Search results for: bacterial infection
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 2358

Search results for: bacterial infection

768 Appearance of Ciguatoxin Fish in Atlantic Europe Waters

Authors: J. Bravo, F. Cabrera Suárez, B. Vega, L. Román, M. Martel, F. Acosta

Abstract:

Ciguatera fish poisoning (CFP) is the most common non-bacterial intoxication in the world caused by ingestion of fish with bio-accumulated ciguatoxins (CTXs). It is typical in tropical and subtropical areas, mainly affecting the Caribbean Sea, Polynesia and other areas in the Pacific and Indian Oceans. Interest in Europe by the CFP is increasing in recent years as more and more cases in European hospitals are appearing, usually by people who have consumed ciguatoxin imported fish or have travelled to areas of risk for this poisoning. Since 2004 a series of poisonings raised the question of a possible occurrence of ciguatoxin in Europe, especially in the area of Macaronesia in the East Atlantic temperate zone. Furthermore, some studies have identified the presence of Gambierdiscus spp. in waters surrounding the Canary Islands and Madeira, a toxic dinoflagellate related to this poisoning. The toxin accumulates and concentrates through the food chain and affects to the end of the chain, the human consumer. Fish were collected from the Canary Islands waters and the toxin has been extracted and purified by using acetone and liquid/liquid partition in order to eliminate the excess of fatty acids that may interfere with the detection of the toxin. The fish extracts were inoculated in Neuroblastoma (neuro-2a) cells. After 24-h cell viability was used as an endpoint for cytotoxic effects measurement. Since 2011 our laboratory is collecting data for species such Seriola spp., Epinephelus spp., Makaira spp., Pomatomus spp., Xiphias spp., and Acantocybium spp., from all islands and including the sports fishing and professional activities, we obtained a 8% of fish that have ciguatoxin in their muscle. With these results, we conclude that the island where fishing and fish size affects the probability of catching a fish with the ciguatoxin.

Keywords: Canary Islands, ciguatera fish poisoning, ciguatoxin, Europe

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767 Portfolio Management for Construction Company during Covid-19 Using AHP Technique

Authors: Sareh Rajabi, Salwa Bheiry

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In general, Covid-19 created many financial and non-financial damages to the economy and community. Level and severity of covid-19 as pandemic case varies over the region and due to different types of the projects. Covid-19 virus emerged as one of the most imperative risk management factors word-wide recently. Therefore, as part of portfolio management assessment, it is essential to evaluate severity of such risk on the project and program in portfolio management level to avoid any risky portfolio. Covid-19 appeared very effectively in South America, part of Europe and Middle East. Such pandemic infection affected the whole universe, due to lock down, interruption in supply chain management, health and safety requirements, transportations and commercial impacts. Therefore, this research proposes Analytical Hierarchy Process (AHP) to analyze and assess such pandemic case like Covid-19 and its impacts on the construction projects. The AHP technique uses four sub-criteria: Health and safety, commercial risk, completion risk and contractual risk to evaluate the project and program. The result will provide the decision makers with information which project has higher or lower risk in case of Covid-19 and pandemic scenario. Therefore, the decision makers can have most feasible solution based on effective weighted criteria for project selection within their portfolio to match with the organization’s strategies.

Keywords: portfolio management, risk management, COVID-19, analytical hierarchy process technique

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766 Development and Evaluation of a Nutraceutical Herbal Summer Drink

Authors: Munish Garg, Vinni Ahuja

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In the past few years, the high consumption of soft drinks has attracted negative attention world-wide due to its possible adverse effects, leading the health conscious people to find alternative nutraceutical or herbal health drinks. In the present study, a nutraceutical soft drink was developed utilizing some easily available and well known traditional herbs having nutritional potential. The key ingredients were selected as bael, amla, lemon juice, ashwagandha and poppy seeds based on their household routine use in the summer with proven refreshing, cooling and energetic feeling since ages. After several trials made, the final composition of nutraceutical summer soft drink was selected as most suitable combination based on the taste, physicochemical, microbial and organoleptic point of view. The physicochemical analysis of the prepared drink found to contain optimum level of titratable acidity, total soluble solids and pH which were in accordance of the commercial recommendations. There were no bacterial colonies found in the product therefore found within limits. During the nine point’s hedonic scale sensory evaluation, the drink was strongly liked for colour, taste, flavour and texture. The formulation was found to contain flavonoids (80mg/100ml), phenolics (103mg/100ml), vitamin C (250mg/100ml) and has antioxidant potential (75.52%) apart from providing several other essential vitamins, minerals and healthy components. The developed nutraceutical drink provides an economical and feasible option for the consumers with very good taste combined with potential health benefits. The present drink is potentially capable to replace the synthetic soft drinks available in the market.

Keywords: herbal drink, summer drink, nutraceuticals, soft drink

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765 Separation, Identification, and Measuring Gossypol in the Cottonseed Oil and Investigating the Performance of Drugs Prepared from the Combination of Plant Extract and Oil in the Treatment of Cutaneous Leishmaniasis Resistant to Drugs

Authors: Sara Taghdisi, M. Mirmohammadi, M. Mokhtarian

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In 2013, the World Health Organization announced the cases of Cutaneous leishmaniasis infection in Iran between 69,000 to 113,000. The most common chemical drugs for Cutaneous leishmaniasis treatment are sodium stibogluconate, and meglumine antimonate, which not only have relatively many side effects, but also some species of the Leishmania genus have become resistant to them .The most prominent compound existing in different parts of the cotton plant is a yellow polyphenol called Gossypol. Gossypol is an extremely valuable compound and has anti-cancer properties. In the current project, Gossypol was extracted with a liquid-liquid extraction method in 120 minutes in the presence of Phosphoric acid from the cotton seed oil of Golestan beach varieties, then got crystallized in darkness using Acetic acid and isolated as Gossypol Acetic acid. The efficiency of the extracted crystal was obtained at 0.12+- 1.28. the cotton plant could be efficient in the treatment of Cutaneous leishmaniasis. The extract of the green-leaf cotton boll of Jargoyeh varieties was tested as an ointment on the target group of patients suffering from Cutaneous leishmaniasis resistant to drugs esistant to drugs by our colleagues in the research team. The results showed the Pearson's correlation coefficient of 0.72 between the two variables of wound diameter and the extract use over time which indicated the positive effect of this extract on the treatment of Cutaneous leishmaniasis was resistant to drugs.

