Search results for: bacterial gene clusters

Authors: Zachary Muthii Rukenya, James Mbaria, Peter Mbaabu, Kiama Stephen Gitahi, Ronald Onzago

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Aloe turkanensis is one of the widely used medicinal shrub and in Kenya the plant is mainly found in Baringo, Isiolo, Laikipia, Turkana and West Pokot Counties where it is used in ethno-medicine and ethno-veterinary medicine. The Turkana community uses the plant products to manage malaria, wounds, stomach ache, constipation, pain, skin infection, poultry diseases and retained afterbirth in cows. This evaluated the efficacy and safety of the plant obtained from Turkana County, Kenya. Preliminary data on the use of the plant in the county was collected through observation, photographing and interviews. A sample of the whole plant was harvested in Natira sublocation, in ex-Turkana west district in February 2012 after identification by Aloe-working group herbalists who voluntarily provided information on its medicinal uses. Botanical identification was done at Kenya Forest Research Centre in Karura where voucher specimen was deposited. Cold maceration using 70% methanol and distilled water was used for extraction. Bioassays were to determine the effects of the plant extracts on brine shrimp and selected bacterial and fungal cultures. The extracts were tested in-vitro activity against standard cultures of B. cereus (ATCC 11778), S. aureus (ATCC25923), P. aeroginosa (ATCC 27853), E. coli (ATCC 25922) and a human infections clinical isolate of C. albicans. The extracts of Aloe turkanensis inhibited the growth B. cereus (100-200 mg/ml), S. aureus (50-100 mg/ml), P. aeroginosa (200mg/ml), E. coli (400mg/ml) while C. albicans was not affected. The extracts also inhibited the growth of S. aureus and B. cereus with mean diameters of inhibition zones being 19.75±1 mm and 18.5±05 mm reapectively. Phytochemical screening showed the presence of alkaloids, tarpenoids, steroids, quinones, saponins and tannins in the plant extracts. The extract was found to be non-toxic at a concentration of 1000µg/ml with a 100% survival of Brine Shrimp larva. It was concluded that Aloe turkanensis growing the study area has metabolites that inhibit the growth of microorganisms and is however, there is need for further studies to validate the in-vivo bioactivity of the plant and more generate adequate toxicological data.to support conservation, value chain addition of its products and widespread use as a herbal remedy.

Keywords: Aloe turkanensis, bioactivity, cultivated, human infections

Procedia PDF Downloads 321

Authors: Matthew Cardeiro, Amalia D. Ardeljan, Lexi Frankel, Dianela Prado Escobar, Catalina Molnar, Omar M. Rashid

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Introduction: Enterococci comprise the natural flora of nearly all animals and are ubiquitous in food manufacturing and probiotics. However, its role in the microbiome remains controversial. The gut microbiome has shown to play an important role in immunology and cancer. Further, recent data has suggested a relationship between gut microbiota and breast cancer. These studies have shown that the gut microbiome of patients with breast cancer differs from that of healthy patients. Research regarding enterococcus infection and its sequala is limited, and further research is needed in order to understand the relationship between infection and cancer. Enterococcus may prevent the development of breast cancer (BC) through complex immunologic and microbiotic adaptations following an enterococcus infection. This study investigated the effect of enterococcus infection and the incidence of BC. Methods: A retrospective study (January 2010- December 2019) was provided by a Health Insurance Portability and Accountability Act (HIPAA) compliant national database and conducted using a Humans Health Insurance Database. International Classification of Disease (ICD) 9th and 10th codes, Current Procedural Terminology (CPT), and National Drug Codes were used to identify BC diagnosis and enterococcus infection. Patients were matched for age, sex, Charlson Comorbidity Index (CCI), antibiotic treatment, and region of residence. Chi-squared, logistic regression, and odds ratio were implemented to assess the significance and estimate relative risk. Results: 671 out of 28,518 (2.35%) patients with a prior enterococcus infection and 1,459 out of 28,518 (5.12%) patients without enterococcus infection subsequently developed BC, and the difference was statistically significant (p<2.2x10⁻¹⁶). Logistic regression also indicated enterococcus infection was associated with a decreased incidence of BC (RR=0.60, 95% CI [0.57, 0.63]). Treatment for enterococcus infection was analyzed and controlled for in both enterococcus infected and noninfected populations. 398 out of 11,523 (3.34%) patients with a prior enterococcus infection and treated with antibiotics were compared to 624 out of 11,523 (5.41%) patients with no history of enterococcus infection (control) and received antibiotic treatment. Both populations subsequently developed BC. Results remained statistically significant (p<2.2x10-16) with a relative risk of 0.57 (95% CI [0.54, 0.60]). Conclusion & Discussion: This study shows a statistically significant correlation between enterococcus infection and a decrease incidence of breast cancer. Further exploration is needed to identify and understand not only the role of enterococcus in the microbiome but also the protective mechanism(s) and impact enterococcus infection may have on breast cancer development. Ultimately, further research is needed in order to understand the complex and intricate relationship between the microbiome, immunology, bacterial infections, and carcinogenesis.

Keywords: breast cancer, enterococcus, immunology, infection, microbiome

Procedia PDF Downloads 173

Authors: Noah Emil Glisik

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Alcohol use disorder (AUD) presents significant health challenges worldwide and is particularly concerning in Slovenia, where high alcohol consumption contributes to elevated rates of comorbidities, including depression and suicide. This review examines emerging evidence linking gut microbiome dysbiosis to AUD, exploring whether gut microbiome alterations merely result from alcohol use or actively contribute to the persistence of addiction. Additionally, it discusses how microbial changes may influence psychological symptoms, including anxiety and depressive states, which are closely associated with suicidality in this population. To address gaps in existing research, a systematic literature search was conducted through PubMed, Web of Science, and ScienceDirect. Inclusion criteria focused on studies examining gut microbiome changes in AUD, particularly those assessing gut-brain axis interactions and microbial species impacting inflammation and neurotransmitter pathways. Studies were excluded if they lacked peer review or did not specifically assess microbiome effects on mental health outcomes. A qualitative literature review approach was applied, synthesizing findings into key themes on microbial changes, neuroinflammatory pathways, and treatment implications. Data were organized into tables to provide a clear comparison of microbiota alterations across studies, highlighting specific bacterial species and their potential effects on AUD. This review emphasizes patterns in AUD patients, where reductions in anti-inflammatory species, such as Faecalibacterium prausnitzii and Roseburia intestinalis, coincide with increases in pro-inflammatory bacteria like Enterococcus faecalisand Lactobacillus rhamnosus. These shifts contribute to increased gut permeability and systemic inflammation, potentially influencing the kynurenine pathway, which is linked to depressive symptoms and elevated alcohol cravings. Furthermore, the review explores the potential of probiotic therapies targeting these microbial imbalances as adjunctive treatments for AUD, particularly those focusing on strains that support anti-inflammatory pathways and gut barrier integrity. Restoring microbial homeostasis through probiotics or fecal microbiota transplantation may not only reduce inflammation but also alleviate mental health symptoms associated with addiction, including suicidality. The findings underscore the need for further clinical trials assessing microbiome-targeted therapies as innovative, multifaceted approaches to AUD treatment in Slovenia and beyond.

Keywords: alcohol use disorder, gut-brain axis, microbiome dysbiosis, probiotic therapy.

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Authors: Maja Benkovic, Dubravka Novotni, Bojana Voucko, Duska Curic, Damir Jezek, Nikolina Cukelj

