Search results for: mutation screening

Authors: Charmi Chande, Ravindra Phadke

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Microfluidics devices are fabricated by using multiple fabrication methods. Photolithography is one of the common methods wherein SU8 is widely used for making master which in turn is used for making working chip by the process of soft lithography. The high-aspect ratio features of SU-8 makes it suitable to be used as micro moulds for injection moulding, hot embossing, and moulds to form polydimethylsiloxane (PDMS) structures for bioMEMS (Microelectromechanical systems) applications. But due to high cost, difficulty in procuring and need for clean room, restricts the use of this polymer especially in developing countries and small research labs. ‘Bisphenol –A’ based polymers in mixture with curing agent are used in various industries like Paints and coatings, Adhesives, Electrical systems and electronics, Industrial tooling and composites. We present the novel use of ‘Bisphenol – A’ based polymer in fabricating micro channels for Lab On Chip(LOC) devices. The present paper describes the prototype for production of microfluidics chips using range of ‘Bisphenol-A’ based polymers viz. GY 250, ATUL B11, DER 331, DER 330 in mixture with cationic photo initiators. All the steps of chip production were carried out using an inexpensive approach that uses low cost chemicals and equipment. This even excludes the need of clean room. The produced chips using all above mentioned polymers were validated with respect to height and the chip giving least height was selected for further experimentation. The lowest height achieved was 7 micrometers by GY250. The cost of the master fabricated was $ 0.20 and working chip was $. 0.22. The best working chip was used for morphological identification and profiling of microorganisms from environmental samples like soil, marine water and salt water pan sites. The current chip can be adapted for various microbiological screening experiments like biochemical based microbial identification, studying uncultivable microorganisms at single cell/community level.

Keywords: bisphenol–A based epoxy, cationic photoinitiators, microfabrication, photolithography

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Authors: I. Wlasny, Z. Klusek, A. Wysmolek

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Hexagonal boron nitride is a representative of a widely popular class of two-dimensional Van Der Waals materials. It finds its uses, among others, in construction of complexly layered heterostructures. Hexagonal boron nitride attracts great interest because of its properties characteristic for wide-gap semiconductors as well as an ultra-flat surface.Van Der Waals heterostructures composed of two-dimensional layered materials, such as transition metal dichalcogenides or graphene give hope for miniaturization of various electronic and optoelectronic elements. In our presentation, we will show the results of our investigations of the not previously reported modification of the hexagonal boron nitride layers with focused laser beam. The electrostatic force microscopy (EFM) images reveal that the irradiation leads to changes of the local electric fields for a wide range of laser wavelengths (from 442 to 785 nm). These changes are also accompanied by alterations of crystallographic structure of the material, as reflected by Raman spectra. They exhibit high stability and remain visible after at least five months. This behavior can be explained in terms of photoionization of the defect centers in h-BN which influence non-uniform electrostatic field screening by the photo-excited charge carriers. Analyzed changes influence local defect structure, and thus the interatomic distances within the lattice. These effects can be amplified by the piezoelectric character of hexagonal boron nitride, similar to that found in nitrides (e.g., GaN, AlN). Our results shed new light on the optical properties of the hexagonal boron nitride, in particular, those associated with electron-phonon coupling. Our study also opens new possibilities for h-BN applications in layered heterostructures where electrostatic fields can be used in tailoring of the local properties of the structures for use in micro- and nanoelectronics or field-controlled memory storage. This work is supported by National Science Centre project granted on the basis of the decision number DEC-2015/16/S/ST3/00451.

Keywords: atomic force microscopy, hexagonal boron nitride, optical properties, raman spectroscopy

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Authors: A. Badran, M. Hollocks, H. Markus

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Background: Common risk factors for cardiovascular disease are associated with age-related cognitive decline. There has been much interest in treating modifiable cardiovascular risk factors in the hope of reducing cognitive decline. However, there is currently no validated neuropsychological test to assess the subclinical cognitive effects of vascular risk. The Brief Memory and Executive Test (BMET) is a clinical screening tool, which was originally designed to be sensitive and specific to Vascular Cognitive Impairment (VCI), an impairment characterised by decline in frontally-mediated cognitive functions (e.g. Executive Function and Processing Speed). Objective: To cross-sectionally assess the validity of the BMET as a measure of the subclinical effects of vascular risk on cognition, in an otherwise healthy elderly cohort. Methods: Data from 346 participants (57 ± 10 years) without major neurological or psychiatric disorders were included in this study, gathered as part of a previous multicentre validation study for the BMET. Framingham Vascular Age was used as a surrogate measure of vascular risk, incorporating several established risk factors. Principal Components Analysis of the subtests was used to produce common constructs: an index for Memory and another for Executive Function/Processing Speed. Univariate General Linear models were used to relate Vascular Age to performance on Executive Function/Processing Speed and Memory subtests of the BMET, adjusting for Age, Premorbid Intelligence and Ethnicity. Results: Adverse vascular risk was associated with poorer performance on both the Memory and Executive Function/Processing Speed indices, adjusted for Age, Premorbid Intelligence and Ethnicity (p=0.011 and p<0.001, respectively). Conclusions: Performance on the BMET reflects the subclinical effects of vascular risk on cognition, in age-related cognitive decline. Vascular risk is associated with decline in both Executive Function/Processing Speed and Memory groups of subtests. Future studies are needed to explore whether treating vascular risk factors can effectively reduce age-related cognitive decline.

Keywords: age-related cognitive decline, vascular cognitive impairment, subclinical cerebrovascular disease, cognitive aging

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Authors: Jose Carlos Tavares Carvalho, Hady Keita, Giovanna Rocha Santana, Igor Victor Ferreira Dos Santos, Jesus Rafael Rodriguez Amado, Ariadna Lafourcade Prada, Adriana Maciel Ferreira, Helison Oliveira

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Summary: As animal model, zebrafish can be a good opportunity to establish a profile of tissue alteration caused by Bothrops alternatus venom and to screen new anti-venom drugs. Objective: To establish tissue biomarkers from zebrafish injected by snake venom and elucidate the use of glucocorticoids in ophidic accidents. Materials and Methods: The Danio rerio fish were randomly divided into four groups: control group, venom group, Dexamethasone1h before venom injected group and Dexamethasone 1 h after venom injected group. The concentration of Bothrops alternatus venom was 0.13 mg/ml and the fish received 20µl/Fish. The Body weight measurement and histological characteristics of gills, kidneys, liver, and intestine were determinate. Results: Physical analysis shows necrosis accompanied by inflammation in animals receiving the Bothrops alternatus venom. Significant difference was observed in the variation of weight between the control group, and the groups received the venom (t student test, p < 0.05). The average histological alterations index of gill, liver, kidney or intestine was statistically higher in animals received the venom (t Student test, p < 0.05). The alterations were lower in the groups that received Dexamethasone 1h before and after venom injected compared to the group that received only the venom. Dexamethasone 1h before venom injected group had minor histopathological alterations. Conclusion: The organs of zebrafish may be a tissue biomarker of alterations from Bothrops alternatus venom and dexamethasone reduced the damage caused by this venom in the organs studied, which may suggest the use of zebrafish as animal model for research related to screening new drug against snake venom.

