Search results for: histological alterations of gill tissue

Authors: Surapong Vongvatcharanon, Fardeela Binalee, Wandee Udomuksorn, Ekkasit Kumarnsit, Uraporn Vongvatcharanon

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Hypogonadism is characterized by a decline in sex hormone levels, especially testosterone. It has been shown to be an important contributor to the decrease in muscle mass, muscle strength and performance, a condition known as sarcopenia. Preparations from Butea superba Roxb. (red Kwao Krua) have been reported to have androgenic properties. The active compounds are proposed to be flavonoids and flavonoid glycosides. Treatment with B. superba has been shown to improve erectile dysfunction in males. Parvalbumin (PV) is a relaxing factor and identified in fast twitch fibers. Alterations of the PV levels affects skeletal muscle functions. This study aimed to investigate the effects of orhchidectomy, testosterone replacement and different doses of Butea superba Roxb. on the structure, performance, levels of parvalbumin, parvalbumin and androgen receptor immunoreactivities in the extensor digitorum longus (EDL) and gastrocnemius muscles of orchidectomized rats. Twelve-week old male Wistar rats were randomly divided into 6 groups; sham-operated (SHAM), orchidectomized (BS-0), orchidectomized group that was treated with testosterone replacement of 6 µg/kg (TP) or an orchidectomized group that was treated with various doses of an extract from Butea superba Roxb.; 5 mg/kg (BS-5), 50 mg/kg (BS-50) and 500 mg/kg (BS-500) all for 90 days. The testosterone level, epididymis, seminal vesicle, prostate gland, vas deference weight, muscle fiber size, strength and endurance in both the EDL and gastrocnemius muscle were decreased in the BS-0 group but increased in the testosterone replacement group. Treatment with the B. superba Roxb. extract replacement group improved muscle fiber size, strength and endurance, but not total testosterone levels, or the epididymis, seminal vesicle, prostate gland, vas deference weight. Furthermore, the parvalbumin level, parvalbumin and androgen receptor immunoreactivities were reduced in the BS-0 group but increased in the testosterone replacement group and the B. superba Roxb. extract groups for both the EDL and gastrocnemius muscle. This study indicated that the reduction of testosterone level led to a decrease of the androgen receptor density resulting in a decline in the muscle mass and parvalbumin levels. The decrease of parvalbumin levels affected muscle performance. Testosterone replacement increased the androgen receptor density and led to an increase of muscle mass and parvalbumin levels. The increase in the parvalbumin levels may result in an improvement of muscle performance. This may explain one mechanism of testosterone on muscle mass and strength in the testosterone dependent sarcopenia. The B. superba Roxb. extract groups also had improved muscle mass, strength and endurance, parvalbumin level, parvalbumin and androgen immunoreactivities compared to the BS-O group . Butea superba Roxb. Extracts contains a flavonoid (3, 7, 3'-Trihydroxy-4'-methoxyflavone), flavonoiglycoside (3, 3'-dihydroxy-4'-methoxyflavone-7-O-β-D-glucopyranoside) and isoflavanolignans (butesuperins A and butesuperins B) all known to inhibit the cAMP phosphodiesterase enzyme. Therefore, cAMP signaling may have adaptive effects on skeletal muscle by increasing muscle mass, strength and endurance.

Keywords: Butea superba, parvalbumin, skeletal muscle, orchidectomy

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Authors: Evgen Prokhorov, Gabriel Luna-Barcenas

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The polysaccharide chitosan (CS) is an attractive biopolymer for the stabilization of several nanoparticles in acidic aqueous media. This is due in part to the presence of abundant primary NH2 and OH groups which may lead to steric or chemical stabilization. Applications of most CS nanocomposites are based upon the interaction of high surface area nanoparticles (NPs) with different substance. Therefore, agglomeration of NPs leads to decreasing effective surface area such that it may decrease the efficiency of nanocomposites. The aim of this work is to measure nanocomposite’s electrical conductivity phenomena that will allow one to formulate optimal concentrations of conductivity NPs in CS-based nanocomposites. Additionally, by comparing the efficiency of such nanocomposites, one can guide applications in the biomedical (antibacterial properties and tissue regeneration) and sensor fields (detection of copper and nitrate ions in aqueous solutions). It was shown that the best antibacterial (CS-AgNPs, CS-AgNPs-carbon nanotubes) and would healing properties (CS-AuNPs) are observed in nanocomposites with concentrations of NPs near the percolation threshold. In this regard, the best detection limit in potentiometric and impedimetric sensors for detection of copper ions (using CS-AuNPs membrane) and nitrate ions (using CS-clay membrane) in aqueous solutions have been observed for membranes with concentrations of NPs near percolation threshold. It is well known that at the percolation concentration of NPs an abrupt increasing of conductivity is observed due to the presence of physical contacts between NPs; above this concentration, agglomeration of NPs takes place such that a decrease in the effective surface and performance of nanocomposite appear. The obtained relationship between electrical percolation threshold and performance of polymer nanocomposites with conductivity NPs is important for the design and optimization of polymer-based nanocomposites for different applications.

Keywords: chitosan, conductivity nanoparticles, percolation threshold, polymer nanocomposites

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Authors: Constant Gillot, Pauline Michaux, Julien Favresse, Jean-Michel Dogné, Jonathan Douxfils

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Introduction: NETosis has emerged as a crucial yet paradoxical factor in severe COVID-19 cases. While neutrophil extracellular traps (NETs) help contain and eliminate viral particles, excessive NET formation can lead to hyperinflammation, exacerbating tissue damage and acute respiratory distress syndrome (ARDS). Aims: This study evaluates the relationship between COVID-19-infected sera and NETosis using an ex-vivo model. Methods: Sera from 8 post-admission COVID-19 patients, after receiving corticoid therapy, were used to induce NETosis in neutrophils from a healthy donor. NET formation was tracked using fluorescent markers for DNA and neutrophil elastase (NE) every 2 minutes for 8 hours. The results were expressed as a percentage of DNA/NE released over time. Key metrics, including T50 (time to 50% release) and AUC (area under the curve), representing total NETosis potential), were calculated. A 27-cytokine screening kit was used to assess the cytokine composition of the sera. Results: COVID-19 sera induced NETosis based on their cytokine profile. The AUC of NE and DNA release decreased with time following corticoid therapy, showing a significant reduction in 6 of the 8 patients (p<0.05). T50 also decreased in parallel with AUC for both markers. Cytokines concentration decrease with time after therapy administration. There is correlation between 14 cytokines concentration and NE release. Conclusion: This ex-vivo model successfully demonstrated the induction of NETosis by COVID-19 sera using two markers. A clear decrease in NETosis potential was observed over time with glucocorticoid therapy. This model can be a valuable tool for monitoring NETosis and investigating potential NETosis inducers and inhibitors.