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Cryo-grinding is an ultra-fine grinding method used in the pharmaceutical industry, production of herbs and spices and in the production and handling of cereals, due to its ability to produce powders with small particle sizes which maintain their favorable bioactive profile. The aim of this study was to determine the particle size distributions of the proso millet (Panicum miliaceum) bran fraction grinded at cryogenic temperature (using liquid nitrogen (LN₂) cooling, T = - 196 °C), in comparison to non-cooled grinding. Proso millet bran is primarily used as an animal feed, but has a potential in food applications, either as a substrate for extraction of bioactive compounds or raw material in the bakery industry. For both applications finer particle sizes of the bran could be beneficial. Thus, millet bran was ground for 2, 4, 8 and 12 minutes using the ball mill (CryoMill, Retsch GmbH, Haan, Germany) at three grinding modes: (I) without cooling, (II) at cryo-temperature, and (III) at cryo-temperature with included 1 minute of intermediate cryo-cooling step after every 2 minutes of grinding, which is usually applied when samples require longer grinding times. The sample was placed in a 50 mL stainless steel jar containing one grinding ball (Ø 25 mm). The oscillation frequency in all three modes was 30 Hz. Particle size distributions of the bran were determined by a laser diffraction particle sizing method (Mastersizer 2000) using the Scirocco 2000 dry dispersion unit (Malvern Instruments, Malvern, UK). Three main effects of the grinding set-up were visible from the results. Firstly, grinding time at all three modes had a significant effect on all particle size parameters: d(0.1), d(0.5), d(0.9), D[3,2], D[4,3], span and specific surface area. Longer grinding times resulted in lower values of the above-listed parameters, e.g. the averaged d(0.5) of the sample (229.57±1.46 µm) dropped to 51.29±1.28 µm after 2 minutes grinding without LN₂, and additionally to 43.00±1.33 µm after 4 minutes of grinding without LN₂. The only exception was the sample ground for 12 minutes without cooling, where an increase in particle diameters occurred (d(0.5)=62.85±2.20 µm), probably due to particles adhering to one another and forming larger particle clusters. Secondly, samples with LN₂ cooling exhibited lower diameters in comparison to non-cooled. For example, after 8 minutes of non-cooled grinding d(0.5)=46.97±1.05 µm was achieved, while the LN₂ cooling enabled collection of particles with average sizes of d(0.5)=18.57±0.18 µm. Thirdly, the application of intermediate cryo-cooling step resulted in similar particle diameters (d(0.5)=15.83±0.36 µm, 12 min of grinding) as cryo-milling without this step (d(0.5)=16.33±2.09 µm, 12 min of grinding). This indicates that intermediate cooling is not necessary for the current application, which consequently reduces the consumption of LN₂. These results point out the potential beneficial effects of millet bran grinding at cryo-temperatures. Further research will show if the lower particle size achieved in comparison to non-cooled grinding could result in increased bioavailability of bioactive compounds, as well as protein digestibility and solubility of dietary fibers of the proso millet bran fraction.

Keywords: ball mill, cryo-milling, particle size distribution, proso millet (Panicum miliaceum) bran

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Authors: Majid Javanmard

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In this study, the effects of coating with whey protein concentrate (7.5% w/v) alone and/or in combination with rice bran oil (0.2, 0.4, 0.6 g in 100 ml coating solution) and Zataria multiflora extract (1 and 2 μL in 100 ml coating solution) on the quality attributes and egg shelf life were carefully observed and analyzed. Weight loss, Haugh index, yolk index, pH, air cell depth, shell strength and the impact of this coating on the microbial load of the eggs surface were studied at the end of each week (during the 4 weeks of storage in a room environment temperature and humidity). After 4 weeks of storage, it was observed that the weight loss in all of the treated eggs with whey protein concentrate and 0.2 gr of rice bran oil (experimental group) was significantly lower than that of the control group(P < 0/05). With regard to Haugh index and yolk index, egg shelf life increased about 4 weeks compared with the control samples. Haugh Index changes revealed that the coated samples remained at grade A after 3 weeks of storage, while the control samples were relegated from grade AA to B after one week. Haugh and yolk Indices in all coated eggs were more than those of the control group. In the coated groups, Haugh and yolk indices of the coated samples with whey protein concentrate and 0.2 g rice bran oil and with whey protein concentrate and 0.2g of rice bran oil and 1 micro liter of Zataria multiflora extract were more than those of the other coated eggs and the control group eggs. PH values of the control group were higher than those of the coated groups during the storage of the eggs. The shell strength of the coated group was more than that of the control group (uncoated) and in coated samples, whey protein concentrate and 0.2 gr of rice bran oil coated samples had high shell strength. In the other treatments, no significant differences were observed. The depth of the air cell of the coated groups was determined to be less than that of the control group during the storage period. The minimum inhibitory concentration was 1 μL of Zataria multiflora extract. The results showed that 1 μL concentration of Zataria multiflora extract reduces the microbial load of the egg shell surface to 87% and 2 μL reduced total bacterial load to zero. In sensory evaluation, from evaluator point of view, the coated eggs had more overall acceptance than the uncoated group (control), and in the treatment group coated eggs, those containing a low percentage of rice bran oil had higher overall acceptability. In conclusion, coating as a practical and cost effective method can maintain the quality parameters of eggs and lead to durability of supply conditions in addition to the product marketability.

Keywords: edible coating, chicken egg, whey protein concentrate, rice bran oil, Zataria multiflora extract, shelf life

Procedia PDF Downloads 302

Authors: Daria Reczynska, Justyna Jarczak, Michal Czopowicz, Danuta Sloniewska, Karina Horbanczuk, Wieslaw Jarmuz, Jaroslaw Kaba, Emilia Bagnicka

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Since people, animals and plants are constantly exposed to pathogens they have developed very complex systems of defense. Among ca. 1000 antimicrobial peptides from different families so far identified, approximately 30 belonging to cathelicidin family can be found in mammals. Cathelicidins probably constitute the first line of defense because they can act at a physiological salt concentration which is present in healthy tissues. Moreover, the low salt concentration which is present in infected tissues inhibits their activity. In goat bactenecin 7.5 (BAC7.5), bactenecin 5 (BAC5), myeloid antimicrobial peptide 28 (MAP28), myeloid antimicrobial peptide 34 (MAP34 A and B), goat bactenecin3.4 (ChBac3.4) were identified. Caprine arthritis-encephalitis (CAE) caused by small ruminant lentivirus (SRLV) is economic problem. The main CAE symptoms are weight loss, arthritis, pneumonia and mastitis (significant elevation of the somatic cell count and deterioration of some technological parameters). The study was conducted on 24 dairy goats. The animals were divided into two groups: experimental (SRLV-infected) and control (non-infected). The blood samples were collected five times: on the 1st, 7th, 30th, 90th and 150thday of lactation. The levels of transcripts of BAC7.5, BAC5, MAP28 and MAP34 genes in blood leucocytes were measured using qPCR method. There were no differences in mRNA levels of studied genes between stages of lactation. The differences were observed in expressions of BAC5, MAP28 and MAP34 genes with lower levels in the experimental group. There was no difference in BAC7.5 expression between groups. The decreased levels of transcripts of cathelicidin genes in blood leucocytes of SRLV-infected goats may indicate the disturbances of homeostasis in organisms. It can be concluded that SRLV infection seems to inhibit expression of cathelicidin genes. The study was financed by a grant from the National Scientific Center No. UMO-2013/09/B/NZ/03514.

Keywords: goat, CAEV, cathelicidins, blood leukocytes, gene expression

Procedia PDF Downloads 283

Authors: A. Zh. Aupova, A. Ulankyzy, A. Sarsenova, A. Kussayin, Sh. Turarbek, N. Moldagulova, A. Kurmanbayev

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One of the methods of organic waste bioconversion into environmentally-friendly fertilizer is composting. Microorganisms that produce hydrolytic enzymes play a significant role in accelerating the process of organic waste composting. We studied the enzymatic potential (amylase, protease, cellulase, lipase, urease activity) of bacteria isolated from the sewage sludge of Nur-Sultan, Rudny, and Fort-Shevchenko cities, the dacha soil of Nur-Sultan city, and freshly cut grass from the dacha for processing organic waste and identifying active strains. Microorganism isolation was carried out by the cultures enrichment method on liquid nutrient media, followed by inoculating on different solid media to isolate individual colonies. As a result, sixty-one microorganisms were isolated, three of which were thermophiles (DS1, DS2, and DS3). The highest number of isolates, twenty-one and eighteen, were isolated from sewage sludge of Nur-Sultan and Rudny cities, respectively. Ten isolates were isolated from the wastewater of the sewage treatment plant in Fort-Shevchenko. From the dacha soil of Nur-Sultan city and freshly cut grass - 9 and 5 isolates were revealed, respectively. The lipolytic, proteolytic, amylolytic, cellulolytic, ureolytic, and oil-oxidizing activities of isolates were studied. According to the results of experiments, starch hydrolysis (amylolytic activity) was found in 2 isolates - CB2/2, and CB2/1. Three isolates - CB2, CB2/1, and CB1/1 were selected for the highest ability to break down casein. Among isolated 61 bacterial cultures, three isolates could break down fats - CB3, CBG1/1, and IL3. Seven strains had cellulolytic activity - DS1, DS2, IL3, IL5, P2, P5, and P3. Six isolates rapidly decomposed urea. Isolate P1 could break down casein and cellulose. Isolate DS3 was a thermophile and had cellulolytic activity. Thus, based on the conducted studies, 15 isolates were selected as a potential for sewage sludge composting - CB2, CB3, CB1/1, CB2/2, CBG1/1, CB2/1, DS1, DS2, DS3, IL3, IL5, P1, P2, P5, P3. Selected strains were identified on a mass spectrometer (Maldi-TOF). The isolate - CB 3 was referred to the genus Rhodococcus rhodochrous; two isolates CB2 and CB1 / 1 - to Bacillus cereus, CB 2/2 - to Cryseobacterium arachidis, CBG 1/1 - to Pseudoxanthomonas sp., CB2/1 - to Bacillus megaterium, DS1 - to Pediococcus acidilactici, DS2 - to Paenibacillus residui, DS3 - to Brevibacillus invocatus, three strains IL3, P5, P3 - to Enterobacter cloacae, two strains IL5, P2 - to Ochrobactrum intermedium, and P1 - Bacillus lichenoformis. Hence, 60 isolates were isolated from the wastewater of the cities of Nur-Sultan, Rudny, Fort-Shevchenko, the dacha soil of Nur-Sultan city, and freshly cut grass from the dacha. Based on the highest enzymatic activity, 15 active isolates were selected and identified. These strains may become the candidates for bio preparation for sewage sludge processing.