Keywords: zebrafish, snake venom, biomarker, drugs

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Authors: Christian Fercher, Jiaul Islam, Simon R. Corrie

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Fluorescence-based immunodiagnostics are an emerging field in biosensor development and exhibit several advantages over traditional detection methods. While various affinity biosensors have been developed to generate a fluorescence signal upon sensing varying concentrations of analytes, reagentless, reversible, and continuous monitoring of complex biological samples remains challenging. Here, we aimed to genetically engineer biosensors based on single-chain antibody fragments (scFv) that are site-specifically labeled with environmentally sensitive fluorescent unnatural amino acids (UAA). A rational design approach resulted in quantifiable analyte-dependent changes in peak fluorescence emission wavelength and enabled antigen detection in vitro. Incorporation of a polarity indicator within the topological neighborhood of the antigen-binding interface generated a titratable wavelength blueshift with nanomolar detection limits. In order to ensure continuous analyte monitoring, scFv candidates with fast binding and dissociation kinetics were selected from a genetic library employing a high-throughput phage display and affinity screening approach. Initial rankings were further refined towards rapid dissociation kinetics using bio-layer interferometry (BLI) and surface plasmon resonance (SPR). The most promising candidates were expressed, purified to homogeneity, and tested for their potential to detect biomarkers in a continuous microfluidic-based assay. Variations of dissociation kinetics within an order of magnitude were achieved without compromising the specificity of the antibody fragments. This approach is generally applicable to numerous antibody/antigen combinations and currently awaits integration in a wide range of assay platforms for one-step protein quantification.

Keywords: antibody engineering, biosensor, phage display, unnatural amino acids

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Authors: Tong Ming Liu

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Human mesenchymal stem cells (MSCs) represent the most used stem cells for clinical application, which have been used in over 1000 clinical trials to treat over 30 diseases due to multilineage differentiation potential, secretome and immunosuppression. Gene therapies of MSCs hold great promise in the treatment of many diseases due to enhanced MSC-based clinical outcomes. To identify genes for gene therapy of MSCs, by comparing gene expression profile before and after MSC differentiation following by functional screening, we have identified ZNF145 that regulated MSC differentiation. Forced expression of ZNF145 resulted in enhanced in vitro chondrogenesis of MSCs as an upstream factor of SOX9 and improved osteochondral repair upon implant into osteochondral defects in rodents. By comparing gene expression profile during differentiation of iPSCs toward MSCs, we also identified gene HOX regulating MSC stemness, which was much downregulated in late-passaged MSCs. Knockdown of this gene greatly compromised MSC stemness including abolished proliferation, decreased CFU-F, promoted senescence and reduced expression of cell surface antigens linked to the MSC phenotype. In addition, multi-linage differentiation was also greatly impaired. Notably, HOX overexpression resulted in improved multi-lineage differentiation. In the mechanism, HOX expression significantly deceased in late passage of MSCs compared with early passage of MSCs, correlating with MSC important genes. ChIP-seq data shown that HOX binds to genes related to MSC self-renewal and differentiation. Most importantly, most HOX binding sites are lost in late passage of MSCs. HOX exerts its effects by directing binding Twist1, one important gene of MSCs. The identification of the genes regulating MSC differentiation and stemness will provide and promising strategies for gene therapy of MSCs in regenerative medicine.

Keywords: mesenchymal stem cell, novel transcription factor, stemness, gene therapy, cartilage repair, signaling pathway

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Authors: Marija Ratkova Manovska, Mirko Prodanov, Dean Jankuloski, Katerina Blagoevska

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Staphylococci, which are widely found in the environment, animals, humans, and food products, include Staphylococcus aureus (S. aureus), the most significant pathogenic species in this genus. The virulence and toxicity of S. aureus are primarily attributed to the presence of specific genes responsible for producing toxins, biofilms, invasive components, and antibiotic resistance. Staphylococcal food poisoning, caused by the production of staphylococcal enterotoxins (SEs) by these strains in food, is a common occurrence. Globally, S. aureus food intoxications are typically ranked as the third or fourth most prevalent foodborne intoxications. For this study, a total of 333 milk samples and 1160 dairy product samples were analyzed between 2016 and 2020. The strains were isolated and confirmed using the ISO 6888-1:1999 "Horizontal method for enumeration of coagulase-positive staphylococci." Molecular analysis of the isolates, conducted using conventional PCR, involved detecting the 23s gene of S. aureus, the nuc gene, the mecA gene, and 11 genes responsible for producing enterotoxins (sea, seb, sec, sed, see, seg, seh, sei, ser, sej, and sep). The 23s gene was found in 93 (75.6%) out of 123 isolates of Staphylococcus spp. obtained from milk. Among the 76 isolates from dairy products, either S. aureus or the 23s gene was detected in 49 (64.5%) of them. The mecA gene was identified in three isolates from raw milk and five isolates from cheese samples. The nuc gene was present in 98.9% of S. aureus strains from milk and 97.9% from dairy products. Other Staphylococcus strains carried the nuc gene in 26.7% of milk strains and 14.8% of dairy product strains. Genes associated with SEs production were detected in 85 (69.1%) strains from milk and 38 (50%) strains from dairy products. In this study, 10 out of the 11 SEs genes were found, with no isolates carrying the see gene. The most prevalent genes detected were seg and sei, with some isolates containing up to five different SEs genes. These findings indicate the presence of enterotoxigenic staphylococci strains in the tested samples, emphasizing the importance of implementing proper sanitation and hygienic practices, utilizing safe raw materials, and ensuring adequate handling of finished products. Continued monitoring for the presence of SEs is necessary to ensure food safety and prevent intoxication.

Keywords: dairy products, milk, Staphylococci, enterotoxins, SE genes

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Authors: Si Jack Chong, Chew Theng Yap, Wan Loong James Mok

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Introduction: Regional burn centres face the spectra of introduced multi-drug resistant organisms (MDRO) from transfer patients resident in MDRO endemic countries. MDRO can cause severe nosocomial infection, which in massive burn patients, will lead to greater morbidity and mortality and strain the institution financially. We aim to highlight 4 key measures that have effectively prevented transmission of imported MDRO. Methods: A case of Candida auris (C. auris) from a massive burn patient transferred from an MDRO endemic country is used to illustrate the measures. C. auris is a globally emerging multi-drug resistant fungal pathogen causing nosocomial transmission. Results: Infection control measures used to mitigate the risk of outbreak from transfer cases are: (1) Multidisciplinary team approach involving Infection Control and Infectious Disease specialists early to ensure appropriate antibiotics use and implementation of barrier measures, (2) aseptic procedures for dressing change with strict isolation and donning of personal protective equipment in the ward, (3) early screening of massive burn patient from MDRO endemic region, (4) hydrogen peroxide vaporization terminal cleaning for operating theatres and rooms. Conclusion: The prevalence of air travel and international transfer to regional burn centres will need effective infection control measures to reduce the risk of transmission from imported massive burn patients. In our centre, we have effectively implemented 4 measures which have reduced the risks of local contamination. We share a recent case report to illustrate successful management of a potential MDRO outbreak resulting from transfer of massive burn patient resident in an MDRO endemic area.