Keywords: cottonseed oil, crystallization, gossypol, green-leaf

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764 Prognostic Value of C-Reactive Protein (CRP) in SARS-CoV-2 Infection: A Simplified Biomarker of COVID-19 Severity in Sub-Saharan Africa

Authors: Teklay Gebrecherkos, Mahmud Abdulkader, Tobias Rinke De Wit, Britta C. Urban, Feyissa Chala, Yazezew Kebede, Dawit Welday

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Background: C-reactive protein (CRP) levels are a reliable surrogate for interleukin-6 bioactivity that plays a pivotal role in the pathogenesis of cytokine storm associated with severe COVID-19. There is a lack of data on the role of CRP as a determinant of COVID-19 severity status in the African context. Methods: We determined the longitudinal kinetics of CRP levels on 78 RT-PCR-confirmed COVID-19 patients (49 non-severe and 29 severe cases) and 50 PCR-negative controls. Results: COVID-19 patients had overall significantly elevated CRP at baseline when compared to PCR-negative controls [median 11.1 (IQR: 2.0-127.8) mg/L vs. 0.9 (IQR: 0.5-1.9) mg/L; p=0.0004)]. Moreover, severe COVID-19 patients had significantly higher median CRP levels than non-severe cases [166.1 (IQR: 48.6-332.5) mg/L vs. 2.4 (IQR: 1.2-7.6) mg/L; p<0.00001)]. In addition, persistently elevated levels of CRP were exhibited among those with comorbidities and higher age groups. Area under receiver operating characteristic curve (AUC) analysis of CRP levels distinguished PCR-confirmed COVID-19 patients from the ones with PCR-negative non-COVID-19 individuals, with an AUC value of 0.77 (95% CI: 0.68-0.84; p=0.001). Moreover, it clearly distinguished severe from non-severe COVID-19 patients, with an AUC value of 0.83 (95% CI: 0.73-0.91). After adjusting for age and the presence of comorbidities, CRP levels above 30 mg/L were significantly associated with an increased risk of developing severe COVID-19 (adjusted relative risk 3.99 (95%CI: 1.35-11.82; p=0.013). Conclusions: Determining CRP levels in COVID-19 patients in African settings may provide a simple, prompt, and inexpensive assessment of the severity status at baseline and monitoring of treatment outcomes.

Keywords: CRP, COVID-19, SARS-CoV-2, biomarker

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763 Knowledge, Attitudes and Preventive Practices of Indigenous Adolescents on Dog Associated Zoonotic Infections

Authors: Fairuz Fadzilah Rahim

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Introduction: Indigenous adolescents are at higher risk of dog associated zoonotic infections (DAZI) as they live closely with free-roaming dogs and have limited access to veterinary care. This study aims to determine the effectiveness of health education interventions towards knowledge, attitudes, and preventive practices (KAP) of adolescents on DAZI. Methods: This one-group pre-and post-intervention study in 5 months period was conducted among Jahai adolescents aged 12 years and above. Jahai is one of the three major tribes of indigenous people in Peninsular Malaysia. Health education intervention programs using posters, slide presentations, comics, video clips, and discussion on DAZI were employed. Repeated measures of within-subjects analysis were used to identify the pre- and post- KAP of the adolescents. Results: There were 54 adolescents participated in this study with a mean age of 15.72 (SD: 2.49) and equal proportions of males (50%) and females (50%). Among the adolescents, 22.2% were married, 5.6% were illiterate, and 44.4% not continuing education at the time of data collection. The majority of them keep dogs as pets (64.8%), and few used dogs for hunting (11.1%). There was significant increase in mean scores of knowledge (F = 40.92, p < 0.001) and attitudes (F = 6.43, p = 0.014) of the adolescents. However, the preventive practices towards DAZI showed non-significant improvement on the intervention. Conclusions: The health education intervention programs showed to be effective in improving the attitudes and practices related to dog associated zoonotic infections. Emphasis on sustained health education programs is important to foster good health and wellbeing of the indigenous community.

Keywords: adolescent health, dog associated infection, zoonotic, KAP, indigenous

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762 Comparison of the Positive and Indeterminate Rates of QuantiFERON-TB Gold In-Tube and T-SPOT. TB According to Age-group

Authors: Kina Kim

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Background: There are two types of interferon-gamma release assays (IGRAs) in use for the detection of latent tuberculosis infection (LTBI), QuantiFERON-TB Gold In-tube (QFT-GIT) and T-SPOT.TB. There are some reports that IGRA results are affected by the patient's age. This study aims to compare the results of both IGRA tests according to age groups. Methods: We reviewed 54,882 samples referred to an independent reference laboratory (Seegene Medical Foundation, Seoul, Korea) for the diagnosis of LTBI from January 1, 2021, to December 31, 2021. This retrospective study enrolled 955 patients tested using QFT-GIT and 53,927 patients tested using T-SPOT.TB. The results of both IGRAs were divided in three age groups (0-9, 10-17, and ≥18-year old). The positive rates and the indeterminate rates between QFT-GIT and T-SPOT.TB were compared. We also evaluated the differences in positive and indeterminate rates by age-group. Results: The positive rate of QFT-GIT was 20.1% (192/955) and that of T-SPOT.TB was 8.7% (4704/53927) in overall patients. The positive rates of QFT-GIT in individuals aged 0-9, 10-17, and over 18-year old were 15.4%, 13.3%, and 22.0%, respectively. The positive rates of T-SPOT.TB were 8.9%, 2.0% and 8.8%,in each agegroup, respectively.The overall prevalence of indeterminate results was 2.1% (20/955) of QFT-GIT and 0.5% (270/53927) of T-SPOT.TB. The indeterminate rates of QFT-GIT in individuals aged 0-9, 10-17, and over 18 years were 0.4%, 6.7%, and 2.6%, respectively. The indeterminate rate of T-SPOT.TB were 0.5%, 0.7% and 0.5%,in each age group, respectively. Conclusion: Our findings suggest that T-SPOT.TB has a lower rate of positive results in overall patients and a lower rate of indeterminate results than those of QFT-GIT. The highest positive rate was found in the over 18 years group for QFT-GIT, but the positive rates of T-SPOT.TB was not significantly different among groups by age. QFT-GIT showed variable and higher indeterminate rates according to age group, but T-SPOT.TB showed lower rates in all age groups(<1%).

