Search results for: viral hemorrhagic fever

Authors: Fatma Sahir- Halouane, Sonia Hamid, Farida Tihar-Benzina, Fatiha Bouhlali, Souad Lourchane

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The Culicidae are biting insects, the most harmful to people, they are almost all bloodsuckers, and they are responsible of the spread of many important diseases such as malaria, yellow fever, and elephantiasis. Entomopathogenic microorganisms occupy an important place among the alternative methods of fighting against pests insect. The fungus Beauveria bassiana is an entomopathogenic agent naturally present in the ecosystems. It offers a very interesting potential for controlling populations of mosquitoes. This study aimed to show the histological changes that occured in Culex pipiens larvae infected with Beauveria bassiana. The 4th instar larvae were infected with B. bassiana in 10-7 spore/ml dilution, the histological section was studied showing that the fungi infected all the body parts specially Cuticle, Epiderms, fat bodies and midgut. After then the insect have a white appearance and covered with a thick coat of hyphea. The obtained results show that the application of Beauveria bassiana on cuticle of the fourth stage larvae of Culex pipiens was dependent of an apparent disturbance on the structure of the cuticle or there has been the degeneration of its different parts, infection of the fungus does not stop at the body walls. Therefore, it affects even the Adipose tissue, epidermal cells and intestine.

Keywords: Culex pipiens, Beauveria bassiana, histological changes, cuticle, intestine and adipose tissue

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Authors: Aliyu Umar, Mohammed Yau, Faruruwa M. Dahiru, Saed Garba

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Evaluation of heavy metals in twelve commonly consumed herbal medicines/preparations in Kano State, Nigeria, was carried out. The samples comprised of five unregistered powdered medicines, namely, Zuwo, (ZW); Rai Dorai, (RD); Miyar Tsanya, (MTS); Bagaruwar Makka, (BM); and Madobiya, (M); five unregistered liquid herbal medicinal concussions for pile (MB), yellow fever (MS), typhoid (MT), stomach pain (MC), sexually transmitted diseases (STDs); and two registered herbal medicines; Alif Powder (AP) and Champion Leaf (CL). The heavy metals evaluation was carried out using Atomic Absorption Spectroscopy (AAS) and the result revealed the concentrations (ppm) ranges of the heavy metals as follows: Cadmium (0.0045 – 0.1601), Chromium (0.0418 – 0.2092), Cobalt (0.0038 – 0.0760), Copper (0.0547 – 0.2465), Iron (0.1197 – 0.3592), Manganese (0.0123 – 1.4462), Nickel (0.0073 – 0.0960), Lead (0.185 – 0.0927) and Zinc (0.0244 – 0.2444). Comparing the results obtained in this work with the standards of the World Health Organization (WHO), the Food and Agricultural Organization (FAO) and permissible limits of other countries, the concentrations of heavy metals in the herbal medicine/preparations are within the allowed permissible limits range in herbal medicines and their use could be safe.

Keywords: Kano state, herbal medicines, registered, unregistered

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Authors: Zengmin Tian, Bin Lv, Rui Hui, Yupeng Liu, Chuan Wang, Qing Liu, Hongyu Li, Yan Qi, Li Song

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Objective Robot was used for replacement of the frame in recent years. The paper is to investigate the safety and effectiveness of frameless stereotactic surgery in the treatment of children with cerebral palsy. Methods Clinical data of 425 children with spastic cerebral palsy were retrospectively analyzed. The patients were treated with robot-assistant frameless stereotactic surgery of nuclear mass destruction. The motor function was evaluated by gross motor function measure-88 (GMFM-88) before the operation, 1 week and 3 months after the operation respectively. The statistical analysis was performed. Results The postoperative CT showed that the destruction area covered the predetermined target in all the patients. Minimal bleeding of puncture channel occurred in 2 patient, and mild fever in 3 cases. Otherwise, there was no severe surgical complication occurred. The GMFM-88 scores were 49.1±22.5 before the operation, 52.8±24.2 and 64.2±21.4 at the time of 1 week and 3 months after the operation, respectively. There was statistical difference between before and after the operation (P<0.01). After 3 months, the total effective rate was 98.1%, and the average improvement rate of motor function was 24.3% . Conclusion Replaced the traditional frame, the robot-assistant frameless stereotactic surgery is safe and reliable for children with spastic cerebral palsy, which has positive significance in improving patients’ motor function.

Keywords: cerebral palsy, robotics, stereotactic techniques, frameless operation

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Authors: Tengku Muhammad Rafi Nazmi Bin Tengku Razali, Habsah Alwi

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This article based on effect of temperature towards extraction of Aquilaria Subintegra. Aquilaria Subintegra which its main habitat is in Asia-tropical and particularly often found in its native which is Thailand. There is claim which is Aquilaria Subintegra contains antipyretic properties that helps fight fever. Research nowadays also shown that paracetamol consumed bring bad effect towards consumers. This sample will first dry using Vacuum Far Infrared which provides better drying than conventional oven. Soxhlet extractor used to extract oil from sample. Gas Chromatography Mass Spectrometer used to analyze sample to determine its compound. Objective from this research was to determine the active ingredients that exist in the Aquilaria Subintegra leaves and to determine whether compound of Acetaminophen exist or not inside the leaves. Moisture content from 400C was 80%, 500C was 620% and 600C was 36%. The greater temperature resulting lower moisture content inside sample leaves. 7 components were identified in sample T=400C while only 5 components were identified in sample at T=50C and T=60C. Four components were commonly identified in three sample which is 1n-Hexadecanoic acid, 9,12,15-Octadecatrienoic acid, methyl ester (z,z,z), Vitamin E and Squalene. Further studies are needed with new series of temperature to refine the best results.

