can use cathelicidin and NO as a biomarker for early diagnosis of pulmonary tuberculosis.

Keywords: vitamin D, VDR, iNOS, tuberculosis

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Authors: Janneth Gonzalez, Andrés Pinzon Velasco, Maria Angarita

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Astrocytes play an important role in various processes in the brain, including pathological conditions such as neurodegenerative diseases. Recent studies have shown that the increase in saturated fatty acids such as palmitic acid (PA) triggers pro-inflammatorypathways in the brain. The use of synthetic neurosteroids such as tibolone has demonstrated neuro-protective mechanisms. However, broad studies with a systemic point of view on the neurodegenerative role of PA and the neuro-protective mechanisms of tibolone are lacking. In this study, we performed the integration of multi-omic data (transcriptome and proteome) into a human astrocyte genomic scale metabolic model to study the astrocytic response during palmitate treatment. We evaluated metabolic fluxes in three scenarios (healthy, induced inflammation by PA, and tibolone treatment under PA inflammation). We also applied a control theory approach to identify those reactions that exert more control in the astrocytic system. Our results suggest that PA generates a modulation of central and secondary metabolism, showing a switch in energy source use through inhibition of folate cycle and fatty acid β‐oxidation and upregulation of ketone bodies formation. We found 25 metabolic switches under PA‐mediated cellular regulation, 9 of which were critical only in the inflammatory scenario but not in the protective tibolone one. Within these reactions, inhibitory, total, and directional coupling profiles were key findings, playing a fundamental role in the (de)regulation of metabolic pathways that may increase neurotoxicity and represent potential treatment targets. Finally, the overall framework of our approach facilitates the understanding of complex metabolic regulation, and it can be used for in silico exploration of the mechanisms of astrocytic cell regulation, directing a more complex future experimental work in neurodegenerative diseases.

Keywords: astrocytes, data integration, palmitic acid, computational model, multi-omics

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Authors: Ricardo Hugo Gonzalez, Amanda Figueiredo Vasconcelos, Francisco Loureiro Neto Monteiro, Yara Luiza Freitas Silva, Ana Cristina Lindsay, Márcia Maria Tavares Machado

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The use of sport as an integration mean is a very important tool regarding the social involvement of children and teenagers in a vulnerability situation. This study aims to report the experiences of a multidisciplinary program that intends to improve the quality of life of children and teenagers in Fortaleza, in the Northeast of Brazil. More than 400 children and teenagers aging 11 and 16 years participated in this study. Poor communities experience many particular difficulties in the urban centers such as violence, poor housing conditions, unemployment, lack in health care and deficient physical education in school. Physical education, physiotherapy, odontology, medicine and pharmacy students are responsible for the activities in the project supervised by a general coordinator and a counselor teacher of each academic unit. There are classes about team sports like basketball and soccer. Lectures about sexual behavior and sexually transmitted diseases are ministered beside the ones about oral health education, basic life support education, first aids, use and care with pharmaceuticals and orientations about healthy nutrition. In order to get the children’s family closer, monthly informative lectures are ministered. There is also the concern about reflecting the actions and producing academic paperwork such as graduation final projects and books. The number of participants has oscillated lately, and one of the causes is the lack of practicing physical activities and sports regularly. However, 250 teenagers have participated regularly for at least two years. These teenagers have shown a healthier lifestyle and a better physical fitness profile. The resources for maintaining the project come from the Pro-Reitoria of Extension, Federal University of Ceara, as well as from the PROEXT/MEC, Federal Government. Actions of this nature need to be done thinking for long periods so the effects results can become effective. Public and private investments are needed due to low socioeconomic families who are most vulnerable and have fewer opportunities to enhance to health prevention services.