Keywords: LTBI, IGRA, QFT-GIT, T-SPOT. TB

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761 Saponins vs Anthraquinones: Different Chemicals, Similar Ecological Roles in Marine Symbioses

Authors: Guillaume Caulier, Lola Brasseur, Patrick Flammang, Pascal Gerbaux, Igor Eeckhaut

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Saponins and quinones are two major groups of secondary metabolites widely distributed in the biosphere. More specifically, triterpenoid saponins and anthraquinones are mainly found in a wide variety of plants, bacteria and fungi. In the animal kingdom, these natural organic compounds are rare and only found in small quantities in arthropods, marine sponges and echinoderms. In this last group, triterpenoid saponins are specific to holothuroids (sea cucumbers) while anthraquinones are the chemical signature of crinoids (feather stars). Depending on the species, they present different molecular cocktails. Despite presenting different chemical properties, these molecules share numerous similarities. This study compares the biological distribution, the pharmacological effects and the ecological roles of holothuroid saponins and crinoid anthraquinones. Both of them have been defined as allomones repelling predators and parasites (i.e. chemical defense) and have interesting pharmacological properties (e.g. anti-bacterial, anti-fungal, anti-cancer). Our study investigates the chemical ecology of two symbiotic associations models; between the snapping shrimp Synalpheus stimpsonii associated with crinoids and the Harlequin crab Lissocarcinus orbicularis associated with holothuroids. Using behavioral experiments in olfactometers, chemical extractions and mass spectrometry analyses, we discovered that saponins and anthraquinones present a second ecological role: the attraction of obligatory symbionts towards their hosts. They can, therefore, be defined as kairomones. This highlights a new paradigm in marine chemical ecology: Chemical repellents are attractants to obligatory symbionts because they constitute host specific chemical signatures.

Keywords: anthraquinones, kairomones, marine symbiosis, saponins, attractant

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760 The Arabian Camel (Camelus dromedarius) as a Major Reservoir of Q Fever in Saudi Arabia

Authors: Mansour F. Hussein, Mohammed A. Alshaikh, Riyadh S. Al-Jumaah, A. GarelNabi, I. Al-Khalifa, Osama B. Mohammed

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Serum samples from 489 male and female camels were tested for antibodies against C. burnetii using indirect enzyme-linked immunosorbent assay (ELISA). Antibodies to C. burnetii were recorded in sera of 252 (51.64%) camels. Significant differences in prevalence were found between male and female camels, juvenile and adult camels, different ecotypes and different sampling locations. 307 camels were simultaneously tested for C. burnetii antibodies by ELISA and indirect immunofluorescence (IFA). Close agreement was found between the results of the two tests. A high prevalence of C. burnetii antibodies was also recorded in milk samples tested by ELISA. Clinical samples from serologically positive camels were subjected to PCR analysis using primers which amplify the repetitive transposon-like and transposase gene regions of C. burnetii. Positive DNA amplification was obtained from both regions, with highest shedding of C. burnetii in faecal samples (27.59%) followed, in descending order, by urine (23.81%), blood (15.85%) and milk (6.5%). The present results indicate that camels are a major reservoir of C. burnetii in Saudi Arabia. The high prevalence of infection in camels, the poor sanitary standards under which the animals are kept and the consumption of raw camel milk indicate that camels could also be a major source of transmission of Q fever to humans in Saudi Arabia.

Keywords: Arabian camel, Camelus dromedarius, Coxiella brunetii, ELISA, immunofluoresence, PCR

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759 Nanosilver Containing Biodegradable Bionanocomposites for Antimicrobial Application: Design, Preparation and Study

Authors: Nino Kupatadze, Shorena Tskhadadze, Mzevinar Bedinashvili, David Tugushi, Ramaz Katsarava

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Surgical device-associated infection and biofilm formation are some of the major problems in biomedicine for today. The losing protection ability of conventional antimicrobial-drugs leads to the challenges in the current antibiotic therapy, the most serious of which is antibiotic resistance. Our strategy to overcome the biofilm formation consists in coating devices with polymeric film containing nanosilver(AgNPs) as a bactericidal agent. Such bionanocomposites are also promising as wound dressing materials. For this purpose, we have developed a new generation of AgNPs containing polymeric composites in which amino acid based biodegradable poly(ester amide)s (PEAs) were served as both matrices and AgNPs stabilizers. The AgNPs were formed by photochemical (daylight) reduction of AgNO3 in ethanol solution. The formation of AgNPs was monitored by coloring the solution in brownish-red and appearance of the absorption maximum at 420-430 nm in UV spectrum. Comparative studies of PEAs with polyvinylpyrrolidone (PVP) as particle stabilizers were carried out. It was found that PVP is better stabilizer in terms of particles yield and stability. Therefore, in subsequent experiments blends of PEAs and PVP were used as stabilizers for fabricating AgNPs. As expected, PVP increased the stabilizing effect and this apparently observed in the UV spectrum of the samples after 7 h daylight irradiation: for pure PVP λmax = 430 nm, D = 2.03, for pure PEA λmax= 420 nm, D = 0.65, and for the blend of PVP and PEA λmax = 435 nm, D = 1.88. Further study of the obtained nanobiocomposites is in progress now.

Keywords: biodegradation, bionanocompositions, polymer, nanosilver

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758 Study of the Genes Involved in the Resistance of Nosocomial Pseudomonas aeruginosa to Fluoroquinolone

Authors: Rosetta Moshirian Farahi, Ahya Abdi Ali, Sara Gharavi

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The major mechanism of Pseudomonas aeruginosa resistance to fluoroquinolones is the alteration of target enzymes, type II and IV topoisomerases due to mutations in the quinolone resistance-determining regions (QRDR) of the gyrA and parC genes coding A subunits of these enzymes. 37 isolates from patients with burn wounds and 20 isolates from blood, urine and sputum specimen were selected to evaluate mutations involved in antibiotic resistance and were subsequently verified for their resistance to ciprofloxacin. QRDRs regions of gyrA and parC were amplified by polymerase chain reaction (PCR) and were subsequently sequenced. 90% of isolates with MIC≥8 µg/ml to ciprofloxacin had a mutation in gyrA gene in which threonine at position 83 changed to isoleucine. 87.5% of isolates had mutation in parC, Serine 87 changed. 75% had Ser87Leu and 12.5% possessed Serin87Trp. Various silent mutations were also detected such as Val103Val, Ala118Ala, Ala136Ala, His132His in gyrA and Ala115Ala in parC. The data indicates that the common mutation in gyrA is Thr83Ile and in parC is Ser87Leu/Trp. No individual parC mutation was observed while mutations in gyrA and parC occurred simultaneously and appears to be the main reason of high-level resistance to fluoroquinolones in patients with burn wounds and urine infection. The vast majority of P.aeruginosa isolates had mutation in parC which can play a crucial role in increased resistance of these isolates. This is a report of parC mutations from resistant P. aeruginosa isolates from Iran, Tehran.