Keywords: aquilaria subintegra, vacuum far infrared, SOXHLET extractor, gas chromatography mass spectrometer, paracetamol

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Authors: Attapon Kamlangdee, Brock Kingstad-Bakke, Tavis K. Anderson, Tony L. Goldberg, Jorge E. Osorio

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A critical failure in our preparedness for an influenza pandemic is the lack of a universal vaccine. Influenza virus strains diverge by 1 to 2% per year, and commercially available vaccines often do not elicit protection from one year to the next, necessitating frequent formulation changes. This represents a major challenge to the development of a cross-protective vaccine that can protect against circulating viral antigenic diversity. We have constructed a recombinant modified vaccinia virus Ankara (MVA) that expresses an H5N1 mosaic hemagglutinin (H5M) (MVA-H5M). This mosaic was generated in silico using 2,145 field-sourced H5N1 isolates. A single dose of MVA-H5M provided 100% protection in mice against clade 0, 1, and 2 avian influenza viruses and also protected against seasonal H1N1 virus (A/Puerto Rico/8/34). It also provided short-term (10 days) and long-term (6 months) protection post vaccination. Both neutralizing antibodies and antigen-specific CD4+and CD8+ T cells were still detected at 5 months post vaccination, suggesting that MVA-H5M provides long-lasting immunity.

Keywords: modified vaccinia Ankara, MVA, H5N1, hemagglutinin, influenza vaccine

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Authors: Thanthrige Thiunuwan Priyathilaka, Don Anushka Sandaruwan Elvitigala, Bong-Soo Lim, Hyung-Bok Jeong, Jehee Lee

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Toll like receptors (TLRs) are a phylogeneticaly conserved family of pattern recognition receptors, which participates in the host immune responses against various pathogens and pathogen derived mitogen. TLR21, a non-mammalian type, is almost restricted to the fish species even though those can be identified rarely in avians and amphibians. Herein, this study was carried out to identify and characterize TLR21 from rock bream (Oplegnathus fasciatus) designated as RbTLR21, at transcriptional and genomic level. In this study, the full length cDNA and genomic sequence of RbTLR21 was identified using previously constructed cDNA sequence database and BAC library, respectively. Identified RbTLR21 sequence was characterized using several bioinformatics tools. The quantitative real time PCR (qPCR) experiment was conducted to determine tissue specific expressional distribution of RbTLR21. Further, transcriptional modulation of RbTLR21 upon the stimulation with Streptococcus iniae (S. iniae), rock bream iridovirus (RBIV) and Edwardsiella tarda (E. tarda) was analyzed in spleen tissues. The complete coding sequence of RbTLR21 was 2919 bp in length which can encode a protein consisting of 973 amino acid residues with molecular mass of 112 kDa and theoretical isoelectric point of 8.6. The anticipated protein sequence resembled a typical TLR domain architecture including C-terminal ectodomain with 16 leucine rich repeats, a transmembrane domain, cytoplasmic TIR domain and signal peptide with 23 amino acid residues. Moreover, protein folding pattern prediction of RbTLR21 exhibited well-structured and folded ectodomain, transmembrane domain and cytoplasmc TIR domain. According to the pair wise sequence analysis data, RbTLR21 showed closest homology with orange-spotted grouper (Epinephelus coioides) TLR21with 76.9% amino acid identity. Furthermore, our phylogenetic analysis revealed that RbTLR21 shows a close evolutionary relationship with its ortholog from Danio rerio. Genomic structure of RbTLR21 consisted of single exon similar to its ortholog of zebra fish. Sevaral putative transcription factor binding sites were also identified in 5ʹ flanking region of RbTLR21. The RBTLR 21 was ubiquitously expressed in all the tissues we tested. Relatively, high expression levels were found in spleen, liver and blood tissues. Upon induction with rock bream iridovirus, RbTLR21 expression was upregulated at the early phase of post induction period even though RbTLR21 expression level was fluctuated at the latter phase of post induction period. Post Edwardsiella tarda injection, RbTLR transcripts were upregulated throughout the experiment. Similarly, Streptococcus iniae induction exhibited significant upregulations of RbTLR21 mRNA expression in the spleen tissues. Collectively, our findings suggest that RbTLR21 is indeed a homolog of TLR21 family members and RbTLR21 may be involved in host immune responses against bacterial and DNA viral infections.

Keywords: rock bream, toll like receptor 21 (TLR21), pattern recognition receptor, genomic characterization

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Authors: V. Baweja, K. K. Gupta, V. Dubey, C. Keshavam

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Antimicrobial activity of six indigenous plants such as Tulsi Ocimum sanctum, Neem Azadirachta indica, Aloe vera, Turmeric Curcuma longa, Lantana Lantana camara, and Clove Syzygium aromaticum was assessed against the gut microbiota of the dengue fever mosquito Aedes aegypti, keeping in view that the presence of midgut bacteria may affect the ability of the vector to transmit pathogens. Eleven different types of bacterial clones were isolated from the midgut of lab-reared fourth instar larvae of Aedes aegypti and were grown on LB agar medium at an optimum temperature of 25 ºC. Identification of these bacteria was done on the basis of their colony characteristic such as colony size, shape, opacity, elevation, consistency, and growth. Light microscopic studies of the gut microbiota revealed dominance of Gram-negative cocci over gram positive cocci and bacilli and Gram-negative bacilli. Identification of species was done by chemical characterization of the colonies. Crude extracts of all test plants were screened for their antimicrobial activities against gut microbiota by disc diffusion assay. The zone of exclusion seen after 24 hr of incubation in different assays revealed the most potent antibacterial activities in neem followed by clove and turmeric. Lantana and Aloe vera were least effective.

