Search results for: clinical laboratory persons

Authors: Chatrabhuti Pipop, Visessri Supattra, Charinpanitkul Tawatchai

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Durian is one of the top agricultural products exported by Thailand. There is the massive market potential for the durian industry. While the global demand for Thai durians, especially the demand from China, is very high, Thailand's durian supply is far from satisfying strong demand. Poor agricultural practices result in low yields and poor quality of fruit. Most irrigation systems currently used by the farmers are fixed schedule or fixed rates that ignore actual weather conditions and crop water requirements. In addition, the technologies emerging are too difficult and complex and prices are too high for the farmers to adopt and afford. Many farmers leave the durian trees to grow naturally. With improper irrigation and nutrient management system, durians are vulnerable to a variety of issues, including stunted growth, not flowering, diseases, and death. Technical development or research for durian is much needed to support the wellbeing of the farmers and the economic development of the country. However, there are a limited number of studies or development projects for durian because durian is a perennial crop requiring a long time to obtain the results to report. This study, therefore, aims to address the problem of durian production by developing an autonomous and precision irrigation system. The system is designed and equipped with an industrial programmable controller, a weather station, and a digital flow meter. Daily water requirements are computed based on weather data such as rainfall and evapotranspiration for daily irrigation with variable flow rates. A prediction model is also designed as a part of the system to enhance the irrigation schedule. Before the system was installed in the field, a simulation model was built and tested in a laboratory setting to ensure its accuracy. Water consumption was measured daily before and after the experiment for further analysis. With this system, the crop water requirement is precisely estimated and optimized based on the data from the weather station. Durian will be irrigated at the right amount and at the right time, offering the opportunity for higher yield and higher income to the farmers.

Keywords: Durian, precision irrigation, precision agriculture, smart farm

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Authors: Alisher Ibragimov, Sofya Senotrusova, Aleksandra Beliaeva, Egor Ushakov, Yuri Markin

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Mammography plays a critical role in screening for breast cancer in women, and artificial intelligence has enabled the automatic detection of diseases in medical images. Many of the current techniques used for mammogram analysis focus on a single view (mediolateral or craniocaudal view), while in clinical practice, radiologists consider multiple views of mammograms from both breasts to make a correct decision. Consequently, computer-aided diagnosis (CAD) systems could benefit from incorporating information gathered from multiple views. In this study, the introduce a method based on a Siamese neural network (SNN) model that simultaneously analyzes mammographic images from tri-view: bilateral and ipsilateral. In this way, when a decision is made on a single image of one breast, attention is also paid to two other images – a view of the same breast in a different projection and an image of the other breast as well. Consequently, the algorithm closely mimics the radiologist's practice of paying attention to the entire examination of a patient rather than to a single image. Additionally, to the best of our knowledge, this research represents the first experiments conducted using the recently released Vietnamese dataset of digital mammography (VinDr-Mammo). On an independent test set of images from this dataset, the best model achieved an AUC of 0.87 per image. Therefore, this suggests that there is a valuable automated second opinion in the interpretation of mammograms and breast cancer diagnosis, which in the future may help to alleviate the burden on radiologists and serve as an additional layer of verification.

Keywords: breast cancer, computer-aided diagnosis, deep learning, multi-view mammogram, siamese neural network

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Authors: Elizabeth Loza-Valerdi, Victor Pardiñas-Rios, Arnulfo Pluma-Pluma, Andres Breton-Toral, Julio Cercado-Jaramillo

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The purification processes for mixtures of isomeric monosaccharides using industrial chromatographic methods poses a serious technical challenge. Mixtures of 2 or 3 monosaccharides are difficult to separate by strictly physical or chemical techniques. Differential fermentation by microbial cultures is an increasingly interesting way of selective enrichment in a particular kind of monosaccharides when a mixture of them is present in the solution, and only one has economical value. Osmotolerant yeast cultures provide an interesting source of biocatalysts for the selective catabolism of monosaccharides in media containing high concentrations of total soluble sugars. A collection of 398 yeast strains has been obtained using endemic and unique sources of fruit juices, industrial syrups, honey, and other high sugar content substrates, either natural or man made, products and by-products from Mexico. The osmotolerance of the strains was assessed by plate assay both in glucose (20-40-60%w/w). Strains were classified according to their osmotolerance in low, medium or highly tolerant to high glucose concentrations. The purified cultures were tested by their ability to growth in a solid plate media or liquid media of Yeas Nitrogen Base (YNB), added with specific monosaccharides as sole carbon source (glucose, galactose, lactose and fructose). Selected strains were subsequently tested in fermentation experiments with mixtures of two monosaccharides (galactose/glucose and glucose/fructose). Their ability to grow and selectively catabolize one monosaccharide was evaluated. Growth, fermentation activity and products of metabolism were determined by plate counts, CO2 production, turbidity and chromatographic analysis by HPLC. Selective catabolism of one monosaccharide in liquid media containing two monosaccharides was confirmed for 8 strains. Ion Exchange chromatographic processes were used in production of high fructose or galactose syrup. Laboratory scale processes for the production of fructose or galactose enriched syrups is now feasible, with important applications in food (like high fructose syrup as edulcorant) and fermentation technology (for GOS production).

Keywords: osmotolerant yeasts, selective metabolism, fructose syrup, GOS

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Authors: Tzu-Hao Li, Yen-Ying Kung, Chang-Youh Tsai

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Oral dryness is a common chief complaint among patients with Sjőgren syndrome (SS), which is a disorder currently known as autoantibodies production; however, to author’s best knowledge, there has been no satisfying pharmacy to relieve the associated symptoms. Hence the effectiveness of other non-pharmacological interventions such as acupuncture should be accessed. We conducted a meta-analysis of randomized clinical trials (RCTs) which evaluated the effectiveness of xerostomia in SS. PubMed, Embase, Cochrane Central Register of Controlled Trials (CENTRAL), Chongqing Weipu Database (CQVIP), China Academic Journals Full-text Database, AiritiLibrary, Chinese Electronic Periodicals Service (CEPS), China National Knowledge Infrastructure (CNKI) Database were searches through May 12, 2018 to select studies. Data for evaluation of subjective and objective xerostomia was extracted and was assessed with random-effects meta-analysis. After searching, a total of 541 references were yielded and five RCTs were included, covering 340 patients dry mouth resulted from SS, among whom 169 patients received acupuncture and 171 patients were control group. Acupuncture group was associated with higher subjective response rate (odds ratio 3.036, 95% confidence interval [CI] 1.828 – 5.042, P < 0.001) and increased salivary flow rate (weighted mean difference [WMD] 3.066, 95% CI 2.969 – 3.164, P < 0.001), as an objective marker. In addition, two studies examined IgG levels, which were lower in the acupuncture group (WMD -166.857, 95% CI -233.138 - -100.576, P < 0.001). Therefore, in the present meta-analysis, acupuncture improves both subjective and objective markers of dry mouth with autoantibodies reduction in patients with SS and is considered as an option of non-pharmacological treatment for SS.