Keywords: NETosis, COVID-19, cytokine storm, biomarkers

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Authors: Huma Shareef, Ghazala H. Rizwani, Ahsana Dar

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In traditional medicine Cardiospermum halicacabum L. (Sapindeaceae) is used against various ailments. In current investigation searching a new remedy that will available easily, non expensive, able to lower hypertension and standardize blood pressure, made us to develop an herbal medicine. Crude ethanol extract of C. halicacabum and its various fractions ethyl acetate and butanol showed a dose-dependent hypotensive effect in anaesthetized rats. The trachea was exposed and freed from connective tissue and incubated by cannula to facilitate spontaneous respiration. The right carotid artery and left jugular vein were cannulated with polyethylene tubing PE-50 for monitoring blood pressure changes via pressure transducer (Gould P23 ID) connected to a Grass model 79D polygraph and for i.v. injection, respectively. Drugs or the plant extracts were administered at a constant volume of 0.5 ml/kg, followed by injection of 0.2 ml of saline that flushed the cannula. Systolic, diastolic and mean arterial blood pressure (MABP) was measured in mm Hg and heart rate in beats/min. Ethanol extract of C. halicacabum showed a significant activity at 50 mg/kg dose. Ethyl acetate fraction (10, 20, 30, 40, and 50 mg/kg) induced dose dependent fall in systolic and diastolic blood pressure, heart rate of rats. At 10-30 mg/kg the hypotensive effect was non significantly reduced by 10 -15%. However, the extract at 40 mg/kg induced significant hypotensive effect calculated as 30.95±3.2% MABP and this effect persists till 50 mg/kg. The higher polar fraction (butanol) of the whole plant failed to produce any significant response against MABP at all the tested doses (10-50 mg/kg). C. halicacabum lowers blood pressure, exerts a dose-dependent hypotensive effect, can be used as hypotensor.

Keywords: cardiospermum halicacabum, calcium channel blocker, hypotensive, various extracts

Procedia PDF Downloads 500

Authors: Abdelkrim Belhaoua, Jean-Pierre Radoux, Mariana Kuras, Vincent Récamier, Martial Piotto, Karim Elbayed, François Proust, Izzie Namer

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This paper proposes an augmented reality system dedicated to neurosurgery in order to assist the surgeon during an operation. This work is part of the ExtempoRMN project (Funded by Bpifrance) which aims at analyzing during a surgical operation the metabolic content of tumoral brain biopsy specimens by HRMAS NMR. Patients affected with a brain tumor (gliomas) frequently need to undergo an operation in order to remove the tumoral mass. During the operation, the neurosurgeon removes biopsy specimens using image-guided surgery. The biopsy specimens removed are then sent for HRMAS NMR analysis in order to obtain a better diagnosis and prognosis. Image-guided refers to the use of MRI images and a computer to precisely locate and target a lesion (abnormal tissue) within the brain. This is performed using preoperative MRI images and the BrainLab neuro-navigation system. With the patient MRI images loaded on the Brainlab Cranial neuro-navigation system in the operating theater, surgeons can better identify their approach before making an incision. The Brainlab neuro-navigation tool tracks in real time the position of the instruments and displays their position on the patient MRI data. The results of the biopsy analysis by 1H HRMAS NMR are then sent back to the operating theater and superimposed on the 3D localization system directly on the MRI images. The method we have developed to communicate between the HRMAS NMR analysis software and Brainlab makes use of a combination of C++, VTK and the Insight Toolkit using OpenIGTLink protocol.

Keywords: neuro-navigation, augmented reality, biopsy, BrainLab, HR-MAS NMR

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Authors: Theophilus Asante, Comfort Nyarko, Daniel Antwi

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Drug-resistant tuberculosis, together with the associated comorbidities like pulmonary fibrosis and silicosis, has been one of the most serious global public health threats that requires immediate action to curb or mitigate it. This prolongs hospital stays, increases the cost of medication, and increases the death toll recorded annually. Crinum asiaticum bulb (CAE) and betulin (BET) are known for their biological and pharmacological effects. Pharmacological effects reported on CAE include antimicrobial, anti-inflammatory, anti-pyretic, anti-analgesic, and anti-cancer effects. Betulin has exhibited a multitude of powerful pharmacological properties ranging from antitumor, anti-inflammatory, anti-parasitic, anti-microbial, and anti-viral activities. This work sought to investigate the anti-tuberculosis and resistant modulatory effects and also assess their effects on mitigating pulmonary fibrosis and silicosis. In the anti-tuberculosis and resistant modulatory effects, both CAE and BET showed strong antimicrobial activities (31.25 ≤ MIC ≤ 500) µg/ml against the studied microorganisms and also produced significant anti-efflux pump and biofilm inhibitory effects (ρ < 0.0001) as well as exhibiting resistance modulatory and synergistic effects when combined with standard antibiotics. Crinum asiaticum bulbs extract and betulin were shown to possess anti-pulmonary fibrosis effects. There was an increased survival rate in the CAE and BET treatment groups compared to the BLM-induced group. There was a marked decrease in the levels of hydroxyproline and collagen I and III in the CAE and BET treatment groups compared to the BLM-treated group. The treatment groups of CAE and BET significantly downregulated the levels of pro-fibrotic and pro-inflammatory cytokine concentrations such as TGF-β1, MMP9, IL-6, IL-1β and TNF-alpha compared to an increase in the BLM-treated groups. The histological findings of the lungs suggested the curative effects of CAE and BET following BLM-induced pulmonary fibrosis in mice. The study showed improved lung functions with a wide focal area of viable alveolar spaces and few collagen fibers deposition on the lungs of the treatment groups. In the anti-silicosis and pulmonoprotective effects of CAE and BET, the levels of NF-κB, TNF-α, IL-1β, IL-6 and hydroxyproline, collagen types I and III were significantly reduced by CAE and BET (ρ < 0.0001). Both CAE and BET significantly (ρ < 0.0001) inhibited the levels of hydroxyproline, collagen I and III when compared with the negative control group. On BALF biomarkers such as macrophages, lymphocytes, monocytes, and neutrophils, CAE and BET were able to reduce their levels significantly (ρ < 0.0001). The CAE and BET were examined for anti-oxidant activity and shown to raise the levels of catalase (CAT) and superoxide dismutase (SOD) while lowering the level of malondialdehyde (MDA). There was an improvement in lung function when lung tissues were examined histologically. Crinum asiaticum bulbs extract and betulin were discovered to exhibit anti-tubercular and resistance-modulatory properties, as well as the capacity to minimize TB comorbidities such as pulmonary fibrosis and silicosis. In addition, CAE and BET may act as protective mechanisms, facilitating the preservation of the lung's physiological integrity. The outcomes of this study might pave the way for the development of leads for producing single medications for the management of drug-resistant tuberculosis and its accompanying comorbidities.

Keywords: fibrosis, crinum, tuberculosis, antiinflammation, drug resistant

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Authors: Allison Herlene Du Plessis, Dalena (R.M.) Van Rooyen, Wilma Ten Ham-Baloyi, Sihaam Jardien-Baboo