Keywords: plant extract, aedes, dengue, antimicrobial activity

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Authors: Mini P. Singh, Manasi Majumdar, Kapil Goyal, Pvm Lakshmi, Deepak Bhatia, Radha Kanta Ratho

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India is endemic for Hepatitis E virus and frequent water borne outbreaks are reported. The conventional diagnosis rests on the detection of serum anti-HEV IgM antibodies which may take 7-10 days to develop. Early diagnosis in such a situation is desirable for the initiation of prompt control measures. The present study compared three diagnostic methods in 60 samples collected during two suspected HEV outbreaks in the vicinity of Chandigarh, India. The anti-HEV IgM, HEV antigen and HEV-RNA could be detected in serum samples of 52 (86.66%), 16 (26.66%) and 18 (30%) patients respectively. The suitability of saliva samples for antibody detection was also evaluated in 21 paired serum- saliva samples. A total of 15 serum samples showed the presence of anti HEV IgM antibodies, out of which 10 (10/15; 66.6%) were also positive for these antibodies in saliva samples (χ2 = 7.636, p < 0.0057), thus showing a concordance of 76.91%. The positivity of reverse transcriptase PCR and HEV antigen detection was 100% within one week of illness which declined to 5-10% thereafter. The outbreak was attributed to HEV Genotype 1, Subtype 1a and the clinical and environmental strains clustered together. HEV antigen and RNA were found to be an early diagnostic marker with 96.66% concordance. The results indicate that the saliva samples can be used as an alternative to serum samples in an outbreak situation.

Keywords: HEV-antigen, outbreak, phylogenetic analysis, saliva

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Authors: Muhammad Asif Rasheed

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Newcastle disease virus (NDV), an avian orthoavulavirus, is a causative agent of Newcastle disease named (NDV) and can cause even the epidemics when the disease is not treated. Previously several vaccines based on attenuated and inactivated viruses have been reported, which are rendered useless with the passage of time due to versatile changes in viral genome. Therefore, we aimed to develop an effective multi-epitope vaccine against the haemagglutinin neuraminidase (HN) protein of 26 NDV strains from Pakistan through a modern immunoinformatic approaches. As a result, a vaccine chimaera was constructed by combining T-cell and B-cell epitopes with the appropriate linkers and adjuvant. The designed vaccine was highly immunogenic, non-allergen, and antigenic; therefore, the potential 3D-structureof multi epitope vaccine was constructed, refined, and validated. A molecular docking study of a multiepitope vaccine candidate with the chicken Toll-like receptor-4 indicated successful binding. An In silico immunological simulation was used to evaluate the candidate vaccine's ability to elicit an effective immune response. According to the computational studies, the proposed multiepitope vaccine is physically stable and may induce immune responses, whichsuggested it a strong candidate against 26 Newcastle disease virus strains from Pakistan. A wet lab study is under process to confirm the results.

Keywords: epitopes, newcastle disease virus, paramyxovirus virus, vaccine

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Authors: Hashaam Akhtar

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IFNλR1 and IL10R2 collectively construct a heterodimer, which is an acknowledged functional receptor for all type III interferons (IFNs). Expression of IFNλR1 is highly tissue specific, which can help in making type III IFNs a drug of choice as comparable to its analogue, type I IFNs, for treating hepatitis C in the near future. Although, expression of IFNλR1 also varies with the concentration of type I IFNs, but in this study it was shown that the expression of IFNλR1 varies with the protein titers of IFN-α, IFN-λ3 and the newly discovered IFN-λ4. High dosage of IFN-α reduces the expression of IFNλR1 in HepG2 cells, which can affect the antiviral activity of type III IFNs in vivo. We premeditated an experimental strategy to differentiate monocytes into dendritic cells (DCs), type I and type II macrophages in vitro and quantified the expression of the IFNλR1 by qPCR. The exposure of newly discovered IFN-λ4 to macrophages and DCs also raised the expression of its own receptor, which shows that expression of IFN-λ4 protein in hepatitis C patient may augment type I treatment and help ease off viral titers. The results of this study may contribute in some understanding towards the mechanisms involved in the selective expression of IFNLR1 and exceptionalities associated with the receptor.

Keywords: IFNLR1, Interferon Lambda 4 (IFN-λ4), Mononuclear Phagocyte Cells (MPCs), expression

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Authors: Jihene Bettaieb, Ghassen kharroubi, Rym mallekh, Ines Cherif, Taoufik Atawa, Kaouther Harrabech

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Brucellosis is a major worldwide zoonosis. It is a reportable condition in Tunisia where the disease remains endemic, especially in rural areas. The aim of this study was to describe the epidemiological, clinical, and bacteriological profile of human brucellosis cases notified in the district of Tunis. It was a retrospective descriptive study of cases reported in the district of Tunis through the national surveillance system between the 1st January and 31th December 2017. During the study period, 133 brucellosis confirmed cases were notified. The mean age was 37.5 ± 18.0 years, and 54.9% of cases were males. More than four-fifths (82.7%) of cases were reported in spring and summer with a peak in the month of May (36 cases). Fever and sweats were the most common symptoms; they occurred in 95% and 72% of cases, respectively. Osteoarticular complications occurred in 10 cases, meningitis in one case and endocarditis in one other case. Wright agglutination test and Rose Bengale test were positive in 100% and 91% of cases, respectively. While blood culture was positive in 9 cases and PCR in 2 cases. Brucella melitensis was the only identified specie (9 cases). Almost all cases (99.2%) reported the habit of consuming raw dairy products. Only 5 cases had a suspect contact with animals; among them, 3 persons were livestock breeders. The transmission was essentially due to raw dairy product consumption. It is important to enhance preventive measures to control animal Brucellosis and to educate the population regarding the risk factors of the disease.