Keywords: acupuncture, meta-analysis, Sjogren syndrome, xerostomia

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Authors: Nitesh Kumar, Adel Elrasheed, Antonio Gagliardilugo

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Introduction and aims: PRP has been increasing in popularity over the past decade. It is used in many facets of medicine and dentistry such as osteoarthritis, hair loss, skin rejunavation, healing of tendons after injury. Due to the increasing popularity of PRP in third molar surgery in dentistry, this study aims to identify the role of platelet rich plasma and its function in third molar surgery. Methodology: Three databases were chosen to source the articles for review: pubmed, science direct, and Cochrane. The keywords “platelet rich plasma”, “third molar extraction” and “wisdom tooth extraction” and literature review were used to search for relevant articles. Articles that were not in English were omitted and only systematic reviews relevant to the study were collected. All systematic reviews abstracts pertinent to the study were read by two reviewers to avoid bias. Results/statistics: 20 review articles were obtained of which 13 fulfilled the criteria. The Amstar tool validified the strength of these review articles. There is strong evidence in the literature that PRP in third molar surgery decreases post op pain, swelling and recovery time. 20 review articles were obtained of which 13 fulfilled the criteria. The Amstar tool validified the strength of these review articles. There is strong evidence in the literature that PRP in third molar surgery decreases post op pain, swelling and recovery time. Conclusions/clinical relevance: Platelet rich plasma plays a crucial role in patient recovery following the extraction of third molars and should be considered and offered as a routine part of third molar therapy.

Keywords: PRP, third molar, extractions, wisdom teeth

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Authors: Mohamed Korany, Azza Gazy, Essam Khamis, Marwa Adel, Miranda Fawzy

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Background: Two new, simple and specific methods; First, a Zero-crossing first-derivative technique and second, a chemometric-assisted spectrophotometric artificial neural network (ANN) were developed and validated in accordance with ICH guidelines. Both methods were used for the simultaneous estimation of the two closely related antioxidant nutraceuticals ; Coenzyme Q10 (Q) ; also known as Ubidecarenone or Ubiquinone-10, and Vitamin E (E); alpha-tocopherol acetate, in their pharmaceutical binary mixture. Results: For first method: By applying the first derivative, both Q and E were alternatively determined; each at the zero-crossing of the other. The D1 amplitudes of Q and E, at 285 nm and 235 nm respectively, were recorded and correlated to their concentrations. The calibration curve is linear over the concentration range of 10-60 and 5.6-70 μg mL-1 for Q and E, respectively. For second method: ANN (as a multivariate calibration method) was developed and applied for the simultaneous determination of both analytes. A training set (or a concentration set) of 90 different synthetic mixtures containing Q and E, in wide concentration ranges between 0-100 µg/mL and 0-556 µg/mL respectively, were prepared in ethanol. The absorption spectra of the training sets were recorded in the spectral region of 230–300 nm. A Gradient Descend Back Propagation ANN chemometric calibration was computed by relating the concentration sets (x-block) to their corresponding absorption data (y-block). Another set of 45 synthetic mixtures of the two drugs, in defined range, was used to validate the proposed network. Neither chemical separation, preparation stage nor mathematical graphical treatment were required. Conclusions: The proposed methods were successfully applied for the assay of Q and E in laboratory prepared mixtures and combined pharmaceutical tablet with excellent recoveries. The ANN method was superior over the derivative technique as the former determined both drugs in the non-linear experimental conditions. It also offers rapidity, high accuracy, effort and money saving. Moreover, no need for an analyst for its application. Although the ANN technique needed a large training set, it is the method of choice in the routine analysis of Q and E tablet. No interference was observed from common pharmaceutical additives. The results of the two methods were compared together

Keywords: coenzyme Q10, vitamin E, chemometry, quantitative analysis, first derivative spectrophotometry, artificial neural network

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Authors: Mary Therese G. Caluna, Mark Justin B. Campanero, Erlin Maris T. Cantiller, Claudine Mae A. Cantillo, Nerissa L. Caño

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Objective: The study determined the effectiveness of cold tomato paste in relieving pain caused by exacerbation of arthritis in the elderly, specifically on clients 60 years old and above. The study focused on alternative, cost-effective and non-pharmacological techniques in relieving pain experienced by the older people with osteoarthritis and rheumatoid arthritis. Methods: Using purposive non-probability sampling, the researchers gathered a total number of 40 subjects that passed the inclusion criteria provided by the researchers. The subjects were divided into two groups, experimental group (20 subjects) and control groups (20 subjects). The Numeric Rating 11-point Scale (NRS-11) was utilized to assess the pain level of the subject prior the application of the treatment and after the application of the treatment. Key findings: There is a significant difference in the pain levels of the experimental group before and after the application of cold tomato paste. This indicates that that the application of cold tomato paste alleviates the pain experienced by elderly clients with exacerbation of arthritis. Conclusion: The effectiveness of cold tomato paste in relieving pain experienced by elderly clients who are in exacerbation of arthritis was proven to be evidence-based. The cold tomato paste application has significant impact in the field of nursing and therefore, can be used in both clinical trials and practices. The effectiveness of cold tomato application promotes innovation in the field of nursing, thus encouraging further researches regarding other uses of tomato and other herbal interventions to relieve the pain caused by osteoarthritis and rheumatoid arthritis.

Keywords: alternative therapy, arthritis, cold tomato paste, elderly clients, exacerbation

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Authors: Anit S. Malhotra, Ajay Duseja, Neelam Chadha

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Endothelial dysfunction is a precursor to atherosclerosis, and flow-mediated dilatation (FMD) in the brachial artery is the commonest method to evaluate endothelial function in humans. Non-alcoholic fatty liver disease (NAFLD) is one of the most common liver disorders encountered in clinical practice. An earlier meta-analysis had quantitatively assessed the degree of endothelial dysfunction using FMD. However, the largest study investigating the relation of FMD with NAFLD was published after that meta-analysis. In addition, that meta-analysis did not include some studies, including one from our centre. Therefore, an updating the previous meta-analysis was considered important. We searched PubMed, Cochrane Library, Embase, Scopus, SCI, Google Scholar, conference proceedings, and references of included studies till June 2017 to identify observational studies evaluating endothelial function using FMD in patients with non-alcoholic fatty liver disease. Data was analyzed using MedCalc. Fourteen studies were found eligible for inclusion in the meta-analysis. Patients with NAFLD had lower brachial artery FMD as compared to controls, standardized mean difference (random effects model) being –1.279%; 95% confidence interval (CI), –1.478 to –0.914. The effect size became smaller after addition of the recent study with the largest sample size was included compared with the earlier meta-analysis. In conclusion, patients with NAFLD had low FMD values indicating that they are at a higher risk of cardiovascular disease although our results suggest the effect size is not as large as reported previously.