Keywords: children and teenagers, health, multidisciplinary program, quality of life

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Authors: Avijit Dey, Shyam S. Paul, Satbir S. Dahiya, Balbir S. Punia, Luciano A. Gonzalez

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Methane emitted from ruminant livestock not only reduces the efficiency of feed energy utilization but also contributes to global warming. Vegetable oils, a source of poly unsaturated fatty acids, have potential to reduce methane production and increase conjugated linoleic acid in the rumen. However, characteristics of oils, level of inclusion and composition of basal diet influences their efficacy. Therefore, this study was aimed to investigate the effects of sunflower (SFL) and cottonseed (CSL) oils on methanogenesis, volatile fatty acids composition and feed fermentation pattern by in vitro gas production (IVGP) test. Four concentrations (0, 0.1, 0.2 and 0.4ml /30ml buffered rumen fluid) of each oil were used. Fresh rumen fluid was collected before morning feeding from two rumen cannulated buffalo steers fed a mixed ration. In vitro incubation was carried out with sorghum hay (200 ± 5 mg) as substrate in 100 ml calibrated glass syringes following standard IVGP protocol. After 24h incubation, gas production was recorded by displacement of piston. Methane in the gas phase and volatile fatty acids in the fermentation medium were estimated by gas chromatography. Addition of oils resulted in increase (p<0.05) in total gas production and decrease (p<0.05) in methane production, irrespective of type and concentration. Although the increase in gas production was similar, methane production (ml/g DM) and its concentration (%) in head space gas was lower (p< 0.01) in CSL than in SFL at corresponding doses. Linear decrease (p<0.001) in degradability of DM was evident with increasing doses of oils (0.2ml onwards). However, these effects were more pronounced with SFL. Acetate production tended to decrease but propionate and butyrate production increased (p<0.05) with addition of oils, irrespective of type and doses. The ratio of acetate to propionate was reduced (p<0.01) with addition of oils but no difference between the oils was noted. It is concluded that both the oils can reduce methane production. However, feed degradability was also affected with higher doses. Cotton seed oil in small dose (0.1ml/30 ml buffered rumen fluid) exerted greater inhibitory effects on methane production without impeding dry matter degradability. Further in vivo studies need to be carried out for their practical application in animal ration.

Keywords: buffalo, methanogenesis, rumen fermentation, vegetable oils

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Authors: Amna Imtiaz

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Aims: To correlate serum ferritin with left ventricular function in beta thalassemia major patients with increased transfusion dependence and to find out whether echocardiography can be used to assess pre clinical cardiac disease in these patients. Methods: The cross sectional study was conducted at Department of Pathology, Shaheed Zulfiqar Ali Bhutto Medical University, Pakistan Institute of Medical Sciences, Islamabad. 60 patients of beta thalassemia major with increased transfusion dependence were enrolled in this study. Serum ferritin levels of all patients were measured by using indirect enzyme linked immunosorbent assay (ELISA). Echocardiography was performed on all patients by a consultant cardiologist by linking conventional echocardiography with tissue Doppler imaging. Ejection fraction and E/A ratio were measured in all patients to assess left ventricular systolic and diastolic function. Results: On the basis of serum ferritin level, patients were divided into three groups. Group I consisted of patients having serum ferritin level equal to or less than 2500 ng/ml. A total of 25 patients were placed in this group. Group II included patients having serum ferritin level between 2500 to 5000 ng/ml. A total of 22 patients were placed in this group. Group III included patients having serum ferritin level more than 5000 ng/ml. This group consisted of 13 patients. All patients having serum ferritin below 2500ng/ml had normal systolic function, and only 16% of the patients in this group had diastolic dysfunction as reflected by abnormal E/A ratio. In group II, 27% of the patients had systolic dysfunction reflected by subnormal ejection fraction while 40% of the patients had diastolic dysfunction. In group III, 62% of the patients had abnormal systolic and diastolic function. Pearson correlation was used to find a correlation between serum ferritin and left ventricular function. A strong negative correlation was found which is reflected by a p value of less than 0.05 which is significant. Chi square test is used to correlate serum ferritin with E/A ratio. P value came out to be less than 0.05 which is significant.