Keywords: brucellosis, risk factors, surveillance system, Tunisia

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Authors: Basavaraj R. Endigeri, Shriharsh Desphande

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Optimization and reduction of stress concentration around holes in a uniaxially loaded plate is one of the important design criteria in many of the engineering applications. These stress risers will lead to failure of the component at the region of high stress concentration which has to be avoided by means of providing auxiliary holes on either side of the parent hole. By literature survey it is known that till date, there is no analytical solution documented to reduce the stress concentration by providing auxiliary holes expect for fever geometries. In the present work, plate with a hole subjected to uniaxial load is analyzed with the numerical method to determine the optimum sizes and locations for the auxillary holes for different center hole diameter to plate width ratios. The introduction of auxiliary holes at a optimum location and radii with its effect on stress concentration is also represented graphically. The finite element analysis package ANSYS 8.0 is used to carry out analysis and optimization is performed to determine the location and radii for optimum values of auxiliary holes to reduce stress concentration. All the results for different diameter to plate width ratio are presented graphically. It is found from the work that introduction of auxiliary holes on either side of central circular hole will reduce stress concentration factor by a factor of 19 to 21 percentage.

Keywords: finite element method, optimization, stress concentration factor, auxiliary holes

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Authors: Nurul Hidayah, Farida Rahmatika, Fathimah Azzahra, Nesty Herennadia

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Aromatherapy candles are one of the insecticide media that have not been much researched. The active ingredient that is proven to have the effect of insecticide is a citrus extract from lemongrass oil (Cymbopogon). Aromatherapy candles are added by citrus compounds to be insecticidal for Ae. aegypti mosquito that was related to the infectious disease such as dengue fever. This research aims to find out if aromatherapy candles of citrus compounds have an insecticidal effect on Ae. aegypti mosquito. We used true experimental design including posttest only with control group design. The samples are 20 male and female Ae. aegypti mosquitos with aged 1-7 days belong to the inclusion criteria. The subjects were divided into 6 groups, consisting of 1 negative control group and 5 treatment groups with variation concentration are 1%; 2%; 3%; 4%; 5%. Each group will be treated for 2 hours and observed death after 24 hours. Replication in each group is done 4 times. The results were then tested statistically using Kruskal-Wallis and probit test. Mean of death in negative control group, and treatment group 1%; 2%; 3%; 4%; 5% respectively 0; 1; 0.25; 0; 1 and 1 mosquito. The Kruskal-Wallis test in the study group found no significant difference (p = 0.178). The probit analysis showed that LC50 and LC90 were 20.069% and 31.557%. The aromatherapy candle of a citrus compound has an insecticidal effect on the Ae aegypti mosquito.

Keywords: Ae. aegypti insecticide, aromatherapy candle, citrus compound, lemongrass oil (Cymbopogon)

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Authors: Ahmad Shahin, Walid Moudani, Ali Bekraki

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This research aims at presenting a predictive data mining model to handle an accurate diagnosis of acute appendicitis with patients for the purpose of maximizing the health service quality, minimizing morbidity/mortality, and reducing cost. However, acute appendicitis is the most common disease which requires timely accurate diagnosis and needs surgical intervention. Although the treatment of acute appendicitis is simple and straightforward, its diagnosis is still difficult because no single sign, symptom, laboratory or image examination accurately confirms the diagnosis of acute appendicitis in all cases. This contributes in increasing morbidity and negative appendectomy. In this study, the authors propose to generate an accurate model in prediction of patients with acute appendicitis which is based, firstly, on the segmentation technique associated to ABC algorithm to segment the patients; secondly, on applying fuzzy logic to process the massive volume of heterogeneous and noisy data (age, sex, fever, white blood cell, neutrophilia, CRP, urine, ultrasound, CT, appendectomy, etc.) in order to express knowledge and analyze the relationships among data in a comprehensive manner; and thirdly, on applying dynamic programming technique to reduce the number of data attributes. The proposed model is evaluated based on a set of benchmark techniques and even on a set of benchmark classification problems of osteoporosis, diabetes and heart obtained from the UCI data and other data sources.

Keywords: healthcare management, acute appendicitis, data mining, classification, decision tree

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Authors: Neda Valizadeh, Mani Mofidi, Sama Haghighi, Ali Hashemaghaee, Soudabeh Shafiee Ardestani

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Background: There are many systems and parameters to evaluate trauma patients in the emergency department. Most of these evaluations are to distinguish patients with worse conditions so that the care systems have a better prediction of condition for a better care-giving. The purpose of this study is to determine the relationship between axillary body temperature and mortality in patients hospitalized in the intensive care unit (ICU) with multiple traumas and with other clinical and para-clinical factors. Methods: All patients between 16 and 75 years old with multiple traumas who were admitted into Emergency Department then hospitalized in the ICU were included in our study. An axillary temperature in the first and the second day of admission, Glasgow cola scale (GCS), systolic blood pressure, Serum glucose levels, and white blood cell counts of all patients at the admission day were recorded and their relationship with mortality were analyzed by SPSS software with suitable statistical tests. Results: Axillary body temperatures in the first and second day were statistically lower in expired traumatic patients (p=0.001 and p<0,001 respectively). Patients with lower GCS had a significantly lower first-day temperature and a significantly higher mortality. (p=0.006 and p=0.006 respectively). Furthermore, the first-day axillary temperature was significantly lower in patients with a lower first-day systolic blood pressure (p=0.014). Conclusion: Our results showed that lower axillary body temperature in the first day is associated with higher mortality, lower GCS, and lower systolic blood pressure. Thus, this could be used as a predictor of mortality in evaluation of traumatic patients in emergency settings.