Keywords: endothelial dysfunction, flow-mediated dilatation, meta-analysis, non-alcoholic fatty liver disease

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Authors: Ester Ato, Maria Angeles Fernández-Vilar, Maria Dolores Galián

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The aim of this work was to analyze the relation between children´s effortful control, marital adjustment and parenting practices in a sample of 345 Spanish children aged between 6 and 8 years. Traditionally, the literature confirms that a higher level of marital conflict has been associated with less effective and less positive parenting, but there are few studies that include the effect that children´s effortful control exert to this relation. To measure marital adjustment, parenting practices and children’s temperament, parents were given the Marital Adjustment Test (MAT), the Spanish version of the PCRI (Parent-Child Relationship Inventory), and the TMCQ (Temperament in Middle Childhood Questionnaire). The results confirmed that higher marital satisfaction predicted more positive parenting practices, whereas lower marital adjustment scores predicted less parenting support and control. Using a statistical modeling approach, we tested a moderation model that revealed the moderating role of effortful control in the relation between marital adjustment and parenting. Concretely, higher marital satisfaction predicts higher parenting communication and involvement, but only in children with low levels of effortful control. Therefore, a difficult temperament interferes in a less negative way in the family system when parents are satisfied and united. And a better self-regulated child predicts more effective parenting practice regardless of the parents´ marital satisfaction. The clinical implications of the present findings should be considered. Specifically, difficult children must be detected and evaluated in community settings, such as school or community programs, in order to take into account the marital adjustment and parenting practices of their parents, and to be able to design adequate family interventions and prevent future pathologizing patterns.

Keywords: effortful control, marital adjustment, parenting, moderation

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Authors: Mariya Dayana, Budhmal Jain

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Kannur International Airport Ltd. (KIAL) is a new Greenfield airport project with airside development on an undulating terrain with an average height of 90m above Mean Sea Level (MSL) and a maximum height of 142m. To accommodate the desired Runway length and Runway End Safety Area (RESA) at both the ends along the proposed alignment, it resulted in 45.5 million cubic meters in cutting and filling. The insufficient availability of land for the construction of free slope embankment at RESA 07 end resulted in the design and construction of Reinforced Soil Slope (RSS) with a maximum slope of 65 degrees. An embankment fill of average 70m height with steep slopes located in high rainfall area is a unique feature of this project. The design and construction was challenging being asymmetrical with curves and bends. The fill was reinforced with high strength Uniaxial geogrids laid perpendicular to the slope. Weld mesh wrapped with coir mat acted as the facia units to protect it against surface failure. Face anchorage were also provided by wrapping the geogrids along the facia units where the slope angle was steeper than 45 degrees. Considering high rainfall received on this table top airport site, extensive drainage system was designed for the high embankment fill. Gabion wall up to 10m height were also designed and constructed along the boundary to accommodate the toe of the RSS fill beside the jeepable track at the base level. The design of RSS fill was done using ReSSA software and verified in PLAXIS 2D modeling. Both slip surface failure and wedge failure cases were considered in static and seismic analysis for local and global failure cases. The site won excavated laterite soil was used as the fill material for the construction. Extensive field and laboratory tests were conducted during the construction of RSS system for quality assurance. This paper represents a case study detailing the design and construction of a very high embankment using geosynthetics for the provision of Runway length and RESA area.

Keywords: airport, embankment, gabion, high strength uniaxial geogrid, kial, laterite soil, plaxis 2d

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Authors: Deepanjali Gurav, Kun Qian

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In-vitro metabolic diagnosis relies on designed materials-based analytical platforms for detection of selected metabolites in biological samples, which has a key role in disease detection and therapeutic evaluation in clinics. However, the basic challenge deals with developing a simple approach for metabolic analysis in bio-samples with high sample complexity and low molecular abundance. In this work, we report a designer plasmonic nanoshells based platform for direct detection of small metabolites in clinical samples for in-vitro metabolic diagnostics. We first synthesized a series of plasmonic core-shell particles with tunable nanoshell structures. The optimized plasmonic nanoshells as new matrices allowed fast, multiplex, sensitive, and selective LDI MS (Laser desorption/ionization mass spectrometry) detection of small metabolites in 0.5 μL of bio-fluids without enrichment or purification. Furthermore, coupling with isotopic quantification of selected metabolites, we demonstrated the use of these plasmonic nanoshells for disease detection and therapeutic evaluation in clinics. For disease detection, we identified patients with postoperative brain infection through glucose quantitation and daily monitoring by cerebrospinal fluid (CSF) analysis. For therapeutic evaluation, we investigated drug distribution in blood and CSF systems and validated the function and permeability of blood-brain/CSF-barriers, during therapeutic treatment of patients with cerebral edema for pharmacokinetic study. Our work sheds light on the design of materials for high-performance metabolic analysis and precision diagnostics in real cases.

Keywords: plasmonic nanoparticles, metabolites, fingerprinting, mass spectrometry, in-vitro diagnostics

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Authors: Shivani Kuttuva, Bridget Fergie, Andrew Mishreki, Shovkat Mir, Fintan Bergin

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Diaphragm disease (DD) of the small bowel is a condition wherein the bowel lumen is divided into a series of short compartments by multiple circumferential membranes of mucosa and submucosa, leading to pinhole lumen and subsequent obstruction. It is a rare condition commonly attributed to non-steroidal anti-inflammatory drugs (NSAIDs) usage. Herein we present a 31-yr-old-female with a history of NSAIDs usage for one year following neurosurgery, who presented with recurrent idiopathic small bowel obstruction, recalcitrant anaemia, and impaction of capsule endoscope on investigating for anaemia. The capsule endoscopy images demonstrated multiple circumferential strictures with ulcers at its tip and villous atrophy in the proximal bowel, suggestive of NSAIDs related damage. However, due to the lack of awareness of the detrimental effects of NSAIDs on bowel mucosa distal to the duodenum, the underlying aetiology of this clinical presentation remained a mystery for a significant duration. The patient had to undergo repeated laparotomies in order to relieve the symptoms of recurring acute small bowel obstruction. Upon examining the resected specimen under microscopy, the histopathological hallmark of expanded, fibrotic, and congested submucosa was picked up, leading to the confirmation of diaphragm disease. Thus, this case report aims to widen the awareness among clinicians and aid surgeons in devising a management plan for young individuals presenting with recurring episodes of obstruction due to Diaphragm disease.