Keywords: beta thalassemia major, left ventricular function, serum ferritin, transfusion dependence

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Authors: Elisa K. Schoenell, Cristiano De Oliveira, Luiz R. H. Dos Santos, Alexandre Giacobbo, Andréa M. Bernardes, Marco A. S. Rodrigues

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Water is an indispensable natural resource, which must be preserved to human activities as well the ecosystems. However, the sewage discharge has been contaminating water resources. Conventional treatment, such as physicochemical treatment followed by biological processes, has not been efficient to the complete degradation of persistent organic compounds, such as medicines and hormones. Therefore, the use of advanced technologies to sewage treatment has become urgent and necessary. The aim of this study was to apply Reverse Osmosis (RO) on sewage tertiary treatment from a Waste Water Treatment Plant (WWTP) in south Brazil. It was collected 200 L of sewage pre-treated by wetland with aquatic macrophytes. The sewage was treated in a RO pilot plant, using a polyamide membrane BW30-4040 model (DOW FILMTEC), with 7.2 m² membrane area. In order to avoid damage to the equipment, this system contains a pleated polyester filter with 5 µm pore size. It was applied 8 bar until achieve 5 times of concentration, obtaining 80% of recovery of permeate, with 10 L.min-1 of concentrate flow rate. Samples of sewage pre-treated on WWTP, permeate and concentrate generated on RO was analyzed for physicochemical parameters and by gas chromatography (GC) to qualitative analysis of organic compounds. The results proved that the sewage treated on WWTP does not comply with the limit of phosphorus and nitrogen of Brazilian legislation. Besides this, it was found many organic compounds in this sewage, such as benzene, which is carcinogenic. Analyzing permeate results, it was verified that the RO as sewage tertiary treatment was efficient to remove of physicochemical parameters, achieving 100% of iron, copper, zinc and phosphorus removal, 98% of color removal, 91% of BOD and 62% of ammoniacal nitrogen. RO was capable of removing organic compounds, however, it was verified the presence of some organic compounds on de RO permeate, showing that RO did not have the capacity of removal all organic compounds of sewage. It has to be considered that permeate showed lower intensity of peaks in chromatogram in comparison to the sewage of WWTP. It is important to note that the concentrate generate on RO needs a treatment before its disposal in environment.

Keywords: organic compounds, reverse osmosis, sewage treatment, tertiary treatment

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Authors: Suveen Kumar

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Oral cancer (OC) is the sixth most death causing cancer in world which includes tumour of lips, floor of the mouth, tongue, palate, cheeks, sinuses, throat, etc. Conventionally, the techniques used for OC detection are toluidine blue staining, biopsy, liquid-based cytology, visual attachments, etc., however these are limited by their highly invasive nature, low sensitivity, time consumption, sophisticated instrument handling, sample processing and high cost. Therefore, we developed biosensing chips for non-invasive detection of OC via CYFRA-21-1 biomarker. CYFRA-21-1 (molecular weight: 40 kDa) is secreted in saliva of OC patients which is a non-invasive biological fluid with a cut-off value of 3.8 ng mL-1, above which the subjects will be suffering from oral cancer. Therefore, in first work, 3-aminopropyl triethoxy silane (APTES) functionalized zirconia (ZrO2) nanoparticles (APTES/nZrO2) were used to successfully detect CYFRA-21-1 in a linear detection range (LDR) of 2-16 ng mL-1 with sensitivity of 2.2 µA mL ng-1. Successively, APTES/nZrO2-RGO was employed to prevent agglomeration of ZrO2 by providing high surface area reduced graphene oxide (RGO) support and much wider LDR (2-22 ng mL-1) was obtained with remarkable limit of detection (LOD) as 0.12 ng mL-1. Further, APTES/nY2O3/ITO platform was used for oral cancer bioseneor development. The developed biosensor (BSA/anti-CYFRA-21-1/APTES/nY2O3/ITO) have wider LDR (0.01-50 ng mL-1) with remarkable limit of detection (LOD) as 0.01 ng mL-1. To improve the sensitivity of the biosensing platform, nanocomposite of yattria stabilized nanostructured zirconia-reduced graphene oxide (nYZR) based biosensor has been developed. The developed biosensing chip having ability to detect CYFRA-21-1 biomolecules in the range of 0.01-50 ng mL-1, LOD of 7.2 pg mL-1 with sensitivity of 200 µA mL ng-1. Further, the applicability of the fabricated biosensing chips were also checked through real sample (saliva) analysis of OC patients and the obtained results showed good correlation with the standard protein detection enzyme linked immunosorbent assay (ELISA) technique.