Keywords: fever, trauma, mortality, emergency

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Authors: Shabnum Shaheen, Nida Haroon, Farah Khan, Sumera Javad, Mehreen Jalal, Samina Sarwar

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Coffee Senna is pharmaceutically very important and used for multiple health disorders such as gastric pains, indigestion, snakebites, asthma and fever, tuberculosis and menstrual problems. However, its immense medicinal value and great demand lead to adulteration issue which could be injurious for users. Some times its adulterant Seena weed (Senna occidentalis L.) is used as its substitute which definitely not as effective as Coffee Senna. Hence, the present study was undertaken to provide some tools for systematic and pharmaceutical authentication of a shrubby plant Coffee Senna (Cassia occidentalis Linn.). These parameters included macro and micro morphological characters, anatomical and palynomorph characterization, solubility, fluorescence and phytochemical analysis. By the application of these parameters acquired results revealed that, these two plants are distinct from each other. The Coffee Seena was found to be an annual shrub with trilobed pollen, diacytic, paracytic and anisocytic stomata whereas the Seena weed stands out as an annual or perennial herb with spheroidal and circular pollen and paracytic type of stomata. The powdered drug of Coffee seena is dark grayish green whereas the powdered drug of Seena weed is light green in color. These findings are constructive in authentic identification of these plants.

Keywords: coffee senna, Senna weed, taxonomic evaluation, pharmaceutical markers

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Authors: C. D. Chen, H. Takaoka, Z. Ya’cob, V. L. Low, K. W. Lau, M. Sofian-Azirun

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Adult black flies (Diptera: Simuliidae) are small (1.5-6.0 mm long), two-winged insects, and are well known as one of the biting flies of medical and veterinary importance. Female of certain species, when they bite and take blood, not only cause severe skin diseases to human and cattle but also play a role as vectors of viral, protozoan and filarial diseases in humans and animals. Black flies also attract environmental biologist and ecologist because their immature states breed only in clean running fresh waters, and larvae are one of the principal processors of plant debris in streams. All these researches on medical and ecological aspects of black flies could not be reliably proceeded without sufficient basic knowledge of the fauna of black flies established by traditional but still important morphotaxonomy. Previously, only 39 species of black flies were recorded from Peninsular Malaysia, all of which are classified into four subgenus (Daviesellum, Gomphostilbia, Nevermannia and Simulium) of the genus Simulium. We carried out faunal surveys and taxonomic works of black flies in Peninsular Malaysia since November 2010. A total of 17 new species and 4 newly recorded species were collected. This increased the number of the described species of black flies in Peninsular Malaysia from 39 to 60. Our results suggest that a much higher diverse nature of black flies in Peninsular Malaysia will be clarified by further extensive surveys.

Keywords: black flies, Simulium, Nevermannia, feuerborni species-group

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Authors: Thanunthon Bowornsakulwong, Charukorn Charukarn, Franck Courtes, Panit Kitsubun, Lalintip Horcharoen

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Vero cell is a continuous cell line that is widely used for the production of viral vaccines. However, due to its adherent characteristic, scaling up strategy in large-scale production remains complicated and thus limited. Consequently, suspension-like Vero cell culture processes based on microcarriers have been introduced and employed while also providing increased surface area per volume unit. However, harvesting Vero cells from microcarriers is a huge challenge due to difficulties in cells detaching, lower recovery yield, time-consuming and dissociation agent carry-over. To overcome these problems, we developed a dissociation-association platform technology for detaching and re-attaching cells during subculturing from microcarriers to microcarriers, which will be conveniently applied to seed trains strategies in large scale bioreactors. Herein, Hillex-2 was used to culture Vero cells in serum-containing media using spinner flasks as a scale-down model. The overall confluency of cells on microcarriers was observed using inverted microscope, and the sample cells were daily detached in order to obtain the kinetics data. The metabolites consumption and by-products formation were determined by Nova Biomedical BioprofileFlex.

Keywords: dissociation-reassociation, microcarrier, scale up, Vero cell

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Authors: Aadil Khan, Yasser Chomayil, P. P. Venugopalan

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Background: Budd-Chiari syndrome is caused by thrombosis of the hepatic veins and/or the thrombosis of the intrahepatic or suprahepatic IVC. The etiology remains idiopathic in 16% -35% of cases. Malignancy, rheumatological disorder, myeloproliferative disease, inheritable coagulopathy, infection or hyperestrogen state can be identified in many cases. Methodology: Review of case records of the patient presented to Aster Medcity, Emergency Department, Cochin. Introduction:17 years old female was presented to ED with fever, jaundice and abdominal distention since 1 week. O/E: Pallor+, icterus+. Abdomen- gross distension+, shifting dullness+, generalized anasarca+. USG abdomen showed hepatomegaly with mild coarse echotexture and moderate to gross ascites. CT abdomen and chest showed hepatomegaly with thrombosis of all three hepatic vein and moderate ascites suggestive of Budd-Chiari syndrome. Patient was taken for catheter vein thrombolysis. Venogram done the next day revealed almost > 50% opening of the right hepatic vein. Concurrent doppler showed colour and doppler signals in middle hepatic veins. She gradually improved and was discharged home on anticoagulant and adviced regular follow up. Conclusion: Being a rare disease in this young population, high suspicion is required when evaluating young patients with abdominal pain and jaundice.

