Search results for: alveolar epithelium
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 160

Search results for: alveolar epithelium

40 Rooibos Extract Antioxidants: In vitro Models to Assess Their Bioavailability

Authors: Ntokozo Dambuza, Maryna Van De Venter, Trevor Koekemoer

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Oxidative stress contributes to the pathogenesis of many diseases and consequently antioxidant therapy has attracted much attention as a potential therapeutic strategy. Regardless of the quantities ingested, antioxidants need to reach the diseased tissues at concentrations sufficient to combat oxidative stress. Bioavailability is thus a defining criterion for the therapeutic efficacy of antioxidants. In addition, therapeutic antioxidants must possess biologically relevant characteristics which can target the specific molecular mechanisms responsible for disease related oxidative stress. While many chemical antioxidant assays are available to quantify antioxidant capacity, they relate poorly to the biological environment and provide no information as to the bioavailability. The present comparative study thus aims to characterise green and fermented rooibos extracts, well recognized for their exceptional antioxidant capacity, in terms of antioxidant bioavailability and efficacy in a disease relevant cellular setting. Chinese green tea antioxidant activity was also evaluated. Chemical antioxidant assays (FRAP, DPPH and ORAC) confirmed the potent antioxidant capacity of both green and fermented rooibos, with green rooibos possessing antioxidant activity superior to that of fermented rooibos and Chinese green tea. Bioavailability was assessed using the PAMPA assay and the results indicate that green and fermented rooibos have a permeation coefficient of 5.7 x 10-6 and 6.9 x 10-6 cm/s, respectively. Chinese green tea permeability coefficient was 8.5 x 10-6 cm/s. These values were comparable to those of rifampicin, which is known to have a high permeability across intestinal epithelium with a permeability coefficient of 5 x 10 -6 cm/s. To assess the antioxidant efficacy in a cellular context, U937 and red blood cells were pre-treated with rooibos and Chinese green tea extracts in the presence of a dye DCFH-DA and then exposed to oxidative stress. Green rooibos exhibited highest activity with an IC50 value of 29 μg/ml and 70 μg/ml, when U937 and red blood cells were exposed oxidative stress, respectively. Fermented rooibos and Chinese green tea had IC50 values of 61 μg/ml and 57 μg/ml for U937, respectively, and 221 μg/ml and 405 μg/ml for red blood cells, respectively. These results indicate that fermented and green rooibos extracts were able to permeate the U937 cells and red blood cell membrane and inhibited oxidation of DCFH-DA to a fluorescent DCF within the cells.

Keywords: rooibos, antioxidants, permeability, bioavailability

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39 Reconstruction of Complex Post Oncologic Maxillectomy Defects

Authors: Vinay Kant Shankhdhar

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Purpose: Maxillary defects are three dimensional and require complex bone and soft tissue reconstruction. Maxillary reconstruction using fibula osteocutaneous flaps in situation requiring orbital floor, orbital wall, palatal defects, and external skin, all at the same time require special planning and multiple osteotomies. We tried to improvise our reconstruction using multiple osteotomies and skin paddle designs for fibula and Flexor Hallucis Longus Muscle. This study aims at discussing the planning and outcome in complex maxillary reconstructions using fibula flaps and soft tissue flaps with or without bone grafts. Material and Methods: From 2011 to 2017 a total of 129 Free fibula flaps were done, 67 required two or more struts, 164 Anterolateral Thigh Flaps, 11 Deep Inferior Epigastric Artery perforator flaps and 3 vertical rectus abdominis muscle flaps with iliac crest bone graft. The age range was 2 to 70 years. The reconstruction was evaluated based on the post-operative rehabilitation including orbital support (prevention of diplopia), oral diet, speech and cosmetic appearance. Results: The follow- up is from 5 years to 1 year. In this series, we observed that the common complications were the de-vascularisation of most distal segment of osteotomised fibula and native skin necrosis. Commonest area of breakdown is the medial canthal region. Plate exposure occurs most commonly at the pyriform sinus. There was extrusion of one non-vascularized bone graft. All these complications were noticed post-radiotherapy. Conclusions: The use of free fibula osteocutaneous flap gives very good results when only alveolar reconstruction is required. The reconstruction of orbital floor with extensive skin loss with post operative radiotherapy has maximum complication rate in long term follow up. A soft tissue flap with non vascularized bone graft may be the best option in such cases.