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Introduction: Women die in pregnancy and childbirth for five main reasons—severe bleeding, infections, unsafe abortions, hypertensive disorders (pre-eclampsia and eclampsia), and medical complications including cardiac disease, diabetes, or HIV/AIDS complicated by pregnancy. In 2015, WHO classified sepsis as the third highest cause for maternal mortalities in the world. Chorioamnionitis is a clinical syndrome of intrauterine infection during any stage of the pregnancy and it refers to ascending bacteria from the vaginal canal up into the uterus, causing infection. While the incidence rates for chorioamnionitis are not well documented, complications related to chorioamnionitis are well documented and midwives still struggle to identify this condition in time due to its complex nature. Few diagnostic methods are available in public health services, due to escalated laboratory costs. Often the affordable biomarkers, such as C-reactive protein CRP, full blood count (FBC) and WBC, have low significance in diagnosing chorioamnionitis. A lack of screening impacts on effective and timeous management of chorioamnionitis, and early identification and management of risks could help to prevent neonatal complications and reduce the subsequent series of morbidities and healthcare costs of infants who are health foci of perinatal infections. Objective: This integrative literature review provides an overview of current best research evidence on the screening of women at risk for chorioamnionitis. Design: An integrative literature review was conducted using a systematic electronic literature search through EBSCOhost, Cochrane Online, Wiley Online, PubMed, Scopus and Google. Guidelines, research studies, and reports in English related to chorioamnionitis from 2008 up until 2020 were included in the study. Findings: After critical appraisal, 31 articles were included. More than one third (67%) of the literature included ranked on the three highest levels of evidence (Level I, II and III). Data extracted regarding screening for chorioamnionitis was synthesized into four themes, namely: screening by clinical signs and symptoms, screening by causative factors of chorioamnionitis, screening of obstetric history, and essential biomarkers to diagnose chorioamnionitis. Key conclusions: There are factors that can be used by midwives to identify women at risk for chorioamnionitis. However, there are a paucity of established sociological, epidemiological and behavioral factors to screen this population. Several biomarkers are available to diagnose chorioamnionitis. Increased Interleukin-6 in amniotic fluid is the better indicator and strongest predictor of histological chorioamnionitis, whereas the available rapid matrix-metalloproteinase-8 test requires further testing. Maternal white blood cells count (WBC) has shown poor selectivity and sensitivity, and C-reactive protein (CRP) thresholds varied among studies and are not ideal for conclusive diagnosis of subclinical chorioamnionitis. Implications for practice: Screening of women at risk for chorioamnionitis by health care providers providing care for pregnant women, including midwives, is important for diagnosis and management before complications arise, particularly in resource-constraint settings.

Keywords: chorioamnionitis, guidelines, best evidence, screening, diagnosis, pregnant women

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Authors: Rashmi Ranade, Neelu Joshi

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Barleria prionitis L. is a well explored Indian medicinal plant valued for its stem and leaf which forms an important ingredient of many Ayurvedic formulations. It is used for the treatment of various disorders like toothache, bleeding gums, strengthening gums, whooping cough, inflammation, arthritis, enlargement of scrotum and sciatica etc. The plant is propagated vegetatively through stem cuttings. Frequent harvesting of this plant has led to the shortage of planting material, and it has acquired the status of vulnerable plant species. Plant tissue culture technology offers a very good alternative for propagation and conservation of such plant species. The present investigation was undertaken to develop in vitro regeneration protocol for B. prionitis L. via callus organogenesis pathway. Stem and leaf explants were used for this purpose. Different media and plant growth regulators were optimized to develop the protocol. The problem of phenol secretion and browning and in vitro cultures at the establishment phase was successfully curbed with the usage of antibrowning agents such as ascorbic acid and activated charcoal. Optimum shoot multiplication was achieved by the use of liquid media and incorporation of silver nitrate and TIBA (triiodobenzoic acid) into the media. High percent rooting (76%) was observed on WPM media supplemented with IBA (2.0 mg/l), IAA (0.5 mg/l), GA3(0.5) and activated charcoal(500 mg/l). The rooted plantlets were subjected to in vitro hardening on sterile potting mix (soil:farmyard manure:compost; 1:2:1) and acclimatized under greenhouse conditions. Around 85% survival of plantlets was recorded upon acclimatization. This lab scale protocol would be tested for in vitro scaling up production of B. prionitis L.

Keywords: explant browning, liquid culture, micropropagation, shoot multiplication, phenolic secretion

Procedia PDF Downloads 279

Authors: Bahar Tunctan, Sefika Pinar Kucukkavruk, Meryem Temiz-Resitoglu, Demet Sinem Guden, Ayse Nihal Sari, Seyhan Sahan-Firat, Mahesh P. Paudyal, John R. Falck, Kafait U. Malik

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We hypothesized that 20-hydroxyeicosatetraenoic acid (20-HETE) mimetics such as N-(20-hydroxyeicosa-5[Z],14[Z]-dienoyl)glycine (5,14-HEDGE) may be beneficial for preventing mortality due to inflammation induced by lipopolysaccharide (LPS). This study aims to assess the effect of 5,14-HEDGE on the LPS-induced changes in nucleotide binding domain and leucine-rich repeat protein 3 (NLRP3)/apoptosis-associated speck-like protein containing a caspase activation and recruitment domain (ASC)/pro-caspase-1 inflammasome. Rats were injected with saline (4 ml/kg) or LPS (10 mg/kg) at time 0. Blood pressure and heart rate were measured using a tail-cuff device. 5,14-HEDGE (30 mg/kg) was administered to rats 1 h after injection of saline or LPS. The rats were sacrificed 4 h after saline or LPS injection and kidney, heart, thoracic aorta, and superior mesenteric artery were isolated for measurement of caspase-1/11 p20, NLRP3, ASC, and β-actin proteins as well as interleukin-1β (IL-1β) levels. Blood pressure decreased by 33 mmHg and heart rate increased by 63 bpm in the LPS-treated rats. In the LPS-treated rats, tissue protein expression of caspase-1/11 p20, NLRP3, and ASC in addition to IL-1β levels were increased. 5,14-HEDGE prevented the LPS-induced changes. Our findings suggest that inhibition of renal, cardiac, and vascular formation/activity of NLRP3/ASC/pro-caspase-1 inflammasome involved in the protective effect of 5,14-HEDGE on LPS-induced septic shock in rats. This work was financially supported by the Mersin University (2015-AP3-1343) and USPHS NIH (PO1 HL034300).

Keywords: 5, 14-HEDGE, lipopolysaccharide, NLRP3, inflammasome, septic shock

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Authors: Andriana Purnama, Reno Rudiman, Kezia Christy

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Anal stenosis which develops due to anoderm scarring usually caused by secondary to surgical trauma, has become common, causing significant decrease patient’s quality of life. Even though mild anal stenosis was treated with non-surgical treatment, but surgical reconstruction in unavoidable for moderate to severe anal stenosis that cause distressing, severe anal pain and inability to defecate. In our study, we intend to share our result with the use of diamond flap in treatment of anal stenosis. This case report illustrates a 57-year-old male patient who presented with difficulty and discomfort in defecation caused by anal stenosis after 2 years of open hemorrhoidectomy surgery. At physical examination, there was requirement of forceful dilatation when the index finger was inserted or precisely 6mm as measured by hegar dilator (moderate anal stenosis). Blood test result was within normal limits. The patient underwent anal repair and diamond flap where the scar tissue at 6 and 9 o’clock directions was excised and diamond graft was incised carefully while paying attention to the vascular supply. Finally, the graft was fixated without any tension to the anal canal, resulting in diameter of 2 cm after operation. After 2 days post operation, the patient was in stable condition, without any complication, and discharged. There was no abnormality concerning the stool. Ten days after the operation, diamond flap was in normal condition and without any complication. He was scheduled for futher follow up at the Digestive Surgery Department. Anal stenosis due to overzealous hemorrhoidectomy is a complication that is preventable when performed in experienced hands. Diamond flap was one of the options for the anal stenosis treatment with less complication.