Keywords: Budd-Chiari syndrome, rare disease, abdominal pain, India

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Authors: Andi Aliah Hidayani, Asmi Citra Malina A. R. Tassakka, Andi Parenrengi

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Vanname Shrimp is one of the high yielding varieties that are more resistant to virus attacks. However, now this shrimp more death due to virus attack such as white spot disease caused by white spot syndrome virus (WSSV). Various efforts have done to prevent the disease, like immunostimulatory, probiotics, and vaccine. White spot syndrome virus (WSSV) envelope protein VP19 gene is important because of its involvement in the system infection of shrimp. This study aimed to isolate and characterize an envelope protein VP19 – encoding gene of WSSV using WSSV infected Vanname Shrimp sample from some areas in South Sulawesi (Pangkep, Barru and Pinrang). The genomic of DNA were isolated from shrimp muscle using DTAB-CTAB method. Isolation of gene encoding envelope protein VP19 WSSV ws successfully performed with the results of the length of DNA fragment was 387 bp. The results of homology analysis using BLASTn homology suggested that these isolates genes from Barru, Pangkep and Pinrang have closest relationship with isolates from Mexican.

Keywords: vanname, shrimp, WSSV, viral protein 19

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Authors: Samina Nazli, Nadia Qamar, Quratulain, Akasha, Saman Jamal

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Objective: The objective is to determine the frequency of iron deficiency anemia among children having febrile seizures. A descriptive Cross-Sectional Study was done in the Pediatric Unit of Allama Iqbal Memorial Teaching Hospital Sialkot from September 2020 to February 2021. Material & Methods: A total of 70 children were studied aged six months to 10 years, with either gender presenting with febrile seizures. All data of the patients was documented, including demographic data like age, gender, residential area, educational status, socioeconomic status and clinical findings at the time of presentation like fever, fits and duration of symptoms etc. Blood hemoglobin and ferritin levels were tested for each patient to evaluate iron deficiency anemia. Results: There were 65.7% male and 34.3% female cases in this study. The age range of the patients was 6 months to 10 years, with a mean age of 4.36 ± 2.71 years. Most of the children (60%) were below three years of age. Most children belonged to low and middle socioeconomic status with a frequency of 42.8% and 45.7%, respectively. Iron deficiency anemia was found in 38.6% of cases. The majority of the mothers were illiterate (65%). There were 44.3% cases from rural areas and 55.7% from urban areas. Conclusion: Iron deficiency anemia is a common problem among children with febrile seizures, younger than 03 years and belonging to rural areas. Illiterate mothers are an important risk factor for iron deficiency anemia in their children.

Keywords: febrile seizure, iron deficiency anemia, illetrate mother, low scioeconomic status, febrile siezure

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Authors: Patricia Abigail B. Miranda, Imelda A. Luna

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Introduction: COVID-19 infection is a novel viral illness that began as a local epidemic in December 2019 in Wuhan, China which quickly emerged into a pandemic by February 2020. The virus causes a spectrum of signs and symptoms, ranging from mild upper respiratory symptoms to acute respiratory distress syndrome, which may lead to death. Among children and neonates, those afflicted with the disease may present asymptomatically or with mild symptoms. To date, there has been limited local data that describes the outcomes of the growing number of COVID-19 cases, specifically in neonates. Methods: A cross-sectional study was conducted to determine the outcomes of neonates born to COVID-19 Positive Mothers from March 2020 until June 2022. The prevalence of COVID-19 among these neonates was also determined. Results: COVID-positive prevalence after 24 hours of life is at 8%, while prevalence after 48 hours among those who still underwent testing was at 13.51%. Moreover, among those COVID-19-negative neonates who had symptoms, they mostly presented with tachypnea (5.7%). The prevalence of complications among COVID-19-negative neonates delivered to COVID-19-positive mothers is 22.7%. Conclusion: Neonates born to COVID-19-positive mothers who yielded positive COVID-19 results are generally asymptomatic. Moreover, there are no associated mortalities among those who yielded positive results.

Keywords: COVID-19, neonates, outcomes, clinical profile

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Authors: Surangrat Srisurapanon, Chatchawal Wongjitrat, Navin Horthongkham, Ruengpung Sutthent

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HBV infection causes a potential serious public health problem. The ability to detect the HBV DNA concentration is of the importance and improved continuously. By using quantitative Polymerase Chain Reaction (qPCR), several factors in standardized; source of material, calibration standard curve and PCR efficiency are inconsistent. Digital PCR (dPCR) is an alternative PCR-based technique for absolute quantification using Poisson's statistics without requiring a standard curve. Therefore, the aim of this study is to compare the data set of HBV DNA generated between dPCR and qPCR methods. All samples were quantified by Abbott’s real time PCR and 54 samples with 2 -6 log10 HBV DNA were selected for comparison with dPCR. Of these 54 samples, there were two outlier samples defined as negative by dPCR. Of these two, samples were defined as negative by dPCR, whereas 52 samples were positive by both the tests. The difference between the two assays was less than 0.25 log IU/mL in 24/52 samples (46%) of paired samples; less than 0.5 log IU/mL in 46/52 samples (88%) and less than 1 log in 50/52 samples (96%). The correlation coefficient was r=0.788 and P-value <0.0001. Comparison to qPCR, data generated by dPCR tend to be the overestimation in the sample with low HBV DNA concentration and underestimated in the sample with high viral load. The variation in DNA by dPCR measurement might be due to the pre-amplification bias, template. Moreover, a minor drawback of dPCR is the large quantity of DNA had to be used when compare to the qPCR. Since the technology is relatively new, the limitations of this assay will be improved.

