Search results for: leaf disease

Authors: Maia Kukhaleishvili, Ekaterine Bulauri, Iveta Megrelishvili, Tamar Shamatava, Tamar Chipashvili

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Plant viruses can cause loss of yield and quality in a lot of important crops. Symptoms of pathogens are variable depending on the cultivars and virus strain. Selection of resistant potato varieties would reduce the risk of virus transmission and significant economic impact. Other way to avoid reduced harvest yields is regular potato seed production sampling and testing for viral infection. The aim of this study was to determine the occurrence and distribution of viral diseases according potato cultivars for further selection of virus-free material in Georgia. During the summer 2015- 2016, 5 potato cultivars (Sante, Laura, Jelly, Red Sonia, Anushka) at 5 different farms located in Akhalkalaki were tested for 6 different potato viruses: Potato virus A (PVA), Potato virus M (PVM), Potato virus S (PVS), Potato virus X (PVX), Potato virus Y (PVY) and potato leaf roll virus (PLRV). A serological method, Double Antibody Sandwich-Enzyme linked Immunosorbent Assay (DASELISA) was used at the laboratory to analyze the results. The result showed that PVY (21.4%) and PLRV (19.7%) virus presence in collected samples was relatively high compared to others. Researched potato cultivars except Jelly and Laura were infected by PVY with different concentrations. PLRV was found only in three potato cultivars (Sante, Jelly, Red Sonia) and PVM virus (3.12%) was characterized with low prevalence. PVX, PVA and PVS virus infection was not reported. It would be noted that 7.9% of samples were containing PVY/PLRV mix infection. Based on the results it can be concluded that PVY and PLRV infections are dominant in all research cultivars. Therefore significant yield losses are expected. Systematic, long-term control of potato viral infection, especially seed-potatoes, must be regarded as the most important factor to increase seed productivity.

Keywords: virus, potato, infection, diseases

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Authors: Eti Dwi Wiraningsih, Folashade Agusto, Lina Aryati, Syamsuddin Toaha, Suzanne Lenhart, Widodo, Willy Govaerts

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This paper considers a deterministic model for the transmission dynamics of rabies virus in the wild dogs-domestic dogs-human zoonotic cycle. The effect of vaccination and culling in dogs is considered on the model, then the stability was analysed to get basic reproduction number. We use the next generation matrix method and Routh-Hurwitz test to analyze the stability of the Disease-Free Equilibrium and Endemic Equilibrium of this model.

Keywords: stability analysis, rabies model, vaccination effect, culling in dogs

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Authors: Tatiana Butkova, Nikolai Kibrik, Kristina Malsagova, Alexander Izotov, Alexander Stepanov, Anna Kaysheva

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Relevance. The genetic risk of developing schizophrenia is determined by two factors: single nucleotide polymorphisms and gene copy number variations. The search for serological markers for early diagnosis of schizophrenia is driven by the fact that the first five years of the disease are accompanied by significant biological, psychological, and social changes. It is during this period that pathological processes are most amenable to correction. The aim of this study was to analyze single nucleotide polymorphisms (SNPs) that are hypothesized to potentially influence the onset and development of the endogenous process. Materials and Methods It was analyzed 73 single nucleotide polymorphism variants. The study included 48 patients undergoing inpatient treatment at "Psychiatric Clinical Hospital No. 1" in Moscow, comprising 23 females and 25 males. Inclusion criteria: - Patients aged 18 and above. - Diagnosis according to ICD-10: F20.0, F20.2, F20.8, F21.8, F25.1, F25.2. - Voluntary informed consent from patients. Exclusion criteria included: - The presence of concurrent somatic or neurological pathology, neuroinfections, epilepsy, organic central nervous system damage of any etiology, and regular use of medication. - Substance abuse and alcohol dependence. - Women who were pregnant or breastfeeding. Clinical and psychopathological assessment was complemented by psychometric evaluation using the PANSS scale at the beginning and end of treatment. The duration of observation during therapy was 4-6 weeks. Total DNA extraction was performed using QIAamp DNA. Blood samples were processed on Illumina HiScan and genotyped for 652,297 markers on the Infinium Global Chips Screening Array-24v2.0 using the IMPUTE2 program with parameters Ne=20,000 and k=90. Additional filtration was performed based on INFO>0.5 and genotype probability>0.5. Quality control of the obtained DNA was conducted using agarose gel electrophoresis, with each tested sample having a volume of 100 µL. Results. It was observed that several SNPs exhibited gender dependence. We identified groups of single nucleotide polymorphisms with a membership of 80% or more in either the female or male gender. These SNPs included rs2661319, rs2842030, rs4606, rs11868035, rs518147, rs5993883, and rs6269.Another noteworthy finding was the limited combination of SNPs sufficient to manifest clinical symptoms leading to hospitalization. Among all 48 patients, each of whom was analyzed for deviations in 73 SNPs, it was discovered that the combination of involved SNPs in the manifestation of pronounced clinical symptoms of schizophrenia was 19±3 out of 73 possible. In study, the frequency of occurrence of single nucleotide polymorphisms also varied. The most frequently observed SNPs were rs4849127 (in 90% of cases), rs1150226 (86%), rs1414334 (75%), rs10170310 (73%), rs2857657, and rs4436578 (71%). Conclusion. Thus, the results of this study provide additional evidence that these genes may be associated with the development of schizophrenia spectrum disorders. However, it's impossible cannot rule out the hypothesis that these polymorphisms may be in linkage disequilibrium with other functionally significant polymorphisms that may actually be involved in schizophrenia spectrum disorders. It has been shown that missense SNPs by themselves are likely not causative of the disease but are in strong linkage disequilibrium with non-functional SNPs that may indeed contribute to disease predisposition.

Keywords: gene polymorphisms, genotyping, single nucleotide polymorphisms, schizophrenia.