Keywords: capsule endoscopy, diaphragm disease, NSAIDs, recurrent small bowel obstruction

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Authors: Kazunori Nomura, Hiromichi Ogi, Masaumi Nakahara, Sou Watanabe, Atsuhiro Shibata

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Chemical Processing Facility of Japan Atomic Energy Agency is a basic research field for advanced back-end technology developments with using actual high-level radioactive materials such as irradiated fuels from the fast reactor, high-level liquid waste from reprocessing plant. In the nature of a research facility, various kinds of chemical reagents have been offered for fundamental tests. Most of them were treated properly and stored in the liquid waste vessel equipped in the facility, but some were not treated and remained at the experimental space as a kind of legacy waste. It is required to treat the waste in safety. On the other hand, we formulated the Medium- and Long-Term Management Plan of Japan Atomic Energy Agency Facilities. This comprehensive plan considers Chemical Processing Facility as one of the facilities to be decommissioned. Even if the plan is executed, treatment of the “legacy” waste beforehand must be a necessary step for decommissioning operation. Under this circumstance, we launched a collaborative research project called the STRAD project, which stands for Systematic Treatment of Radioactive liquid waste for Decommissioning, in order to develop the treatment processes for wastes of the nuclear research facility. In this project, decomposition methods of chemicals causing a troublesome phenomenon such as corrosion and explosion have been developed and there is a prospect of their decomposition in the facility by simple method. And solidification of aqueous or organic liquid wastes after the decomposition has been studied by adding cement or coagulants. Furthermore, we treated experimental tools of various materials with making an effort to stabilize and to compact them before the package into the waste container. It is expected to decrease the number of transportation of the solid waste and widen the operation space. Some achievements of these studies will be shown in this paper. The project is expected to contribute beneficial waste management outcome that can be shared world widely.

Keywords: chemical processing facility, medium- and long-term management plan of JAEA facilities, STRAD project, treatment of radioactive waste

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Authors: Raad Nari, Maura Moriaty, Maha T. Barakat

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Introduction: Sodium-glucose co-transporter-2 (SGLT2) inhibitors are a new class of oral anti-diabetic drugs with a unique mechanism of action. They are used to improve glycaemic control in adults with type 2 diabetes by enhancing urinary glucose excretion. In the UAE, there has been certainly an increased use of these medications. As with any new medication, there are safety considerations related to their use in patients with type two diabetes. A retrospective study was conducted at the three main centres of the Imperial College London Diabetes Centre. Methodology: All patients in electronic database (Diamond) from October 2014 to October 2017 were included with a minimum of six months usage of sodium glucose co-transporter inhibitors that comprise canagliflozin, dapagliflozin and empagliflozin. There were 15 paired sample biochemical and clinical correlations. The analysis was done at the start of the study, three months and six months apart. SPSS version 24 was used for this study. Conclusion: This study of sodium glucose co-transporter-2 inhibitors used showed significant reductions in weight, glycated haemoglobin A1C, systolic and diastolic blood pressures. As the case with systematic reviews, there were similar changes in liver enzymes, raised total cholesterol, low density lipopoptein and high density lipoprotein. There was slight improvement in estimated glomerular filtration rate too. Our analysis also showed that they increased in the incidence of urinary tract symptoms and incidence of urinary tract infections.

Keywords: SGLT2 inhibitors dapagliflozin empagliflozin canagliflozin, adverse effects, amputation diabetic ketoacidosis DKA, urinary tract infection

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Authors: Anan Jarab, Riham Almrayat, Salam Alqudah, Maher Khdour, Tareq Mukattash, Sharell Pinto

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Background: The prevalence of diabetes in Jordan is among the highest in the world, making it a particularly alarming health problem there. It has been indicated that poor adherence to the prescribed therapy lead to poor glycemic control and enhance the development of diabetes complications and unnecessary hospitalization. Purpose: To explore factors associated with medication non-adherence in patients with type 2 diabetes in Jordan. Materials and Methods: Variables including socio-demographics, disease and therapy factors, diabetes knowledge, and health-related quality of life in addition to adherence assessment were collected for 171 patients with type 2 diabetes using custom-designed and validated questionnaires. Logistic regression was performed to develop a model with variables that best predicted medication non-adherence in patients with type 2 diabetes in Jordan. Results: The majority of the patients (72.5%) were non-adherent. Patients were found four times less likely to adhere to their medications with each unit increase in the number of prescribed medications (OR = 0.244, CI = 0.08-0.63) and nine times less likely to adhere to their medications with each unit increase in the frequency of administration of diabetic medication (OR = 0.111, CI = 0.04-2.01). Patients in the present study were also approximately three times less likely (OR = 0.362, CI = 0.24-0.87) to adhere to their medications if they reported having concerns about side effects and twice more likely to adhere to medications (OR = 0.493, CI = 0.08-1.16) if they had one or more micro-vascular complication. Conclusion: The current study revealed low adherence rate to the prescribed therapy among Jordanians with type 2 diabetes. Simplifying dosage regimen, selecting treatments with lower side effects along with an emphasis on diabetes complications should be taken into account when developing care plans for patients with type 2 diabetes.

Keywords: type 2 diabetes, adherence, glycemic control, clinical pharmacist, Jordan

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Authors: A. Ferasat, S. Rostampour Yasouri

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Seasonal flu is a highly contagious infection caused by influenza viruses. These viruses undergo genetic changes that result in new epidemics across the globe. Medical attention is crucial in severe cases, particularly for the elderly, frail, and those with chronic illnesses, as their immune systems are often weaker. The purpose of this study was to detect new subtypes of the influenza A virus rapidly using a specific RT-PCR method based on the HA gene (hemagglutinin). In the winter and spring of 2022_2023, 120 embryonated egg samples were cultured, suspected of seasonal influenza. RNA synthesis, followed by cDNA synthesis, was performed. Finally, the PCR technique was applied using a pair of specific primers designed based on the HA gene. The PCR product was identified after purification, and the nucleotide sequence of purified PCR products was compared with the sequences in the gene bank. The results showed a high similarity between the sequence of the positive samples isolated from the patients and the sequence of the new strains isolated in recent years. This RT-PCR technique is entirely specific in this study, enabling the detection and multiplication of influenza and its subspecies from clinical samples. The RT-PCR technique based on the HA gene, along with sequencing, is a fast, specific, and sensitive diagnostic method for those infected with influenza viruses and its new subtypes. Rapid molecular diagnosis of influenza is essential for suspected people to control and prevent the spread of the disease to others. It also prevents the occurrence of secondary (sometimes fatal) pneumonia that results from influenza and pathogenic bacteria. The critical role of rapid diagnosis of new strains of influenza is to prepare a drug vaccine against the latest viruses that did not exist in the community last year and are entirely new viruses.