Keywords: P. aeruginosa, fluoroquinolones, gyrA, parC, antibiotic resistance

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757 Evaluation of Video Development about Exclusive Breastfeeding as a Nutrition Education Media for Posyandu Cadre

Authors: Ari Istiany, Guspri Devi Artanti, M. Si

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Based on the results Riskesdas, it is known that breastfeeding awareness about the importance of exclusive breastfeeding is still low at only 15.3 %. These conditions resulted in a very infant at risk for infectious diseases, such as diarrhea and acute respiratory infection. Therefore, the aim of this study to evaluate the video development about exclusive breastfeeding as a nutrition education media for posyandu cadre. This research used development methods for making the video about exclusive breastfeeding. The study was conducted in urban areas Rawamangun, East Jakarta. Respondents of this study were 1 media experts from the Department of Educational Technology - UNJ, 2 subject matter experts from Department of Home Economics - UNJ and 20 posyandu cadres to assess the quality of the video. Aspects assessed include the legibility of text, image display quality, color composition, clarity of sound, music appropriateness, duration, suitability of the material and language. Data were analyzed descriptively likes frequency distribution table, the average value, and deviation standard. The result of this study showed that the average score assessment according to media experts, subject matter experts, and posyandu cadres respectively was 3.43 ± 0.51 (good), 4.37 ± 0.52 (very good) and 3.6 ± 0.73 (good). The conclusion is on exclusive breastfeeding video as feasible as a media for nutrition education. While suggestions for the improvement of visual media is multiply illustrations, add material about the correct way of breastfeeding and healthy baby pictures.

Keywords: exclusive breastfeeding, posyandu cadre, video, nutrition education

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756 Retrospective Evaluation of Vector-borne Infections in Cats Living in Germany (2012-2019)

Authors: I. Schäfer, B. Kohn, M. Volkmann, E. Müller

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Introduction: Blood-feeding arthropods transmit parasitic, bacterial, or viral pathogens to domestic animals and wildlife. Vector-borne infections are gaining significance due to the increase of travel, import of domestic animals from abroad, and the changing climate in Europe. Aims of the study: The main objective of this retrospective study was to assess the prevalence of vector-borne infections in cats in which a ‘Feline Travel Profile’ had been conducted. Material and Methods: This retrospective study included test results from cats for which a ‘Feline Travel Profile’ established by LABOKLIN had been requested by veterinarians between April 2012 and December 2019. This profile contains direct detection methods via polymerase chain reaction (PCR) for Hepatozoon spp. and Dirofilaria spp. as well as indirect detection methods via immunofluorescence antibody test (IFAT) for Ehrlichia spp. and Leishmania spp. This profile was expanded to include an IFAT for Rickettsia spp. from July 2015 onwards. The prevalence of the different vector-borne infectious agents was calculated. Results: A total of 602 cats were tested using the ‘Feline Travel Profile’. Positive test results were as follows: Rickettsia spp. IFAT 54/442 (12.2%), Ehrlichia spp. IFAT 68/602 (11.3%), Leishmania spp. IFAT 21/602 (3.5%), Hepatozoon spp. PCR 51/595 (8.6%), and Dirofilaria spp. PCR 1/595 cats (0.2%). Co-infections with more than one pathogen could be detected in 22/602 cats. Conclusions: 170/602 cats (28.2%) were tested positive for at least one vector-borne pathogen. Infections with multiple pathogens could be detected in 3.7% of the cats. The data emphasizes the importance of considering vector-borne infections as potential differential diagnoses in cats.

Keywords: arthopod-transmitted infections, feline vector-borne infections, Germany, laboratory diagnostics

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755 Prevalence and Pattern of Abnormalities Pap Smear Examination in Women Attending Fertility Clinic in Uniosun Teaching Hospitals, Osun State, Nigeria

Authors: Ayodeji Blessing Ajileye

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Introduction: Infertility is described as failure to conceive after one year of unprotected sexual intercourse. One of the causes of female infertility is caused by cervical abnormalities which may be due to bacterial and parasitological infections, hormonal imbalances of Lentinizing hormone, follicular stimulating hormone, oestrogen hormone and progesterone hormone. Aim of the Study: This study aimed to determine the prevalence and pattern of abnormal cervical Pap smear in women with infertility attending fertility clinics at Uniosun Teaching Hospitals Osogbo, Osun State. Methods: This study was conducted at the fertility clinic of University of Osun Teaching Hospital, Osogbo, Osun State. The study population comprised of 50 infertile women and 50 fertile women who are attending the gynecology clinic of University of Osun Teaching Hospital, Osogbo, Osun State. Questionnaire was used to obtain relevant data. Cervical sample was collected using Ayre’s spatula, two smears were prepared and stained with Papanicolaous and H&E staining techniques. Results were analyzed using frequency table. Results: This study observed the prevalence of abnormal cervical smear among infertility women to be 16(30%), while only 03(6%) were observed among the control group (fertile women). Atypical squamous cells of undetermined significance have the highest abnormalities observed in this study with 30%, about 28% of the Pap smear results were negative for inflammation, while total inflammation observed was 72% among the infertility women. Conclusion: This study concluded that abnormal pap smears in this study is significantly more often in women with infertility as compared with fertile women.

Keywords: infertility, oestrogen hormone, pap smears, progesterone hormone

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754 M. J. Rodríguez, F. M. Sánchez, B. Velardo, P. Calvo, M. J. Serradilla, J. Delgado, J. M. López

Authors: Q. Rzina, M. Lahrouni, S. Rida, N. Saadaoui, Y. Almossaid, K. Oufdou, K. Fares

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Many organic solid wastes are produced in the world. Poultry manure (PM), municipal organic wastes (MOW) and sugar beet lime sludge (LS) are produced in large quantities in Morocco. The co-composting of these organic wastes was investigated. The recycling and the valorization of such wastes is environmentally and economically beneficial especially for PM which is known source of bacterial pathogens. The aerobic biodegradation process was carried out by using three windrows of variable compositions: C1 prepared without LS (only MOW were composted with PM), C2 prepared from MOW plus PM and10% LS; and the last one C3 from MOW plus PM and 20% LS. The main process physico-chemical parameters (temperature, pH, humidity and C/N) and microbiological populations (mesophilic and thermophilic flora, total coliform, fecal coliform, Streptococci, Staphylococcus aureus and mesophilic fungi) were monitored over three months to ascertain the compost maturity and to ensure the compost hygienic aspect. The final products were characterized by their relatively high organic matter content, and low C/N ratio of 10.6-10.9. The organic matter degradation was reached approximately 59% for C2 and C3. In addition, the monitoring of the microbial population showed that the produced composts are mature and hygienic. The agronomic valorization of the final composts was tested on radish plant with tree level of composts and poultry manure without composting. The primary results of field trial showed a growth of radish plant biomass and root development without any phytotoxicity detected which reflects the quality of the composts produced. As for poultry manure it allowed to have a better results than other composts because of its readily available nitrogen.