Keywords: maxilla reconstruction, fibula maxilla, post cancer maxillary reconstruction

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38 Strategies For Management Of Massive Intraoperative Airway Haemorrhage Complicating Surgical Pulmonary Embolectomy

Authors: Nicholas Bayfield, Liam Bibo, Kaushelandra Rathore, Lucas Sanders, Mark Newman

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INTRODUCTION: Surgical pulmonary embolectomy is an established therapy for acute pulmonary embolism causing right heart dysfunction and haemodynamic instability. Massive intraoperative airway haemorrhage is a rare complication of pulmonary embolectomy. We present our institutional experience with massive airway haemorrhage complicating pulmonary embolectomy and discuss optimal therapeutic strategies. METHODS: A retrospective review of emergent surgical pulmonary embolectomy patients was undertaken. Cases complicated by massive intra-operative airway haemorrhage were identified. Intra- and peri-operative management strategies were analysed and discussed. RESULTS: Of 76 patients undergoing emergent or salvage pulmonary embolectomy, three cases (3.9%) of massive intraoperative airway haemorrhage were identified. Haemorrhage always began on weaning from cardiopulmonary bypass. Successful management strategies involved intraoperative isolation of the side of bleeding, occluding the affected airway with an endobronchial blocker, institution of veno-arterial (VA) extracorporeal membrane oxygenation (ECMO) and reversal of anticoagulation. Running the ECMO without heparinisation allows coagulation to occur. Airway haemorrhage was controlled within 24 hours of operation in all patients, allowing re-institution of dual lung ventilation and decannulation from ECMO. One case in which positive end-expiratory airway pressure was trialled initially was complicated by air embolism. Although airway haemorrhage was controlled successfully in all cases, all patients died in-hospital for reasons unrelated to the airway haemorrhage. CONCLUSION: Massive intraoperative airway haemorrhage during pulmonary embolectomy is a rare complication with potentially catastrophic outcomes. Re-perfusion alveolar and capillary injury is the likely aetiology. With a systematic approach to management, airway haemorrhage can be well controlled intra-operatively and often resolves within 24 hours. Stopping blood flow to the pulmonary arteries and support of oxygenation by the institution of VA ECMO is important. This management has been successful in our 3 cases.

Keywords: pulmonary embolectomy, cardiopulmonary bypass, cardiac surgery, pulmonary embolism

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37 Spontaneous Eruption of Impacted Teeth While Awaiting Surgical Intervention

Authors: Alison Ryan, Himani Chhabra, Mohammed Dungarwalla, Judith Jones

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Background: Impacted and ectopic teeth present in 1-2% of orthodontic patients and often require joint surgical and orthodontic management. The authors present two patients undergoing orthodontic treatment, where the impacted teeth, in a hopeless position, spontaneously erupted during the period of cessation of general anaesthetic lists during the COVID-19 pandemic. Patient information: A healthy 11-year-old boy was referred to the Department of Oral and Maxillofacial Surgery for the management of a mesioangular impacted LR7. The patient was seen by the joint oral surgery/orthodontic team, who planned for the removal of the LR7 under general anaesthetic. A healthy 13-year-old boy was referred to the same Department and team for surgical extraction of unerupted and buccally impacted UL3 and UR3 under general anaesthetic. Management and outcome: The majority of elective dental-alveolar work ceased as a result of the global pandemic. On resumption of activity, the first patient was reviewed in July 2021. The LR7 had spontaneously erupted in a favourable position, and following MDT review, a decision was made to forgo any further surgical intervention. The second patient was reviewed in July 2021. The UL3 had clinically erupted, and there was radiographic evidence of favourable movement of UR3. Due to the nature of the patient’s malocclusion, the decision was made to proceed with the extractions as previously planned. Key Learning Points: Severely impacted teeth do have a prospect of spontaneous eruption or alignment without clinical intervention, and current literature states the initial location, axial inclination, degree of root formation, and relation of the impacted tooth to adjacent teeth roots may influence spontaneous eruption. There is potential to introduce a period of observation to account for this possibility in the developing dentition, with the aim of reducing the unnecessary need for surgical intervention. This could help prevent episodes of general anaesthetic and allocate theatre space more appropriately.