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Authors: Abu S. Mustafa, Mohammad W. Khan, Faraz Shaheed Khan, Nazima Habibi

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Brucellosis is a zoonotic disease of worldwide prevalence. There are at least four species and several strains of Brucella that cause human disease. Brucella genomes have very limited variation across strains, which hinder strain identification using classical molecular techniques, including PCR and 16 S rDNA sequencing. The aim of this study was to perform whole genome sequencing of clinical isolates of Brucella and perform bioinformatics and comparative genomics analyses to determine the existence of genetic differences across the isolates of a single Brucella species and strain. The draft sequence data were generated from 15 clinical isolates of Brucella melitensis (biovar 2 strain 63/9) using MiSeq next generation sequencing platform. The generated reads were used for further assembly and analysis. All the analysis was performed using Bioinformatics work station (8 core i7 processor, 8GB RAM with Bio-Linux operating system). FastQC was used to determine the quality of reads and low quality reads were trimmed or eliminated using Fastx_trimmer. Assembly was done by using Velvet and ABySS softwares. The ordering of assembled contigs was performed by Mauve. An online server RAST was employed to annotate the contigs assembly. Annotated genomes were compared using Mauve and ACT tools. The QC score for DNA sequence data, generated by MiSeq, was higher than 30 for 80% of reads with more than 100x coverage, which suggested that data could be utilized for further analysis. However when analyzed by FastQC, quality of four reads was not good enough for creating a complete genome draft so remaining 11 samples were used for further analysis. The comparative genome analyses showed that despite sharing same gene sets, single nucleotide polymorphisms and insertions/deletions existed across different genomes, which provided a variable extent of diversity to these bacteria. In conclusion, the next generation sequencing, bioinformatics, and comparative genome analysis can be utilized to find variations (point mutations, insertions and deletions) across different genomes of Brucella within a single strain. This information could be useful in surveillance and epidemiological studies supported by Kuwait University Research Sector grants MI04/15 and SRUL02/13.

Keywords: brucella, bioinformatics, comparative genomics, whole genome sequencing

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Authors: Farideh Doostan, Sahar Mohseni Taklloo, Mohammad Nosrati

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Chronic diseases are non-communicable and largely preventable by lifestyle changes including healthy diet consumption. They are the most common cause of death in the world and projected to increase by 15% globally between 2010 and 2020.The hazardous effects of behavioral and dietary risk factors on chronic disease have been established in prospective cohort studies and randomized trials. Because of some changes occur in college students’ lifestyle, assessment of dietary risk factors is important in these populations. Objective: This research was the first study that conducted to evaluate dietary intakes of Kermanian college students with WHO/FAO nutritional objectives. Material and Methods: In this descriptive cross-sectional study, 229 healthy college students of health faculty in Kerman University of Medical Sciences that do not intake any medical drugs were recruited using multistage sampling in 2013.Usual dietary intake was collected using a valid Food Frequency Questionnaire (FFQ) and diet quality was calculated based on WHO nutrient goals. To analysis of data between two groups, independent sample t. test and man whitney were applied. Results: Two hundred and twenty-nine college students; 151 females (65.9%) and 78 males (34.1%), the mean age of 21.9 years were studied. The mean of the Body Mass Index (Kg/m2) and Waist Circumference (cm) in males were 22.34 ±3.52 and 80.76±11.16 and in females were 21.19±2.62 and 73.67±7.65 respectively. Mean of daily cholesterol intake in males was significantly more than females (305±101 VS 268±98; P=0.008) and more than WHO/FAO recommendation (less than 300 mg/day). The mean of daily sodium intake in men and women were 10.4±1 and 10.9±5.3 respectively. These amounts were more than WHO/FAO recommendation (less than 2g/day). In addition, women were consumed fruit and vegetables more than men (839±336 VS 638±281; p ‹ 0.001) and these amounts were more than WHO/FAO recommendation (more than 400g/day) in both groups. Other intake indices were in the range of WHO/FAO recommendations, So that Percent of calories intake from total fat, saturated fatty acids, polyunsaturated fatty acids and added sugar were in compliance with WHO/FAO recommendations. Conclusion: Cholesterol intake in men and sodium intake in all participants were more than WHO/FAO recommendation. These dietary components are the most important causes of cardiovascular disease (one of the main causes of death in our population). These results indicated that proper nutritional education and interventions are needed in this population.

Keywords: college students, food intake, WHO /FAO nutrient intake goals, Kerman

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Authors: A. Mirzaei, S. Zolghadri, M. Athari-Allaf, H. Yousefnia, A. R. Jalilian

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Rheumatoid arthritis is the most common autoimmune disease, leading to the destruction of the joints. The aim of this study was the preparation of 90Y-chitosan complex as a novel agent for radiosynovectomy. The complex was prepared in the diluted acetic acid solution. At the optimized condition, the radiochemical purity of higher than 99% was obtained by ITLC method on Whatman No. 1 and by using a mixture of methanol/water/acetic acid (4:4:2) as the mobile phase. The complex was stable in acidic media (pH=3) and its radiochemical purity was above 98% even after 48 hours. The biodistribution data in rats showed that there was no significant leakage of the injected activity even after 48 h. Considering all of the excellent features of the complex, 90Y-chitosan can be used to manipulate synovial inflammation effectively.

Keywords: chitosan, Y-90, radiosynovectomy, biodistribution

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Authors: Sara Zelman, Courtney Meyer, Hiren Patel, Lisa Philpotts, Sue Lahey, Thomas Burke

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Background: As the global burden of disease shifts from infectious diseases to noncommunicable diseases, there is growing urgency to provide treatment for time-sensitive illnesses, such as ST-Elevation Myocardial Infarctions (STEMIs). The standard of care for STEMIs in developed countries is Percutaneous Coronary Intervention (PCI). However, this is inaccessible in resource-limited settings. Before the discovery of PCI, Streptokinase (STK) and other thrombolytic drugs were first-line treatments for STEMIs. STK has been recognized as a cost-effective and safe treatment for STEMIs; however, in settings which lack access to PCI, it has not become the established second-line therapy. A systematic literature review was conducted to geographically map the use of STK for STEMIs in resource-limited settings. Methods: Our literature review group searched the databases Cinhal, Embase, Ovid, Pubmed, Web of Science, and WHO’s Index Medicus. The search terms included ‘thrombolytics’ AND ‘myocardial infarction’ AND ‘resource-limited’ and were restricted to human studies and papers written in English. A considerable number of studies came from Latin America; however, these studies were not written in English and were excluded. The initial search yielded 3,487 articles, which was reduced to 3,196 papers after titles were screened. Three medical professionals then screened abstracts, from which 291 articles were selected for full-text review and 94 papers were chosen for final inclusion. These articles were then analyzed and mapped geographically. Results: This systematic literature review revealed that STK has been used for the treatment of STEMIs in 33 resource-limited countries, with 18 of 94 studies taking place in India. Furthermore, 13 studies occurred in Pakistan, followed by Iran (6), Sri Lanka (5), Brazil (4), China (4), and South Africa (4). Conclusion: Our systematic review revealed that STK has been used for the treatment of STEMIs in 33 resource-limited countries, with the highest utilization occurring in India. This demonstrates that even though STK has high utility for STEMI treatment in resource-limited settings, it still has not become the standard of care. Future research should investigate the barriers preventing the establishment of STK use as second-line treatment after PCI.