Keywords: sewage sludge, composting, bacteria, enzymatic activity

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Authors: N. N. Pandey, Raghvendra Singh, Biju S. Kamlam, Bipin K. Vishwakarma, Preetam Kala

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The rainbow trout (Oncorhynchus mykiss) is an exotic salmonid fish, well known for its fast growth, tremendous ability to thrive in diverse conditions, delicious flesh and hard fighting nature in Europe and other countries. Rainbow trout farming has a great potential for its contribution to the mainstream economy of Himalayan states in India and other temperate countries. These characteristics establish them as one of the most widely introduced and cultured fish across the globe, and its farming is also prominent in the cold water regions of India. Nevertheless, genetic fatigue, slow growth, early maturity, and low productivity are limiting the expansion of trout production. Moreover, farms adjacent to natural streams or other water sources are subject to escape of domesticated rainbow trout into the wild, which is a serious environmental concern as the escaped fish is subject to contaminate and disrupt the receiving ecosystem. A decline in production traits due to early maturity prolongs the culture duration and affects the profit margin of rainbow trout farms in India. A viable strategy that could overcome these farming constraints in large scale operation is the production of triploid fish that are sterile and more heterozygous. For better triploidy induction rate (TR), heat shock at 28°C for 10 minutes and pressure shock 9500 psi pressure for 5 minutes is applied to green eggs with 90-100% of triploidy success and 72-80% survival upto swim-up fry stage. There is 20% better growth in aquaculture with triploids rainbow trout over diploids. As compared to wild diploid fish, larger sized and fitter triploid rainbow trout in natural waters attract to trout anglers, and support the development of recreational fisheries by state fisheries departments without the risk of contaminating existing gene pools and disrupting local fish diversity. Overall, enhancement of productivity in rainbow trout farms and trout production in coldwater regions, development of lucrative trout angling and better ecological management is feasible with triploid rainbow trout.

Keywords: rainbow trout, triploids fish, heat shock, pressure shock, trout angling

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Authors: Basma Hassabo, Sarah Ahmed, Aisha Hameed

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Aims and Objectives: To determine the incidence of maternal mortality amongst pregnant women with sickle cell disease (SCD) in the United Kingdom and to determine exact cause of death in these women. Background: SCD is caused by the ‘sickle’ gene and is characterized by episodes of severe bone pain and other complications like acute chest syndrome, chronic pulmonary hypertension, stroke, retinopathy, chronic renal failure, hepato-splenic crises, avascular bone necrosis, sepsis and leg ulcers. SCD is a continual cause of maternal mortality and fetal complications, and it comprises 1.5% of all Direct and Indirect deaths in the UK. Sepsis following premature rupture of membranes with ascending infection, post-partum infection and pre-labour overwhelming septic shock is one of its leading causes of death. Over the last fifty years of maternal mortality reports in UK, between 1 to 4 pregnant women died in each triennium. Material and Method: This is a retrospective study that involves pregnant women who died from SCD complications in the UK between 1952-2012. Data were collected from the UK Confidential Enquiries into Maternal Death and its causes between 1952–2012. Prior to 1985, exact cause of death in this cohort was not recorded. Results: 33 deaths reported between 1964 and 1984. 17 deaths were reported due to sickle cell disease between 1985 and 2012. Five women in this group died of sickle cell crisis, one woman had liver sequestration crisis, two women died of venous thromboembolism, two had myocardial fibrosis and three died of sepsis. Remaining women died of amniotic fluid embolism, SUDEP, myocardial ischemia and intracranial haemorrhage. Conclusion: The leading causes of death in sickle cell sick pregnant women are sickle cell crises, sepsis, venous thrombosis and thromboembolism. Prenatal care for women with SCD should be managed by a multidisciplinary team that includes an obstetrician, nutritionist, primary care physician, and haematologist. In every sick Sickle Cell woman Sickle Cell crises should be on the top of the list of differential diagnosis. Aggressive treatment of complications with low threshold to commence broad-spectrum antibiotics and LMWH contribute to better outcomes.

Keywords: incidence, maternal mortality, sickle cell disease (SCD), uk

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Authors: Isabel Brás, Artur Figueirinha, Bruno Esteves, Luísa P. Cruz-Lopes

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The agriculture lignocellulosic by-products are receiving increased attention, namely in the search for filter materials that retain contaminants from water. These by-products, specifically almond and hazelnut shells are abundant in Portugal once almond and hazelnuts production is a local important activity. Hazelnut and almond shells have as main constituents lignin, cellulose and hemicelluloses, water soluble extractives and tannins. Along the adsorption of heavy metals from contaminated waters, water soluble compounds can leach from shells and have a negative impact in the environment. Usually, the chemical characterization of treated water by itself may not show environmental impact caused by the discharges when parameters obey to legal quality standards for water. Only biological systems can detect the toxic effects of the water constituents. Therefore, the evaluation of toxicity by biological tests is very important when deciding the suitability for safe water discharge or for irrigation applications. The main purpose of the present work was to assess the potential impacts of waters after been treated for heavy metal removal by hazelnut and almond shells adsorption systems, with short term acute toxicity tests. To conduct the study, water at pH 6 with 25 mg.L-1 of lead, was treated with 10 g of shell per litre of wastewater, for 24 hours. This procedure was followed for each bark. Afterwards the water was collected for toxicological assays; namely bacterial resistance, seed germination, Lemna minor L. test and plant grow. The effect in isolated bacteria strains was determined by disc diffusion method and the germination index of seed was evaluated using lettuce, with temperature and humidity germination control for 7 days. For aquatic higher organism, Lemnas were used with 4 days contact time with shell solutions, in controlled light and temperature. For terrestrial higher plants, biomass production was evaluated after 14 days of tomato germination had occurred in soil, with controlled humidity, light and temperature. Toxicity tests of water treated with shells revealed in some extent effects in the tested organisms, with the test assays showing a close behaviour as the control, leading to the conclusion that its further utilization may not be considered to create a serious risk to the environment.

Keywords: lignocellulosic wastes, adsorption, acute toxicity tests, risk assessment

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Authors: Rafael Mineiro, André Furtado, Isabel Gonçalves, Cecília Santos, Telma Quintela

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The choroid plexus (CP) epithelial cells form the blood-cerebrospinal fluid barrier. This barrier is highly important for brain protection by physically separating the blood from the cerebrospinal fluid, controlling the trafficking of molecules, including therapeutic drugs, from blood to the brain. The control is achieved by tight junctions between epithelial cells, membrane receptors and transport proteins from the solute carrier and ATP-binding cassette superfamily on the choroid plexus epithelial cells membrane. Previous research of our group showed a functional molecular clock in the CP. The key findings included a rhythmic expression of Bmal1, Per2, and Cry2 in female rat CP. and a rhythmic expression of Cry2 and Per2 in male rat CP. Furthermore, in cultured rat CP epithelial cells we already showed that 17β-estradiol upregulates the expression of Bmal1 and Per1, where the Per1 and Per2 upregulation was abrogated in the presence of the estrogen receptors antagonist ICI. These findings, together with the fact that the CP produces robust rhythms, prompt us to understand the impact of sex hormones and circadian rhythms in CP drug transporters expression, which is a step towards the development and optimization of therapeutic strategies for efficiently delivering drugs to the brain. For that, we analyzed the circadian rhythmicity of the Abcb1, Abcc2, Abcc4 Abcg2, and Oat3 drug transporters at the CP of male and female rats. This analysis was performed by accessing the gene expression of the mentioned transporters at 4 time points by RT-qPCR and the presence of rhythms was evaluated by the CircWave software. Our findings showed a rhythmic expression of Abcc1 in the CP of male rats, of Abcg2 in female rats, and of Abcc4 and Oat3 in both male and female rats with an almost antiphasic pattern between male and female rats for Abcc4. In conclusion, these findings translated to a functional point of view may account for daily variations in brain permeability for several therapeutic drugs, making our findings important data for the future establishment and development of therapeutic strategies according to daytime.