Keywords: influenza, molecular diagnosis, patients, RT-PCR technique

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Authors: Shu-Jen Chen, Yi-Chien Wan, Ruey-Chi Wang

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Hydroxyapatite (Ca10(PO4)6(OH)2, HAp) can be applied to the fabrication of bone replacement materials, the composite of dental filling, and the adsorption of biomolecules and dyes. The integration of HAp and magnetic materials would offer several advantages for bio-separation process because the magnetic adsorbents is capable of recovered by applied magnetic field. C-phycocyanin (C-PC) and Allophycocyanin (APC), isolated from Spirulina platensis, can be used in fluorescent labeling probes, health care foods and clinical diagnostic reagents. Although the purification of C-PC and APC are reported by HAp adsorption, the adsorption of C-PC and APC by magnetic HAp composites was not reported yet. Therefore, the fabrication of HAp with magnetic silica nanoparticles for proteins adsorption was investigated in this work. First, the magnetic silica particles were prepared by covering silica layer on Fe3O4 nanoparticles with a reverse micelle method. Then, the Fe3O4@SiO2 nanoparticles were mixed with calcium carbonate to obtain magnetic silica/calcium carbonate composites (Fe3O4@SiO2/CaCO3). The Fe3O4@SiO2/CaCO3 was further reacted with K2HPO4 for preparing the magnetic silica/hydroxyapatite composites (Fe3O4@SiO2/HAp). The adsorption experiments indicated that the adsorption capacity of Fe3O4@SiO2/HAp toward C-PC and APC were highest at pH 6. The adsorption of C-PC and APC by Fe3O4@SiO2/HAp could be correlated by the pseudo-second-order model, indicating chemical adsorption dominating the adsorption process. Furthermore, the adsorption data showed that the adsorption of Fe3O4@SiO2/HAp toward C-PC and APC followed the Langmuir isotherm. The isoelectric points of C-PC and APC were around 5.0. Additionally, the zeta potential data showed the Fe3O4@SiO2/HAp composite was negative charged at pH 6. Accordingly, the adsorption mechanism of Fe3O4@SiO2/HAp toward C-PC and APC should be governed by hydrogen bonding rather than electrostatic interaction. On the other hand, as compared to C-PC, the Fe3O4@SiO2/HAp shows higher adsorption affinity toward APC. Although the Fe3O4@SiO2/HAp cannot recover C-PC and APC from Spirulina platensis homogenate, the Fe3O4@SiO2/HAp can be applied to separate C-PC and APC.

Keywords: hydroxyapatite, magnetic, C-phycocyanin, allophycocyanin

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Authors: Shraddha Acharya, Sharad Kumar Sharma, Ratna Bhattarai, Bhagwan Maharjan, Deepak Dahal, Serpahine Kaminsa

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Background: Drug-resistant (DR) tuberculosis (TB) continues to be a threat in Nepal, with an estimated 2800 new cases every year. The treatment of DR-TB with second line TB drugs is complex and takes longer time with comparatively lower treatment success rate than drug-susceptible TB. Delay in treatment initiation for DR-TB patients might further result in unfavorable treatment outcomes and increased transmission. This study thus aims to determine median time taken to initiate second-line treatment among Rifampicin Resistant (RR) diagnosed TB patients and to assess the proportion of treatment delays among various type of DR-TB cases. Method: A retrospective cohort study was done using national routine electronic data (DRTB and TB Laboratory Patient Tracking System-DHIS2) on drug resistant tuberculosis patients between January 2020 and December 2022. The time taken for treatment initiation was computed as– days from first diagnosis as RR TB through Xpert MTB/Rif test to enrollment on second-line treatment. The treatment delay (>7 days after diagnosis) was calculated. Results: Among total RR TB cases (N=954) diagnosed via Xpert nationwide, 61.4% were enrolled under shorter-treatment regimen (STR), 33.0% under longer treatment regimen (LTR), 5.1% for Pre-extensively drug resistant TB (Pre-XDR) and 0.4% for Extensively drug resistant TB (XDR) treatment. Among these cases, it was found that the median time from diagnosis to treatment initiation was 6 days (IQR:2-15.8). The median time was 5 days (IQR:2.0-13.3) among STR, 6 days (IQR:3.0-15.0) among LTR, 30 days (IQR:5.5-66.8) among Pre-XDR and 4 days (IQR:2.5-9.0) among XDR TB cases. The overall treatment delay (>7 days after diagnosis) was observed in 42.4% of the patients, among which, cases enrolled under Pre-XDR contributed substantially to treatment delay (72.0%), followed by LTR (43.6%), STR (39.1%) and XDR (33.3%). Conclusion: Timely diagnosis and prompt treatment initiation remain fundamental focus of the National TB program. The findings of the study, however suggest gaps in timeliness of treatment initiation for the drug-resistant TB patients, which could bring adverse treatment outcomes. Moreover, there is an alarming delay in second line treatment initiation for the Pre-XDR TB patients. Therefore, this study generates evidence to identify existing gaps in treatment initiation and highlights need for formulating specific policies and intervention in creating effective linkage between the RR TB diagnosis and enrollment on second line TB treatment with intensified efforts from health providers for follow-ups and expansion of more decentralized, adequate, and accessible diagnostic and treatment services for DR-TB, especially Pre-XDR TB cases, due to the observed long treatment delays.