Keywords: burns, burn unit, cross infection, infection control

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Authors: D. Sigirli

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The Receiver Operating Characteristic (ROC) curve is a commonly used statistical tool for evaluating the diagnostic performance of screening and diagnostic test with continuous or ordinal scale results which aims to predict the presence or absence probability of a condition, usually a disease. When the test results were measured as numeric values, sensitivity and specificity can be computed across all possible threshold values which discriminate the subjects as diseased and non-diseased. There are infinite numbers of possible decision thresholds along the continuum of the test results. The ROC curve presents the trade-off between sensitivity and the 1-specificity as the threshold changes. The empirical ROC curve which is a non-parametric estimator of the ROC curve is robust and it represents data accurately. However, especially for small sample sizes, it has a problem of variability and as it is a step function there can be different false positive rates for a true positive rate value and vice versa. Besides, the estimated ROC curve being in a jagged form, since the true ROC curve is a smooth curve, it underestimates the true ROC curve. Since the true ROC curve is assumed to be smooth, several smoothing methods have been explored to smooth a ROC curve. These include using kernel estimates, using log-concave densities, to fit parameters for the specified density function to the data with the maximum-likelihood fitting of univariate distributions or to create a probability distribution by fitting the specified distribution to the data nd using smooth versions of the empirical distribution functions. In the present paper, we aimed to propose a smooth ROC curve estimation based on the boundary corrected kernel function and to compare the performances of ROC curve smoothing methods for the diagnostic test results coming from different distributions in different sample sizes. We performed simulation study to compare the performances of different methods for different scenarios with 1000 repetitions. It is seen that the performance of the proposed method was typically better than that of the empirical ROC curve and only slightly worse compared to the binormal model when in fact the underlying samples were generated from the normal distribution.

Keywords: empirical estimator, kernel function, smoothing, receiver operating characteristic curve

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Authors: Ekaterina Sánchez Romero, Emily Gabriela Aguirre Herrera, Sandra Luz Espinoza Esquerra, Jorge García Campos

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Female, 7 months old, daughter of a mother with anemia during pregnancy, with no history of atopy in the family, since birth she presents with recurrent dermatological and gastrointestinal infections, chronically treated for recurrent diaper dermatitis. At 6 months of age, she begins with generalized pallor, hyperpigmentation in hands and feet, smooth tongue, psychomotor retardation with lack of head support, sedation, and hypoactivity. She was referred to our hospital for a fever of 38°C, severe diaper rash, and pancytopenia with HB 9.3, platelets 38000, neutrophils 0.39 MCV: 86.80 high for her age. The approach was initiated to rule out myeloproliferative syndrome, with negative immunohistochemical results of bone marrow aspirate; during her stay, she presented neurological regression, lack of sucking, and focal seizures. CT scan showed cortical atrophy. The patient was diagnosed with primary immunodeficiency due to history; gamma globulin was administered without improvement with normal results of immunoglobulins and metabolic screening. When dermatological and neurological diagnoses were ruled out as the primary cause, a nutritional factor was evaluated, and a therapeutic trial was started with the administration of vitamin B12 and zinc, presenting clinical neurological improvement and resolution of pancytopenia in 2 months. It was decided to continue outpatient management. Discussion: We present a patient with neurological, dermatological involvement, and pancytopenia, so the most common differential diagnoses in this population were ruled out. Vitamin B12 deficiency is an uncommon entity. Due to maternal and clinical history, a therapeutic trial was started resulting in an improvement. Conclusion: VitaminB12 deficiency should be considered one of the differential diagnoses in the approach to pancytopenia with megaloblastic anemia associated with dermatologic and neurologic manifestations. Early treatment can reduce irreversible damage in these patients.

Keywords: vitamin B12 deficiency, pediatrics, pancytopenia, diaper dermatitis

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Authors: Yi-Fan Lin, Peter P. Howley, Frank A. Tuyl

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Quality measurement and reporting systems are used in healthcare internationally. In Australia, the Australian Council on Healthcare Standards records and reports hundreds of clinical indicators (CIs) nationally across the healthcare system. These CIs are measures of performance in the clinical setting, and are used as a screening tool to help assess whether a standard of care is being met. Existing analysis and reporting of these CIs incorporate Bayesian methods to address sampling variation; however, such assessments are retrospective in nature, reporting upon the previous six or twelve months of data. The use of Bayesian methods within statistical process control for monitoring systems is an important pursuit to support more timely decision-making. Our research has developed and assessed a new graphical monitoring tool, similar to a control chart, based on the beta-binomial posterior predictive (BBPP) distribution to facilitate the real-time assessment of health care organizational performance via CIs. The BBPP charts have been compared with the traditional Bernoulli CUSUM (BC) chart by simulation. The more traditional “central” and “highest posterior density” (HPD) interval approaches were each considered to define the limits, and the multiple charts were compared via in-control and out-of-control average run lengths (ARLs), assuming that the parameter representing the underlying CI rate (proportion of cases with an event of interest) required estimation. Preliminary results have identified that the BBPP chart with HPD-based control limits provides better out-of-control run length performance than the central interval-based and BC charts. Further, the BC chart’s performance may be improved by using Bayesian parameter estimation of the underlying CI rate.

Keywords: average run length (ARL), bernoulli cusum (BC) chart, beta binomial posterior predictive (BBPP) distribution, clinical indicator (CI), healthcare organization (HCO), highest posterior density (HPD) interval

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Authors: Atefeh Abedini, Pegah Soltani, Arda Kiani

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Background: Sarcoidosis and Tuberculosis are both considered granulomatous chronic diseases with some similar pulmonary and extra-pulmonary manifestations. It is hypothesized that given these morphological similarities, the genome of mycobacterium could have an impact on the development of Sarcoidosis. Identifying the potential correlation of these diseases may assist in the management of sarcoidosis. Herein, we aimed to inspect the lymph node biopsy of sarcoidosis patients for the existence of the HSP-65 mycobacterium DNA sequence. Methods: This cross-sectional survey was conducted on 1188 Sarcoidosis patients without active/latent tuberculosis infection who were diagnosed in Masih Daneshvari Hospital in Tehran, Iran, from January 2020 to January 2022. Trans-bronchial needle aspiration (TBNA) was performed due to bilateral hilar lymphadenopathy to take a specimen. Results: The under-evaluated patients were mainly women (N=815 (68.6%)), none-smoker (N=1016 (85.5%)), and middle-aged (50.1 (SD=4.22)) with average angiotensin-converting enzyme (ACE) index of 75.6 (SD=6.42). Dyslipidemias (n=314 (26.4%), Hypertension (n=295 (24.8%)), Diabetes mellitus (n=131 (11.0%)), and chronic heart diseases (n=97 (8.2%)) had the highest prevalence between comorbidities. Skin lesions (n= 655 (55.1%)), ophthalmic (n=341 (28.7%)), and cardiac involvement (n=229 (19.3%)) were obtained as the most common extra-pulmonary characteristics of the patients. Amongst 1188 enrolled patients who were not afflicted with Mycobacterium tuberculosis based on smear/culture essay, clinical symptoms, and Chest x-ray screening, 121 (10.2%) cases had detectable amplified DNA for Mycobacterium Tuberculosis extracted from mediastinal lung lymph nodes. Conclusion: In this survey, the mycobacterium genome was detected in almost 1 per 10 case biopsies of sarcoidosis. The remarkable number of cases (n=1188) evaluated in this study was the strength of this study which supported the hypothesis regarding sarcoidosis and mycobacterium genome correlation. Further investigation, such as case-control surveys, is required to better clarify this association.