Keywords: compost, municipal organic wastes, poultry manure, radish crop, sugar beet lime sludge

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753 Development and Efficacy Assessment of an Enteric Coated Porous Tablet Loaded with F4 Fimbriae for Oral Vaccination against Enterotoxigenic Escherichia coli Infections

Authors: Atul Srivastava, D. V. Gowda

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Enterotoxigenic Escherichia coli (ETEC) infection is one of the major causes contributing to the development of diarrhoea in adults and children in developing countries. To date, no preventive/treatment strategy showed promising results, which could be due to the lack of potent vaccines, and/or due to the development of resistance of ETEC to antibiotics. Therefore, in the present investigation, a novel porous Sodium Alginate (SA) tablet formulation loaded with F4 fimbriae antigen was developed and tested for efficacy against ETEC infections in piglet models. Pre-compression parameters of the powder mixes and post compression parameters of tablets have been evaluated and results were found to be satisfactory. Loading of F4 fimbrial antigens in to the tablets was achieved by inducing pores in the tablets via the sublimation of camphor followed by incubation with purified F4 fimbriae. The loaded tablets have been coated with Eudragit L100 to protect the F4 fimbriae from (a) highly acidic gastric environment; (b) proteolytic cleavage by pepsin; and (c) to promote subsequent release in the intestine. Evaluation of developed F4 fimbrial tablets in a Pig model demonstrated induction of mucosal immunity, and a significant reduction of F4+ E. coli in faeces. Therefore, F4 fimbriae loaded porous tablets could be a novel oral vaccination candidate to induce mucosal and systemic immunity against ETEC infections.

Keywords: porous tablets, sublimation, f4 fimbriae, eudragit l100, vaccination

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752 Quartz Crystal Microbalance Based Hydrophobic Nanosensor for Lysozyme Detection

Authors: F. Yılmaz, Y. Saylan, A. Derazshamshir, S. Atay, A. Denizli

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Quartz crystal microbalance (QCM), high-resolution mass-sensing technique, measures changes in mass on oscillating quartz crystal surface by measuring changes in oscillation frequency of crystal in real time. Protein adsorption techniques via hydrophobic interaction between protein and solid support, called hydrophobic interaction chromatography (HIC), can be favorable in many cases. Some nanoparticles can be effectively applied for HIC. HIC takes advantage of the hydrophobicity of proteins by promoting its separation on the basis of hydrophobic interactions between immobilized hydrophobic ligands and nonpolar regions on the surface of the proteins. Lysozyme is found in a variety of vertebrate cells and secretions, such as spleen, milk, tears, and egg white. Its common applications are as a cell-disrupting agent for extraction of bacterial intracellular products, as an antibacterial agent in ophthalmologic preparations, as a food additive in milk products and as a drug for treatment of ulcers and infections. Lysozyme has also been used in cancer chemotherapy. The aim of this study is the synthesis of hydrophobic nanoparticles for Lysozyme detection. For this purpose, methacryoyl-L-phenylalanine was chosen as a hydrophobic matrix. The hydrophobic nanoparticles were synthesized by micro-emulsion polymerization method. Then, hydrophobic QCM nanosensor was characterized by Attenuated total reflection Fourier transform infrared (ATR-FTIR) spectroscopy, atomic force microscopy (AFM) and zeta size analysis. Hydrophobic QCM nanosensor was tested for real-time detection of Lysozyme from aqueous solution. The kinetic and affinity studies were determined by using Lysozyme solutions with different concentrations. The responses related to a mass (Δm) and frequency (Δf) shifts were used to evaluate adsorption properties.

Keywords: nanosensor, HIC, lysozyme, QCM

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751 Identifying Pathogenic Mycobacterium Species Using Multiple Gene Phylogenetic Analysis

Authors: Lemar Blake, Chris Oura, Ayanna C. N. Phillips Savage

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Improved DNA sequencing technology has greatly enhanced bacterial identification, especially for organisms that are difficult to culture. Mycobacteriosis with consistent hyphema, bilateral exophthalmia, open mouth gape and ocular lesions, were observed in various fish populations at the School of Veterinary Medicine, Aquaculture/Aquatic Animal Health Unit. Objective: To identify the species of Mycobacterium that is affecting aquarium fish at the School of Veterinary Medicine, Aquaculture/Aquatic Animal Health Unit. Method: A total of 13 fish samples were collected and analyzed via: Ziehl-Neelsen, conventional polymerase chain reaction (PCR) and real-time PCR. These tests were carried out simultaneously for confirmation. The following combination of conventional primers: 16s rRNA (564 bp), rpoB (396 bp), sod (408 bp) were used. Concatenation of the gene fragments was carried out to phylogenetically classify the organism. Results: Acid fast non-branching bacilli were detected in all samples from homogenized internal organs. All 13 acid fast samples were positive for Mycobacterium via real-time PCR. Partial gene sequences using all three primer sets were obtained from two samples and demonstrated a novel strain. A strain 99% related to Mycobacterium marinum was also confirmed in one sample, using 16srRNA and rpoB genes. The two novel strains were clustered with the rapid growers and strains that are known to affect humans. Conclusions: Phylogenetic analysis demonstrated two novel Mycobacterium strains with the potential of being zoonotic and one strain 99% related to Mycobacterium marinum.