Keywords: drug-resistant, tuberculosis, treatment initiation, Nepal, treatment delay

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Authors: Marwan Khalil Qader, Arshad Mohammad Abdullah

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Antimicrobial resistance is a major cause of significant morbidity and mortality globally. Seven plant extracts (Plantago mediastepposa, Quercusc infectoria, Punic granatum, Thymus lcotschyana, Ginger officeinals, Rhus angustifolia and Cinnamon) were collected from different regions of Kurdistan region of Iraq. These plants’ extracts were dissolved in absolute ethanol and distillate water, after which they were assayed in vitro as an antimicrobial activity against Candida tropicalis, Candida albicanus, Candida dublinensis, Candida krusei and Candida glabrata also against 2 Gram-positive (Bacillus subtilis and Staphylococcus aureus) and 3 Gram-negative bacteria (Escherichia coli, Pseudomonas aeruginosa and Klebsilla pneumonia). The antimicrobial activity was determined in ethanol extracts and distilled water extracts of these plants. The ethanolic extracts of Q. infectoria showed the maximum activity against all species of Candida fungus. The minimum inhibition zone of the Punic granatum ethanol extracts was 0.2 mg/ml for all microorganisms tested. Klebsilla pneumonia was the most sensitive bacterial strain to Quercusc infectoria and Rhus angustifolia ethanol extracts. Among both Gram-positive and Gram-negative bacteria tested with MIC of 0.2 mg/ml, the minimum inhibition zone of Ginger officeinals D. W. extracts was 0.2 mg/mL against Pseudomonas aeruginosa and Klebsilla pneumonia. The most sensitive bacterial strain to Thymus lcotschyana and Plantago mediastepposa D.W. extracts was S. aureus and E. coli.

Keywords: antimicrobial activity, pathogenic bacteria, plant extracts, chemical systems engineering

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Authors: T. Qasim, B. El Masoud, D. Ailabouni

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This in vitro study focused on investigating the fatigue resistance of veneered zirconia molar crowns with different veneering ceramic thicknesses, simulating the relative wear depths under simulated cyclic loading. A mandibular first molar was prepared and then scanned using computer-aided design/computer-aided manufacturing (CAD/CAM) technology to fabricate 32 zirconia copings of uniform 0.5 mm thickness. The manufactured copings then veneered with 1.5 mm, 1.0 mm, 0.5 mm, and 0.0 mm representing 0%, 33%, 66%, and 100% relative wear of a normal ceramic thickness of 1.5 mm. All samples were thermally aged to 6000 thermo-cycles for 2 minutes with distilled water between 5 ˚C and 55 ˚C. The samples subjected to cyclic fatigue and fracture testing using SD Mechatronik chewing simulator. These samples are loaded up to 1.25x10⁶ cycles or until they fail. During fatigue, testing, extensive cracks were observed in samples with 0.5 mm veneering layer thickness. Veneering layer thickness 1.5-mm group and 1.0-mm group were not different in terms of resisting loads necessary to cause an initial crack or final failure. All ceramic zirconia-based crown restorations with varying occlusal veneering layer thicknesses appeared to be fatigue resistant. Fracture load measurement for all tested groups before and after fatigue loading exceeded the clinical chewing forces in the posterior region. In general, the fracture loads increased after fatigue loading and with the increase in the thickness of the occlusal layering ceramic.

Keywords: all ceramic, cyclic loading, chewing simulator, dental crowns, relative wear, thermally ageing

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Authors: A. Shirazibeheshti, T. Radwan, A. Ettefaghian, G. Wilson, C. Luca, Farbod Khanizadeh

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Anticholinergic medication has been associated with events such as falls, delirium, and cognitive impairment in older patients. To further assess this, anticholinergic burden scores have been developed to quantify risk. A risk model based on clustering was deployed in a healthcare management system to cluster patients into multiple risk groups according to anticholinergic burden scores of multiple medicines prescribed to patients to facilitate clinical decision-making. To do so, anticholinergic burden scores of drugs were extracted from the literature, which categorizes the risk on a scale of 1 to 3. Given the patients’ prescription data on the healthcare database, a weighted anticholinergic risk score was derived per patient based on the prescription of multiple anticholinergic drugs. This study was conducted on over 300,000 records of patients currently registered with a major regional UK-based healthcare provider. The weighted risk scores were used as inputs to an unsupervised learning algorithm (mean-shift clustering) that groups patients into clusters that represent different levels of anticholinergic risk. To further evaluate the performance of the model, any association between the average risk score within each group and other factors such as socioeconomic status (i.e., Index of Multiple Deprivation) and an index of health and disability were investigated. The clustering identifies a group of 15 patients at the highest risk from multiple anticholinergic medication. Our findings also show that this group of patients is located within more deprived areas of London compared to the population of other risk groups. Furthermore, the prescription of anticholinergic medicines is more skewed to female than male patients, indicating that females are more at risk from this kind of multiple medications. The risk may be monitored and controlled in well artificial intelligence-equipped healthcare management systems.

Keywords: anticholinergic medicines, clustering, deprivation, socioeconomic status

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Authors: Usha Barahmand, Maria Stalias, Abdul Haq, Esther Rotlevi, Ying Xiang

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Misophonia is a condition in which specific repetitive oral, nasal, or other sounds and movements made by humans trigger impulsive aversive reactions of irritation or disgust that instantly become anger. A few measures exist for the assessment of misophonia, but each has some limitations, and evidence for a formal diagnosis is still lacking. The objective of this study was to develop a reliable and valid measure of misophonia for use in the general population. Adopting a purely descriptive approach, this study focused on developing a self-report measure using all triggers and reactions identified in previous studies on misophonia. A measure with two subscales, one assessing the aversive quality of various triggers and the other assessing reactions of individuals, was developed. Data were gathered from a large sample of both men and women ranging in age from 18 to 65 years. Exploratory factor analysis revealed three main triggers: oral/nasal sounds, hand and leg movements, and environmental sounds. Two clusters of reactions also emerged: nonangry attempts to avoid the impact of the aversive stimuli and angry attempts to stop the aversive stimuli. The examination of the psychometric properties of the scale revealed its internal consistency and test-retest reliability to be excellent. The scale was also found to have very good concurrent and convergent validity. Significant annoyance and disgust in response to the triggers were reported by 12% of the sample, although for some specific triggers, rates as high as 31% were also reported. These findings have implications for the delineation of the criteria for identifying misophonia as a clinical condition.