Keywords: choroid plexus, circadian rhythm, membrane transporters, sex hormones

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Authors: Ben Morrow, Patrick O'Hara, Natalie Ban, Tunai Marques, Molly Fraser, Christopher Bone

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Relative to commercial fisheries, recreational fisheries are often poorly understood and pose various challenges for monitoring frameworks. In British Columbia (BC), Canada, Pacific salmon are heavily targeted by recreational fishers while also being a key source of nutrient flow and crucial prey for a variety of marine and terrestrial fauna, including endangered Southern Resident killer whales (Orcinus orca). Although commercial fisheries were historically responsible for the majority of salmon retention, recreational fishing now comprises both greater effort and retention. The current monitoring scheme for recreational salmon fisheries involves aerial-roving creel surveys. However, this method has been identified as costly and having low predictive power as it is often limited to sampling fragments of fluid and temporally dynamic fisheries. This study used imagery from two shore-based autonomous cameras in a highly active recreational fishery around Sooke, BC, and evaluated their efficacy in supplementing existing aerial-roving surveys for monitoring a recreational salmon fishery. This study involved continuous monitoring and high temporal resolution (over one million images analyzed in a single fishing season), using a deep learning-based vessel detection algorithm and a custom image annotation tool to efficiently thin datasets. This allowed for the quantification of peak-season effort from a busy harbour, species-specific retention estimates, high levels of detected fishing events at a nearby popular fishing location, as well as the proportion of the fishery management area represented by cameras. Then, this study demonstrated how it could substantially enhance the temporal resolution of a fishery through diel activity pattern analyses, scaled monthly to visualize clusters of activity. This work also highlighted considerable off-season fishing detection, currently unaccounted for in the existing monitoring framework. These results demonstrate several distinct applications of autonomous cameras for providing enhanced detail currently unavailable in the current monitoring framework, each of which has important considerations for the managerial allocation of resources. Further, the approach and methodology can benefit other studies that apply shore-based camera monitoring, supplement aerial-roving creel surveys to improve fine-scale temporal understanding, inform the optimal timing of creel surveys, and improve the predictive power of recreational stock assessments to preserve important and endangered fish species.

Keywords: cameras, monitoring, recreational fishing, stock assessment

Procedia PDF Downloads 122

Authors: Vanessa Castro, Liria H. Okuda, Daniela P. Chiebao, Adriana H. C. N. Romaldini, Harumi Hojo, Marina Grandi, Joao Paulo A. Silva, Alessandra F. C. Nassar

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The Biological Institute of Sao Paulo, in partnership with a private company, develops an Animal Health Family Farming Program (Prosaf) to enable communication among smallholder farmers and scientists, on-farm consulting and lectures, solving health questions that will benefit agricultural productivity. In Vale do Paraiba region, a dairy region of Sao Paulo State, southern Brazil, many of these types of farms are found with several milk quality problems. Most of these farms are profit-based business; however, with non-technified cattle rearing systems and uncertain veterinary assistance. Feedback from Prosaf showed that the biggest complaints from farmers were low milk production, sick animals and, mainly, loss of selling price due to a high somatic cell count (SCC) and a total bacterial count (TBC). The aims of this study were to improve milk quality, animal hygiene and herd health status by adjustments into general management practices and introducing techniques of sanitary control and milk monitoring in five dairy farms from Sao Jose do Barreiro municipality, Sao Paulo State, Brazil, to increase their profits. A total of 119 milk samples from 56 animals positive for California Mastitis Test (CMT) were collected. The positive CMT indicates subclinical mastitis, therefore laboratorial exams were performed in the milk (microbiological, biochemical and antibiogram test) detect the presence of Staphylococcus aureus (41.8%), Bacillus sp. (11.8%), Streptococcus sp. (2.1%), nonfermenting, motile and oxidase-negative Gram-negative Bacilli (2.1%) and Enterobacter (2.1%). Antibiograms revealed high resistance to gentamicin and streptomycin, probably due to indiscriminate use of antibiotics without veterinarian prescription. We suggested the improvement of hygiene management in the complete milking and cooling tanks system. Using the results of the laboratory tests, animals were properly treated, and the effects observed were better CMT outcomes, lower SCCs, and TBCs leading to an increase in milk pricing. This study will have a positive impact on the family farmers from Sao Paulo State dairy region by improving their market milk competitiveness.

Keywords: milk, family farming, food quality, antibiogram, profitability

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Authors: Pedro B. Antunes, Paulo J. Ramísio

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Road runoff is a linear source of diffuse pollution that can cause significant environmental impacts. During rainfall events, pollutants from both stationary and mobile sources, which have accumulated on the road surface, are dragged through the superficial runoff. Road runoff in coastal zones may present high levels of salinity and chlorides due to the proximity of the sea and transported marine aerosols. Appearing to be correlated to this process, organic matter concentration may also be significant. This study assesses this phenomenon with the purpose of identifying the relationships between monitored water quality parameters and intrinsic site variables. To achieve this objective, an extensive monitoring program was conducted on a Portuguese coastal highway. The study included thirty rainfall events, in different weather, traffic and salt deposition conditions in a three years period. The evaluations of various water quality parameters were carried out in over 200 samples. In addition, the meteorological, hydrological and traffic parameters were continuously measured. The salt deposition rates (SDR) were determined by means of a wet candle device, which is an innovative feature of the monitoring program. The SDR, variable throughout the year, appears to show a high correlation with wind speed and direction, but mostly with wave propagation, so that it is lower in the summer, in spite of the favorable wind direction in the case study. The distance to the sea, topography, ground obstacles and the platform altitude seems to be also relevant. It was confirmed the high salinity in the runoff, increasing the concentration of the water quality parameters analyzed, with significant amounts of seawater features. In order to estimate the correlations and patterns of different water quality parameters and variables related to weather, road section and salt deposition, the study included exploratory data analysis using different techniques (e.g. Pearson correlation coefficients, Cluster Analysis and Principal Component Analysis), confirming some specific features of the investigated road runoff. Significant correlations among pollutants were observed. Organic matter was highlighted as very dependent of salinity. Indeed, data analysis showed that some important water quality parameters could be divided into two major clusters based on their correlations to salinity (including organic matter associated parameters) and total suspended solids (including some heavy metals). Furthermore, the concentrations of the most relevant pollutants seemed to be very dependent on some meteorological variables, particularly the duration of the antecedent dry period prior to each rainfall event and the average wind speed. Based on the results of a monitoring case study, in a coastal zone, it was proven that SDR, associated with the hydrological characteristics of road runoff, can contribute for a better knowledge of the runoff characteristics, and help to estimate the specific nature of the runoff and related water quality parameters.

Keywords: coastal zones, monitoring, road runoff pollution, salt deposition

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Authors: Mike Yaw Osei-Atweneboana, John Asiedu Larbi, Edward Jenner Tettevi

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Background: The lymphatic filariasis (LF) control programme has been on-going in Ghana since 2000. This community-wide approach involves the use of ivermectin (IVM) and albendazole (ALB). Soil-transmitted helminth (STH) infections control is augmented within this programme; however, in areas where LF is not prevalent, albendazole alone is administered to school children. The purpose of this study was therefore, to determine the efficacy of albendazole against soils transmitted helminths and the impact of mass drug administration of albendazole and ivermectin on the health status of children of school going age and pregnant women. Material/Methods: This was a twelve months longitudinal study. A total of 412 subjects including school children (between the ages of 2-17 years) and pregnant women were randomly selected from four endemic communities in Kpandai district of the Northern region. Coprological assessment for parasites was based on the Kato–Katz technique in both dry and rainy seasons at baseline, 21 days and 3 months post-treatment. Single-dose albendazole treatment was administered to all patients at baseline. Preserved samples are currently under molecular studies to identify possible single nucleotide polymorphism (SNP) within the beta tubulin gene which is associated with benzimidazole resistance. Results: Of all the parasites found (hookworm, Trichuris trichiura, Hymenolepis nana, and Taenia sp.); hookworm was the most prevalent. In the dry season, the overall STHs prevalence at pre-treatment was 29%, while 9% and 13% prevalence was recorded at 21 days, and three months after treatment respectively. However, in the rainy season, the overall STHs prevalence was 8%, while 4% and 12% was recorded at 21 days and three months respectively after ALB treatment. In general, ALB treatment resulted in an overall hookworm egg count reduction rate of 89% in the dry season and 93% in the rainy season, while the T. trichiura egg count reduction rate was 100% in both seasons. Conclusions: STH infections still remains a significant public health burden in Ghana. Hookworm infection seems to respond poorly or sub-optimally to ALB, raising concerns of possible emergence of resistance which may lead to a major setback for the control and elimination of STH infections, especially hookworm infections.