Keywords: hepatitis B virus, real time PCR, digital PCR, DNA quantification

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Authors: Yudi Zhao, Ziyun Zhou, Yueting Yao, Shuying Dai, Zhiling Yan, Longyu Yang, Chuanyin Li, Li Shi, Yufeng Yao

Abstract:

HPV16 E4 gene plays an important role in viral genome amplification and release. Therefore, a variation of the E4 gene nucleic acid sequence may affect the carcinogenicity of HPV16. In order to understand the relationship between the variation of HPV16 E4 gene and cervical cancer, this study was to amplify and sequence the DNA sequences of E4 genes in 118 HPV16-positive cervical cancer patients and 151 HPV16-positive asymptomatic individuals. After obtaining E4 gene sequences, the phylogenetic trees were constructed by the Neighbor-joining method for gene variation analysis. The results showed that: 1) The distribution of HPV16 variants between the case group and the control group differed greatly (P = 0.015)，and the Asian-American（AA）variant was likely to relate to the occurrence of cervical cancer. 2) DNA sequence analysis showed that there were significant differences in the distribution of 8 variants between the case group and the control group (P < 0.05). And 3) In European (EUR) variant, two variations, C3384T (L18L) and A3449G (P39P), were associated with the initiation and development of cervical cancer. The results suggested that the variation of HPV16 E4 gene may be a contributor affecting the occurrence as well as the development of cervical cancer, and different HPV16 variants may have different carcinogenic capability.

Keywords: cervical cancer, HPV16, E4 gene, variations

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Authors: Nabil Ghedadba, Abdessemed Samira, Leila Hambaba, Sidi Mohamed Ould Mokhtar, Nassima Fercha, Houas Bousselsela

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The overall objective of this study was to evaluate in vitro the hemostatic activity of secondary metabolites (polyphenols, flavonoids, and tannins) of Marrubium vulgare leaves, aromatic plant widely used in traditional medicine for the treatment of asthma, cough, diabetes (by its effect on the pancreas to secrete insulin), heart disease, fever has a high efficiency as against inflammation. Qualitative analysis of the aqueous extract (AQE) by thin layer chromatography revealed the presence of quercetin, kaempferol and rutin. Quantification of total phenols by Folin Ciocalteu method and flavonoids by AlCl3 method gave high values with AQE: 175±0.80 mg GAE per 100g of the dry matter, 23.86±0.36 mg QE per 100g of dry matter. Moreover, the assay of condensed tannins by the vanillin method showed that AQE contains the highest value: 16.55±0.03 mg e-catechin per 100 g of dry matter. Assessment of hemostatic activity by the plasma recalcification method (time of Howell) has allowed us to discover the surprising dose dependent anticoagulant effect of AQE lyophilized from leaves of M. vulgare. A positive linear correlation between the two parameters studied: the content of condensed tannins and hemostatic activity (r=0.96) were used to highlight a possible role of these compounds that are potent vasoconstrictor activity in hemostatic. From these results we can see that Marrubium vulgre could be used for the treatment of health.

Keywords: Marrubium vulgare L., aqueous extract, phenolic compounds dosing, hemostatic activity, condensed tannins

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Authors: Vanja Misic, Mohamed El-Mogy, Yousef Haj-Ahmad

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Endonuclease G (EndoG) is a well conserved mitochondrio-nuclear nuclease with dual lethal and vital roles in the cell. The aim of our study was to examine whether EndoG exerts its nuclease activity on exogenous DNA substrates such as plasmid DNA (pDNA), considering their importance in gene therapy applications. The effects of EndoG knockdown on pDNA stability and levels of encoded reporter gene expression were evaluated in the cervical carcinoma HeLa cells. Transfection of pDNA vectors encoding short-hairpin RNAs (shRNAs) reduced levels of EndoG mRNA and nuclease activity in HeLa cells. In physiological circumstances, EndoG knockdown did not have an effect on the stability of pDNA or the levels of encoded transgene expression as measured over a four day time-course. However, when endogenous expression of EndoG was induced by an extrinsic stimulus, targeting of EndoG by shRNA improved the perceived stability and transgene expression of pDNA vectors. Therefore, EndoG is not a mediator of exogenous DNA clearance, but in non-physiological circumstances it may non-specifically cleave intracellular DNA regardless of its origin. These findings make it unlikely that targeting of EndoG is a viable strategy for improving the duration and level of transgene expression from non-viral DNA vectors in gene therapy efforts.

Keywords: EndoG, silencing, exogenous DNA stability, HeLa cells

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Authors: Shweh Fern Loo, Jun Yin Ong, Than Zaw Oo

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Acute respiratory distress syndrome (ARDS) is a common serious condition of bilateral lung infiltrates that develops secondary to various underlying conditions such as diseases or injuries. ARDS with severe hypercapnia is associated with higher ICU mortality and morbidity. Venovenous Extracorporeal membrane oxygenation (VV-ECMO) support has been established to avert life-threatening hypoxemia and hypercapnic respiratory failure despite optimal conventional mechanical ventilation. However, VV-ECMO is relatively not advisable in particular groups of patients, especially in multi-organ failure, advanced age, hemorrhagic complications and irreversible central nervous system pathology. We presented a case of a 79-year-old Chinese lady without any pre-existing lung disease admitted to our hospital intensive care unit (ICU) after acute presentation of breathlessness and chest pain. After extensive workup, she was diagnosed with rapidly progressing acute interstitial pneumonia with ARDS and hypercapnia respiratory failure. The patient received lung protective strategies of mechanical ventilation and neuromuscular blockage therapy as per clinical guidelines. However, hypercapnia respiratory failure was refractory, and she was deemed not a good candidate for VV-ECMO support given her advanced age and high vasopressor requirements from shock. Alternative therapy with extracorporeal CO2 removal (ECCO2R) was considered and implemented. The patient received 12 days of ECCO2R paired with muscle paralysis, optimization of lung-protective mechanical ventilation and dialysis. Unfortunately, the patient still had refractory hypercapnic respiratory failure with dual vasopressor support despite prolonged therapy. Given failed and futile medical treatment, the family opted for withdrawal of care, a conservative approach, and comfort care, which led to her demise. The effectivity of extracorporeal CO2 removal may depend on disease burden, involvement and severity of the disease. There is insufficient data to make strong recommendations about its benefit-risk ratio for ECCO2R devices, and further studies and data would be required. Nonetheless, ECCO2R can be considered an alternative treatment for refractory hypercapnic respiratory failure patients who are unsuitable for initiating venovenous ECMO.