Keywords: adults, factor analysis, misophonia, psychometric properties, scale

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Authors: Samantha Meyr, Eman Mirdamadi, Martha Wang, Tao Lowe, Ryan Smith, Quinn Burke

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Critical-sized bone defects (CSD) treatment options remain a major clinical orthopedic challenge. They are uniquely contoured diseased or damaged bones and can be defined as those that will not heal spontaneously and require surgical intervention. Autografts are the current gold standard CSD treatment, which are histocompatible and provoke a minimal immunogenic response; however, they can cause donor site morbidity and will not suffice for the size required for replacement. As an alternative to traditional surgical methods, bone tissue engineering will be implemented via 3D printing methods. A freeze-dried bone matrix (FDBM) is a type of graft material available but will only function as desired when in the presence of bone growth factors. Polycaprolactone (PCL) is a known biodegradable material with good biocompatibility that has been proven manageable in 3D printing as a medical device. A 3D-extrusion printing strategy is introduced to print these materials into scaffolds for bone grafting purposes, which could be more accessible and rapid than the current standard. Mechanical, thermal, cytotoxic, and physical properties were investigated throughout a degradation period of 6 months using fibroblasts and dental pulp stem cells. PCL-FDBM scaffolds were successfully printed with high print fidelity in their respective pore sizes and allograft content. Additionally, we have created a method for evaluating PCL using differential scanning calorimetry (DSC) and have evaluated PCL degradation over roughly 6 months.

Keywords: 3D printing, bone tissue engineering, cytotoxicity, degradation, scaffolds

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Authors: Mwafaq Ibdah, Mossab, Yahyaa, Dor Rachmany, Yoram Gerchman, Doron Holland, Liora Shaltiel-Harpaz

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Pear Psylla is the most important pest of pear in all pear-growing regions, in Asian, European, and the USA. Pear psylla damages pears in several ways: high-density populations of these insects can cause premature leaf and fruit drop, diminish plant growth, and reduce fruit size. In addition, their honeydew promotes sooty mold on leaves and russeting on fruit. Pear psyllas are also considered vectors of pear pathogens such as Candidatus Phytoplasma pyri causing pear decline that can lead to loss of crop and tree vigor, and sometimes loss of trees. Psylla control is a major obstacle to efficient integrated pest management. Recently we have identified two naturally resistance pear accessions (Py.760-261 and Py.701-202) in the Newe Ya’ar live collection. GC-MS volatile metabolic profiling identified several volatile compounds common in these accessions but lacking, or much less common, in a sensitive accession, the commercial Spadona variety. Among these volatiles were styrene and its derivatives. When the resistant accessions were used as inter-stock, the volatile compounds appear in commercial Spadona scion leaves, and it showed reduced susceptibility to pear psylla. Laboratory experiments and applications of some of these volatile compounds were very effective against psylla eggs, nymphs, and adults. The genes and enzymes involved in the specific reactions that lead to the biosynthesis of styrene in plant are unknown. We have identified a phenolic acid decarboxylase that catalyzes the formation of p-hydroxystyrene, which occurs as a styrene analog in resistant pear genotypes. The His-tagged and affinity chromatography purified E. coli-expressed pear PyPAD1 protein could decarboxylate p-coumaric acid and ferulic acid to p-hydroxystyrene and 3-methoxy-4-hydroxystyrene. In addition, PyPAD1 had the highest activity toward p-coumaric acid. Expression analysis of the PyPAD gene revealed that its expressed as expected, i.e., high when styrene levels and psylla resistance were high.

Keywords: pear Psylla, volatile, GC-MS, resistance

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Authors: Jožef Ritonja

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The biochemical industry is one of the most important modern industries. Biochemical reactors are crucial devices of the biochemical industry. The essential bioprocess carried out in bioreactors is the fermentation process. A thorough insight into the fermentation process and the knowledge how to control it are essential for effective use of bioreactors to produce high quality and quantitatively enough products. The development of the control system starts with the determination of a mathematical model that describes the steady state and dynamic properties of the controlled plant satisfactorily, and is suitable for the development of the control system. The paper analyses the fermentation process in bioreactors thoroughly, using existing mathematical models. Most existing mathematical models do not allow the design of a control system for controlling the fermentation process in batch bioreactors. Due to this, a mathematical model was developed and presented that allows the development of a control system for batch bioreactors. Based on the developed mathematical model, a control system was designed to ensure optimal response of the biochemical quantities in the fermentation process. Due to the time-varying and non-linear nature of the controlled plant, the conventional control system with a proportional-integral-differential controller with constant parameters does not provide the desired transient response. The improved adaptive control system was proposed to improve the dynamics of the fermentation. The use of the adaptive control is suggested because the parameters’ variations of the fermentation process are very slow. The developed control system was tested to produce dairy products in the laboratory bioreactor. A carbon dioxide concentration was chosen as the controlled variable. The carbon dioxide concentration correlates well with the other, for the quality of the fermentation process in significant quantities. The level of the carbon dioxide concentration gives important information about the fermentation process. The obtained results showed that the designed control system provides minimum error between reference and actual values of carbon dioxide concentration during a transient response and in a steady state. The recommended control system makes reference signal tracking much more efficient than the currently used conventional control systems which are based on linear control theory. The proposed control system represents a very effective solution for the improvement of the milk fermentation process.

Keywords: biochemical reactor, fermentation process, modelling, adaptive control

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Authors: Nadeen Eltoukhy, Sahar S. Sheta, Walaa Elnaggar, Karim Bakr

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Purpose: The aim of this study is to compare the clinical efficacy and side effects of oral versus intralesional propranolol treatment of infantile capillary hemangiomas in infants. Methods: The study enrolled 40 infants diagnosed with infantile capillary hemangiomas. Patients were divided into 2 groups: Group A (Non-invasive group) included 20 infants who received oral propranolol hydrochloride starting at a dose of 1 mg/kg/day BID, then increased to a max of 2 mg/kg/day BID gradually over 2 weeks for 3 months. Group B (Invasive group) included 20 infants who received intralesional propranolol injection at a dose of 1 mg/mL; the volume of the injected drug depended on the size of the lesion (0.2 mL injected per cm of lesion diameter), with a maximum volume of 1 mL for a lesion of 5 cm diameter under complete aseptic conditions in the operating theater. Results: At three months after initiating treatment, the circumferential size of the hemangioma showed a statistically significant decrease in both groups; in Group A from 3.66±2.89 cm to 1.56±1.26 cm with p-value <0.05 and in Group B from 2.99±2.73 cm to 1.32±1.18 cm with p-value <0.05. There is no statistically significant comparative difference between the two groups (p-value = 0.538 = insignificant). Regarding the complications of oral propranolol, one patient (5%) had bradycardia, and one patient (5%) had diarrhea. In the injection group, 20 patients (100%) had local edema, and one patient (5%) had a local infection. Conclusions: Both oral non-invasive and intralesional invasive propranolol are safely used to successfully treat and decrease the size of infantile hemangioma while showing no statistically comparative difference between both treatment techniques.