Keywords: polymerase chain reaction, phylogenetic, DNA sequencing, zoonotic

Procedia PDF Downloads 136
750 Retrospective Study of Bronchial Secretions Cultures Carried out in the Microbiology Department of General Hospital of Ioannina in 2017

Authors: S. Mantzoukis, M. Gerasimou, P. Christodoulou, N. Varsamis, G. Kolliopoulou, N. Zotos

Abstract:

Purpose: Patients in Intensive Care Units (ICU) are exposed to a different spectrum of microorganisms relative to the hospital. Due to the fact that the majority of these patients are intubated, bronchial secretions should be examined. Material and Method: Bronchial secretions should be taken with care so as not to be mixed with sputum or saliva. The bronchial secretions are placed in a sterile container and then inoculated into blood, Mac Conkey No2, Chocolate, Mueller Hinton, Chapman and Saboureaud agar. After this period, if any number of microbial colonies are detected, gram staining is performed and then the isolated organisms are identified by biochemical techniques in the automated Microscan system (Siemens) followed by a sensitivity test in the same system using the minimum inhibitory concentration MIC technique. The sensitivity test is verified by a Kirby Bauer test. Results: In 2017 the Laboratory of Microbiology received 365 samples of bronchial secretions from the Intensive Care Unit. 237 were found positive. S. epidermidis was identified in 1 specimen, A. baumannii in 60, K. pneumoniae in 42, P. aeruginosa in 50, C. albicans in 40, P. mirabilis in 4, E. coli in 4, S. maltophilia in 6, S. marcescens in 6, S. aureus in 12, S. pneumoniae in 1, S. haemolyticus in 4, P. fluorescens in 1, E. aerogenes in 1, E. cloacae in 5. Conclusions: The majority of ICU patients appear to be a fertile ground for the development of infections. The nature of the findings suggests that a significant part of the bacteria found comes from the unit (nosocomial infection).

Keywords: bronchial secretions, cultures, infections, intensive care units

Procedia PDF Downloads 183
749 Polymer Composites Of MOF-5 For Efficient and Sustained Delivery of Cephalexin and Metronidazole

Authors: Anoff Anim, Lila Mahmoud, Maria Katsikogianni, Sanjit Nayak

Abstract:

Sustained and controlled delivery of antimicrobial drugs have been largely studied recently using metal organic frameworks (MOFs)and different polymers. However, much attention has not been given to combining both MOFs and biodegradable polymers, which would be a good strategy in providing a sustained gradual release of the drugs. Herein, we report a comparative study of the sustained and controlled release of widely used antibacterial drugs, cephalexin and metronidazole, from zinc-based MOF-5 incorporated in biodegradable polycaprolactone (PCL) and poly-lactic glycolic acid (PLGA) membranes. Cephalexin and metronidazole were separately incorporated in MOF-5 post-synthetically, followed by their integration into biodegradable PLGA and PCL membranes. The pristine MOF-5 and the loaded MOFs were thoroughly characterized by FT-IR, SEM, TGA and PXRD. Drug release studies were carried out to assess the release rate of the drugs in PBS and distilled water for up to 48 hours using UV-Vis Spectroscopy. Four bacterial strains from both the Gram-positive and Gram-negative types, Staphylococus aureus, Staphylococuss epidermidis, Escherichia coli, Acinetobacter baumanii, were tested against the pristine MOF, pure drugs, loaded MOFs and the drug-loaded MOF-polymer composites. Metronidazole-loaded MOF-5 composite of PLGA (PLGA-Met@MOF-5) was found to show highest efficiency to inhibit the growth of S. epidermidis compared to the other bacteria strains while maintaining a sustained minimum inhibitory concentration (MIC). This study demonstrates that the combination of biodegradable MOF-polymer composites can provide an efficient platform for sustained and controlled release of antimicrobial drugs and can be a potential strategy to integrate them in biomedical devices.

Keywords: antimicrobial resistance, biodegradable polymers, cephalexin, drug release metronidazole, MOF-5, PCL, PLGA

Procedia PDF Downloads 126
748 Forecasting Unusual Infection of Patient Used by Irregular Weighted Point Set

Authors: Seema Vaidya

Abstract:

Mining association rule is a key issue in data mining. In any case, the standard models ignore the distinction among the exchanges, and the weighted association rule mining does not transform on databases with just binary attributes. This paper proposes a novel continuous example and executes a tree (FP-tree) structure, which is an increased prefix-tree structure for securing compacted, discriminating data about examples, and makes a fit FP-tree-based mining system, FP enhanced capacity algorithm is used, for mining the complete game plan of examples by illustration incessant development. Here, this paper handles the motivation behind making remarkable and weighted item sets, i.e. rare weighted item set mining issue. The two novel brightness measures are proposed for figuring the infrequent weighted item set mining issue. Also, the algorithm are handled which perform IWI which is more insignificant IWI mining. Moreover we utilized the rare item set for choice based structure. The general issue of the start of reliable definite rules is troublesome for the grounds that hypothetically no inciting technique with no other person can promise the rightness of influenced theories. In this way, this framework expects the disorder with the uncommon signs. Usage study demonstrates that proposed algorithm upgrades the structure which is successful and versatile for mining both long and short diagnostics rules. Structure upgrades aftereffects of foreseeing rare diseases of patient.

Keywords: association rule, data mining, IWI mining, infrequent item set, frequent pattern growth

Procedia PDF Downloads 392
747 GeneNet: Temporal Graph Data Visualization for Gene Nomenclature and Relationships

Authors: Jake Gonzalez, Tommy Dang

Abstract:

This paper proposes a temporal graph approach to visualize and analyze the evolution of gene relationships and nomenclature over time. An interactive web-based tool implements this temporal graph, enabling researchers to traverse a timeline and observe coupled dynamics in network topology and naming conventions. Analysis of a real human genomic dataset reveals the emergence of densely interconnected functional modules over time, representing groups of genes involved in key biological processes. For example, the antimicrobial peptide DEFA1A3 shows increased connections to related alpha-defensins involved in infection response. Tracking degree and betweenness centrality shifts over timeline iterations also quantitatively highlight the reprioritization of certain genes’ topological importance as knowledge advances. Examination of the CNR1 gene encoding the cannabinoid receptor CB1 demonstrates changing synonymous relationships and consolidating naming patterns over time, reflecting its unique functional role discovery. The integrated framework interconnecting these topological and nomenclature dynamics provides richer contextual insights compared to isolated analysis methods. Overall, this temporal graph approach enables a more holistic study of knowledge evolution to elucidate complex biology.