Keywords: cardiovascular disease, global health, resource-limited setting, ST-Elevation Myocardial Infarction, Streptokinase

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Authors: Marko Jankovic, Aleksandra Knezevic, Maja Cupic, Dragana Vujic, Zeljko Zecevic, Borko Gobeljic, Marija Simic, Tanja Jovanovic

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The human cytomegalovirus (CMV) is a notorious pathogen in the pediatric transplant setting. Although studies on factors in complicity with CMV infection abound, the role of age, gender, allogeneic hematopoietic stem cell transplantation (alloHSCT) modality, and underlying disease as regards CMV infection and viral load in children are poorly explored. We examined the significance of various factors related to the risk of CMV infection and viral load in Serbian children and adolescents undergoing alloHSCT. This was a prospective single centre study of thirty two pediatric patients in receipt of alloHSCT for various malignant and non-malignant disorders. Screening for active viral infection was performed by regular weekly monitoring. The Real-Time PCR method was used for CMV DNA detection and quantitation. Statistical analysis was performed using the IBM SPSS Statistics v20 software. Chi-square test was used to evaluate categorical variables. Comparison between scalar and nominal data was done by Wilcoxon-Mann-Whitney test. Pearson correlation was applied for studying the association between patient age and viral load. CMV was detected in 23 (71.9%) patients. Infection occurred significantly more often (p=0.015) in patients with haploidentical donors. The opposite was noted for matched sibling grafts (p=0.006). The viral load was higher in females (p=0.041) and children in the aftermath of alloHSCT with malignant diseases (p=0.019). There was no significant relationship between the viral infection dynamics and overt medical consequences. This is the first study of risk factors for CMV infection in Serbian pediatric alloHSCT patients. Transplanted patients presented with a high incidence of CMV viremia. The HLA compatibility of donated graft is associated with the frequency of CMV positive events. Age, gender, underlying disease, and medically relevant events were not conducive to occurrences of viremia. Notably, substantial viral burdens were evidenced in females and patients with neoplastic diseases. Studies comprising larger populations are clearly needed to scrutinize current results.

Keywords: allogeneic hematopoietic stem cell transplantation, children, cytomegalovirus, risk factors, viral load

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Authors: Gabriella Jorge, Carlos A. Pérez, Hanna Friberg, Sara Söderlund, Jan Lagerlöf

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Fusarium Head Blight (FHB) is one of the most important Fusarium-caused diseases, which affects cereals with serious detrimental effects on yield and grain quality worldwide. Earthworms have been suggested as an alternative to control this disease, which requires a combination of preventive methods to reduce level of damage, although it has been proven that their effect is species dependent. Our objective was to evaluate the effect of the earthworms Aporrectodea longa and Lumbricus rubellus, on the inoculum of Fusarium graminearum on wheat straw. To test this we kept earthworms in vessels with soil, and F. graminearum-inoculated straw covering the surface, under controlled conditions for 6 weeks. Two factors were evaluated with a complete factorial design: earthworms (three levels: without earthworms, A. longa, and L. rubellus), and straw (two levels: inoculated with the pathogen, and sterile). The presence of L. rubellus significantly (P<0.05) reduced the amount of inoculated straw at the soil surface 31% after 6 weeks, while the presence of A. longa, most found in quiescence, did not have any significant effect on the amount of straw when compared to the control. After incubation, F. graminearum was detected by qPCR, only in the surface straw in those treatments inoculated with the pathogen but without earthworms. None of the treatments showed presence of Fusarium in the buried straw, soil or earthworm casts. Both earthworm species decreased in body weight during incubation, most likely due to the decrease in soil water content during the experiment, from 25% to 20%, and/or inadequate food supply, since no other source of food was added. However, this reduction in weight occurred indistinctly of the presence or not of Fusarium (P<0.05). This indicates that both species, of different ecological groups, anecic and epigeic, can reduce F. graminearum inoculum present in wheat straw, while their growth is not negatively affected by this pathogen. These promising results place A. longa, and L. rubellus as potential biocontrol agents of this fungal plant pathogen responsible for Fusarium Head Blight disease in wheat, although further ongoing experiments are needed to confirm the repeatability of these results.

Keywords: Aporrectodea longa, biological control, fungal plant pathogen, Lumbricus rubellus, qPCR, wheat straw

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Authors: Maha M. Eissa, Safia M. Ali, Sonia R. Allam, Ezzat A. Badawy, Nahed M. Baddour

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The obligate follicle mite Demodex folliculorum (Df) has been demonstrated in increased numbers in patients with rosacea. The significance of this finding is still not completely understood because many people free of skin disease may also have mites. So, the aim of this work was to study the possible role of Df in patients with rosacea. Facial punch biopsy specimens were taken from patients with rosacea (either telangiectatic, papulopustular, squamous, granulomatous form or with rhinophyma) and age and sex-matched healthy subjects. Results of this study showed that 63.3% of the rosacea patients were harbouring the mite, with a higher incidence in patients with papulopustular form.

Keywords: Demodex folliculorum, histopathology, punch biopsy, rosacea

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Authors: Stephanie Khater, Ali Chehade, Lamis Chalak

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The progressive replacement of the Lebanese autochthonous grapevine cultivars during the last decade by the imported foreign varieties almost resulted in the genetic erosion of the local germplasm and the confusion with cultivars' names. Hence there is a need to characterize these local cultivars and to assess the possible existing variability at the cultivar level. This work was conducted in an attempt to evaluate the intra-varietal diversity within Lebanese traditional cultivars 'Aswad', 'Maghdoushe', 'Maryame', 'Merweh', 'Meksese' and 'Obeide'. A total of 50 accessions distributed over five main geographical areas in Lebanon were collected and submitted to both ampelographic description and ISSR DNA analysis. A set of 35 ampelographic descriptors previously established by the International Office of Vine and Wine and related to leaf, bunch, berry, and phenological stages, were examined. Variability was observed between accessions within cultivars for blade shape, density of prostrate and erect hairs, teeth shape, berry shape, size and color, cluster shape and size, and flesh juiciness. At the molecular level, nine ISSR (inter-simple sequence repeat) primers, previously developed for grapevine, were used in this study. These primers generated a total of 35 bands, of which 30 (85.7%) were polymorphic. Totally, 29 genetic profiles were differentiated, of which 9 revealed within 'Obeide', 6 for 'Maghdoushe', 5 for 'Merweh', 4 within 'Maryame', 3 for 'Aswad' and 2 within 'Meksese'. Findings of this study indicate the existence of several genotypes that form the basis of the main indigenous cultivars grown in Lebanon and which should be further considered in the establishment of new vineyards and selection programs.

Keywords: ampelography, autochthonous cultivars, ISSR markers, Lebanon, Vitis vinifera L.