Keywords: hookworm, sub-optimal response, albendazole, trichuriasis, soil-transmitted helminths

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Authors: Lubna M. Al-Otaibi

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Mucocele is a common benign neoplasm of the oral cavity and the most common after fibroma. The lesion develops as a result of retention or extravasation of mucous material from minor salivary glands. Extravasation type of mucocele results from trauma and mostly occurs in the lower lip of young patients. The various treatment options available for the treatment of mucocele are associated with a relatively high incidence of recurrence making surgical intervention necessary for a permanent cure. The conventional surgical procedure, however, arouses apprehension in the patient and is associated with bleeding and postoperative pain. Recently, treatment of mucocele with lasers has become a viable treatment option. Various types of lasers are being used and are preferable over the conventional surgical procedure as they provide good hemostasis, reduced postoperative swelling and pain, reduced bacterial population, lesser need for suturing, faster healing and low recurrence rates. Er,Cr:YSGG is a solid-state laser with great affinity to water molecule. Its hydrokinetic cutting action allows it to work effectively on hydrated tissues without any thermal damage. However, up to date, only a few studies have reported its use in the removal of lip mucocele, especially in children. In this case, a 6 year old female patient with history of trauma to the lower lip presented with a soft, sessile, whitish-bluish 4 mm papule. The lesion was present for approximately four months and was fluctuant in size. The child developed a habit of biting the lesion causing injury, bleeding and discomfort. Surgical excision under local anaesthesia was performed using 2780 nm Er,Cr:YSGG Laser (WaterLase iPlus, Irvine, CA) with a Gold handpiece and MZ6 tip (3.5w, 50 Hz, 20% H2O, 20% Air, S mode). The tip was first applied in contact mode with focused beam using the Circumferential Incision Technique (CIT) to excise the tissue followed by the removal of the underlying causative minor salivary gland. Bleeding was stopped using Laser Dry Bandage setting (0.5w, 50 Hz, 1% H2O, 20% Air, S mode) and no suturing was needed. Safety goggles were worn and high-speed suction was used for smoke evacuation. Mucocele excision using 2780 nm Er,Cr:YSGG laser was rapid, easy to perform with excellent precision and allowed for histopathological examination of the excised tissue. The patient was comfortable and there were minimum bleeding and no sutures, postoperative pain, scarring or recurrence. Laser assisted mucocele excision appears to have efficient and reliable benefits in young patients and should be considered as an alternative to conventional surgical and non-surgical techniques.

Keywords: Erbium, excision, laser, lip, mucocele

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Authors: Sanjay Shukla

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Excessive use of antibiotics is a main problem in the treatment of wounds and other chronic infections and antibiotic treatment is frequently non-curative, thus alternative treatment is necessary. Phage therapy is considered one of the most effective approaches to treat multi-drug resistant bacterial pathogens. Infections caused by Staphylococcus aureus are very efficiently controlled with phage cocktails, containing a different individual phages lysate infecting a majority of known pathogenic S. aureus strains. The aim of current study was to investigate the efficiency of a purified phage cocktail for prophylactic as well as therapeutic application in mouse model and in large animals with chronic septic infection of wounds. A total of 150 sewage samples were collected from various livestock farms. These samples were subjected for the isolation of bacteriophage by double agar layer method. A total of 27 sewage samples showed plaque formation by producing lytic activity against S. aureus in double agar overlay method out of 150 sewage samples. In TEM recovered isolates of bacteriophages showed hexagonal structure with tail fiber. In the bacteriophage (ØVS) had an icosahedral symmetry with the head size 52.20 nm in diameter and long tail of 109 nm. Head and tail were held together by connector and can be classified as a member of the Myoviridae family under the order of Caudovirale. Recovered bacteriophage had shown the antibacterial activity against the S. aureus in vitro. Cocktail (ØVS1, ØVS5, ØVS9 and ØVS 27) of phage lysate were tested to know in vivo antibacterial activity as well as the safety profile. Result of mice experiment indicated that the bacteriophage lysate was very safe, did not show any appearance of abscess formation which indicates its safety in living system. The mice were also prophylactically protected against S. aureus when administered with cocktail of bacteriophage lysate just before the administration of S. aureus which indicates that they are good prophylactic agent. The S. aureus inoculated mice were completely recovered by bacteriophage administration with 100% recovery which was very good as compere to conventional therapy. In present study ten chronic cases of wound were treated with phage lysate and follow up of these cases was done regularly up to ten days (at 0, 5 and 10 d). Result indicated that the six cases out of ten showed complete recovery of wounds within 10 d. The efficacy of bacteriophage therapy was found to be 60% which was very good as compared to the conventional antibiotic therapy in chronic septic wounds infections. Thus, the application of lytic phage in single dose proved to be innovative and effective therapy for treatment of septic chronic wounds.

Keywords: phage therapy, phage lysate, antimicrobial resistance, S. aureus

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Authors: Rupinder Bakshi, Geeta Walia, Anita Gupta

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Introduction: Urinary tract infections (UTI) are considered to be one of the most common bacterial infections with an estimated annual global incidence of 150 million. Antimicrobial drug resistance is one of the major threats due to widespread usage of uncontrolled antibiotics. Materials and Methods: A total number of 9149 urine samples were collected from R.H Patiala and processed in the Department of Microbiology G.M.C Patiala. Urine samples were inoculated on MacConkey’s and blood agar plates by using calibrated loop delivering 0.001 ml of sample and incubated at 37 °C for 24 hrs. The organisms were identified by colony characters, gram’s staining and biochemical reactions. Antimicrobial susceptibility of the isolates was determined against various antimicrobial agents (Hi – Media Mumbai India) by Kirby-Bauer disk diffusion method on Muller Hinton agar plates. Results: Maximum patients were in the age group of 21-30 yrs followed by 31-40 yrs. Males (34%) are less prone to urinary tract infections than females (66%). Out of 9149 urine sample, the culture was positive in 25% (2290) samples. Esch. coli was the most common isolate 60.3% (n = 1378) followed by Klebsiella pneumoniae 13.5% (n = 310), Proteus spp. 9% (n = 209), Staphylococcus aureus 7.6 % (n = 173), Pseudomonas aeruginosa 3.7% (n = 84), Citrobacter spp. 3.1 % (70), Staphylococcus saprophyticus 1.8 % (n = 142), Enterococcus faecalis 0.8%(n=19) and Acinetobacter spp. 0.2%(n=5). Gram negative isolates showed higher sensitivity towards, Piperacillin +Tazobactum (67%), Amikacin (80%), Nitrofurantoin (82%), Aztreonam (100%), Imipenem (100%) and Meropenam (100%) while gram positive showed good response towards Netilmicin (69%), Nitrofurantoin (79%), Linezolid (98%), Vancomycin (100%) and Teicoplanin (100%). 465 (23%) isolates were resistant to Penicillins, 1st generation and 2nd generation Cehalosporins which were further tested by double disk approximation test and combined disk method for ESBL production. Out of 465 isolates, 375 were ESBLs consisting of n 264 (70.6%) Esch.coli and 111 (29.4%) Klebsiella pneumoniae. Susceptibility of ESBL producers to Imipenem, Nitrofurantoin and Amikacin were found to be 100%, 76%, and 75% respectively. Conclusion: Uropathogens are increasingly showing resistance to many antibiotics making empiric management of outpatients UTIs challenging. Ampicillin, Cotrimoxazole, and Ciprofloxacin should not be used in empiric treatment. Nitrofurantoin could be used in lower urinary tract infection. Knowledge of uropathogens and their antimicrobial susceptibility pattern in a geographical region will help inappropriate and judicious antibiotic usage in a health care setup.