Keywords: non-invasive, oral cancer, nanomaterials, biosensor, biochip

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Authors: Mauricio A. Sarabia-Vallejos, Carmen M. Gonzalez-Henriquez, Adolfo Del Campo-Garcia, Aitzibier L. Cortajarena, Juan Rodriguez-Hernandez

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In this study, it was explored the formation and the morphological differences between wrinkled hydrogel patterns obtained via generation of surface instabilities. The slight variations in the polymerization conditions produce important changes in the material composition and pattern structuration. The compounds were synthesized using three main components, i.e. an amphiphilic monomer, hydroxyethyl methacrylate (HEMA), a hydrophobic monomer, trifluoroethyl methacrylate (TFMA), and a hydrophilic crosslinking agent, poly(ethylene glycol) diacrylate (PEGDA). The first part of this study was related to the formation of wrinkled surfaces using only HEMA and PEGDA and varying the amount of water added in the reaction. The second part of this study involves the gradual insertion of TFMA into the hydrophilic reaction mixture. Interestingly, the manipulation of the chemical composition of this hydrogel affects both surface morphology and physicochemical characteristics of the patterns, inducing transitions from one particular type of structure (wrinkles or ripples) to different ones (creases, folds, and crumples). Contact angle measurements show that the insertion of TFMA produces a slight decrease in surface wettability of the samples, remaining however highly hydrophilic (contact angle below 45°). More interestingly, by using confocal Raman spectroscopy, important information about the wrinkle formation mechanism is obtained. The procedure involving two consecutive thermal and photopolymerization steps lead to a “pseudo” two-layer system. Thus, upon photopolymerization, the surface is crosslinked to a higher extent than the bulk and water evaporation drives the formation of wrinkled surfaces. Finally, cellular, and bacterial proliferation studies were performed to the samples, showing that the amount of TFMA included in each sample slightly affects the proliferation of both (bacteria and cells), but in the case of bacteria, the morphology of the sample also plays an important role, importantly reducing the bacterial proliferation.

Keywords: antibiofouling properties, hydrophobic/hydrophilic balance, morphologic characterization, wrinkled hydrogel patterns

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Authors: Rozeena Baig, R. Rehana Rehman, Rifat Ahmed

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Background: Osteoporosis, characterized by low bone mineral density, poses a global health concern. Diagnosis increases the likelihood of developing osteoporosis, a multifactorial disorder marked by low bone mass, elevating the risk of fractures in the lumbar spine, femoral neck, hip, vertebras, and distal forearm, particularly in postmenopausal women due to bone loss influenced by various pathophysiological factors. Objectives: The aim is to investigate the association of serum cytokine, bone turnover marker, bone mineral density and TLR4 gene polymorphism in pre and post-menopausal women and to find if any of these can be the potential predictor of osteoporosis in postmenopausal women. Material and methods: The study participants consisted of Group A (n=91) healthy pre-menopausal women and Group B (n=102) healthy postmenopausal women having ≥ 5 years’ history of menopause. ELISA was performed for cytokine (TNFα) and bone turnover markers (carboxytelopeptides), respectively. Bone Mineral Density (BMD)was measured through a dual X-ray absorptiometry (DEXA) scan. Toll-like Receptors 4 (TLR4) gene polymorphisms (A896G; Asp299Gly) and (C1196T; Thr399Ile) were investigated by PCR and Sanger sequencing. Results: Statistical analysis reveals a positive correlation of age and BMI with T scores in the premenopausal group, whereas in post-menopausal group found a significant negative correlation between age and T-score at hip (r = - 0.352**), spine (r = - .306**), and femoral neck (r = - 0.344**) and a significant negative correlation of BMI with TNF-α (- 0.316**). No association and significant differences were observed for TLR4 genotype and allele frequencies among studied groups However, both SNPs exhibited significant association with each other. Conclusions: This study concludes that BMI, BMD and TNF-α are the potential predictors of osteoporosis in post-menopausal women. However, CTX and TLR4 gene polymorphism did not appear as potential predictors of bone loss in this study and apparently cannot help in predicting bone loss in post-menopausal women.