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Authors: Mustafa Kucukoduk, Evren Yildiztugay, A. Hediye Sekmen, Ismail Turkan, Yavuz Bagci

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In this study, physiological and biochemical responses of Centaurea tuzgoluensis, a Turkish endemic halophyte, to salinity were studied. Therefore, the changes in shoot growth, leaf relative water content (RWC), ion concentrations, lipid peroxidation, hydroxyl (OH.) radical scavenging activity, proline (Pro) content, and antioxidant system [superoxide dismutase (SOD), catalase (CAT), ascorbate peroxidase (APX) and glutathione reductase (GR)] were investigated. The 60 days (d) old C. tuzgoluensis seedlings were subjected to 0, 150 and 300 mM NaCl for 7 d and 14 d. The relative shoot growth was generally did not change in the 150 mM NaCl, but reduced with 300 mM NaCl stress at 7 d and 14 d. RWC was higher in 150 mM NaCl-treated leaves than that of 300 mM NaCl. Salinity decreased K+/Na+ ratio, but increased Na+, Cl, Ca+2 and Na+/Cl ratio in the leaves. On the other hand, it did not change or increase the K+ content at 150 and 300 mM NaCl, respectively. MDA content in the 150 and 300 mM NaCl-treated leaves remained close to control at 7 d. This was related to enhanced activities of SOD, CAT, APX and GR enzymes, and their isoenzymes especially Fe-SOD in the leaves. On the other hand, the higher sensitivity to 300 mM NaCl at 14 d was associated with inadequate increase in antioxidant enzymes and the decreased OH radical scavenging activity. All these results suggest that C. tuzgoluensis has different antioxidant metabolisms between short- (7 d) and long-term (14 d) salt treatments and salinity tolerance of C. tuzgoluensis might be closely related to increased capacity of antioxidative system to scavenge reactive oxygen species (ROS) and accumulation of osmoprotectant proline under salinity conditions.

Keywords: antioxidant enzymes, endemic halophyte, ion exchange, lipid peroxidation, antioxidant, enzymes, endemic halophyte, ion exchange, lipid peroxidation, proline, Centaurea tuzgoluensis

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Authors: Kumari Rekha, Mathur K Dhananjay, Maheshwari Deepanshu, Nautiyal Nidhi, Shubham Smriti, Laal Deepika, Sinha Swati, Kumar Anupam, Biswas Subhrajit, Shiv Kumar Sarin

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Background: Mesenchymal stem cells are multipotent stem cells derived from mesoderm and are used for therapeutic purposes because of their self-renewal, homing capacity, Immunomodulatory capability, low immunogenicity and mitochondrial transfer signaling. MSCs have the ability to regulate the mechanism of both innate as well as adaptive immune responses through the modulation of cellular response and the secretion of inflammatory mediators. Different sources of MSC are UC MSC, BM MSC, Dental Pulp, and Adipose MSC. The most frequent source used is umbilical cord tissue due to its being easily available and free of limitations of collection procedures from respective hospitals. The immunosuppressive role of MSCs is particularly interesting for clinical use since it confers resistance to rejection by the host immune response. Methodology: In this study, T helper cells (TH4), Cytotoxic T cells (CD-8), immunoregulatory cells (CD25 +FOXP3+) are compared from isolated MSC from three different methods, UC Dissociation Kit (Miltenyi), Explant Culture and Collagenase Type-IV. To check the immunomodulatory property, these MSCs were seeded with PBMC(Coculture) in CD3 coated 24 well plates. Cd28 antibody was added in coculture for six days. The coculture was analyzed in FACS Verse flow cytometry. Results: From flow cytometry analysis of coculture, it found that All over T helper cells (CD4+) number p<0.0264 increases in (All Enzymes) MSC rather than explant MSC(p>0.0895) as compared to Collagenase(p>0.7889) in a coculture of Activated T cell and Mesenchymal Stem Cell. Similar T reg cells (CD25+, FOXP3+) expression p<0.0234increases in All Enzymes), decreases in Explant and Collagenase. Experiments have shown that MSCs can also directly prevent the cytotoxic activity of CD8 lymphocytes mainly by blocking their proliferation rather than by inhibiting the cytotoxic effect. And promoting the t-reg cells, which helps in the mediation of immune response in various diseases. Conclusion: MSC suppress Cytotoxic CD8 T cell and Enhance immunoregulatory T reg (CD4+, CD25+, FOXP3+) Cell expression. Thus, MSC maintains a proper balance(ratio) between CD4 T cells and Cytotoxic CD8 T cells.

Keywords: MSC, disease, T cell, T regulatory

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Authors: Ledi Neçaj

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Introduction: Cardio-respiratory rehabilitation is the set of coordinated interventions needed to provide the best physical, psychological, and social conditions so that patients with chronic or post-acute cardiopulmonary disease, with their efforts, maintain or resume optimal functioning in society through improved health behaviors. Purpose: To study the effectiveness of the application of Cardio-Respiratory Rehabilitation in the typology of patients with chronic or post-acute cardiomyopathy and chronic respiratory diseases in order to facilitate their therapeutic use and to improve the overall quality of life. Material and Method: This is a prospective study including patients with COPD and cardiac disease who were included in the rehabilitation program during the period January 2019 - November 2021. The study was conducted at the University Hospital Center "Mother Teresa" in Tirana, University Hospital "SHEFQET NDROQI", AMERICAN Hospital, HYGEA Hospital, and "Our Lady of Good Counsel, Tirana". An individual chart was used to collect sociodemographic, physical, clinical, and functional examinations for each patient. Results: The study included 253 patients, with a mean age of 62.1 (± 7.9) years, ranging from 48 to 82 years. (67.6%) of the patients were males, and (32.4%) female. Male patients predominated in all age groups, with a statistically significant difference with females (p<0.01). The most common cardiac pathologies are coronary artery bypass (24%), cerebral stroke (9%), myocardial infarction (17%), Stent placement (8%) (p<0.01). Correlation matrix of risk factors found a significant correlation of alcohol consumption with diabetes, smoking, dyslipidemia, sedentary life, obesity, AVC, and hypertension. Functional capacity estimated by change in metabolic equivalents (MET) improved by 46% from 4. ±2.2 to 7.2± .8 METs (p<0.01). Duration of exercise after rehabilitation was increased by 21% compared to baseline (p<0.01). The mean score of all three subscales of the questionnaire: symptoms (p=0.03), activity (p<0.01), and impact (p<0.01) after rehabilitation, was lower compared to pre-rehabilitation. Conclusions: The rehabilitation program has impacted on improving the quality of life, reducing symptoms, reducing the impact of negative factors on daily life, and reducing dyspnea during daily activities.