Keywords: Urinary Tract Infection, UTI, antibiotic susceptibility pattern, ESBL

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Authors: Jitka Rychtarikova

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Regional differences in mortality in the Czech Republic (CR) may be moderate from a broader European perspective, but important discrepancies in life expectancy can be found between smaller territorial units. In this study territorial units are based on Administrative Districts of Municipalities with Extended Powers (MEP). This definition came into force January 1, 2003. There are 205 units and the city of Prague. MEP represents the smallest unit for which mortality patterns based on life tables can be investigated and the Czech Statistical Office has been calculating such life tables (every five-years) since 2004. MEP life tables from 2009-2013 for males and females allowed the investigation of three main life cycles with the use of temporary life expectancies between the exact ages of 0 and 35; 35 and 65; and the life expectancy at exact age 65. The results showed regional survival inequalities primarily in adult and older ages. Consequently, only mortality indicators for adult and elderly population were related to census 2011 unlinked data for the same age groups. The most relevant socio-economic factors taken from the census are: having a partner, educational level and unemployment rate. The unemployment rate was measured for adults aged 35-64 completed years. Exploratory spatial data analysis methods were used to detect regional patterns in spatially contiguous units of MEP. The presence of spatial non-stationarity (spatial autocorrelation) of mortality levels for male and female adults (35-64), and elderly males and females (65+) was tested using global Moran’s I. Spatial autocorrelation of mortality patterns was mapped using local Moran’s I with the intention to depict clusters of low or high mortality and spatial outliers for two age groups (35-64 and 65+). The highest Moran’s I was observed for male temporary life expectancy between exact ages 35 and 65 (0.52) and the lowest was among women with life expectancy of 65 (0.26). Generally, men showed stronger spatial autocorrelation compared to women. The relationship between mortality indicators such as life expectancies and socio-economic factors like the percentage of males/females having a partner; percentage of males/females with at least higher secondary education; and percentage of unemployed males/females from economically active population aged 35-64 years, was evaluated using multiple regression (OLS). The results were then compared to outputs from geographically weighted regression (GWR). In the Czech Republic, there are two broader territories North-West Bohemia (NWB) and North Moravia (NM), in which excess mortality is well established. Results of the t-test of spatial regression showed that for males aged 30-64 the association between mortality and unemployment (when adjusted for education and partnership) was stronger in NM compared to NWB, while educational level impacted the length of survival more in NWB. Geographic variation and relationships in mortality of the CR MEP will also be tested using the spatial Durbin approach. The calculations were conducted by means of ArcGIS 10.6 and SAS 9.4.

Keywords: Czech Republic, mortality, municipality, socio-economic factors, spatial analysis

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Authors: Daniela E. Marin, Cornelia Braicu, Gina C. Pistol, Roxana Cojocneanu-Petric, Ioana Berindan Neagoe, Mihail A. Gras, Ionelia Taranu

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Aristolochic acid (AA) is a carcinogenic, mutagenic, and nephrotoxic compound commonly found in the Aristolochiaceae family of plants. AA is frequently associated with urothelial carcinoma of the upper urinary tract in human and animals and is considered as being responsible for Balkan Endemic Nephropathy. The pig provides a good animal model because the porcine urological system is very similar to that of humans, both in aspects of physiology and anatomy. MicroRNA (miRNA) are small non-coding RNAs that have an impact on a wide range of biological processes by regulating gene expression at post-transcriptional level. The objective of this study was to analyze the miRNA profiling in the kidneys of AA intoxicated swine. For this purpose, ten TOPIGS-40 crossbred weaned piglets, 4-week-old, male and females with an initial average body weight of 9.83 ± 0.5 kg were studied for 28 days. They were given ad libitum access to water and feed and randomly allotted to one of the following groups: control group (C) or aristolochic acid group (AA). They were fed a maize-soybean-meal-based diet contaminated or not with 0.25mgAA/kg. To profile miRNA in the kidneys of pigs, microarrays and bioinformatics approaches were applied to analyze the miRNA in the kidney of control and AA intoxicated pigs. After normalization, our results have shown that a total of 5 known miRNAs and 4 novel miRNAs had different profiling in the kidney of intoxicated animals versus control ones. Expression of miR-32-5p, miR-497-5p, miR-423-3p, miR-218-5p, miR-128-3p were up-regulated by 0.25mgAA/kg feed, while the expression of miR-9793-5p, miR-9835-3p, miR-9840-3p, miR-4334-5p was down-regulated. The microRNA profiling in kidney of intoxicated animals was associated with modified expression of target genes as: RICTOR, LASP1, SFRP2, DKK2, BMI1, RAF1, IGF1R, MAP2K1, WEE1, HDGF, BCL2, EIF4E etc, involved in cell division cycle, apoptosis, cell differentiation and cell migration, cell signaling, cancer etc. In conclusion, this study provides new data concerning the microRNA profiling in kidney after aristolochic acid intoxications with important implications for human and animal health.

Keywords: aristolochic acid, kidney, microRNA, swine

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Authors: Madhusudana S. N., Reeta Mani, Ashwini S. Satishchandra P., Netravati, Udhani V., Fiaz A., Karande S.

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Rabies is an acute encephalitis which is considered 100% fatal despite occasional reports of survivors. However, in recent times more cases of human rabies survivors are being reported. In the last 5 years, there are six laboratories confirmed human rabies survivors in India alone. All cases were children below 15 years and all contracted the disease by dog bites. All of them also had received the full or partial course of rabies vaccination and 4 out of 6 had also received rabies immunoglobulin. All cases were treated in intensive care units in hospitals at Bangalore, Mumbai, Chandigarh, Lucknow and Goa. We report here the results of immunological and virological studies conducted at our laboratory on these patients. The clinical samples that were obtained from these patients were Serum, CSF, nuchal skin biopsy and saliva. Serum and CSF samples were subjected to standard RFFIT for estimation of rabies neutralizing antibodies. Skin biopsy, CSF and saliva were processed by TaqMan real-time PCR for detection of viral RNA. CSF, saliva and skin homogenates were also processed for virus isolation by inoculation of suckling mice. The PBMCs isolated from fresh blood was subjected to ELISPOT assay to determine the type of immune response (Th1/Th2). Both CSF and serum were also investigated for selected cytokines by Luminex assay. The level of antibodies to virus G protein and N protein were determined by ELISA. All survivors had very high titers of RVNA in serum and CSF 100 fold higher than non-survivors and vaccine controls. A five-fold rise in titer could be demonstrated in 4 out of 6 patients. All survivors had a significant increase in antibodies to G protein in both CSF and serum when compared to non-survivors. There was a profound and robust Th1 response in all survivors indicating that interferon gamma could play an important factor in virus clearance. We could isolate viral RNA in only one patient four years after he had developed symptoms. The partial N gene sequencing revealed 99% homology to species I strain prevalent in India. Levels of selected cytokines in CSF and serum did not reveal any difference between survivors and non-survivors. To conclude, survival from rabies is mediated by virus-specific immune responses of the host and clearance of rabies virus from CNS may involve the participation of both Th2 and Th1 immune responses.

Keywords: rabies, rabies treatment, rabies survivors, immune reponse in rabies encephalitis

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Authors: Gachara Grace, Kebira Anthony, Harvey Jagger, Wainaina James

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Aflatoxin contamination of maize in Africa poses a major threat to food security and the health of many African people. In Kenya, aflatoxin contamination of maize is high due to the environmental, agricultural and socio-economic factors. Many studies have been conducted to understand the scope of the problem, especially at pre-harvest level. This research was carried out to gather scientific information on the fungi population, diversity and aflatoxin level during the post-harvest period. The study was conducted in three geographical locations of; Kitui, Kitale and Nakuru. Samples were collected from storage structures of farmers and transported to the Biosciences eastern and central Africa (BecA), International Livestock and Research Institute (ILRI) hub laboratories. Mycoflora was recovered using the direct plating method. A total of five fungal genera (Aspergillus, Penicillium, Fusarium, Rhizopus and Bssyochlamys spp.) were isolated from the stored maize samples. The most common fungal species that were isolated from the three study sites included A. flavus at 82.03% followed by A.niger and F.solani at 49% and 26% respectively. The aflatoxin producing fungi A. flavus was recovered in 82.03% of the samples. Aflatoxin levels were analysed on both the maize samples and in vitro. Most of the A. flavus isolates recorded a high level of aflatoxin when they were analysed for presence of aflatoxin B1 using ELISA. In Kitui, all the samples (100%) had aflatoxin levels above 10ppb with a total aflatoxin mean of 219.2ppb. In Kitale, only 3 samples (n=39) had their aflatoxin levels less than 10ppb while in Nakuru, the total aflatoxin mean level of this region was 239.7ppb. When individual samples were analysed using Vicam fluorometer method, aflatoxin analysis revealed that most of the samples (58.4%) had been contaminated. The means were significantly different (p=0.00<0.05) in all the three locations. Genetic relationships of A. flavus isolates were determined using 13 Simple Sequence Repeats (SSRs) markers. The results were used to generate a phylogenetic tree using DARwin5 software program. A total of 5 distinct clusters were revealed among the genotypes. The isolates appeared to cluster separately according to the geographical locations. Principal Coordinates Analysis (PCoA) of the genetic distances among the 91 A. flavus isolates explained over 50.3% of the total variation when two coordinates were used to cluster the isolates. Analysis of Molecular Variance (AMOVA) showed a high variation of 87% within populations and 13% among populations. This research has shown that A. flavus is the main fungal species infecting maize grains in Kenya. The influence of aflatoxins on human populations in Kenya demonstrates a clear need for tools to manage contamination of locally produced maize. Food basket surveys for aflatoxin contamination should be conducted on a regular basis. This would assist in obtaining reliable data on aflatoxin incidence in different food crops. This would go a long way in defining control strategies for this menace.