Keywords: mycobacterium tuberculosis, sarcoidosis, genome, DNA, trans-bronchial needle aspiration

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Authors: Mohammed Shaath

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Aims Technological advancements in teaching and learning have made significant improvements over the past decade and have been incorporated in institutions to aid the learner’s experience. This review aims to assess whether Technology Enhanced Learning (TEL) pedagogy is more effective at improving students’ attitude and knowledge retention in orthodontic training than traditional methods. Methodology The searches comprised Systematic Reviews (SRs) related to the comparison of TEL and traditional teaching methods from the following databases: PubMed, SCOPUS, Medline, and Embase. One researcher performed the screening, data extraction, and analysis and assessed the risk of bias and quality using A Measurement Tool to Assess Systematic Reviews 2 (AMSTAR-2). Kirkpatrick’s 4-level evaluation model was used to evaluate the educational values. Results A sum of 34 SRs was identified after the removal of duplications and irrelevant SRs; 4 fit the inclusion criteria. On Level 1, students showed positivity to TEL methods, although acknowledging that the harder the platforms to use, the less favourable. Nonetheless, the students still showed high levels of acceptability. Level 2 showed there is no significant overall advantage of increased knowledge when it comes to TEL methods. One SR showed that certain aspects of study within orthodontics deliver a statistical improvement with TEL. Level 3 was the least reported on. Results showed that if left without time restrictions, TEL methods may be advantageous. Level 4 shows that both methods are equally as effective, but TEL has the potential to overtake traditional methods in the future as a form of active, student-centered approach. Conclusion TEL has a high level of acceptability and potential to improve learning in orthodontics. Current reviews have potential to be improved, but the biggest aspect that needs to be addressed is the primary study, which shows a lower level of evidence and heterogeneity in their results. As it stands, the replacement of traditional methods with TEL cannot be fully supported in an evidence-based manner. The potential of TEL methods has been recognized and is already starting to show some evidence of the ability to be more effective in some aspects of learning to cater for a more technology savvy generation.

Keywords: TEL, orthodontic, teaching, traditional

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Authors: Ajmal Khan Kassi, Humayun Javed, Muhammad Asif Aziz

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Response of different aubergine cultivars against Brinjal shoot and fruit borer (Leucinodes orbonalis Guenee.) was evaluated at research farm of PMAS, Arid Agriculture University, Rawalpindi, during 2013. Field trials were conducted in randomized completed block design with four replications for the screening of five cultivars of Brinjal (Solanum melongena L) (Short Purpal, Singhnath 666, Brinjal long 6275, Round Brinjal 86602, Round Egg Plant White). Cultivar Round White Brinjal showed maximum fruit infestation (54.44%) followed by Singhnath 666 (53.19%), while minimum fruit infestation was observed in Round Brinjal 86602 (42.39%). Cultivar Short Purpal showed maximum larval population (0.43) followed by Round White Brinjal (0.39), while the minimum larval population was observed in Round Brinjal 86602 with (0.27). It was observed that Round Brinjal 86602 cultivar showed comparatively minimum (L. orbonalis) larval population per leaf. The correlation of Brinjal fruit infestation and larval population of (L. orbonalis) with the different environmental factors showed that, the average relative humidity was positively and significantly correlated with fruit infestation on cultivars average precipitation showed positive but non- significant correlation on all the cultivars except Singhnath 666 with the value of (0.79) which was positive and significant. The average temperature showed non-significant and negative correlation with Brinjal long 6275, Round Brinjal 86602 and Singhnath 666, but significant negative correlation with Short Purpal and Round White Brinjal. Maximum temperature also showed the significant and negative correlation on all the five Brinjal cultivars which were significant and highly significant. Minimum temperature showed negative correlation and not significant correlation with all the cultivars. Consequently, based on the (L. orbonalis) larval density and Brinjal fruit infestation, the Round Brinjal 86602 proved least susceptible and Short Purpal highly susceptible cultivar.

Keywords: evaluation, Brinjal (Solanum melongena L), Cultivars, L. orbonalis

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Authors: Ranjot Kaur, Om P. Katare, Anupama Sharma, Sarah R. Dennison, Kamalinder K. Singh, Bhupinder Singh

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Respiratory microbial infections being among the top leading cause of death worldwide are difficult to treat as the microbes reside deep inside the airways, where only a small fraction of drug can access after traditional oral or parenteral routes. As a result, high doses of drugs are required to maintain drug levels above minimum inhibitory concentrations (MIC) at the infection site, unfortunately leading to severe systemic side-effects. Therefore, delivering antimicrobials directly to the respiratory tract provides an attractive way out in such situations. In this context, current study embarks on the systematic development of lung lia pid-modified chitosan nanoparticles for inhalation of voriconazole. Following the principles of quality by design, the chitosan nanoparticles were prepared by ionic gelation method and further coated with major lung lipid by precipitation method. The factor screening studies were performed by fractional factorial design, followed by optimization of the nanoparticles by Box-Behnken Design. The optimized formulation has a particle size range of 170-180nm, PDI 0.3-0.4, zeta potential 14-17, entrapment efficiency 45-50% and drug loading of 3-5%. The presence of a lipid coating was confirmed by FESEM, FTIR, and X-RD. Furthermore, the nanoparticles were found to be safe upto 40µg/ml on A549 and Calu-3 cell lines. The quantitative and qualitative uptake studies also revealed the uptake of nanoparticles in lung epithelial cells. Moreover, the data from Spraytec and next-generation impactor studies confirmed the deposition of nanoparticles in lower airways. Also, the interaction of nanoparticles with DPPC monolayers signifies its biocompatibility with lungs. Overall, the study describes the methodology and potential of lipid-coated chitosan nanoparticles in futuristic inhalation nanomedicine for the management of pulmonary aspergillosis.

Keywords: dipalmitoylphosphatidylcholine, nebulization, DPPC monolayers, quality-by-design

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Authors: Marja A. Bernardo, Coreena A. Bueser, Cybele Lara R. Abad, Raul V. Destura

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Pneumocystis jirovecii pneumonia (PCP) is the most common opportunistic infection among people with HIV. Early consideration of PCP should be made even in patients whose HIV status is unknown as delay in treatment may be fatal. The use of absolute lymphocyte count (ALC) has been suggested as an alternative predictor of PCP especially in resource limited settings where PCR testing is costly or delayed. Objective: To determine whether the absolute lymphocyte count (ALC) can be used as a screening tool to predict Pneumocystis pneumonia in patients with unknown HIV status admitted at a private tertiary hospital. Methods: A retrospective cross-sectional study was conducted at a private tertiary medical center. Inpatient medical records of patients aged 18 years old and above from January 2012 to May 2014, in whom a clinical diagnosis of Pneumocystis jirovecii pneumonia was made were reviewed for inclusion. Demographic data, clinical features, hospital course, PCP PCR and HIV results were recorded. Independent t-test and chi-square analysis was used to determine any statistical difference between PCP-positive and PCP-negative groups. Mann-Whitney U-test was used for comparison of hospital stay. Results: There were no statistically significant differences in baseline characteristics between PCP positive and negative groups. While both the percent lymphocyte count (0.14 ± 0.13 vs 0.21 ± 0.16) and ALC (1160 ± 528.67 vs 1493.70 ± 988.61) were lower for the PCP-positive group, only the percent lymphocyte count reached a statistically significant difference (p= 0.067 vs p= 0.042). Conclusion: A quick determination of the ALC may be useful as an additional parameter to help screen for and diagnose pneumocystis pneumonia. In our study, the ALC of patients with PCP appear to be lower than in patients without PCP. A low ALC (e.g. below 1200) may help with the decision regarding empiric treatment. However, it should be used in conjunction with the patient’s clinical presentation, as well as other diagnostic tests. Larger, prospective studies incorporating the ALC with other clinical predictors are necessary to optimally predict those who would benefit from empiric or expedited management for potential PCP.