Keywords: anal stenosis, diamond flap, post hemorrhoidectomy, anal repair

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Authors: Alireza Jafarzadeh, Samad Mosaferi, Mansour Khakpour

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Mastitis is an inflammation of the parenchyma of mammary gland regardless of the causes. Mastitis is characterized by a range of physical and chemical changes in the glandular tissue. The most important change in milk includes discoloration, the presence of clots and large number of leucocytes. There is swelling, heat, pain and edema in mammary gland in many clinical cases. Positive coagulase S. aureus is a major pathogen of the bovine mammary gland and a common cause of contagious mastitis in cattle. The aim of this study was to evaluate the outbreaks of Staphylococcus aureus mastitis. This study is conducted in ten dairy herds about one thousand cows. After doing CMT and identifying infected cows, the milk samples obtained from infected teats and transported to microbiological laboratories. After microbial culture of milk samples and isolating S. aureus, antimicrobial, sensitivity test was performed with disk diffusion method by ciprofloxacin, co-amoxiclav, erythromycin, penicillin, oxytetracyclin, sulfonamides, lincomycin and cefquinome. The study defined that the outbreak of subclinical positive coagulase Staphylococcus mastitis in dairy herd was 13.11% (5.6% S. aureus and 7.51% S. intermedicus). The antimicrobial sensitivity test shown that 87.23% of Staphylococcus aureus isolated from bovine mastitis in dairy herd was susceptible to ciprofloxacin, 93.9% to cefquinome, 4.67% to co-amoxiclav, 12.16% to erythromycin 86.11% to sulfonamides (co-trimoxazole), 3.35% lincomycin, 12.7% to oxytetracyclin and 5.98% to penicillin. Results of present defined that ciprofloxacin has a great effect on Staphylococcus aureus isolated from subclinical bovine mastitis dairy herd. It seems that cefquinome sulfonamides has a great effect on isolated Staphylococcus aureus in vivo.

Keywords: ciprofloxacin, mastitis, Staphylococcus aureus, dairy herd

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Authors: Steven Arcidiacono, Robert Stote, Erin Anderson, Molly Richards

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There are numerous standard test methods for antimicrobial textiles that measure activity against specific microorganisms. However, many times these results do not translate to the performance of treated textiles when worn by individuals. Standard test methods apply a single target organism grown under optimal conditions to a textile, then recover the organism to quantitate and determine activity; this does not reflect the actual performance environment that consists of polymicrobial communities in less than optimal conditions or interaction of the textile with the skin substrate. Here we propose the development of in vitro skin model method to bridge the gap between lab testing and wear studies. The model will consist of a defined polymicrobial community of 5-7 commensal microbes simulating the skin microbiome, seeded onto a solid tissue platform to represent the skin. The protocol would entail adding a non-commensal test organism of interest to the defined community and applying a textile sample to the solid substrate. Following incubation, the textile would be removed and the organisms recovered, which would then be quantitated to determine antimicrobial activity. Important parameters to consider include identification and assembly of the defined polymicrobial community, growth conditions to allow the establishment of a stable community, and choice of skin surrogate. This model could answer the following questions: 1) is the treated textile effective against the target organism? 2) How is the defined community affected? And 3) does the textile cause unwanted effects toward the skin simulant? The proposed model would determine activity under conditions comparable to the intended application and provide expanded knowledge relative to current test methods.

Keywords: antimicrobial textiles, defined polymicrobial community, in vitro skin model, skin microbiome

Procedia PDF Downloads 129

Authors: Felice Elefant, Akanksha Bhatnaghar, Keegan Krick, Elizabeth Heller

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Context: The severity of Alzheimer’s Disease (AD) progression involves an interplay of genetics, age, and environmental factors orchestrated by histone acetyltransferase (HAT) mediated neuroepigenetic mechanisms. While disruption of Tip60 HAT action in neural gene control is implicated in AD, alternative mechanisms underlying Tip60 function remain unexplored. Altered RNA splicing has recently been highlighted as a widespread hallmark in the AD transcriptome that is implicated in the disease. Research Aim: The aim of this study was to identify a novel RNA binding/splicing function for Tip60 in human hippocampus and impaired in brains from AD fly models and AD patients. Methodology/Analysis: The authors used RNA immunoprecipitation using RNA isolated from 200 pooled wild type Drosophila brains for each of the 3 biological replicates. To identify Tip60’s RNA targets, they performed genome sequencing (DNB-SequencingTM technology, BGI genomics) on 3 replicates for Input RNA and RNA IPs by Tip60. Findings: The authors' transcriptomic analysis of RNA bound to Tip60 by Tip60-RNA immunoprecipitation (RIP) revealed Tip60 RNA targets enriched for critical neuronal processes implicated in AD. Remarkably, 79% of Tip60’s RNA targets overlap with its chromatin gene targets, supporting a model by which Tip60 orchestrates bi-level transcriptional regulation at both the chromatin and RNA level, a function unprecedented for any HAT to date. Since RNA splicing occurs co-transcriptionally and splicing defects are implicated in AD, the authors investigated whether Tip60-RNA targeting modulates splicing decisions and if this function is altered in AD. Replicate multivariate analysis of transcript splicing (rMATS) analysis of RNA-Seq data sets from wild-type and AD fly brains revealed a multitude of mammalian-like AS defects. Strikingly, over half of these altered RNAs were bonafide Tip60-RNA targets enriched for in the AD-gene curated database, with some AS alterations prevented against by increasing Tip60 in fly brain. Importantly, human orthologs of several Tip60-modulated spliced genes in Drosophila are well characterized aberrantly spliced genes in human AD brains, implicating disruption of Tip60’s splicing function in AD pathogenesis. Theoretical Importance: The authors' findings support a novel RNA interaction and splicing regulatory function for Tip60 that may underlie AS impairments that hallmark AD etiology. Data Collection: The authors collected data from RNA immunoprecipitation experiments using RNA isolated from 200 pooled wild type Drosophila brains for each of the 3 biological replicates. They also performed genome sequencing (DNBSequencingTM technology, BGI genomics) on 3 replicates for Input RNA and RNA IPs by Tip60. Questions: The question addressed by this study was whether Tip60 has a novel RNA binding/splicing function in human hippocampus and whether this function is impaired in brains from AD fly models and AD patients. Conclusions: The authors' findings support a novel RNA interaction and splicing regulatory function for Tip60 that may underlie AS impairments that hallmark AD etiology.

Keywords: Alzheimer's disease, cognition, aging, neuroepigenetics

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Authors: Ibrahima Farikou, Kolontchang Gatchou Alberic Lionel, Tsiagadgui Jean Gustave, Ngo Yamben Marie-Ange, Handy Eone Daniel

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Introduction: It was in 1956 that Ilizarov pioneered the concept of osteogenesis in distraction by the device that bears his name to help produce bone and soft tissue regeneration and bone consolidation. This technique is not widely used in Africa where, however, its applications are numerous (loss of bone substances, congenital or acquired malformations). Our goal is to bring the indications of Ilizarov's device back to our practice conditions. Methods: Our study was conducted in 2 hospitals over a period of 20 years. For the retrospective phase, this study included all complete usable records of patients operated on in the Ilizarov external fixator department, and for the prospective phase, all patients operated on in the departments with complete usable records. Our sample was consecutive and not exhaustive. Data were analyzed by SPSS software version 23.0. Results: A total of 52 patients were reviewed. The average age of our patients was 14.7 years. The sex ratio was 1.6 in favor of men. The lower limb was the most affected (49), with a predominance of the tibia (62.4%). The average elongation was 6.4 cm. Traumatic acquired pathologies (delayed union, malunion) represented 60.6%. The mean time to union was seven months. Correction of the limb length discrepancy or filling of loss of bone substance was obtained in 75% of cases. Functionally, 80.8% of the patients treated had regained autonomy at the end of treatment, but in 17.3% of the patients, pain and limping persisted. Conclusion: This technique should be popularized in Africa because the benefit that would accrue to patients is invaluable and would be an attractive alternative to many amputations sometimes carried out in Africa by despair.