Keywords: hemangioma, oral beta blocker, intralesional beta blocker, infants

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Authors: Mannan Mridha, Mohammad S. Islam

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Innovative medical technologies offer more effective medical care, with less risk to patient and healthcare personnel. Medical technology and devices when properly used provide better data, precise monitoring and less invasive treatments and can be more targeted and often less costly. The Intensive Care Unit (ICU) equipped with patient monitoring, respiratory and cardiac support, pain management, emergency resuscitation and life support devices is particularly prone to medical errors for various reasons. Many people in the developing countries now wonder whether their visit to hospital might harm rather than help them. This is because; clinicians in the developing countries are required to maintain an increasing workload with limited resources and absence of well-functioning safety system. A team of experts from the medical, biomedical and clinical engineering in Sweden and Bangladesh have worked together to study the incidents, adverse events at the ICU in Bangladesh. The study included both public and private hospitals to provide a better understanding for physical structure, organization and practice in operating processes of care, and the occurrence of adverse outcomes the errors, risks and accidents related to medical devices at the ICU, and to develop a ICT based support system in order to reduce hazards and errors and thus improve the quality of performance, care and cost effectiveness at the ICU. Concrete recommendations and guidelines have been made for preparing appropriate ICT related tools and methods for improving the routine for use of medical devices, reporting and analyzing of the incidents at the ICU in order to reduce the number of undetected and unsolved incidents and thus improve the patient safety.

Keywords: intensive care units, medical errors, medical devices, patient care and safety

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Authors: Mona Hejazi, Ali Motie Nasrabadi

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Epilepsy is a persistent neurological disorder that affects more than 50 million people worldwide. Hence, there is a necessity to introduce an efficient prediction model for making a correct diagnosis of the epileptic seizure and accurate prediction of its type. In this study we consider how the Effective Connectivity (EC) patterns obtained from intracranial Electroencephalographic (EEG) recordings reveal information about the dynamics of the epileptic brain and can be used to predict imminent seizures, as this will enable the patients (and caregivers) to take appropriate precautions. We use this definition because we believe that effective connectivity near seizures begin to change, so we can predict seizures according to this feature. Results are reported on the standard Freiburg EEG dataset which contains data from 21 patients suffering from medically intractable focal epilepsy. Six channels of EEG from each patients are considered and effective connectivity using Directed Transfer Function (DTF) and Granger Causality (GC) methods is estimated. We concentrate on effective connectivity standard deviation over time and feature changes in five brain frequency sub-bands (Alpha, Beta, Theta, Delta, and Gamma) are compared. The performance obtained for the proposed scheme in predicting seizures is: average prediction time is 50 minutes before seizure onset, the maximum sensitivity is approximate ~80% and the false positive rate is 0.33 FP/h. DTF method is more acceptable to predict epileptic seizures and generally we can observe that the greater results are in gamma and beta sub-bands. The research of this paper is significantly helpful for clinical applications, especially for the exploitation of online portable devices.

Keywords: effective connectivity, Granger causality, directed transfer function, epilepsy seizure prediction, EEG

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Authors: Rabia Ashfaq, Saeed Iqbal, Atiq ur Rehman, Irfanullah Khan

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An extensive series of radiopharmaceuticals have been explored in order to discover a better brain tumour diagnostic agent. Tc99m labelling with cysteine and its derivatives in liposomes shows effective tagging of about 70% to 80 %. Due to microscopic size it successfully crossed the brain barrier in 2 minutes which gradually decreases in 5 to 15 minutes. HMPAO labelled with Tc99m is another important radiopharmaceutical used to study brain perfusion but it comes with a flaw that it’s only functional during epilepsy. 1, 1 ECD is purely used in Tc99m ECD formulation; because it not only tends to cross the blood brain barrier but it can be metabolized which can be easily entrapped in human brain. Radio labelling of Cysteine with Tc99m at room temperature was performed which yielded no good results. Hence cysteine derivatives with salicylaldehyde were prepared that produced about 75 % yield for ligand. In order to perform it’s radio labelling a suitable solvent DMSO was selected and physical parameters were performed. Elemental analyser produced remarkably similar results for ligand as reported in literature. IR spectra of Ligand in DMSO concluded in the absence of SH stretch and presence of N-H vibration. Thermal analysis of the ligand further suggested its decomposition pattern with no distinct curve for a melting point. Radio labelling of ligand was performed which produced excellent results giving up to 88% labelling at pH 5.0. Clinical trials using Rabbit were performed after validating the products reproducibility. The radiopharmaceutical prepared was injected into the rabbit. Dynamic as well as static study was performed under the SPECT. It showed considerable uptake in the kidneys and liver considering it suitable for the Hypatobilliary study.

Keywords: marcapto compounds, 99mTc - radiolabeling, salicylaldicysteine, thiozolidine

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Authors: Sumeeta Khurana, Kirti Megha, Amit Gupta, Rakesh Sehgal

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Background: Amoebic keratitis is a painful vision threatening infection caused by a free living pathogenic amoeba Acanthamoeba. It can be misdiagnosed and very difficult to treat if not suspected early. The epidemiology of Acanthamoeba genotypes causing infection in our geographical area is not yet known to the best of our knowledge. Objective: To characterize Acanthamoeba isolates from amoebic keratitis patients. Methods: A total of 19 isolates obtained from patients with amoebic keratitis presenting to the Advanced Eye Centre at Postgraduate Institute of Medical Education and Research, a tertiary care centre of North India over a period of last 10 years were included. Their corneal scrapings, lens solution and lens case (in case of lens wearer) were collected for microscopic examination, culture and molecular diagnosis. All the isolates were maintained in the Non Nutrient agar culture medium overlaid with E.coli and 13 strains were axenised and maintained in modified Peptone Yeast Dextrose Agar. Identification of Acanthamoeba genotypes was based on amplification of diagnostic fragment 3 (DF3) region of the 18srRNA gene followed by sequencing. Nucleotide similarity search was performed by BLAST search of sequenced amplicons in GenBank database (http//www.ncbi.nlm.nih.gov/blast). Multiple Sequence alignments were determined by using CLUSTAL X. Results: Nine out of 19 Acanthamoeba isolates were found to belong to Genotype T4 followed by 6 isolates of genotype T11, 3 T5 and 1 T3 genotype. Conclusion: T4 is the predominant Acanthamoeba genotype in our geographical area. Further studies should focus on differences in pathogenicity of these genotypes and their clinical significance.

Keywords: Acanthamoeba, free living amoeba, keratitis, genotype, ocular

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