Keywords: Pneumocystis carinii pneumonia, Absolute Lymphocyte Count, infection, PCP

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Authors: Jaco Oosthuizen, Nerina C. Van Der Merwe

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The National Health Laboratory Services in Bloemfontien has been the diagnostic testing facility for 1830 patients for familial breast cancer since 1997. From the cohort, 540 were comprehensively screened using High-Resolution Melting Analysis or Next Generation Sequencing for the presence of point mutations and/or indels. Approximately 90% of these patients stil remain undiagnosed as they are BRCA1/2 negative. Multiplex ligation-dependent probe amplification was initially added to screen for copy number variation detection, but with the introduction of next generation sequencing in 2017, was substituted and is currently used as a confirmation assay. The aim was to investigate the viability of utilizing internationally designed copy number variation detection assays based on mostly European/Caucasian genomic data for use within a South African context. The multiplex ligation-dependent probe amplification technique is based on the hybridization and subsequent ligation of multiple probes to a targeted exon. The ligated probes are amplified using conventional polymerase chain reaction, followed by fragment analysis by means of capillary electrophoresis. The experimental design of the assay was performed according to the guidelines of MRC-Holland. For BRCA1 (P002-D1) and BRCA2 (P045-B3), both multiplex assays were validated, and results were confirmed using a secondary probe set for each gene. The next generation sequencing technique is based on target amplification via multiplex polymerase chain reaction, where after the amplicons are sequenced parallel on a semiconductor chip. Amplified read counts are visualized as relative copy numbers to determine the median of the absolute values of all pairwise differences. Various experimental parameters such as DNA quality, quantity, and signal intensity or read depth were verified using positive and negative patients previously tested internationally. DNA quality and quantity proved to be the critical factors during the verification of both assays. The quantity influenced the relative copy number frequency directly whereas the quality of the DNA and its salt concentration influenced denaturation consistency in both assays. Multiplex ligation-dependent probe amplification produced false positives due to ligation failure when ligation was inhibited due to a variant present within the ligation site. Next generation sequencing produced false positives due to read dropout when primer sequences did not meet optimal multiplex binding kinetics due to population variants in the primer binding site. The analytical sensitivity and specificity for the South African population have been proven. Verification resulted in repeatable reactions with regards to the detection of relative copy number differences. Both multiplex ligation-dependent probe amplification and next generation sequencing multiplex panels need to be optimized to accommodate South African polymorphisms present within the genetically diverse ethnic groups to reduce the false copy number variation positive rate and increase performance efficiency.

Keywords: familial breast cancer, multiplex ligation-dependent probe amplification, next generation sequencing, South Africa

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Authors: H. Hammoudi, S. Bendenia, I. Batonneau-Gener, A. Khelifa

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Applying growing zeolites is due to their intrinsic physicochemical properties: a porous structure, regular, generating a large free volume, a high specific surface area, acidic properties of interest to the origin of their activity, selectivity energy and dimensional, leading to a screening phenomenon, hence the name of molecular sieves is generally attributed to them. Most of the special properties of zeolites have been valued as direct applications such as ion exchange, adsorption, separation and catalysis. Due to their crystalline structure stable, their large pore volume and their high content of cation X zeolites are widely used in the process of adsorption and separation. The acidic properties of zeolites X and interesting selectivity conferred on them their porous structure is also have potential catalysts. The study presented in this manuscript is devoted to the chemical modification of an X zeolite by cation exchange. Ion exchange of zeolite NaX by Zn 2 + cations and / or Cu 2 + is gradually conducted by following the evolution of some of its characteristics: crystallinity by XRD, micropore volume by nitrogen adsorption. Once characterized, the different samples will be used for the adsorption of propane and propylene. Particular attention is paid thereafter, on the modeling of adsorption isotherms. In this vein, various equations of adsorption isotherms and localized mobile, some taking into account the adsorbate-adsorbate interactions, are used to describe the experimental isotherms. We also used the Toth equation, a mathematical model with three parameters whose adjustment requires nonlinear regression. The last part is dedicated to the study of acid properties of Cu (x) X, Zn (x) X and CuZn (x) X, with the adsorption-desorption of pyridine followed by IR. The effect of substitution at different rates of Na + by Cu2 + cations and / or Zn 2 +, on the crystallinity and on the textural properties was treated. Some results on the morphology of the crystallites and the thermal effects during a temperature rise, obtained by scanning electron microscopy and DTA-TGA thermal analyzer, respectively, are also reported. The acidity of our different samples was also studied. Thus, the nature and strength of each type of acidity are estimated. The evaluation of these various features will provide a comparison between Cu (x) X, Zn (x) X and CuZn (x) X. One study on adsorption of C3H8 and C3H6 in NaX, Cu (x) X , Zn (x) x and CuZn (x) x has been undertaken.

Keywords: adsorption, acidity, ion exchange, zeolite

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Authors: S. Ben-Shabat, T. Turgeman, O. Leubinski, N. Roth-Bejerano, V. Kagan-Zur, Y. Sitrit

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The ectomycorrhizal (ECM) desert truffle Terfezia boudieri produces edible fruit bodies and forms symbiosis with its host plant Helianthemum sessiliflorum (Cistaceae) in the Negev desert of Israel. The symbiosis is vital for both partners' survival under desert conditions. Under desert habitat conditions, ECMs must form symbiosis before entering the dry season. To secure a successful encounter, in the course of evolution, both partners have responded by evolving special signals exchange that facilitates recognition. Members of the Cistaceae family serve as host plants for many important truffles. Conceivably, during evolution a common molecule present in Cistaceae plants was recruited to facilitate successful encounter with ectomycorrhizas. Arbuscular vesicular fungi (AM) are promiscuous in host preferences, in contrast, ECM fungi show specificity to host plants. Accordingly, we hypothesize that H. sessiliflorum secretes a chemotrophic-signaling, which is common to plants hosting ECM fungi belonging to the Pezizales. However, thus far no signaling molecules have been identified in ECM fungi. We developed a bioassay for chemotrophic activity. Fractionation of root exudates revealed a substance with chemotrophic activity and molecular mass of 534. Following the above concept, screening the transcriptome of Terfezia, grown under chemoattraction, discovered genes showing high homology to G proteins-coupled receptors of plant pathogens involved in positive chemotaxis and chemotaxis suppression. This study aimed to identify the active molecule using analytical methods (LC-MS, NMR etc.). This should contribute to our understanding of how ECM fungi communicate with their hosts in the rhizosphere. In line with the ability of Terfezia to form also endomycorrhizal symbiosis like AM fungi, analysis of the mechanisms may likewise be applicable to AM fungi. Developing methods to manipulate fungal growth by the chemoattractant can open new ways to improve inoculation of plants.