Keywords: ilizarov, external fixator, limb lengthening, bone regeneration, africa

Procedia PDF Downloads 98

Authors: Seema Ahlawat, Parul Saxena, Malik Zainul Abdin

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Malaria is a major health problem in many developing countries. The parasite responsible for the vast majority of fatal malaria infections is Plasmodium falciparum. Unfortunately, most Plasmodium strains including P. falciparum have become resistant to most of the antimalarials including chloroquine, mefloquine, etc. To combat this problem, WHO has recommended the use of artemisinin and its derivatives in artemisinin based combination therapy (ACT). Due to its current use in artemisinin based-combination therapy (ACT), its global demand is increasing continuously. But, the relatively low yield of artemisinin in A. annua L. plants and unavailability of economically viable synthetic protocols are the major bottlenecks for its commercial production and clinical use. Chemical synthesis of artemisinin is also very complex and uneconomical. The hairy root system, using the Agrobacterium rhizogenes LBA 9402 strain to enhance the production of artemisinin in A. annua L., is developed in our laboratory. The transgenic nature of hairy root lines and the copy number of trans gene (rol B) were confirmed using PCR and Southern Blot analyses, respectively. The effect of different concentrations of Piriformospora indica on artemisinin production in hairy root cultures were evaluated. 3% P. indica has resulted 1.97 times increase in artemisinin production in comparison to control cultures. The effects of P. indica on artemisinin production was positively correlated with regulatory genes of MVA, MEP and artemisinin biosynthetic pathways, viz. hmgr, ads, cyp71av1, aldh1, dxs, dxr and dbr2 in hairy root cultures of A. annua L. Mass scale cultivation of A. annua L. hairy roots by plant tissue culture technology may be an alternative route for production of artemisinin. A comprehensive investigation of the hairy root system of A. annua L. would help in developing a viable process for the production of artemisinin. The efficiency of the scaling up systems still needs optimization before industrial exploitation becomes viable.

Keywords: A. annua L., artemisinin, hairy root cultures, malaria

Procedia PDF Downloads 413

Authors: Keundong Lee, Dongha Yoo, Youngbin Tchoe, Gyu-Chul Yi

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Light emitting diode (LED) array is an ideal optical stimulation tool for optogenetics, which controls inhibition and excitation of specific neurons with light-sensitive ion channels or pumps. Although a fiber-optic cable with an external light source, either a laser or LED mechanically connected to the end of the fiber-optic cable has widely been used for illumination on neural tissue, a new approach to use micro LEDs (µLEDs) has recently been demonstrated. The LEDs can be placed directly either on the cortical surface or within the deep brain using a penetrating depth probe. Accordingly, this method would not need a permanent opening in the skull if the LEDs are integrated with miniature electrical power source and wireless communication. In addition, multiple color generation from single µLED cell would enable to excite and/or inhibit neurons in localized regions. Here, we demonstrate flexible and color tunable µLEDs for the optogenetic device applications. The flexible and color tunable LEDs was fabricated using multifaceted gallium nitride (GaN) nanorod arrays with GaN nanorods grown on InxGa1−xN/GaN single quantum well structures (SQW) anisotropically formed on the nanorod tips and sidewalls. For various electroluminescence (EL) colors, current injection paths were controlled through a continuous p-GaN layer depending on the applied bias voltage. The electric current was injected through different thickness and composition, thus changing the color of light from red to blue that the LED emits. We believe that the flexible and color tunable µLEDs enable us to control activities of the neuron by emitting various colors from the single µLED cell.

Keywords: light emitting diode, optogenetics, graphene, flexible optoelectronics

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Authors: Luísa P. Cruz-Lopes, Hugo Costa e Silva, Idalina Domingos, José Ferreira, Luís Teixeira de Lemos, Bruno Esteves

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The liquefaction process of cork based tree barks has led to an increase of interest due to its potential innovation in the lumber and wood industries. In this particular study the bark of Quercus cerris (Turkish oak) is used due to its appreciable amount of cork tissue, although of inferior quality when compared to the cork provided by other Quercus trees. This study aims to optimize alkaline catalysis liquefaction conditions, regarding several parameters. To better comprehend the possible chemical characteristics of the bark of Quercus cerris, a complete chemical analysis was performed. The liquefaction process was performed in a double-jacket reactor heated with oil, using glycerol and a mixture of glycerol/ethylene glycol as solvents, potassium hydroxide as a catalyst, and varying the temperature, liquefaction time and granulometry. Due to low liquefaction efficiency resulting from the first experimental procedures a study was made regarding different washing techniques after the filtration process using methanol and methanol/water. The chemical analysis stated that the bark of Quercus cerris is mostly composed by suberin (ca. 30%) and lignin (ca. 24%) as well as insolvent hemicelluloses in hot water (ca. 23%). On the liquefaction stage, the results that led to higher yields were: using a mixture of methanol/ethylene glycol as reagents and a time and temperature of 120 minutes and 200 ºC, respectively. It is concluded that using a granulometry of <80 mesh leads to better results, even if this parameter barely influences the liquefaction efficiency. Regarding the filtration stage, washing the residue with methanol and then distilled water leads to a considerable increase on final liquefaction percentages, which proves that this procedure is effective at liquefying suberin content and lignocellulose fraction.

Keywords: liquefaction, Quercus cerris, polyalcohol liquefaction, temperature

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Authors: David Karasek, Veronika Kubickova, Ondrej Krystynik, Dominika Goldmannova, Lubica Cibickova, Jan Schovanek

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Introduction: Adiponectin, adipocyte-fatty acid-binding protein (A-FABP), and Wnt1 inducible signaling pathway protein-1 (WISP-1) are adipokines particularly associated with insulin resistance. The aim of the study was to compare their levels in women with gestational diabetes (GDM), type 2 diabetes mellitus (T2DM) and healthy controls and determine their relation with metabolic parameters. Methods: Fifty women with GDM, 50 women with T2DM, and 35 healthy women were included in the study. In addition to adipokines, anthropometric, lipid parameters, and markers, insulin resistance, and glucose control were assessed in all participants. Results: Compared to healthy controls only significantly lower levels of adiponectin were detected in women with GDM, whereas lower levels of adiponectin, higher levels of A-FABP and of WISP-1 were present in women with T2DM. Women with T2DM had also lower levels of adiponectin and higher levels of A-FABP compared to women with GDM. In women with GDM or T2DM adiponectin correlated negatively with body mass index (BMI), triglycerides (TG), C-peptide and positively with HDL-cholesterol; A-FABP positively correlated with BMI, TG, waist, and C-peptide. Moreover, there was a positive correlation between WISP-1 and C-peptide in women with T2DM. Conclusion: Adverse adipokines production detecting dysfunctional fat tissue is in women with GDM less presented than in women with T2DM, but more expressed compared to healthy women. Acknowledgment: Supported by AZV NV18-01-00139 and MH CZ DRO (FNOl, 00098892).