Keywords: osteoporosis, post-menopausal, pre-menopausal woemn, genetics mutaiont, TLR4 genepolymorphsum

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Authors: Yusuf E. Mustapha, Inioluwa A Akindoyeni, Oluwatoyin G. Ezekiel, Ifeoluwa O. Awogbindin, Ebenezer O. Farombi

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Background: Parkinson's disease (PD) is the most prevalent motor disorder. Available therapies are palliative with no effect on disease progression. Kolaviron (KV), a natural anti-inflammatory and antioxidant agent, has been reported to possess neuroprotective effects in Parkinsonian flies and rats. Objective: The present study investigates the neuroprotective effect of KV, focusing on the DJ1/Nrf2 signaling pathway. Methodology: All-trans retinoic acid (ATRA, 10 mg/kg, i.p.) was used to inhibit Nrf2. Murine model of PD was established with four doses of MPTP (20 mg/kg i.p.) at 2 hours interval. MPTP mice were pre-treated with either KV (200 mg/kg/day p.o), ATRA, or both conditions for seven days before PD induction. Motor behaviour was evaluated, and markers of oxidative stress/damage and its regulators were assessed with immunofluorescence and ELISA techniques. Results: MPTP-treated mice covered less distance with reduced numbers of anticlockwise rotations, heightened freezing, and prolonged immobility when compared to control. However, KV significantly attenuated these deficits. Pretreatment of MPTP mice with KV upregulated Nrf2 expression beyond MPTP level with a remarkable reduction in Keap1 expression and marked elevation of DJ-1 level, whereas co-administration with ATRA abrogated these effects. KV treatment restored MPTP-mediated depletion of endogenous antioxidant, striatal oxidative stress, oxidative damage, and inhibition of acetylcholinesterase activity. However, ATRA treatment potentiated acetylcholinesterase inhibition and attenuated the protective effect of KV on the level of nitric oxide and activities of catalase and superoxide dismutase. Conclusion: Kolaviron protects Parkinsonian mice by stabilizing and activating the Nrf2 signaling pathway. Thus, kolaviron can be explored as a pharmacological lead in PD management.

Keywords: Garcinia kola, Kolaviron, Parkinson Disease, Nrf2, behavioral incompetence, neurodegeneration

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Authors: Zeb Hussain, Muhammad Asif Qureshi, Shaheen Sharafat

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Background: Measles is a contagious viral infection common in childhood. Vaccination against measles is included in the expanded program of immunization (EPI). However, and alarmingly, a high mortality rate is observed due to measles infection in Pakistan. Moreover a recent outbreak of measles in various areas of Pakistan further highlights the problem. It is therefore important to investigate measles specific IgG (antibody) levels in our population. Objective: To quantify measles specific IgG antibodies amongst vaccinated children in district Qamber Shahdadkot, Sindh. Methodology: This cross-sectional study was conducted at the Microbiology section of the Dow-Diagnostic-Research-and-Reference-Laboratory (DDRRL), DUHS after Institutional Review Board approval (IRB-516/DUHS/-14) during August-December-2014. A total of 173 participants (residents of district Qamber Shahdadkot, Sindh) aged between 1-5 years were recruited in the study. Blood samples were collected as per standard phlebotomy guidelines. Blood was stored at 4 °C overnight. Samples were subsequently spun at a speed of 10000rpm to separate sera, which were divided into small aliquots to be frozen at -20 °C. Frozen sera were transported to the DDRRL on dry ice. Measles specific IgG (antibody) titers were quantified using enzyme linked immunosorbant assay (ELISA). Results: Blood was collected from a total of 173 individuals ranging between 1-5 years of age. Of these, a total of 88 participants were males and 85 were females. Of the 173 investigated samples, only 53 (30.6%) showed protective IgG titers against measles while 120 (69%) were sero-negative. Measles specific IgG antibodies titers were higher in female participants compared to the males. Conclusion: Our data demonstrate that a substantial percentage of vaccinated children in district Qamber-Shahdadkot did not have protective antibody titres against measles. It is therefore extremely important to investigate measles specific IgG levels in various parts of Pakistan in order to implement appropriate protective measures.