Keywords: chemotrophic signal, Helianthemum sessiliflorum, Terfezia boudieri, ECM

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Authors: Rajesh Kumar Suman, Ipseeta Ray Mohanty, Manjusha K. Borde, Ujjawala maheswari, Y. A. Deshmukh

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Background: Metabolic syndrome encompasses cluster of risk factors for cardiovascular disease which includes abdominal obesity, dyslipidemia, hypertension, and hyperglycemia. The incidence of metabolic syndrome is on the rise globally. Objective: The present study was designed to develop a unique animal model that will mimic the pathological features seen in a large pool of individuals with diabetes and metabolic syndrome; suitable for pharmacological screening of drugs beneficial in this condition. Material and Methods: A combination of high fat diet (HFD) and low dose of streptozotocin (STZ) at 30, 35 and 40 mg/kg was used to induce metabolic syndrome co-existing with diabetes mellitus in Wistar rats. Results: The 40 mg/kg STZ produced sustained hyperglycemia and the dose was thus selected for our study to induce diabetes mellitus. Rat fed HFD (HF-DC) group showed significant (p < 0.001) increase in body weight on 4th and 7th week as compared with NC (Normal Control) group rats. However, the increase in body weight of HF-DC group rats was not sustained at the end of 10th weeks. Various components of metabolic syndrome such as dyslipidemia {(Increased Triglyceride, total Cholesterol, LDL Cholesterol and decreased HDL Cholesterol)}, diabetes mellitus (Blood Glucose, HbA1c, Serum Insulin, C-peptide), hypertension {Systolic Blood pressure (p < 0.001)} were mimicked in the developed model of metabolic syndrome co existing with diabetes mellitus. In addition significant cardiac injury as indicated by CPK-MB levels, artherogenic index, hs-CRP. The decline in hepatic function {(p < 0.01) increase in the level of SGPT (U/L)} and renal function {(increase in creatinine levels (p < 0.01)} when compared to NC group rats. The histopathological assessment confirmed presence of edema, necrosis and inflammation in Heart, Pancreas, Liver and Kidney of HFD-DC group as compared to NC. Conclusion: The present study has developed a unique rodent model of metabolic syndrome; with diabetes as an essential component.

Keywords: diabetes, metabolic syndrome, high fat diet, streptozotocin, rats

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Authors: Samar Saadeldin Abdelmotalab Omer, Ikram Mohammed Eltayeb Elsiddig, Amna Beshir Medani Ahmed, Saad Mohammed Hussein Ayoub

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Many herbal medicinal products are considered as potential hypocholesterolemic agents with encouraging safety profiles, however, only a limited amount of clinical research exists to support their efficacy. The present study was designed to compare the antihypercholesterolaemic and antioxidant activities of the crude ethanolic extracts of Citrus reticulata peel, Zingiber officinale rhizome, and Sesamum indicum seeds. Forty-five rats were used throughout the experiment, which were divided into nine groups, five rats in each as follows; normal control group (normal rats fed with standard normal diet), rats fed hypercholesterolemic diet consisting of 1% cholesterol and 10% saturated animal fat, which were further divided into eight groups; hypercholesterolemic control group (rats only fed hypercholesterolemic diet), groups 3,4,5,6,7, and 8 were given Citrus reticulata, Zingiber officinale, and Sesamum indicum ethanolic extracts at doses of (250mg/kg and 500mg/kg, respectively) orally; and group 9 rats were given atorvastatin (0.18mg/kg) orally as a reference antihypercholesterolaemic drug. Blood samples were obtained four weeks following treatment from the retro-orbital venous plexus after fasting overnight from all groups and the lipid profile (serum total cholesterol (TC), high-density-lipoprotein-cholesterol (HDL-C), low-density lipoprotein-cholesterol (LDL-C), and triglycerides levels) was measured and the risk ratio (TC/HDL-C) was assessed. The antioxidant activity of the three plant extracts was determined using DPPH free-radical assay. Results of in vivo and in vitro antihypercholesterolaemic and antioxidant assay, respectively, revealed that the three extracts possess comparable antioxidant and anti-hypercholesterolaemic activities.

Keywords: anti hypercholesterolemic effects, antioxidant activity, HDL, LDL, TC, TGs, citrus reticulata, sesamum indicum, zingiber officinale

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Authors: Sachiko Hosoya

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Objectives: The purpose of this paper is to investigate the social policy of preventive genetic medicine in Iran, by following the legalization process of abortion law and the factors affecting the process in wider Iranian contexts. In this paper, ethical discussions of prenatal diagnosis and selective abortion in Iran will be presented, by exploring Iranian social policy to control genetic diseases, especially a genetic hemoglobin disorder called Thalassemia. The ethical dilemmas in application of genetic medicine into social policy will be focused. Method: In order to examine the role of the policy for prevention of genetic diseases and selective abortion in Iran, various resources have been sutudied, not only academic articles, but also discussion in the Parliament and documents related to a court case, as well as ethnographic data on living situation of Thalassemia patients. Results: Firstly, the discussion on prenatal diagnosis and selective abortion is overviewed from the viewpoints of ethics, disability rights activists, and public policy for lower-resources countries. As a result, it should be noted that the point more important in the discussion on prenatal diagnosis and selective abortion in Iran is the allocation of medical resources. Secondly, the process of implementation of national thalassemia screening program and legalization of ‘Therapeutic Abortion Law’ is analyzed, through scrutinizing documents such as the Majlis record, government documents and related laws and regulations. Although some western academics accuse that Iranian policy of selective abortion seems to be akin to eugenic public policy, Iranian government carefully avoid to distortions of the policy as ‘eugenic’. Thirdly, as a comparative example, discussions on an Iranian court case of patient’s ‘right not to be born’ will be introduced. Along with that, restrictive living environments of people with Thalassemia patients and the carriers are depicted, to understand some disabling social factors for people with genetic diseases in the local contexts of Iran.

Keywords: abortion, Iran, prenatal diagnosis, public health ethics, Thalassemia prevention program

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Authors: Soumaya Mrabet, Taieb Ach, Imen Akkari, Amira Atig, Neirouz Ghannouchi, Koussay Ach, Elhem Ben Jazia

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Introduction: Celiac disease (CD) is an immune-mediated small intestinal disorder that occurs in genetically susceptible people. It is significantly associated with other autoimmune disorders represented mainly by type 1 diabetes and autoimmune dysthyroidism. The aim of our study is to determine the prevalence and the type of the various autoimmune diseases associated with CD in adult patients. Material and methods: This is a retrospective study including patients diagnosed with CD, explored in Internal Medicine, Gastroenterology and Endocrinology and Diabetology Departments of the Farhat Hached University Hospital, between January 2005 and January 2016. The diagnosis of CD was confirmed by serological tests and duodenal biopsy. The screening of autoimmune diseases was based on physical examination, biological and serological tests. Results: Sixty five patients with a female predominance were included, 48women (73.8%) and 17 men (26.2%). The mean age was 31.8 years (17-75). A family history of CD or other autoimmune diseases was present in 5 and 10 patients respectively. Clinical presentation of CD was made by recurrent abdominal pain in 49 cases, diarrhea in 29 cases, bloating in 17 cases, constipation in 25 cases and vomiting in 8 cases. Autoimmune diseases associated with CD were found in 30 cases (46.1%): type 1 diabetes in 15 patients attested by the positivity of anti-GAD antibodies in 11 cases and anti-IA2 in 4 cases, Hashimoto thyroiditis in 8 cases confirmed by the positivity of anti-TPO antibodies, Addison's disease in 2 patients, Anemia of Biermer in 2 patients, autoimmune hepatitis, Systemic erythematosus lupus, Gougerot Sjögren syndrome, rheumatoid arthritis, Vitiligo and antiphospholipid syndrome in one patient each. CD was associated with more than one autoimmune disease defining multiple autoimmune syndrome in 2 female patients. The first patient had Basedow disease, Addison disease and type 1 diabetes. The second patient had systemic erythematosus lupus and Gougerot Sjögren syndrome. Conclusion: In our study autoimmune diseases were associated with CD in 46.1% of cases and were dominated by diabetes and dysthroidism. After establishing the diagnosis of CD the search of associated autoimmune diseases is necessary in order to avoid any therapeutic delay which can alter the prognosis of the patient.