Keywords: adiponectin, adipocyte-fatty acid binding protein, wnt1 inducible signaling pathway protein-1, gestational diabetes, type 2 diabetes mellitus

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Authors: Recep Kesli, Merih Simsek, Cengiz Demir, Onur Turkyilmaz

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Objective: Goal of this study was to determine the antibiotic resistance of Staphylococcus aureus (S. aureus) and Methicillin resistant staphylococcus aureus (MRSA) strains isolated at Medical Microbiology Laboratory of ANS Application and Research Hospital, Afyon Kocatepe University, Turkey. Methods: S. aureus strains isolated between October 2012 and September 2016, from various clinical specimens were evaluated retrospectively. S. aureus strains were identified by both the conventional methods and automated identification system -VITEK 2 (bio-Mérieux, Marcy l’etoile, France), and Meticillin resistance was verified using oxacillin disk with disk-diffusion method. Antibiotic resistance testing was performed by Kirby-Bauer disc diffusion method according to CLSI criteria, and intermediate susceptible strains were considered as resistant. Results: Seven hundred S.aureus strains which were isolated from various clinical specimens were included in this study. These strains were mostly isolated from blood culture, tissue, wounds and bronchial aspiration. All of 306 (43,7%) were oxacillin resistant. While all the S.aureus strains were found to be susceptible to vancomycin, teicoplanin, daptomycin and linezolid, 38 (9.6 %), 77 (19.5 %), 116 (29.4 %), 152 (38.6 %) and 28 (7.1 %) were found to be resistant aganist to clindamycin, erythromycin, gentamicin, tetracycline and sulfamethoxazole/trimethoprim, retrospectively. Conclusions: Comparing to the Methicillin sensitive staphylococcus aureus (MSSA) strains, increased resistance rates of, trimethoprim-sulfamethoxazole, clindamycin, erythromycin, gentamicin, and tetracycline were observed among the MRSA strains. In this study, the most effective antibiotic on the total of strains was found to be trimethoprim-sulfamethoxazole, the least effective antibiotic on the total of strains was found to be tetracycline.

Keywords: antibiotic resistance, MRSA, Staphylococcus aureus, VITEK 2

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Authors: Amber C. W. Vandepoele, Michael A. Marciano

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Biological analyses frequently focus on single organisms, however many times, the biological sample consists of more than the target organism; for example, human microbiome research targets bacterial DNA, yet most samples consist largely of human DNA. Therefore, there would be an advantage to removing these contaminating organisms. Conversely, some analyses focus on a single organism but would greatly benefit from the additional information regarding the other organismal components of the sample. Forensic analysis is one such example, wherein most forensic casework, human DNA is targeted; however, it typically exists in complex non-pristine sample substrates such as soil or unclean surfaces. These complex samples are commonly comprised of not just human tissue but also microbial and plant life, where these organisms may help gain more forensically relevant information about a specific location or interaction. This project aims to optimize a ‘phylogenetic’ differential extraction method that will separate mammalian, bacterial and plant cells in a mixed sample. This is accomplished through the use of size exclusion separation, whereby the different cell types are separated through multiple filtrations using 5 μm filters. The components are then lysed via differential enzymatic sensitivities among the cells and extracted with minimal contribution from the preceding component. This extraction method will then allow complex DNA samples to be more easily interpreted through non-targeting sequencing since the data will not be skewed toward the smaller and usually more numerous bacterial DNAs. This research project has demonstrated that this ‘phylogenetic’ differential extraction method successfully separated the epithelial and bacterial cells from each other with minimal cell loss. We will take this one step further, showing that when adding the plant cells into the mixture, they will be separated and extracted from the sample. Research is ongoing, and results are pending.

Keywords: DNA isolation, geolocation, non-human, phylogenetic separation

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Authors: Somaiya Mateen, Shagufta Moin, Mohammad Owais, Abdul Khan, Atif Zafar

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In rheumatoid arthritis (RA), impaired oxidative metabolism and imbalance between pro-and anti-inflammatory cytokines are responsible for causing inflammation and the degradation of cartilage and bone. The present study was done to evaluate the level and hence the role of reactive oxygen species (ROS) and inflammatory cytokines in the pathogenesis of RA. The present study was performed in the blood of 80 RA patients and 55 age and sex-matched healthy controls. The level of ROS (in 5% hematocrit) and the plasma level of pro-inflammatory cytokines [TNF-α, interleukin-6 (IL-6), IL-22] and anti-inflammatory cytokines (IL-4 and IL-5) were monitored in healthy subjects and RA patients. For evaluating the role of rheumatoid factor (RF) in the pathogenesis of RA, patients were sub-divided on the basis of presence or absence of RF. Reactive species and inflammatory cytokines were correlated with disease activity measure-Disease Activity Score for 28 joints (DAS28). The level of ROS, TNF-α, IL-6 and IL-22 were found to be significantly higher in RA patients as compared to the healthy controls, with the increase being more significant in patients positive for rheumatoid factor and those having high disease severity. On the other hand, a significant decrease in the level of IL-4 and IL-10 were observed in RA patients compared with healthy controls, with the decrease being more prominent in severe cases of RA. Higher ROS (indicative of impaired anti-oxidant defence system) and pro-inflammatory cytokines level in RA patients may lead to the damage of biomolecules which in turn contributes to tissue damage and hence to the development of more severe RA. The imbalance between pro-and anti-inflammatory cytokines may lead to the development of multi-system immune complications. ROS and inflammatory cytokines may also serve as a potential biomarker for assessing the disease severity.

Keywords: rheumatoid arthritis, reactive oxygen species, pro-inflammatory cytokines, anti-inflammatory cytokines

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Authors: Ahmed Ghachem, Johann Colomba, Denis Prud'homme, Martin Brochu

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Background: Inactive obese postmenopausal women, are at greater risk for metabolic complications. On the other hand, high levels of cardiorespiratory fitness (CRF) are associated with a lower risk of metabolic complications. Objective: To compare inactive obese postmenopausal women displaying ‘lower’ vs ‘higher’ levels of CRF for body composition, metabolic profile, inflammatory profile and measures of energy expenditure. Methods: 132 women (age: 57.6 ± 4.8 yrs; BMI: 32.3 ± 4.6 kg/m2; Peak VO2: 17.81 ± 3.02 ml O2•kg-1•min-1) were studied. They were first divided into tertiles based on their CRF. Then, women in the first (< 16.51 ml O2•min-1•kg-1) and second tertiles (16.51 to 19.22 ml O2•min-1•kg-1) were combined (N= 88), and compared with those in the third tertile (> 19.22 ml O2•min-1•kg-1) (N= 44). Variables of interest were: Peak VO2 (stationary bike), body composition (DXA), body fat distribution (CT scan), glucose homeostasis (fasting state and euglycemic/ hyperinsulinemic clamp), fasting lipids, resting blood pressure, inflammatory profile and energy expenditure (DLW). Results: Both CRF groups (lower= 16.0 ± 2.0 ml O2•kg-1•min-1 vs higher= 21.2 ± 1.7 ml O2•kg-1•min-1; p < 0.001) were similar for age. Significant differences were observed between groups for body composition; with lower values for body weight, BMI, fat mass and visceral fat in women with higher CRF (p between 0.001 and 0.005). Also, women with higher CRF had lower values for fasting insulin (13.4 ± 4.5 vs 15.6 ± 6.6 μU/ml; p = 0.03) and CRP levels (2.31 ± 1.97 vs 3.83 ± 3.24 mg/liter; p = 0.001); and higher values for glucose disposal (6.71 ± 1.78 vs 5.92 ± 1.67 mg/kg/min; p = 0.01). However, these differences were no longer significant after controlling for visceral adipose tissue accumulations. Finally, no significant difference was observed between groups for the other variables of interest. Conclusion: Our results suggest that, among inactive overweight/obese postmenopausal women, those with higher CRF levels have a better metabolic profile; which is caused by lower visceral fat accumulations.