Keywords: temporal graph, gene relationships, nomenclature evolution, interactive visualization, biological insights

Procedia PDF Downloads 56
746 Inhibition of Streptococcus Mutans Biofilm Development of Dental Caries In Vitro and In Vivo by Trachyspermum ammi Seeds: An Approach of Alternative Medicine

Authors: Mohd Adil, Rosina Khan, Danishuddin, Asad U. Khan

Abstract:

The aim of this study was to evaluate the influence of the crude and active solvent fraction of Trachyspermum ammi on S. mutans cariogenicity, effect on expression of genes involved in biofilm formation and caries development in rats. GC–MS was carried out to identify the major components present in the crude and the active fraction of T. ammi. The crude extract and the solvent fraction exhibiting least MIC were selected for further experiments. Scanning electron microscopy was carried out to observe the effect of the extracts on S. mutans biofilm. Comparative gene expression analysis was carried out for nine selected genes. 2-Isopropyl-5-methyl-phenol was found as major compound in crude and the active fraction. Binding site of this compound within the proteins involved in biofilm formation was mapped with the help of docking studies. Real-time RT-PCR analyses revealed significant suppression of the genes involved in biofilm formation. All the test groups showed reduction in caries (smooth surface as well as sulcal surface caries) in rats. Moreover, it also provides new insight to understand the mechanism influencing biofilm formation in S. mutans. Furthermore, the data suggest the putative cariostatic properties of T. Ammi and hence can be used as an alternative medicine to prevent caries infection.

Keywords: bio-film, Streptococcus mutans, dental caries, bio-informatic

Procedia PDF Downloads 471
745 Spontaneous Generation of Wrinkled Patterns on pH-Sensitive Smart-Hydrogel Films

Authors: Carmen M. Gonzalez-Henriquez, Mauricio A. Sarabia-Vallejos, Juan Rodriguez-Hernandez

Abstract:

DMAEMA, as a monomer, has been widely studied and used in several application fields due to their pH-sensitive capacity (tertiary amine protonation), being relevant in the biomedical area as a potential carrier for drugs focused on the treatment of genetic or acquired diseases (efficient gene transfection), among others. Additionally, the inhibition of bacterial growth and, therefore, their antimicrobial activity, can be used as dual-functional antifogging/antimicrobial polymer coatings. According to their interesting physicochemical characteristics and biocompatible properties, DMAEMA was used as a monomer to synthesize a smart pH-sensitive hydrogel, namely poly(HEMA-co-PEGDA575-co-DMAEMA). Thus, different mole ratios (ranging from 5:1:0 to 0:1:5, according to the mole ratio between HEMA, PEGDA, and DEAEMA, respectively) were used in this research. The surface patterns formed via a two-step polymerization (redox- and photo-polymerization) were first chemically studied via 1H-NMR and elemental analysis. Secondly, the samples were morphologically analyzed by using Field-Emission Scanning Electron Microscopy (FE-SEM) and Atomic Force Microscopy (AFM) techniques. Then, a particular relation between HEMA, PEGDA, and DEAEMA (0:1:5) was also characterized at three different pH (5.4, 7.4 and 8.3). The hydrodynamic radius and zeta potential of the micro-hydrogel particles (emulsion) were carried out as a possible control for morphology, exploring the effect that produces hydrogel micelle dimensions in the wavelength, height, and roughness of the wrinkled patterns. Finally, contact angle and cross-hatch adhesion test was carried out for the hydrogels supported on glass using TSM-silanized surfaces in order to measure their mechanical properties.

Keywords: wrinkled patterns, smart pH-sensitive hydrogels, hydrogel micelle diameter, adhesion tests

Procedia PDF Downloads 199
744 Enhanced Production of Endo-β-1,4-Xylanase from a Newly Isolated Thermophile Geobacillus stearothermophilus KIBGE-IB29 for Prospective Industrial Applications

Authors: Zainab Bibi, Afsheen Aman, Shah Ali Ul Qader

Abstract:

Endo-β-1,4-xylanases [EC 3.2.1.8] are one of the major groups of enzymes that are involved in degradation process of xylan and have several applications in food, textile and paper processing industries. Due to broad utility of endo-β-1,4-xylanase, researchers are focusing to increase the productivity of this hydrolase from various microbial species. Harsh industrial condition, faster reaction rate and efficient hydrolysis of xylan with low risk of contamination are critical requirements of industry that can be fulfilled by synthesizing the enzyme with efficient properties. In the current study, a newly isolated thermophile Geobacillus stearothermophilus KIBGE-IB29 was used in order to attain the maximum production of endo-1,4-β-xylanase. Bacterial culture was isolated from soil, collected around the blast furnace site of a steel processing mill, Karachi. Optimization of various nutritional and physical factors resulted the maximum synthesis of endo-1,4-β-xylanase from a thermophile. High production yield was achieved at 60°C and pH-6.0 after 24 hours of incubation period. Various nitrogen sources viz. peptone, yeast extract and meat extract improved the enzyme synthesis with 0.5%, 0.2% and 0.1% optimum concentrations. Dipotassium hydrogen phosphate (0.25%), potassium dihydrogen phosphate (0.05%), ammonium sulfate (0.05%) and calcium chloride (0.01%) were noticed as valuable salts to improve the production of enzyme. The thermophilic nature of isolate, with its broad pH stability profile and reduced fermentation time indicates its importance for effective xylan saccharification and for large scale production of endo-1,4-β-xylanase.

Keywords: geobacillus, optimization, production, xylanase

Procedia PDF Downloads 306
743 RNA Antisense Coat Protein Showing Promising Effects against Cotton Leaf Curl Disease in Pakistani Cotton

Authors: Zunnu Raen Akhtar

Abstract:

Cotton Leaf Curl Disease (CLCuD) is from Gemini virus and is transmitted through whiteflies in cotton. Transgenic cotton containing Antisense Coat Protein (ACP) has been found to show better results against CLCuD in cotton. In current research, Antisense Coat Protein was inserted in cotton plants to observe resistance developed in the cotton plants against CLCuD. T1 generation of plants were observed for its expression in plants. Tests were carried out to observe the expression of Antisense Coat Protein using Polymerase Chain Reaction (PCR) technique and by southern blotting. Whiteflies showing positive Cotton Leaf Curl Virus (CLCV) were reared and released in bioassay on ACP expressing cotton plants under laboratory as well as confined semi-field conditions. Results confirmed the expression of AC protein in PCR and southern blotting. Further laboratory results showed that cotton plants expressing AC protein showed rare incidence of CLCuD infection as compared to control. In the confined semi-field, similar results were observed in AC protein expressing cotton as compared to control. These results explicitly show that ACP can help to tackle the CLCuD issue in the future and further studies on biochemical processes involved in these plants and effects of ACP induction on non-target organisms should also be studied for eco-system.