Keywords: association, autoimmune thyroiditis, celiac disease, diabetes

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Authors: Thomas Louis, Irina Primac, Florent Morfoisse, Tania Durre, Silvia Blacher, Agnes Noel

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In vitro and in vivo lymphangiogenesis assays are essential for the identification of potential lymphangiogenic agents and the screening of pharmacological inhibitors. In the present study, we analyse three biological models: in vitro lymphatic endothelial cell spheroids, in vivo ear sponge assay, and in vivo lymph node colonisation by tumour cells. These assays provide suitable 3D models to test pro- and anti-lymphangiogenic factors or drugs. 3D images were acquired by confocal laser scanning and light sheet fluorescence microscopy. Virtual scan microscopy followed by 3D reconstruction by image aligning methods was also used to obtain 3D images of whole large sponge and ganglion samples. 3D reconstruction, image segmentation, skeletonisation, and other image processing algorithms are described. Fixed and time-lapse imaging techniques are used to analyse lymphatic endothelial cell spheroids behaviour. The study of cell spatial distribution in spheroid models enables to detect interactions between cells and to identify invasion hierarchy and guidance patterns. Global measurements such as volume, length, and density of lymphatic vessels are measured in both in vivo models. Branching density and tortuosity evaluation are also proposed to determine structure complexity. Those properties combined with vessel spatial distribution are evaluated in order to determine lymphangiogenesis extent. Lymphatic endothelial cell invasion and lymphangiogenesis were evaluated under various experimental conditions. The comparison of these conditions enables to identify lymphangiogenic agents and to better comprehend their roles in the lymphangiogenesis process. The proposed methodology is validated by its application on the three presented models.

Keywords: 3D image segmentation, 3D image skeletonisation, cell invasion, confocal microscopy, ear sponges, light sheet microscopy, lymph nodes, lymphangiogenesis, spheroids

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Authors: Andrea Parisi, Samantha Vilkins, Luis Furuya-Kanamori, John A. Crump, Benjamin P. Howden, Darren Gray, Kathryn Glass, Martyn Kirk

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Objectives: Salmonella is a leading cause of foodborne enterocolitis worldwide. Nontyphoidal Salmonella (NTS) infections that are Multi-Drug Resistant (MDR) (non-susceptible to ≥1 agent in ≥3 antimicrobial categories) may result in more severe outcomes, although these effects have not been systematically examined. We conducted a systematic review and meta-analysis to examine impacts of MDR NTS on health in high-income settings. Methods: We systematically reviewed the literature from scientific databases, including PubMed, Scopus and grey literature sources, using PRISMA guidelines. We searched for data from case-control studies, cohorts, outbreaks, reports and theses, imposing no language restriction. We included only publications from January 1990 to September 2016 from high income countries as classified by World Bank. We extracted data from papers on duration of illness, hospitalisation rates, morbidity and mortality for MDR and non-MDR NTS strains. Results: After removing duplicates, the initial search revealed 4258 articles. After further screening, we identified 16 eligible studies for the systematic review, and 9 of these were included in meta-analysis. NTS serotypes differed among the reported studies but serotype Typhimurium, Enteritidis, Newport and Heidelberg were among the most often reported as MDR pathogens. Salmonella infections that were MDR were associated with excess bloodstream infections (OR 1.63; 95%CI 1.18-2.26), excess hospitalisations (OR 2.77; 95%CI 1.47-5.21) and higher mortality (OR 3.54; 95%CI 1.10-11.40). Conclusions: MDR NTS infections are a serious public health concern. With the emergence of MDR Salmonella strains in the high-income countries, it is crucial to restrict the use of antimicrobials both in animals and humans, and intervene to prevent foodborne infections.

Keywords: Antimicrobial Resistance, Bloodstream Infection, Health Outcomes, Hospitalisation, Invasive Disease, Multi-Drug Resistance (MDR), Mortality, Nontyphoidal Salmonella

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Authors: Maftunakhon Latipova, Feruza Khaydarova

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Obesity is considered a chronic metabolic disease that occurs at any age. Regulation of body weight in the body is carried out through complex interaction of a complex of interrelated systems that control the body's energy system. Energy imbalance is the cause of obesity and overweight, in which the supply of energy from food exceeds the energy needs of the body. Obesity is closely related to impaired appetite regulation, and a hypothalamus is a key place for neural regulation of food consumption. The nucleus of the hypothalamus is connected and interdependent on receiving, integrating and sending hunger signals to regulate appetite. Purpose of the study: to identify markers of food behavior. Materials and methods: The screening was carried out to identify eating disorders in 200 men and women aged 18 to 35 years with overweight and obesity and to check the effects of Orexin A and Neuropeptide Y markers. A questionnaire and questionnaires were conducted with over 200 people aged 18 to 35 years. Questionnaires were for eating disorders and hidden depression (on the Zang scale). Anthropometry is measured by OT, OB, BMI, Weight, and Height. Based on the results of the collected data, 3 groups were divided: People with obesity, People with overweight, Control Group of Healthy People. Results: Of the 200 analysed persons, 86% had eating disorders. Of these, 60% of eating disorders were associated with childhood. According to the Zang test result: Normal condition was about 37%, mild depressive disorder 20%, moderate depressive disorder 25% and 18% of people suffered from severe depressive disorder without knowing it. One group of people with obesity had eating disorders and moderate and severe depressive disorder, and group 2 was overweight with mild depressive disorder. According to laboratory data, the first group had the lowest concentration of Orexin A and Neuropeptide U in blood serum. Conclusions: Being overweight and obese are the first signal of many diseases, and prevention and detection of these disorders will prevent various diseases, including type 2 diabetes. Obesity etiology is associated with eating disorders and signal transmission of the orexinorghetic system of the hypothalamus.

Keywords: obesity, endocrinology, hypothalamus, overweight

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Authors: Artem Kim, Clara Savary, Christele Dubourg, Wilfrid Carre, Houda Hamdi-Roze, Valerie Dupé, Sylvie Odent, Marie De Tayrac, Veronique David