Keywords: cardiorespiratory fitness, metabolic profile, menopause, obesity

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Authors: Bryony Davies

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Many cell and tissue culture practices ignore the effects of gravity on cell biology, and little is known about how cell components may move in response to gravitational forces. Starfish oocytes provide an excellent model for interrogating the movement of cell components due to their unusually large size, ease of handling, and high transparency. Chromosomes from starfish oocytes can be visualised by microinjection of the histone-H2B-mCherry plasmid into the oocytes. The movement of the chromosomes can then be tracked by live-cell fluorescence microscopy. The results from experiments using these methods suggest that there is a replicable downward movement of centrally located chromosomes at a median velocity of 0.39 μm/min. Chromosomes nearer the nuclear boundary showed more restricted movement. Chromosome density and shape could also be altered by microinjection of restriction enzymes, primarily Alu1, before imaging. This was found to alter the speed of chromosome movement, with chromosomes from Alu1-injected nuclei showing a median downward velocity of 0.60 μm/min. Overall, these results suggest that there is a non-negligible movement of chromosomes in response to gravitational forces and that this movement can be altered by enzyme activity. Future directions based on these results could interrogate if this observed downward movement extends to other cell components and to other cell types. Additionally, it may be important to understand whether gravitational orientation and vertical positioning of cell components alter cell behaviour. The findings here may have implications for current cell culture practices, which do not replicate cell orientations or external forces experienced in vivo. It is possible that a failure to account for gravitational forces in 2D cell culture alters experimental results and the accuracy of conclusions drawn from them. Understanding possible behavioural changes in cells due to the effects of gravity would therefore be beneficial.

Keywords: starfish, oocytes, live-cell imaging, microinjection, chromosome dynamics

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Authors: Gulcin Yildiz

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Drying (dehydration) is the process of removing water from food in order to preserve the food and an alternative to reduce post-harvest loss of fruits. Different pre-treatment methods have been developed for fruit drying, such as ultrasound. If no pre-treatment is done, the fruits will continue to darken after they are dried. However, the effects of ultrasound as pre-treatment on drying of apples has not been well documented. This study was undertaken to investigate the effect of ultrasound as pre-treatment before oven drying of red delicious and golden delicious apples. Red delicious and golden delicious apples were dried in different temperatures. Before performing drying experiments in an oven at 50, 75 and 100 °C, ultrasound as pretreatment was applied in 5, 10, and 15 minutes. Colors of the dried apples were measured with a Minolta Chroma Meter CR-300 (Minolta Camera Co. Ltd., Osaka, Japan) by directly holding the device vertically to the surface of the samples. Content of total phenols was determined spectrophotometrically with the FolinCiocalteau assay, and the antioxidant capacity was evaluated by using 1,1-diphenyl-2-picrylhydrazyl (DPPH) assay. The samples (both red delicious and golden delicious apples) with longer ultrasound treatment produced higher weight loss due to the changes in tissue structure. However less phenolic content and antioxidant capacity were observed for the samples with longer ultrasound pre-treatment. The highest total phenolic content (TPC) was determined in dried apples at 75 °C with 5 minutes pre-treatment ultrasound and the lowest TPC was determined in dried apples at 50 °C with 15 minutes pre-treatment ultrasound which was subjected to the longest ultrasound pre-treatment and drying. The combination of 5 min of ultrasound pre-treatment and 75 °C of oven-drying showed to be the best combination for an energy efficient process. This combination exhibited good antioxidant properties as well. The present study clearly demonstrated that applying ultrasound as pre-treatment for drying of apples is an effective process in terms of quality of dried products, time, and energy.

Keywords: golden delicious apples, red delicious apples, total phenolic content, Ultrasound

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Authors: Seyed Reza Aghili, Tahereh Shokohi, Lotfollah Davoodi, Zahra Kashi, Azam Moslemi, Mahdi Abastabar, Iman Haghani, Sabah Mayahi, Asoudeh A.

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Introduction: In diabetic foot ulcers, if fungal agents such as Candida species penetrate into the cutaneous or depth of ulcer, can increase the degree of the wound and cause Candia infection and make it more difficult to heal. Material & Methods: A cross-sectional study was performed on 100 diabetic foot ulcer patients in 2020 in Sari, Iran. patient's data and wound grade were recorded in a questionnaire. Candida infection was diagnosed with direct microscopic examination and culture of samples. Colony-PCR molecular method was used for ITS region of DNA and then PCR-RFLP with Msp1 enzyme and using HWP1 specific gene to determine species of Candida agent. Results: Of 100 patients, the mean age 62.1 ± 10.8 years, 95% type 2 diabetes, 83%>10 years duration diabetes, 59% male, 66%> poor education level, 99% married, 52% rural, 95% neuropathic symptoms, 88% using antibiotics, 69%HbA1C >9%, and mean ulcer degree 2.6±1.05 were. Candida infection was seen in 13% of the deep tissue of the wound and 7% cutaneous around the wound. The predominant Candida isolated was C. parapsilosis (71.5%), C .albicans (14.3%). Fungal infections caused by mold fungi were not detected. There was a statistically significant relationship between yeast infection and gender, rural, HbA1C and ulcer degree. Conclusion: Mycological evaluations often are ignored. Candida parapsilosis is the most common infectious agent in these patients and may require specific treatment. Therefore, more attention of physicians to Candida infections particularly, early diagnosis and effective treatment can help faster recovery and prevent amputation.

Keywords: diabetic foot ulcer, candida infection, risk factors, c. parapsilosis

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Authors: Fatemeh Rasooli, Behzad Simiari, Pooya Payandemehr, Amir Nejati, Maryam Bahreini, Atefeh Abdollahi

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Introduction: Pain is a complex physiological reaction to tissue injury. In the course of painful procedures such as nerve block, ice has been shown to be a feasible and inexpensive material to control pain. It delays nerve conduction, actives other senses and reduces inflammatory and painful responses. This study assessed the effect of ice in reducing pain caused by needling and infiltration during digital block. Patient satisfaction recorded as a secondary outcome. Methods: This study was designed as a non-blinded randomized clinical trial approved by Tehran University of Medical Sciences Ethical Committee. Informed consent was taken from all the participants who were then randomly divided into two groups. Digital block performed by standard approach in selected patients. Tubes of ice were prepared in gloves and were fragmented at a time of application for circling around the finger. Tubes were applied for 6 minutes before digital nerve block in the site of needling in the case group. Patients in the control group underwent digital nerve block with the conventional method without ice administration. Numeric Rating Scale (NRS) used for grading pain. 0 used for no pain and 10 for the worst pain that patient had experienced until now. Scores were analyzed by Wilcoxon Rank Sum test and compared in case and control groups. Results: 100 patients aged 16-50 years were enrolled. Mean NRS scores with and without ice were 1.5 mm (S.D ± 1.44) and 6.8 mm (S.D ± 1.40) for needling pain and for infiltration pain were 2.7mm ( S.D ±1.65) and 8.5mm ( S.D ± 1.47), respectively (p<0.001). Besides, patients’ satisfactions were significantly higher in the ice group (p<0.001). Conclusion: Application of ice for 6 minutes significantly reduced pain of needling and infiltration in digital nerve block; thus, it seems to be a feasible and inexpensive material which acts effectively to decrease pain and stress before the procedure.