Keywords: aflatoxin, Aspergillus flavus, genotyping, Kenya

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Authors: Gabriela Wyszogrodzka, Przemyslaw Dorozynski, Barbara Gil, Maciej Strzempek, Bartosz Marszalek, Piotr Kulinowski, Wladyslaw Piotr Weglarz, Elzbieta Menaszek

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MOFs (Metal-Organic Frameworks) belong to a new group of porous materials with a hybrid organic-inorganic construction. Their structure is a network consisting of metal cations or clusters (acting as metallic centers, nodes) and the organic linkers between nodes. The interest in MOFs is primarily associated with the use of their well-developed surface and large porous. Possibility to build MOFs of biocompatible components let to use them as potential drug carriers. Furthermore, forming MOFs structure from cations possessing paramagnetic properties (e.g. iron cations) allows to use them as MRI (Magnetic Resonance Imaging) contrast agents. The concept of formation of particles that combine the ability to transfer active substance with imaging properties has been called theranostic (from words combination therapy and diagnostics). By building MOF structure from iron cations it is possible to use them as theranostic agents and monitoring the distribution of the active substance after administration in real time. In the study iron-MOF: Fe-MIL-101-NH2 was chosen, consisting of iron cluster in nodes of the structure and amino-terephthalic acid as a linker. The aim of the study was to investigate the possibility of applying Fe-MIL-101-NH2 as inhalable theranostic particulate system for the first-line anti-tuberculosis antibiotic – isoniazid. The drug content incorporated into Fe-MIL-101-NH2 was evaluated by dissolution study using spectrophotometric method. Results showed isoniazid encapsulation efficiency – ca. 12.5% wt. Possibility of Fe-MIL-101-NH2 application as the MRI contrast agent was demonstrated by magnetic resonance tomography. FeMIL-101-NH2 effectively shortening T1 and T2 relaxation times (increasing R1 and R2 relaxation rates) linearly with the concentrations of suspended material. Images obtained using multi-echo magnetic resonance imaging sequence revealed possibility to use FeMIL-101-NH2 as positive and negative contrasts depending on applied repetition time. MOFs micronization via ultrasound was evaluated by XRD, nitrogen adsorption, FTIR, SEM imaging and did not influence their crystal shape and size. Ultrasonication let to break the aggregates and achieve very homogeneously looking SEM images. MOFs cytotoxicity was evaluated in in vitro test with a highly sensitive resazurin based reagent PrestoBlue™ on L929 fibroblast cell line. After 24h no inhibition of cell proliferation was observed. All results proved potential possibility of application of ironMOFs as an isoniazid carrier and as MRI contrast agent in inhalatory treatment of tuberculosis. Acknowledgments: Authors gratefully acknowledge the National Science Center Poland for providing financial support, grant no 2014/15/B/ST5/04498.

Keywords: imaging agents, metal-organic frameworks, theranostics, tuberculosis

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Authors: Viviane A. O. Silva, Angela M. Costa, Glaucia N. M. Hajj, Ana Preto, Aline Tansini, Martin Roffé, Peter Jordan, Rui M. Reis

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Autophagy is a recycling and degradative system suggested to be a major cell death pathway in cancer cells. Autophagy pathway is interconnected with the endocytosis pathways sharing the same ultimate lysosomal destination. Lysosomes are crucial regulators of cell homeostasis, responsible to downregulate receptor signalling and turnover. It seems highly likely that derailed endocytosis can make major contributions to several hallmarks of cancer. WNK2, a member of the WNK (with-no-lysine [K]) subfamily of protein kinases, had been found downregulated by its promoter hypermethylation, and has been proposed to act as a specific tumour-suppressor gene in brain tumors. Although some contradictory studies indicated WNK2 as an autophagy modulator, its role in cancer cell death is largely unknown. There is also growing evidence for additional roles of WNK kinases in vesicular traffic. Aim: To evaluate the role of WNK2 in autophagy and endocytosis on glioma context. Methods: Wild-type (wt) A172 cells (WNK2 promoter-methylated), and A172 transfected either with an empty vector (Ev) or with a WNK2 expression vector, were used to assess the cellular basal capacities to promote autophagy, through western blot and flow-cytometry analysis. Additionally, we evaluated the effect of WNK2 on general endocytosis trafficking routes by immunofluorescence. Results: The re-expression of ectopic WNK2 did not interfere with autophagy-related protein light chain 3 (LC3-II) expression levels as well as did not promote mTOR signaling pathway alteration when compared with Ev or wt A172 cells. However, the restoration of WNK2 resulted in a marked increase (8 to 92,4%) of Acidic Vesicular Organelles formation (AVOs). Moreover, our results also suggest that WNK2 cells promotes delay in uptake and internalization rate of cholera toxin B and transferrin ligands. Conclusions: The restoration of WNK2 interferes in vesicular traffic during endocytosis pathway and increase AVOs formation. This results also suggest the role of WNK2 in growth factor receptor turnover related to cell growth and homeostasis and associates one more time, WNK2 silencing contribution in genesis of gliomas.

Keywords: autophagy, endocytosis, glioma, WNK2

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Authors: Giorgio Bertolazzi, Panayiotis Benos, Michele Tumminello, Claudia Coronnello

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MicroRNAs are small non-coding RNAs that post-transcriptionally regulate the expression levels of messenger RNAs. MicroRNA regulation activity depends on the recognition of binding sites located on mRNA molecules. ComiR (Combinatorial miRNA targeting) is a user friendly web tool realized to predict the targets of a set of microRNAs, starting from their expression profile. ComiR incorporates miRNA expression in a thermodynamic binding model, and it associates each gene with the probability of being a target of a set of miRNAs. ComiR algorithms were trained with the information regarding binding sites in the 3’UTR region, by using a reliable dataset containing the targets of endogenously expressed microRNA in D. melanogaster S2 cells. This dataset was obtained by comparing the results from two different experimental approaches, i.e., inhibition, and immunoprecipitation of the AGO1 protein; this protein is a component of the microRNA induced silencing complex. In this work, we tested whether including coding region binding sites in the ComiR algorithm improves the performance of the tool in predicting microRNA targets. We focused the analysis on the D. melanogaster species and updated the ComiR underlying database with the currently available releases of mRNA and microRNA sequences. As a result, we find that the ComiR algorithm trained with the information related to the coding regions is more efficient in predicting the microRNA targets, with respect to the algorithm trained with 3’utr information. On the other hand, we show that 3’utr based predictions can be seen as complementary to the coding region based predictions, which suggests that both predictions, from 3'UTR and coding regions, should be considered in a comprehensive analysis. Furthermore, we observed that the lists of targets obtained by analyzing data from one experimental approach only, that is, inhibition or immunoprecipitation of AGO1, are not reliable enough to test the performance of our microRNA target prediction algorithm. Further analysis will be conducted to investigate the effectiveness of the tool with data from other species, provided that validated datasets, as obtained from the comparison of RISC proteins inhibition and immunoprecipitation experiments, will be available for the same samples. Finally, we propose to upgrade the existing ComiR web-tool by including the coding region based trained model, available together with the 3’UTR based one.