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Holoprosencephaly (HPE) is a rare congenital brain malformation resulting from the incomplete separation of the two cerebral hemispheres. It is characterized by a wide phenotypic spectrum and a high degree of locus heterogeneity. Genetic defects in 16 genes have already been implicated in HPE, but account for only 30% of cases, suggesting that a large part of genetic factors remains to be discovered. HPE has been recently redefined as a complex multigenic disorder, requiring the joint effect of multiple mutational events in genes belonging to one or several developmental pathways. The onset of HPE may result from accumulation of the effects of multiple rare variants in functionally-related genes, each conferring a moderate increase in the risk of HPE onset. In order to decipher the genetic basis of HPE, unconventional patterns of inheritance involving multiple genetic factors need to be considered. The primary objective of this study was to uncover possible disease causing combinations of multiple rare variants underlying HPE by performing trio-based Whole Exome Sequencing (WES) of familial cases where no molecular diagnosis could be established. 39 families were selected with no fully-penetrant causal mutation in known HPE gene, no chromosomic aberrations/copy number variants and without any implication of environmental factors. As the main challenge was to identify disease-related variants among a large number of nonpathogenic polymorphisms detected by WES classical scheme, a novel variant prioritization approach was established. It combined WES filtering with complementary gene-level approaches: transcriptome-driven (RNA-Seq data) and clinically-driven (public clinical data) strategies. Briefly, a filtering approach was performed to select variants compatible with disease segregation, population frequency and pathogenicity prediction to identify an exhaustive list of rare deleterious variants. The exome search space was then reduced by restricting the analysis to candidate genes identified by either transcriptome-driven strategy (genes sharing highly similar expression patterns with known HPE genes during cerebral development) or clinically-driven strategy (genes associated to phenotypes of interest overlapping with HPE). Deeper analyses of candidate variants were then performed on a family-by-family basis. These included the exploration of clinical information, expression studies, variant characteristics, recurrence of mutated genes and available biological knowledge. A novel bioinformatics pipeline was designed. Applied to the 39 families, this final integrated workflow identified an average of 11 candidate variants per family. Most of candidate variants were inherited from asymptomatic parents suggesting a multigenic inheritance pattern requiring the association of multiple mutational events. The manual analysis highlighted 5 new strong HPE candidate genes showing recurrences in distinct families. Functional validations of these genes are foreseen.

Keywords: complex genetic disorder, holoprosencephaly, multiple rare variants, whole exome sequencing

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Authors: Zahraa Albasry, Hadeel D. Najim, Anmar Al-Taie

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Background: Hypertension is considered as one of the most important causes of cardiovascular complications and one of the leading causes of worldwide mortality. Identifying the potential risk factors associated with this medical health problem plays an important role in minimizing its incidence and related complications. The objective of this study is to explore the prevalence of receptor sensitivity regarding assess and understand the perception of specific predictive confounding factors on the prevalence of hypertension (HT) among a sample of Iraqi population in Baghdad, Iraq. Materials and Methods: A randomized cross sectional study was carried out on 100 adult subjects during their visit to the outpatient clinic at a certain sector of Baghdad Province, Iraq. Demographic, clinical and health records alongside specific screening and laboratory tests of the participants were collected and analyzed to detect the potential of confounding factors on the prevalence of HT. Results: 63% of the study participants suffered from HT, most of them were female patients (P < 0.005). Patients aged between 41-50 years old significantly suffered from HT than other age groups (63.5%, P < 0.001). 88.9% of the participants were obese (P < 0.001) and 47.6% had diabetes with HT. Positive family history and sedentary lifestyle were significantly higher among all hypertensive groups (P < 0.05). High salt and fatty food intake was significantly found among patients suffered from isolated systolic hypertension (ISHT) (P < 0.05). A significant positive correlation between packed cell volume (PCV) and systolic blood pressure (SBP) (r = 0.353, P = 0.048) found among normotensive participants. Among hypertensive patients, a positive significant correlation found between triglycerides (TG) and both SBP (r = 0.484, P = 0.031) and diastolic blood pressure (DBP) (r = 0.463, P = 0.040), while low density lipoprotein-cholesterol (LDL-c) showed a positive significant correlation with DBP (r = 0.443, P = 0.021). Conclusion: The prevalence of HT among Iraqi populations is of major concern. Further consideration is required to detect the impact of potential risk factors and to minimize blood pressure (BP) elevation and reduce the risk of other cardiovascular complications later in life.

Keywords: Correlation, Hypertension, Iraq, Risk factors

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Authors: Masoumeh Ghorbania, Falk Liebnerb, Hendrikus W.G. van Herwijnenc, Johannes Konnertha

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Phenol formaldehyde (PF) resins are widely used as wood adhesives for variety of industrial products such as plywood, laminated veneer lumber and others. Lignin as a main constituent of wood has become well-known as a potential substitute for phenol in PF adhesives because of their structural similarity. During the last decades numerous research approaches have been carried out to substitute phenol with pulping-derived lignin, whereby the lower reactivity of resins synthesized with shares of lignin seem to be one of the major challenges. This work reports about a systematic screening of different types of lignin (plant origin and pulping process) for their suitability to replace phenol in phenolic resins. Lignin from different plant sources (softwood, hardwood and grass) were used, as these should differ significantly in their reactivity towards formaldehyde of their reactive phenolic core units. Additionally a possible influence of the pulping process was addressed by using the different types of lignin from soda, kraft, and organosolv process and various lignosulfonates (sodium, ammonium, calcium, magnesium). To determine the influence of lignin on the adhesive performance beside others the rate of viscosity development, bond strength development of varying hot pressing time and other thermal properties were investigated. To evaluate the performance of the cured end product, a few selected properties were studied at the example of solid wood-adhesive bond joints, compact panels and plywood. As main results it was found that lignin significantly accelerates the viscosity development in adhesive synthesis. Bonding strength development during curing of adhesives decelerated for all lignin types, while this trend was least for pine kraft lignin and spruce sodium lignosulfonate. However, the overall performance of the products prepared with the latter adhesives was able to fulfill main standard requirements, even after exposing the products to harsh environmental conditions. Thus, a potential application can be considered for processes where reactivity is less critical but adhesive cost and product performance is essential.

Keywords: phenol formaldehyde resin, lignin phenol formaldehyde resin, ABES, DSC

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Authors: Sachin, Shantanu Arya, Gunjan Mehta, Md. Shamim Ansari

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The cross-modal influence of visual information on speech perception can be illustrated by the McGurk effect which is an illusion of hearing of syllable /ta/ when a listener listens one syllable, e.g.: /pa/ while watching a synchronized video recording of syllable, /ka/. The McGurk effect is an excellent tool to investigate multisensory integration in speech perception in both normal hearing and hearing impaired populations. As the visual cue is unaffected by noise, individuals with hearing impairment rely more than normal listeners on the visual cues.However, when non congruent visual and auditory cues are processed together, audiovisual interaction seems to occur differently in normal and persons with hearing impairment. Therefore, this study aims to observe the audiovisual interaction in speech perception in Cochlear Implant users compares the same with normal hearing children. Auditory stimuli was routed through calibrated Clinical audiometer in sound field condition, and visual stimuli were presented on laptop screen placed at a distance of 1m at 0 degree azimuth. Out of 4 presentations, if 3 responses were a fusion, then McGurk effect was considered to be present. The congruent audiovisual stimuli /pa/ /pa/ and /ka/ /ka/ were perceived correctly as ‘‘pa’’ and ‘‘ka,’’ respectively by both the groups. For the non- congruent stimuli /da/ /pa/, 23 children out of 35 with normal hearing and 9 children out of 35 with cochlear implant had a fusion of sounds i.e. McGurk effect was present. For the non-congruent stimulus /pa/ /ka/, 25 children out of 35 with normal hearing and 8 children out of 35 with cochlear implant had fusion of sounds.The children who used cochlear implants for less than three years did not exhibit fusion of sound i.e. McGurk effect was absent in this group of children. To conclude, the results demonstrate that consistent fusion of visual with auditory information for speech perception is shaped by experience with bimodal spoken language during early life. When auditory experience with speech is mediated by cochlear implant, the likelihood of acquiring bimodal fusion is increased and it greatly depends on the age of implantation. All the above results strongly support the need for screening children for hearing capabilities and providing cochlear implants and aural rehabilitation as early as possible.

Keywords: cochlear implant, congruent stimuli, mcgurk effect, non-congruent stimuli

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