Keywords: digital block, ice, needle, pain

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Authors: Yara Mohamed Taha, Mohamed Ali El Desouky, Heba Kamal Abdel Hakim, Maha Hanafy Mahmoud

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Hepatotoxicity due to drugs and toxic chemicals constitutes a crucial health problem nowadays. Medicinal plants are widely used recently for protecting against many liver disorders and inflammatory conditions. This study aims to evaluate hepatoprotective impact of green tea extract (GTE), rosemary extract (RE) and rosmarinic acid (RA) against hepatotoxins; ferric nitrilotriacetate (Fe-NTA) and diethylnitrosamine (DEN) in rats. Five groups of male Wistar rats were included; one control negative, while the other groups were treated intraperitoneally with DEN as 160 mg.kg-1 b.w. on 15th day and Fe-NTA as 5 mg.kg-1 b.w. on 33rd day. One of them was control positive. The other three groups were pre-administered with daily protective oral doses of either 200 mg.kg-1 b.w. of RE or 1 g.kg- 1 b.w. of GTE or 50 mg.kg-1 b.w. of RA two weeks prior to DEN exposure and continued till the end of the experimental period. The obtained data revealed a highly significant increase of MDA, 8-OHdG, DNA damage percent, a significant depletion of GSH and elevated Gr-1 protein expression in hepatocytes with liver tissue histopathological changes of rats exposed to DEN+Fe-NTA. Pre-administration of protective doses of RE, GTE and RA to DEN+Fe-NTA treated rats could normalize the altered biochemical, histopathological and immunohistochemical parameters. In conclusion, RE, GTE and RA showed a hepatoprotective effect against liver toxicity induced by DEN+Fe-NTA, with the best antioxidant and anti-inflammatory impact were for RA and GTE. Therefore, the current study declared that rosemary, green tea and products enriched with rosmarinic acid should be involved daily in diet of people who are exposed to chemicals and environmental toxins to protect themselves from hepatotoxicity.

Keywords: hepatotoxicity, diethylnitrosamine and ferric nitrilotriacetate, rosemary extract (RE), green tea extract (GTE), rosmarinic acid (RA)

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Authors: Esra Turker, Umit Hakan Yildiz, Ahu Arslan Yildiz

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The key of regenerative medicine is mimicking natural three dimensional (3D) microenvironment of tissues by utilizing appropriate biomaterials. In this study, a synthetic biodegradable polymer; poly (L-lactide-co-ε-caprolactone) (PLLCL) and a natural polymer; collagen was used to mimic the biochemical structure of the natural extracellular matrix (ECM), and by means of electrospinning technique the real physical structure of ECM has mimicked. PLLCL/Collagen biocomposite scaffolds enables cell attachment, proliferation and nutrient transport through fabrication of micro to nanometer scale nanofibers. Biocomposite materials are commonly preferred due to limitations of physical and biocompatible properties of natural and synthetic materials. Combination of both materials improves the strength, degradation and biocompatibility of scaffold. Literature studies have shown that collagen is mostly solved with heavy chemicals, which is not suitable for cell culturing. To overcome this problem, a new approach has been developed in this study where polyvinylpyrrolidone (PVP) is used as co-electrospinning agent. PVP is preferred due to its water solubility, so PLLCL/collagen biocomposite scaffold can be easily and rapidly produced. Hydrolytic and enzymatic biodegradation as well as mechanical strength of scaffolds were examined in vitro. Cell adhesion, proliferation and cell morphology characterization studies have been performed as well. Further, on-chip drug screening analysis has been performed over 3D tumor models. Overall, the developed biocomposite scaffold was used for 3D tumor model formation and obtained results confirmed that developed model could be used for drug screening studies to predict clinical efficacy of a drug.

Keywords: biomaterials, 3D cell culture, drug screening, electrospinning, lab-on-a-chip, tissue engineering

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Authors: Hiba T. Allah ALtieb Gusm ALsied, Amna Beshir Medani Ahmed, Ikram Mohamed ELtayeb, Saad Mohamed Hussein Ayoub

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This study was carried out to determine the hepatoactivity and the antioxidant activity of Coiradrum sativum L. aerial part and fruit extracts against CCL4 induced acute liver damages in Wistar rats. The aerial parts and fruits part of the plant were extracted 96% ethanol with soxhlet apparatus. Hepatic injury was achieved by subcutaneous injection of 3 ml/kg of CCL4 diluted with olive oil with ratio 1:1. The extracts were mixed together 1:1 ratio and given in different doses 100,200,400 mg/kg/day for 5 days under CCL4 induction at 3rd day. The significance of differences between means by using T-test was compared among the groups. The mixture of the two extracts didn’t show any significant result in protecting liver injury (antagonistic effects), it shows high level of liver enzyme like alkaline phosphatase (ALP), glutamate oxaloacetate transaminase (SGOT) and glutamate pyruvate transaminase (SGPT). Serological studies further confirmed the results. The results obtained were compared with silymarin (70 mg/kg/day) orally, the standard drug for hepatoprotection which show recovery close to normalization almost like that of silymarin; therefore, further studies on this plant with different ratios especially in isolated tissue to spot more light on antagonistic effects of the two extracts. Antioxidant activity of the extracts was determined by the DPPH method. The results obtained show high anti-oxidant activity for fruits extract while slight or moderate antioxidant activity to aerial extracts.

Keywords: antioxidant, aerial part, Coriadrum sativum L., fruity, hepatoprotection, Silymarin, phytochemical screening

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Authors: Jungkyun Im

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Nonsteroidal anti-inflammatory drugs (NSAIDs) are effective for relieving pain and reducing inflammation. They are nonselective inhibitors of two isoforms of COX, cyclooxygenase-1 (COX-1) and cyclooxygenase-2 (COX-2), and thereby inhibiting the production of hormone-like lipid compounds such as, prostaglandins and thromboxanes which cause inflammation, pain, fever, platelet aggregation, etc. In addition, recently there are many research articles reporting the neuroprotective effect of NSAIDs in neurodegenerative diseases, such as Alzheimer’s disease (AD) and Parkinson’s disease (PD). However, the clinical use of NSAIDs in these diseases is limited by low brain distribution. Therefore, in order to assist the in-depth investigation on the pharmaceutical mechanism of flurbiprofen in neuroprotection and to make flurbiprofen a more potent drug to prevent or alleviate neurodegenerative diseases, delivery of flurbiprofen to brain should be effective and sufficient amount of flurbiprofen must penetrate the BBB thus gaining access into the patient’s brain. We have recently developed several types of guanidine-rich molecular carriers with high molecular weights and good water solubility that readily cross the blood-brain barrier (BBB) and display efficient distributions in the mouse brain. The G8 (having eight guanidine groups) molecular carrier based on D-sorbitol was found to be very effective in delivering anticancer drugs to a mouse brain. In the present study, employing the same molecular carrier, we prepared the flurbiprofen conjugate and studied its BBB permeation by mouse tissue distribution study. Flurbiprofen was attached to a molecular carrier with a fluorescein probe and multiple terminal guanidiniums. The conjugate was found to internalize into live cells and readily cross the BBB to enter the mouse brain. Our novel synthetic flurbiprofen conjugate will hopefully delivery NSAIDs into brain, and is therefore applicable to the neurodegenerative diseases treatment or prevention.

Keywords: flurbiprofen, drug delivery, molecular carrier, organic synthesis

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