Keywords: cardio-respiratory rehabilitation, physical exercise, quality of life, diseases

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Authors: Mohamed Ahmed Tony, Mohamed Hamoud

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The present study was conducted to determine the effect of commercially coated slow-release sodium butyrate (CM3000®) as a feed additive on zootechnical performance, immune status and Clostridium perfringens severity after experimental infection. Three hundred 1-d-old broiler chicks (Cobb 500) were randomly distributed into 3 treatment groups (4 replicates each) using 25 chicks per replicate on floor pens. Control (C) birds were offered non-supplemented basal diets. Treatments 1 and 2 (T1 and T2) were fed diets containing CM3000® at 300 and 500 g/ton feed, respectively, during the entire experimental period (35 days). Feed and water were offered ad-libitum. Feed consumption and body weight were recorded weekly to calculate body weight gain and feed conversion. Blood samples were collected to evaluate the immune status of the birds against Newcastle disease vaccines using HI test. At the end of the experimental period, 20 birds were chosen randomly from each group (5 birds from each pen) to compare carcass yield. At day 16 of age 20 birds from each group (5 birds/replicate) were bacteriologically examined and proved to be free from Clostridium perfringens. The isolated birds were challenged orally with 1 ml buffer containing 106 CFU/ml Clostridium perfringens local isolate and prepared from necrotic enteritis (NE) diseased farms. Birds were observed on a regular basis daily for any signs of NE. Birds that died in the challenged group were necropsied to determine the cause of death. On day 28 of age, the surviving chickens were killed by cervical dislocation and necropsied immediately. Intestinal tracts were removed and intestinal lesions were scored. Tissue samples of the duodenum, jejunum, ileum and cecum for histopathological examination were collected. All collected data were statistically analyzed using IBM SPSS® version 19 software for personal computers. Means were compared by one-way ANOVA (P<0.05) followed by the Duncan Post Hoc test. The results revealed that body weight gain was significantly (P<0.05) improved in chicks fed on both doses of CM3000® compared to the control one. Final body weight gain in T1 and T2 were 2064.94 and 2141.37 g/bird, respectively, while in the control group, the weight gain showed 1952.78 g/bird. In addition, supplementation of diets with CM3000® increased significantly feed intake (P<0.05). Total feed intake in T1 and T2 were 3186.32 and 3273.29 g/bird, respectively; however, feed intake in the control group recorded 3081.95 g/bird. The best feed conversion was recorded in T2 group (1.53). Feed conversion in the control and T1 groups were 1.58 and 1.54, respectively. Dressing percentage, liver weights and the other carcasses yields were not different between treatments. The butyrate significantly enhanced immune responses measured against Newcastle disease vaccines. Sodium butyrate significantly reduced NE lesions and healthy improved the intestinal tissues in the samples collected from T1 and T2-challenged chickens versus those collected from the control group. In conclusion, exogenous administration of slow-release butyrate (CM3000®) is capable of improving performance, enhancing immunity and NE disease resistance in broiler chickens.

Keywords: sodium butyrate, broiler chicken, zootechnical performance, immunity, necrotic enteritis

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Authors: Ouzid Yasmina, Aiche-Iratni Ghenima, Harchaoui Lina, Saadoun Noria, Houali Karim

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Medicinal plants are an important source of bioactive molecules with biological activities such as anticancer, antioxidant, anti-inflammatory, antibacterial, antimitotic.... These molecules include alkaloids, polyphenols and terpenes. The latter can be extracted by different solvents, namely: water, ethanol, methanol, butanol, acetone... This is why it seemed interesting to us to evaluate in vitro the antimitotic and genotoxic effect of these secondary metabolites contained in the aqueous extract of the leaves of Peganum harmala L. by the Allium cepa L. test on meristematic cells by calculating the mitotic parameters (The mitotic index, the aberration index and the limit value of cytotoxicity).A spectrophotometric determination of secondary metabolites, namely alkaloids and flavonoids in the aqueous extract of this essence, was performed. As a result, the alkaloid content is estimated to be 28.42 μg EC/mg extract, and the flavonoid content is 12.52 μg EQ/mg extract. The determination of the mitotic index revealed disturbances in cell division with a highly significant difference between the negative control (distilled water) and the different samples (aqueous extracts, colchicine and quecetin). The exposure of meristematic cells to our samples resulted in a large number of chromosomal, nuclear and cellular aberrations with an aberration index reaching 16.21±1.28% for the 4mg/ml aqueous extract and 11.71±3.32% for the 10mg/ml aqueous extract. The limit value of cytotoxicity revealed that our samples are sublethal on Allium cepa L. meristematic cells.

Keywords: allium cepa l., antimitotic and genotoxic effect, aqueous leaf extract, laghouat (algeria), peganum harmala l., secondary metabolites

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Authors: Lok Raj Joshi, Naveen Prakash Shah, Sharad Kumar Sharma, I. Ratna Bhattarai, Rajendra Basnet, Deepak Dahal, Bahagwan Maharjan, Seraphine Kaminsa

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Context: Tuberculosis (TB) is a significant health issue in Nepal, a country with a high burden of the disease. Despite efforts to control TB, there is still a gap in the notification of TB cases, which hinders effective control and treatment. This paper aims to address this notification gap and proposes strategies to improve TB control in Nepal. Research Aim: The aim of this research is to understand and address the tuberculosis notification gap in Nepal. The focus is on enhancing the healthcare system, involving the private sector and communities, raising awareness, and addressing social determinants to achieve sustainable TB control. Methodology: The research methodology involved a review of existing epidemiological data and research studies related to TB in Nepal. Additionally, consultation with an expert group from the TB control program in Nepal provided insights into the current state of TB control and challenges in addressing the notification gap. Findings: The findings reveal that only 55% of TB cases were reported in 2022, indicating a significant notification gap. Of the reported cases, only 32% and 19% were referred by the private sector and community, respectively. Furthermore, 20% of diagnosed cases were not treated in the initial phase. The estimated number of cases of multidrug-resistant TB (MDR TB) was 2,800, suggesting a low diagnosis rate. Among the diagnosed MDR TB cases, only 60% were receiving treatment. Additionally, it was observed that 20% of diagnosed MDR TB cases were from India and not enrolling in TB treatment in Nepal, indicating a high rate of defaulters. Theoretical Importance: The study highlights the importance of adopting a holistic strategy to address the notification gap in TB cases in Nepal. It emphasizes the need to enhance healthcare infrastructure, raise awareness, involve the private sector and local communities, establish effective methods to trace initial defaulters, implement TB interventions in border regions, and mitigate the social stigma associated with the disease. Data Collection and Analysis Procedures: Data for this study was collected through a review of existing epidemiological data and research studies. The data were then analyzed to identify patterns, trends, and gaps in TB case notification in Nepal.