Keywords: extracorporeal CO2 removal (ECCO2R), acute respiratory distress syndrome (ARDS), acute interstitial pneumonia (AIP), hypercapnic respiratory failure

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Authors: Anupalli Roja Rani, Saraswathi Jaggali

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Rose-scented geranium (Pelargonium graveolens L’Hér.) is an erect, much-branched shrub. It is indigenous to various parts of southern Africa, and it is often called Geranium. Pelargonium species are widely used by traditional healers in the areas of Southern Africa by Sotho, Xhosa, Khoi-San and Zulus for its curative and palliative effects in the treatment of diarrhea, dysentery, fever, respiratory tract infections, liver complaints, wounds, gastroenteritis, haemorrhage, kidney and bladder disorders. We have used Plant materials for extracting active compounds from analytical grades of solvents methanol, ethyl acetate, chloroform and water by a soxhlet apparatus. The phytochemical screening reveals that extracts of Pelargonium graveolens contains alkaloids, glycosides, steroids, tannins, saponins and phenols in ethyl acetate solvent. The antioxidant activity was determined using 1, 1-diphenyl-2-picrylhydrazyl (DPPH) bleaching method and the total phenolic content in the extracts was determined by the Folin–Ciocalteu method. Due to the presence of different phytochemical compounds in Pelargonium the anti-microbial activity against different micro-organisms like E.coli, Streptococcus, Klebsiella and Bacillus. Fractionation of plant extract was performed by column chromatography and was confirmed with HPLC analysis, NMR and FTIR spectroscopy for the compound identification in different organic solvent extracts.

Keywords: Pelargonium graveolens L’Hér, DPPH, micro-organisms, HPLC analysis, NMR, FTIR spectroscopy

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Authors: Emmanuel Ademola Anigilaje, Ibraheem Ishola

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Background: Childhood steroid-sensitive idiopathic nephrotic syndrome (SSINS) is plagued with relapses that contribute to its morbidity and the cost of treatment. Materials and Methods: This is a retrospective review of relapses among children with SSINS at the University of Abuja Teaching Hospital from January 2016 to July 2020. Triggers related to relapse incidents were noted. Chi-square test was deployed for predictors (factors at the first clinical presentations that associate with subsequent relapses) of relapses. Predictors with p-values of less than 0.05 were considered significant and 95% confidence intervals (CI) and odd ratio (OR) were described. Results: Sixty SSINS comprising 52 males (86.7%), aged 23 months to 18 years, with a mean age of 7.04±4.16 years were studied. Thirty-eight (63.3%) subjects had 126 relapses including infrequent relapses in 30 (78.9%) and frequent relapses in 8 (21.1%). The commonest triggers were acute upper respiratory tract infections (68, 53.9%) and urinary tract infections (UTIs) in 25 (19.8%) relapses. In 4 (3.2%) relapses, no trigger was identified. The time-to-first relapse ranged 14 days to 365 days with a median time of 60 days. The significant predictors were hypertension (OR=3.4, 95% CI; 1.04-11.09, p=0.038), UTIs (OR=9.9, 95% CI; 1.16-80.71, p= 0.014), malaria fever (OR=8.0, 95% CI; 2.45-26.38, p˂0.001), micro-haematuria (OR=4.9, 95% CI; 11.58-15.16, p=0.004), elevated serum creatinine (OR=12.3, 95%CI; 1.48-101.20, p=0.005) and hypercholesterolaemia (OR=4.1, 95%CI; 1.35-12.63, p=0.011). Conclusion: While the pathogenesis of relapses remains unknown, it is prudent to consider relapse-specific preventive strategies against triggers and predictors of relapses in our setting.

Keywords: Patterns, triggers, predictors, steroid-sensitive idiopathic nephrotic syndrome, relapses, Nigeria

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Authors: Beryl Apondi Obola

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Introduction: Mosquitoes are known to transmit various diseases such as malaria, dengue fever, and Zika virus. Mosquito repellents are commonly used to prevent mosquito bites. However, some of these repellents contain chemicals that can be harmful to human health and the environment. Therefore, there is a need to find alternative mosquito repellents that are safe and effective. Objective: The objective of this research is to investigate the effectiveness of banana leaves as an alternative mosquito repellent on Plasmodium falciparum and Plasmodium vivax. Methodology: The research will be conducted in two phases. In the first phase, the repellent properties of banana leaves will be tested in a laboratory setting. The leaves will be crushed and mixed with water to extract the active ingredients. The extract will be tested against mosquitoes in a controlled environment. The number of mosquitoes that are repelled by the extract will be recorded. In the second phase, the effectiveness of the banana leaf extract will be tested in the field. The extract will be applied to the skin of human volunteers, and the number of mosquito bites will be recorded. The results will be compared to a commercially available mosquito repellent. Expected Outcomes: The expected outcome of this research is to determine whether banana leaves can be used as an effective mosquito repellent. If the results are positive, banana leaves could be used as an alternative to chemical-based mosquito repellents. Conclusion: Banana leaves have been used for various purposes in traditional medicine. This research aims to investigate the potential of banana leaves as an alternative mosquito repellent. The results of this research could have significant implications for public health and the environment

Keywords: banana leaf extract, mosquito repellant, plasmodium falciparum, public health

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