Keywords: AGO1, coding region, Drosophila melanogaster, microRNA target prediction

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Authors: Sumedha Weerasekara, Morgan Miles, Mark Morrison, Branka Krivokapic-Skoko

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This paper is aimed at exploring the interrelationships among entrepreneurial ecosystem factors and entrepreneurial cognition in regional and metropolitan ecosystems. Entrepreneurial ecosystem factors examined include: culture, infrastructure, access to finance, informal networks, support services, access to universities, and the depth and breadth of the talent pool. Using a multivariate approach we explore the impact of these ecosystem factors or elements on entrepreneurial cognition. In doing so, the existing body of knowledge from the literature on entrepreneurial ecosystem and cognition have been blended to explore the relationship between entrepreneurial ecosystem factors and cognition in a way not hitherto investigated. The concept of the entrepreneurial ecosystem has received increased attention as governments, universities and communities have started to recognize the potential of integrated policies, structures, programs and processes that foster entrepreneurship activities by supporting innovation, productivity and employment growth. The notion of entrepreneurial ecosystems has evolved and grown with the advancement of theoretical research and empirical studies. Importance of incorporating external factors like culture, political environment, and the economic environment within a single framework will enhance the capacity of examining the whole systems functionality to better understand the interaction of the entrepreneurial actors and factors within a single framework. The literature on clusters underplays the role of entrepreneurs and entrepreneurial management in creating and co-creating organizations, markets, and supporting ecosystems. Entrepreneurs are only one actor following a limited set of roles and dependent upon many other factors to thrive. As a consequence, entrepreneurs and relevant authorities should be aware of the other actors and factors with which they engage and rely, and make strategic choices to achieve both self and also collective objectives. The study uses stratified random sampling method to collect survey data from 12 different regions in regional and metropolitan regions of NSW, Australia. A questionnaire was administered online among 512 Small and medium enterprise owners operating their business in selected 12 regions in NSW, Australia. Data were analyzed using descriptive analyzing techniques and partial least squares - structural equation modeling. The findings show that even though there is a significant relationship between each and every entrepreneurial ecosystem factors, there is a weak relationship between most entrepreneurial ecosystem factors and entrepreneurial cognition. In the metropolitan context, the availability of finance and informal networks have the largest impact on entrepreneurial cognition while culture, infrastructure, and support services having the smallest impact and the talent pool and universities having a moderate impact on entrepreneurial cognition. Interestingly, in a regional context, culture, availability of finance, and the talent pool have the highest impact on entrepreneurial cognition, while informal networks having the smallest impact and the remaining factors – infrastructure, universities, and support services have a moderate impact on entrepreneurial cognition. These findings suggest the need for a location-specific strategy for supporting the development of entrepreneurial cognition.

Keywords: academic achievement, colour response card, feedback

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Authors: Ezzati Givi M., Hoveizi E., Nezhad Marani N.

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Platinum-based anticancer therapeutics are the most widely used drugs in clinical chemotherapy but have major limitations and various side effects in clinical applications. Gonadotoxicity and sterility is one of the most common complications for cancer survivors, which seem to be drug-specific and dose-related. Therefore, many efforts have been dedicated to discovering a new structure of platinum-based anticancer agents with improved therapeutic index, fewer side effects. In this regard, new Pt(II)-phosphane complexes containing heterocyclic thionate ligands (PCTL) have been synthesized, which show more potent antitumor activities in comparison to cisplatin. Cisplatin, the best leading metal-based antitumor drug in the field, induces testicular toxicity on Leydig and Sertoli cells leading to serious side effects such as azoospermia and infertility. Therefore in the present study, we aimed to investigate the cytotoxicity effect of PCTL on mice TM4 Sertoli cells with particular emphasis on the role of autophagy in comparison to cisplatin. In this study, an MTT assay was performed to evaluate the IC50 of PCTL and to analyze the TM3 Leydig cell's viability. Cells morphology was evaluated via invert microscope and Changing in morphology for nuclei swelling or autophagic vacuoles formation were assessed by DAPI and MDC staining. Testosterone production in the culture medium was measured using an ELISA kit. Finally, the expression of Autophagy-related genes, Atg5, Beclin1 and p62, were analyzed by qPCR. Based on the obtained results by MTT, the IC50 value of PCTL was 50 μM in TM3 cells and cytotoxic effects was in a dose- and time-dependent manner. Cells morphological changes investigated by inverted microscopy, DAPI, and MDC staining which showed the cytotoxic concentrations of PCTL was significantly higher than cisplatin in the treated TM3 Leydig cells. The results of PCR showed a lack of expression of the p62, Atg5 and Beclin1 gene in TM3 cells treated with PCTL in comparison to cisplatin and control groups. It should be noted that the effects of 25 μM PCTL concentration on TM3 cells have been associated with increased testosterone production and secretion, which requires further study to explain the possible causes and involved molecular mechanisms. The results of the study showed that the PCTL had less-lethal effects on TM3 cells in comparison to cisplatin and probably did not induce autophagy in TM3 cells.

Keywords: platinum-based anticancer agents, cisplatin, Leydig TM3 cells, autophagy

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Authors: Mohammadjavad Sotoudeheian

Abstract:

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

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

Procedia PDF Downloads 86

Authors: Aisha Maroof

Abstract:

Introduction: Coronavirus Disease 2019 (COVID-19) is spreading rapidly around the world with devastating consequences on patients, health care workers and health systems. Severe 2019 novel coronavirus infectious disease (COVID-19) with pneumonia is associated with high rates of admission to the intensive care unit (ICU) and they are at high risk to obtain the hospital acquire bloodstream infection (HAIs) such as central line associated bloodstream infection (CLABSI), catheter associated urinary tract infections (CAUTI) and laboratory confirm bloodstream infection (LCBSI). The chances of infection transmission increase when healthcare worker’s (HCWs) practice is inappropriate. Risk related to hand hygiene (HH) and personal protective equipment (PPE) as regards multidrug-resistant organism transmission: use of multiple gloving instead of HH and incorrect use of PPE can lead to a significant increase of device-related infections. As it reaches low- and middle-income countries, its effects could be even more, because it will be difficult for them to react aggressively to the pandemic. HAIs are one of the biggest medical concerns, resulting in increased mortality rates. Objective: To assess the effect of intervention on compliance of hand hygiene and PPE among HCWs reduce the rate of HAI in COVID-19 patients. Method: An interventional study was done between July to December, 2020. CLABSI, CAUTI and LCBSI data were collected from the medical record and direct observation. There were total of 50 Nurses, 18 doctors and all patients with laboratory-confirmed severe COVID-19 admitted to the hospital were included in this research study. Respiratory tract specimens were obtained after the first 48 h of ICU admission. Practices were observed after and before intervention. Education was provided based on WHO guidelines. Results: During the six months of study July to December, the rate of CLABSI, CAUTI and LCBSI pre and post intervention was reported. CLABSI rate decreasedd from 22.7 to 0, CAUTI rate was decreased from 1.6 to 0, LCBSI declined from 3.3 to 0 after implementation of intervention. Conclusion: HAIs are an important cause of morbidity and mortality. Most of the device related infections occurs due to lack of correct use of PPE and hand hygiene compliance. Hand hygiene and PPE is the most important measure to protect patients, through education it can be improved the correct use of PPE and hand hygiene compliance and can reduce the bacterial infection in COVID-19 patients.

Keywords: hospital acquire infection, healthcare workers, hand hygiene, personal protective equipment

Procedia PDF Downloads 129

Authors: Xiaoyu Sun

Abstract:

In recent years, the rapid development of Artificial Intelligence (AI) has extensively promoted medicine, pharmaceutical, and other related fields. The medical research and development of artificial intelligence by scientific and commercial organizations are on the fast track. The ethics review is one of the critical procedures of registration to get the products approved and launched. However, the SOPs for ethics review is not enough to guide the healthy and rapid development of artificial intelligence in healthcare in China. Ethical Review Measures for Biomedical Research Involving Human Beings was enacted by the National Health Commission of the People's Republic of China (NHC) on December 1st, 2016. However, from a legislative design perspective, it was neither updated timely nor in line with the trends of AI international development. Therefore, it was great that NHC published a consultation paper on the updated version on March 16th, 2021. Based on the most updated laws and regulations in the States and EU, and in-depth-interviewed 11 subject matter experts in China, including lawmakers, regulators, and key members of ethics review committees, heads of Regulatory Affairs in SaMD industry, and data scientists, several suggestions were proposed on top of the updated version. Although the new version indicated that the Ethics Review Committees need to be created by National, Provincial and individual institute levels, the review authorities of different levels were not clarified. The suggestion is that the precise scope of review authorities for each level should be identified based on Risk Analysis and Management Model, such as the complicated leading technology, gene editing, should be reviewed by National Ethics Review Committees, it will be the job of individual institute Ethics Review Committees to review and approve the clinical study with less risk such as an innovative cream to treat acne. Furthermore, to standardize the research and development of artificial intelligence in healthcare in the age of AI, more clear guidance should be given to data security in the layers of data, algorithm, and application in the process of ethics review. In addition, transparency and responsibility, as two of six principles in the Rome Call for AI Ethics, could be further strengthened in the updated version. It is the shared goal among all countries to manage well and develop AI to benefit human beings. Learned from the other countries who have more learning and experience, China could be one of the most advanced countries in artificial intelligence in healthcare.

Keywords: biomedical research involving human beings, data security, ethics committees, ethical review, medical artificial intelligence

Procedia PDF Downloads 168