Keywords: TB, tuberculosis, private sector, community, migrants, nepal

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Authors: Witthaya Mekhum, Waleerak Sittisom

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This research investigated the effect on occupational health safety and environment at work from metal handicraft using Rattanakosin local wisdom focusing on pollution, accidents, and injuries from work. The sample group in this study included 48 metal handicraft workers in 5 communities by using questionnaires and interview to collect data. The evaluation form TISI 18001 was used to analyze job safety analysis (JSA). The results showed that risk at work reduced after applying the developed model. Banbu Community produces alloy bowl rubbed with stone. The high risk process is melting and hitting process. Before the application, the work risk was 82.71%. After the application of the developed model, the work risk was reduced to 50.61%. Banbart Community produces monk’s food bowl. The high risk process is blow pipe welding. Before the application, the work risk was 93.59%. After the application of the developed model, the work risk was reduced to 48.14%. Bannoen Community produces circle gong. The high risk process is milling process. Before the application, the work risk was 85.18%. After the application of the developed model, the work risk was reduced to 46.91%. Teethong Community produces gold leaf. The high risk process is hitting and spreading process. Before the application, the work risk was 86.42%. After the application of the developed model, the work risk was reduced to 64.19%. Ban Changthong Community produces gold ornament. The high risk process is gold melting process. Before the application, the work risk was 67.90%. After the application of the developed model, the work risk was reduced to 37.03%. It can be concluded that with the application of the developed model, the work risk of 5 communities was reduced in the 3 main groups: (1) Work illness reduced by 16.77%; (2) Pollution from work reduced by 10.31%; (3) Accidents and injuries from work reduced by 15.62%.

Keywords: occupational health, safety, local wisdom, Rattanakosin

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Authors: Maha M. Eissa, Safia M. Ali, Sonia R. Allam, Ezzat A. Badawy, Nahid M. Baddour

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The obligate follicle mite Demodex folliculorum (Df) has been demonstrated in increased numbers in patients with rosacea. The significance of this finding is still not completely understood, because many people free of skin disease may also have mites. So, the aim of this work was to study the possible role of Df in patients with rosacea. Facial punch biopsy specimens were taken from patients with rosacea (either telangiectatic, papulopustular, squamous, granulomatous form or with rhinophyma) and age and sex matched healthy subjects. Results of this study showed that 63.3% of the rosacea patients were harbouring the mite with higher incidence in patients with papulopustular form.

Keywords: Demodex folliculorum, rosacea, papulopustular, punch biopsy

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Authors: Djamila Benhaddouche, Abdelkader Benyettou

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In this paper, we used data mining to extract biomedical knowledge. In general, complex biomedical data collected in studies of populations are treated by statistical methods, although they are robust, they are not sufficient in themselves to harness the potential wealth of data. For that you used in step two learning algorithms: the Decision Trees and Support Vector Machine (SVM). These supervised classification methods are used to make the diagnosis of thyroid disease. In this context, we propose to promote the study and use of symbolic data mining techniques.

Keywords: biomedical data, learning, classifier, algorithms decision tree, knowledge extraction

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Authors: Sujata S. Kulkarni

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Effective and efficient texture feature extraction and classification is an important problem in image understanding and recognition. This paper gives a review on effective texture classification method. The objective of the problem of texture representation is to reduce the amount of raw data presented by the image, while preserving the information needed for the task. Texture analysis is important in many applications of computer image analysis for classification include industrial and biomedical surface inspection, for example for defects and disease, ground classification of satellite or aerial imagery and content-based access to image databases.

Keywords: compressed sensing, feature extraction, image classification, texture analysis

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Authors: Ji Jiazheng, Wang Jingrao, Jin Xin, Zhang Hong

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Diabetes mellitus is a metabolic disease characterized by high blood sugar. A long-standing hyperglycemic state can lead to various tissue damage. Diabetic retinopathy is the most common and widely studied ocular complication and has become the leading cause of blindness in my country. At the same time, diabetes has profound clinically relevant effects on the cornea, leading to keratopathy and vision-threatening. The cornea is an avascular tissue and is sensitive to hyperglycemia, Keratopathy caused by diabetes is usually chronic, they are called diabetic keratopathy or diabetic neurotrophic keratopathy, leading to several diabetic corneal complications including delayed epithelial wound healing, recurrent erosions, neuropathy, loss of sensitivity. Corneal stem cell dysfunction in diabetic patients as an important influencing factor of diabetic keratopathy. The consequences of this condition are often underestimated. The limbus is located between the cornea and the sclera tissue. The limbal stroma consists of a series of radial elevations with fibrovascular centers known as palisades of Vogt (POV). Previous studies have shown that palisades of Vogt (POV), as the main site of limbal stem cells, plays an important role in the homeostasis of the corneal epithelium. Therefore, POV plays a vital role in the healing of corneal epithelial surgery and postoperative evaluation. IVCM can observe the condition of the corneal epithelium at the cellular level. It has profound significance and guidance for the evaluation of limbal and limbal stem cells. We have previously observed structural changes in POV in HSK and HZO patients on IVCM. At present, there have been reports involving limbal stem cell dysfunction in diabetic patients, but the specific pathogenesis is still unclear. However, there are no studies on POV morphological changes in patients with DM. Therefore, we performed statistics and compared the correlation between POV morphological changes and corneal epithelial basal cell density, corneal nerves, and length of disease in DM patients and normal humans using IVCM studies. At the same time, fundoscopy was used to observe the correlation between the thickness of RNFL and the thickness of GCC and POV in diabetic patients. And to observe the correlation between SVD, DVD and POV for research.