Keywords: sero-surveillance, measles, vaccinated children, Pakistan

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Authors: Q. B. Maria de la Luz Ortega Juárez, Saúl Rojas Hernández, Itzel Berenice Rodríguez Mera, María Maricela Carrasco Yépez, Mara Gutierrez Sánchez

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Naegleria fowleri is a free-living amoeba found mainly in temperate freshwater and is the etiologic agent of primary amebic meningoencephalitis (PAM), a fatal acute disease with a mortality rate greater than 95%. At present, there are no treatments available for MAP, and the development of effective vaccines that generate long-term immunological memory allowing protection against MAP would be of great importance. The objective of this work was to analyze the effector and memory immune response in BALB/c mice immunized with total extract of N. fowleri co-administered with cholera toxin. In this study, BALB/c mice were immunized four times intranasally with ET of N. fowleri adjuvanted with CT with or without booster at three months and were challenged or not with the lethal dose of N. fowleri, determining survival, the humoral, effector and memory response, by ELISA and flow cytometry techniques. The results obtained showed that the survival of mice immunized with booster had 60% protection compared to the group without booster, which obtained 20% protection. Evaluating the humoral response, it was found that both IgG and IgA levels were higher in sera than in nasal washes in both treatments. In the cellular response, the increase in the percentage of positive cells was found for effector T and B lymphocytes in the nasal passages (NP) in the group with boost and nasopharynx-associated lymphoid tissue (NALT) in the group without boost and lymphocytes only. B in both treatments, as well as in memory cells treatment with boost T lymphocytes in PN and NALT and without boost in cervical lymph nodes (CG) with respect to B lymphocytes, in PN, GC and NALT in treatment with boost and NALT in treatment without booster. Therefore, the involvement of the effector immune response and memory play a fundamental role for protection against N. fowleri and for the development of vaccine candidates.

Keywords: immune response, immunological memory, naegleria fowleri, primary amebic meningoencephalitis

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Authors: Joseph L. Mathew, Shourjendra N. Banerjee, R. K. Ratho, Sourabh Dutta, Vanita Suri

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Background: In India and many other developing countries, a single dose of measles vaccine is administered to infants at 9 months of age. This is based on the assumption that maternal transplacentally transferred antibodies will protect infants until that age. However, our previous data showed that most infants lose maternal anti-measles antibodies before 6 months of age, making them susceptible to measles before vaccination at 9 months. Objective: This prospective study was designed to compare susceptibility in pre-term vs term infants, at different time points. Material and Methods: Following Institutional Ethics Committee approval and a formal informed consent process, venous blood was drawn from a cohort of 45 consecutive term infants and 45 consecutive pre-term infants (both groups delivered by the vaginal route); at birth, 3 months, 6 months and 9 months (prior to measles vaccination). Serum was separated and anti-measles IgG antibody levels were measured by quantitative ELISA kits (with sensitivity and specificity > 95%). Susceptibility to measles was defined as antibody titre < 200mIU/ml. The mean antibody levels were compared between the two groups at the four time points. Results: The mean gestation of term babies was 38.5±1.2 weeks; and pre-term babies 34.7±2.8 weeks. The respective mean birth weights were 2655±215g and 1985±175g. Reliable maternal vaccination record was available in only 7 of the 90 mothers. Mean anti-measles IgG antibody (±SD) in terms babies was 3165±533 IU/ml at birth, 1074±272 IU/ml at 3 months, 314±153 IU/ml at 6 months, and 68±21 IU/ml at 9 months. The corresponding levels in pre-term babies were 2875±612 IU/ml, 948±377 IU/ml, 265±98 IU/ml, and 72±33 IU/ml at 9 months (p > 0.05 for all inter-group comparisons). The proportion of susceptible term infants at birth, 3months, 6months and 9months was 0%, 16%, 67% and 96%. The corresponding proportions in the pre-term infants were 0%, 29%, 82%, and 100% (p > 0.05 for all inter-group comparisons). Conclusion: Majority of infants are susceptible to measles before 9 months of age suggesting the need to anticipate measles vaccination, but there was no statistically significant difference between the proportion of susceptible term and pre-term infants, at any of the four-time points. A larger study is required to confirm these findings and compare sero-protection if vaccination is anticipated to be administered between 6 and 9 months.

Keywords: measles, preterm, susceptibility, term infant

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