Keywords: cotton, white flies, antisense coat protein, CLCV

Procedia PDF Downloads 179
742 Biodegradable Polymer Composites of MOF-5 for Efficient and Sustained Delivery of Cephalexin and Metronidazole

Authors: Anoff Anim, Lila A. M. Mahmoud, Maria Katsikogianni, Sanjit Nayak

Abstract:

Sustained and controlled delivery of antimicrobial drugs have been largely studied recently using metal organic frameworks (MOFs)and different polymers. However, much attention has not been given to combining both MOFs and biodegradable polymers, which would be a good strategy in providing a sustained gradual release of the drugs. Herein, we report a comparative study of the sustained and controlled release of widely used antibacterial drugs, cephalexin and metronidazole, from zinc-based MOF-5 incorporated in biodegradable polycaprolactone (PCL) and poly-lactic glycolic acid (PLGA) membranes. Cephalexin and metronidazole were separately incorporated in MOF-5 post-synthetically, followed by their integration into biodegradable PLGA and PCL membranes. The pristine MOF-5 and the loaded MOFs were thoroughly characterized by FT-IR, SEM, TGA and PXRD. Drug release studies were carried out to assess the release rate of the drugs in PBS and distilled water for up to 48 hours using UV-Vis Spectroscopy. Four bacterial strains from both the Gram-positive and Gram-negative types, Staphylococus aureus, Staphylococuss epidermidis, Escherichia coli, Acinetobacter baumanii, were tested against the pristine MOF, pure drugs, loaded MOFs and the drug-loaded MOF-polymer composites. Metronidazole-loaded MOF-5 composite of PLGA (PLGA-Met@MOF-5) was found to show highest efficiency to inhibit the growth of S. epidermidis compared to the other bacteria strains while maintaining a sustained minimum inhibitory concentration (MIC). This study demonstrates that the combination of biodegradable MOF-polymer composites can provide an efficient platform for sustained and controlled release of antimicrobial drugs and can be a potential strategy to integrate them in biomedical devices.

Keywords: antimicrobial resistance, biodegradable polymers, cephalexin, drug release metronidazole, MOF-5, PCL, PLGA

Procedia PDF Downloads 129
741 Apollo Clinical Excellence Scorecard (ACE@25): An Initiative to Drive Quality Improvement in Hospitals

Authors: Anupam Sibal

Abstract:

Whatever is measured tends to improve. With a view to objectively measuring and improving clinical quality across the Apollo Group Hospitals, the initiative of ACE @ 25 (Apollo Clinical Excellence@25) was launched on Jan 09. ACE @ 25 is a clinically balanced scorecard incorporating 25 clinical quality parameters involving complication rates, mortality rates, one-year survival rates and average length of stay after major procedures like liver and renal transplant, CABG, TKR, THR, TURP, PTCA, endoscopy, large bowel resection and MRM covering all major specialties. Also included are hospital acquired infection rates, pain satisfaction and medication errors. Benchmarks have been chosen from the world’s best hospitals. There are weighted scores for outcomes color coded green, orange and red. The cumulative score is 100. Data is reported monthly by 43 Group Hospitals online on the Lighthouse platform. Action taken reports for parameters falling in red are submitted quarterly and reviewed by the board. An audit team audits the data at all locations every six months. Scores are linked to appraisal of the medical head and there is an “ACE @ 25” Champion Award for the highest scorer. Scores for different parameters were variable from green to red at the start of the initiative. Most hospitals showed an improvement in scores over the last four years for parameters where they had showed scores in red or orange at the start of the initiative. The overall scores for the group have shown an increase from 72 in 2010 to 81 in 2015.

Keywords: benchmarks, clinical quality, lighthouse, platform, scores

Procedia PDF Downloads 293
740 Contactless Attendance System along with Temperature Monitoring

Authors: Nalini C. Iyer, Shraddha H., Anagha B. Varahamurthy, Dikshith C. S., Ishwar G. Kubasad, Vinayak I. Karalatti, Pavan B. Mulimani

Abstract:

The current scenario of the pandemic due to COVID-19 has led to the awareness among the people to avoid unneces-sary contact in public places. There is a need to avoid contact with physical objects to stop the spreading of infection. The contactless feature has to be included in the systems in public places wherever possible. For example, attendance monitoring systems with fingerprint biometric can be replaced with a contactless feature. One more important protocol followed in the current situation is temperature monitoring and screening. The paper describes an attendance system with a contactless feature and temperature screening for the university. The system displays a QR code to scan, which redirects to the student login web page only if the location is valid (the location where the student scans the QR code should be the location of the display of the QR code). Once the student logs in, the temperature of the student is scanned by the contactless temperature sensor (mlx90614) with an error of 0.5°C. If the temperature falls in the range of the desired value (range of normal body temperature), then the attendance of the student is marked as present, stored in the database, and the door opens automatically. The attendance is marked as absent in the other case, alerted with the display of temperature, and the door remains closed. The door is automated with the help of a servomotor. To avoid the proxy, IR sensors are used to count the number of students in the classroom. The hardware system consisting of a contactless temperature sensor and IR sensor is implemented on the microcontroller, NodeMCU.

Keywords: NodeMCU, IR sensor, attendance monitoring, contactless, temperature

Procedia PDF Downloads 181
739 Optimization of Hepatitis B Surface Antigen Purifications to Improving the Production of Hepatitis B Vaccines on Pichia pastoris

Authors: Rizky Kusuma Cahyani

Abstract:

Hepatitis B is a liver inflammatory disease caused by hepatitis B virus (HBV). This infection can be prevented by vaccination which contains HBV surface protein (sHBsAg). However, vaccine supply is limited. Several attempts have been conducted to produce local sHBsAg. However, the purity degree and protein yield are still inadequate. Therefore optimization of HBsAg purification steps is required to obtain high yield with better purification fold. In this study, optimization of purification was done in 2 steps, precipitation using variation of NaCl concentration (0,3 M; 0,5 M; 0,7 M) and PEG (3%, 5%, 7%); ion exchange chromatography (IEC) using NaCl 300-500 mM elution buffer concentration.To determine HBsAg protein, bicinchoninic acid assay (BCA) and enzyme-linked immunosorbent assay (ELISA) was used in this study. Visualization of HBsAg protein was done by SDS-PAGE analysis. Based on quantitative analysis, optimal condition at precipitation step was given 0,3 M NaCl and PEG 3%, while in ion exchange chromatography step, the optimum condition when protein eluted with NaCl 500 mM. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis indicates that the presence of protein HBsAg with a molecular weight of 25 kDa (monomer) and 50 kDa (dimer). The optimum condition for purification of sHBsAg produced in Pichia pastoris gave a yield of 47% and purification fold 17x so that it would increase the production of hepatitis B vaccine to be more optimal.

Keywords: hepatitis B virus, HBsAg, hepatitis B surface antigen, Pichia pastoris, purification

Procedia PDF Downloads 148