Keywords: confocal microscopy, fundus, limbal stem cells, diabetes

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Authors: Farhad Farahvash, Parya Mobaseri

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Weeds control based on their critical period leads to less production costs and risks of wide chemical application of weeds control methods. The present study considered effect of weeds control time (weeds interference after 20, 40 and 60 days, weeds full control, weeds interference and weeds control after 20, 40 and 60 days) on growth and yield of forage maize 454. The experiment based on full-randomized blocks design with three replications was conducted at research farm of Islamic Azad University of Tabriz located at 15th km of East Tabriz in 2013. According to the results, weeds interference after 40 and 60 days as well as weeds control after 20 days prevented from decrease of maize biomass resulted from weeds presence while weeds interference after 20 days, weeds interference and weeds control after 40 and 60 days led respectively to 41.2%, 35%, 25% and 32.5% decrease of forage maize biomass. The weeds-influenced decrease was manifested at different parts of the plant depending on presence period of weeds. Decrease of fresh weight of ear and fresh weight of leaf and stem was observed due to weeds interference after 20 days and weeds interference. If weeds are controlled after 60 days, decrease of ear weight and fresh weight of stem will lead to biomass decrease. Also, if weeds are controlled after 40 days, decrease of fresh weight of maize stems will result in biomass decrease. Ear traits were affected by weeds control treatment. Being affected by treatments of weeds interference after 20 days, weeds non-interference, weeds control after 40 and 60 days, ear length was shortened 29.9 %, 41.4 %, 27.6 % and 37.2 %, respectively. The stem diameter demonstrated a significant decrease although it was only affected by treatments of weeds interference and weeds control after 60 days. Considering results of the present study, generally, it is suggested to control weeds during initial 20-60 days of maize growth in order to prevent undesirable effect of weeds on growth, production and production biomass of maize and decrease of production costs.

Keywords: maize, competition, weed, biomass

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Authors: Xiaobo Wang, Zhen-Yu Chen

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Higher plasma total cholesterol (TC) and low-density lipoprotein cholesterol (LDL-C) are independent risk factors of cardiovascular disease while high-density lipoprotein cholesterol (HDL-C) is protective. Vitamin D is well-known for its regulatory role in calcium homeostasis. Its potential important role in cardiovascular disease has recently attracted much attention. This study was conducted to investigate effects of different dosage of calcitriol on plasma lipoprotein profile and the underlying mechanism. Sixty male Syrian Golden hamsters were randomly divided into 6 groups: no-cholesterol control (NCD), high-cholesterol control (HCD), groups with calcitriol supplementation at 10/20/40/80ng/kg body weight (CA, CB, CC, CD), respectively. Calcitriol in medium-chain triacylglycerol (MCT) oil was delivered to four experimental groups via oral gavage every other day, while NCD and HCD received MCT oil in the equivalent amount. NCD hamsters were fed with non-cholesterol diet while other five groups were maintained on diet containing 0.2% cholesterol to induce a hypercholesterolemic condition. The treatment lasts for 6 weeks followed by sample collection after hamsters sacrificed. Four experimental groups experienced a reduction in average food intake around 11% compared to HCD with slight decrease in body weight (not exceeding 10%). This reduction reflects on the deceased relative weights of testis, epididymal and perirenal adipose tissue in a dose-dependent manner. Plasma calcitriol levels were measured and was corresponding to oral gavage. At the end of week 6, lipoprotein profiles were improved with calcitriol supplementation with TC, non-HDL-C and plasma triglyceride (TG) decreased in a dose-dependent manner (TC: r=0.373, p=0.009, non-HDL-C: r=0.479, p=0.001, TG: r=0.405, p=0.004). Since HDL-C of four experiment groups showed no significant difference compared to HCD, the ratio of nHDL-C to HDL-C and HDL-C to TC had been restored in a dose-dependent manner. For hamsters receiving the highest level of calcitriol (80ng/kg) showed a reduction of TC by 11.5%, nHDL-C by 24.1% and TG by 31.25%. Little difference was found among six groups on the acetylcholine-induced endothelium-dependent relaxation or contraction of thoracic aorta. To summarize, calcitriol supplementation in hamster at maximum 80ng/kg body weight for 6 weeks lead to an overall improvement in plasma lipoprotein profile with decreased TC and TG level. The molecular mechanism of its effects is under investigation.

Keywords: cholesterol, vitamin D, calcitriol, hamster

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Authors: Ann Enright

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More than ever, animal shelters need to identify ways to reduce the number of animals entering shelter facilities and the incidence of euthanasia. Managing animal overpopulation using euthanasia can have detrimental health and emotional consequences for the shelter staff involved. There are also community expectations with moral and financial implications to consider. To achieve the goals of reducing animal intake and the incidence of euthanasia, shelter best practice involves combining programs, procedures and partnerships to increase live release rates (LRR), reduce the incidence of disease, length of stay (LOS) and shelter intake whilst overall remaining financially viable. Analysing daily processes, tracking outcomes and implementing simple strategies enabled shelter staff to more effectively focus their efforts and achieve amazing results. The objective of this retrospective study was to assess the effect of implementing the capacity for care (C4C) management model. Data focusing on the average daily number of animals on site for a two year period (2016 – 2017) was exported from a shelter management system, Customer Logic (CL) Vet to Excel for manipulation and comparison. Following the implementation of C4C practices the average daily number of animals on site was reduced by >50%, (2016 average 103 compared to 2017 average 49), average LOS reduced by 50% from 8 weeks to 4 weeks and incidence of disease reduced from ≥ 70% to less than 2% of the cats on site at the completion of the study. The total number of stray cats entering the shelter due to council contracts reduced by 50% (486 to 248). Improved cat outcomes were attributed to strategies that increased adoptions and reduced euthanasia of poorly socialized cats, including foster programs. To continue to achieve improvements in LRR and LOS, strategies to decrease intake further would be beneficial, for example, targeted sterilisation programs. In conclusion, the study highlighted the benefits of using C4C as a management tool, delivering a significant reduction in animal intake and euthanasia with positive emotional, financial and community outcomes.

Keywords: animal welfare, capacity for care, cat, euthanasia, length of stay, managed intake, shelter

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Authors: Mihaela Munteanu (Pila), Silvius Stanciu

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This paper presents the aspects of the official control of fishery in the Republic of Moldova. Currently, the regulations and the activity of national institutions with responsibilities in the field of food quality are in a process of harmonization with the European rules, aiming at European integration, quality improvement and providing a higher level of food safety. The National Agency for Food Safety is the main national body with responsibilities in the field of food safety. In the field of fishery products, the Agency carries out an intensive activity of informing the citizen and controlling the products marketed. The paper presents the dangers related to the consumption of fish and fishery products traded on the national market, the sanitary-veterinary inspections conducted by the profile institution and the improper situations identified. The national market of fishery products depends largely on imports, mainly focused on ocean fish. The research carried out has shown that during the period 2011-2018, following the inspections carried out on fishery products traded on the national market, a number of inconsistencies have been identified. Thus, indigenous products were frequently detected with sensory characteristics unfit for consumption, and being commercialized in inappropriate locations or contaminated with chemical pollutants. On import products controlled, the most frequent inconsistent situations have been represented by inconsistent sensory aspects and by parasite contamination. Taking into account the specific aspects of aquatic products, including the high level of alterability, special conditions of growth, marketing, culinary preparation and consumption are necessary in order to decrease the risk of disease over the population. Certificates, attestations and other documents certifying the quality of batches, completed by additional laboratory examinations, are necessary in order to increase the level of confidence on the quality of products marketed in the Republic. The implementation of various control procedures and mechanisms at national level, correlated with the focused activity of the specialized institutions, can decrease the risk of contamination and avoid cases of disease on the population due to the consumption of fishery products.

Keywords: fishery products, food safety, quality control, Republic of Moldova

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Authors: Agnieszka Rolinska, Marta Makara-Studzinska

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Introduction: In recent years, there has been an intensive development of research into the physiological relationships between the experienced stress and obesity. Moreover, strong chronic stress leads to the disorganization of a person’s activeness on various levels of functioning, including the behavioral and cognitive sphere (also in one’s diet). Aim: The present work addresses the following research questions: Is there a relationship between an increase in stress related to the disease and the need for the cyclicality of diabetologic education in gestational diabetes? Are there any differences in terms of the experienced stress during the last three months of pregnancy in women with gestational diabetes and in normal pregnancy between the patients with normal weight gains and those with abnormal weight gains? Are there any differences in terms of stress coping styles in women with gestational diabetes and in normal pregnancy between the patients with normal weight gains and those with abnormal weight gains? Method: The study involved pregnant women with gestational diabetes (treated with diet, without insulin therapy) and in normal pregnancy – 206 women in total. The following psychometric tools were employed: Perceived Stress Scale (PSS; Cohen, Kamarck, Mermelstein), Coping Inventory for Stressful Situations (CISS; Endler, Parker) and authors’ own questionnaire. Gestational diabetes mellitus was diagnosed on the basis of the results of fasting oral glucose tolerance test (75 g OGTT). Body weight measurements were confirmed in a diagnostic interview, taking into account medical data. Regularities in weight gains in pregnancy were determined according to the recommendations of the Polish Gynecological Society and American norms determined by the Institute of Medicine (IOM). Conclusions: An increase in stress related to the disease varies in patients with differing requirements for the cyclical nature of diabetologic education (i.e. education which is systematically repeated). There are no differences in terms of recently experienced stress and stress coping styles between women with gestational diabetes and those in normal pregnancy. There is a relationship between weight gains in pregnancy and the stress experienced in life as well as stress coping styles – both in pregnancy complicated by diabetes and in physiological pregnancy. In the discussion of the obtained results, the authors refer to scientific reports from English-language magazines of international range.

Keywords: diabetologic education, gestational diabetes, stress, weight gain in pregnancy

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Authors: B. O. Oyebanji, I. O. Dudusola, C. T. Ademola, S. A. Olaniyan

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A 56 day study was conducted to evaluate the effect of dietary inclusion of Spondias mombin on hematological, growth, egg parameters and egg shell quality of Japanese quails, Cortunix cortunix japonica. One hundred birds were used for this study, and they were allocated randomly into 5 groups and replicated twice. Group 1 animals served as control without inclusion of extract, groups 2, 3, and 4 had 200 mg/kg, 400 mg/kg and 800 mg/kg inclusion of SM, group 5 had 600 mg/kg of vitamin C respectively. The birds were weighed weekly to determine weight change, the blood parameters analyzed at the completion of the experiment were PCV, Hb, RBC WBC, differential WBC count, MCH, MCH, and MCV were afterwards calculated from these parameters. 5 eggs were collected from each group and egg weight, eggshell weight, eggshell diameter, yolk weight, albumen weight, yolk diameter, yolk height, albumen percentage, yolk percentage and shell percentage were determined. There was no significant difference among the group for the hematological parameters measured and calculated. The egg weight and albumen weight of quails on 800 mg/kg was highest of all the groups, all other egg parameters measured showed no significant difference. The birds supplemented with Vitamin C had the highest weight gain (40.8±2.5 g) and the lowest feed conversion ratio (2.25). There was no mortality recorded in all the groups except in the SM800 group with 10% mortality. It can be concluded from this experiment that Vitamin C supplementation has positive effect on quail production in humid tropics and the inclusion of Spondias mombin leaf extract has a dose-dependent toxicity in quails.

Keywords: hematology, quails, Spondias mombin, vitamin C

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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: Marielle Leijten, Catherine Meulemans, Sven De Maeyer, Luuk Van Waes

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For the diagnosis of Alzheimer's disease (AD), a large variety of neuropsychological tests are available. In some of these tests, linguistic processing - both oral and written - is an important factor. Language disturbances might serve as a strong indicator for an underlying neurodegenerative disorder like AD. However, the current diagnostic instruments for language assessment mainly focus on product measures, such as text length or number of errors, ignoring the importance of the process that leads to written or spoken language production. In this study, it is our aim to describe and test differences between cognitive and impaired elderly on the basis of a selection of writing process variables (inter- and intrapersonal characteristics). These process variables are mainly related to pause times, because the number, length, and location of pauses have proven to be an important indicator of the cognitive complexity of a process. Method: Participants that were enrolled in our research were chosen on the basis of a number of basic criteria necessary to collect reliable writing process data. Furthermore, we opted to match the thirteen cognitively impaired patients (8 MCI and 5 AD) with thirteen cognitively healthy elderly. At the start of the experiment, participants were each given a number of tests, such as the Mini-Mental State Examination test (MMSE), the Geriatric Depression Scale (GDS), the forward and backward digit span and the Edinburgh Handedness Inventory (EHI). Also, a questionnaire was used to collect socio-demographic information (age, gender, eduction) of the subjects as well as more details on their level of computer literacy. The tests and questionnaire were followed by two typing tasks and two picture description tasks. For the typing tasks participants had to copy (type) characters, words and sentences from a screen, whereas the picture description tasks each consisted of an image they had to describe in a few sentences. Both the typing and the picture description tasks were logged with Inputlog, a keystroke logging tool that allows us to log and time stamp keystroke activity to reconstruct and describe text production processes. The main rationale behind keystroke logging is that writing fluency and flow reveal traces of the underlying cognitive processes. This explains the analytical focus on pause (length, number, distribution, location, etc.) and revision (number, type, operation, embeddedness, location, etc.) characteristics. As in speech, pause times are seen as indexical of cognitive effort. Results. Preliminary analysis already showed some promising results concerning pause times before, within and after words. For all variables, mixed effects models were used that included participants as a random effect and MMSE scores, GDS scores and word categories (such as determiners and nouns) as a fixed effect. For pause times before and after words cognitively impaired patients paused longer than healthy elderly. These variables did not show an interaction effect between the group participants (cognitively impaired or healthy elderly) belonged to and word categories. However, pause times within words did show an interaction effect, which indicates pause times within certain word categories differ significantly between patients and healthy elderly.

Keywords: Alzheimer's disease, keystroke logging, matching, writing process

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