Keywords: spontaneous eruption, impaction, observation, hopeless position, surgical, orthodontic, change in treatment plan

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36 In Silico Study of Cell Surface Structures of Parabacteroides distasonis Involved in Its Maintain Within the Gut Microbiota and Its Potential Pathogenicity

Authors: Jordan Chamarande, Lisiane Cunat, Corentine Alauzet, Catherine Cailliez-Grimal

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Gut microbiota (GM) is now considered a new organ mainly due to the microorganism’s specific biochemical interaction with its host. Although mechanisms underlying host-microbiota interactions are not fully described, it is now well-defined that cell surface molecules and structures of the GM play a key role in such relation. The study of surface structures of GM members is also fundamental for their role in the establishment of species in the versatile and competitive environment of the digestive tract and as a potential virulence factor. Among these structures are capsular polysaccharides (CPS), fimbriae, pili and lipopolysaccharides (LPS), all well-described for their central role in microorganism colonization and communication with host epithelium. The health-promoting Parabacteroides distasonis, which is part of the core microbiome, has recently received a lot of attention, showing beneficial properties for its host and as a new potential biotherapeutic product. However, to the best of the authors’ knowledge, the cell surface molecules and structures of P. distasonis that allow its maintain within the GM are not identified. Moreover, although P. distasonis is strongly recognized as intestinal commensal species with benefits for its host, it has also been recognized as an opportunistic pathogen. In this study, we reported gene clusters potentially involved in the synthesis of the capsule, fimbriae-like and pili-like cell surface structures in 26 P. distasonis genomes and applied the new RfbA-Typing classification in order to better understand and characterize the beneficial/pathogenic behaviour related to P. distasonis strains. In context, 2 different types of fimbriae, 3 of pilus and up to 14 capsular polysaccharide loci, have been identified over the 26 genomes studied. Moreover, the addition of data to the rfbA-Type classification modified the outcome by rearranging rfbA genes and adding a fifth group to the classification. In conclusion, the strain variability in terms of external proteinaceous structure could explain the inter-strain differences previously observed in P. distasonis adhesion capacities and its potential pathogenicity.

Keywords: gut microbiota, Parabacteroides distasonis, capsular polysaccharide, fimbriae, pilus, O-antigen, pathogenicity, probiotic, comparative genomics

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35 Uranoplasty Using Tongue Flap for Bilateral Clefts

Authors: Saidasanov Saidazal Shokhmurodovich, Topolnickiy Orest Zinovyevich, Afaunova Olga Arturovna

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Relevance: Bilateral congenital cleft is one of the most complex forms of all clefts, which makes it difficult to choose a surgical method of treatment. During primary operations to close the hard and soft palate, there is a shortage of soft tissues and their lack during standard uranoplasty, and these factors aggravate the period of rehabilitation of patients. Materials and methods: The results of surgical treatment of children with bilateral cleft, who underwent uranoplasty using a flap from the tongue, were analyzed. The study used methods: clinical and statistical, which allowed us to solve the tasks, based on the principles of evidence-based medicine. Results and discussion: in our study, 15 patients were studied, who underwent surgical treatment in the following volume: uranoplasty using a flap from the tongue in two stages. Of these, 9 boys and 6 girls aged 2.5 to 6 years. The first stage was surgical treatment in the volume: veloplasty. The second stage was a surgical intervention in volume: uranoplasty using a flap from the tongue. In all patients, the width of the cleft ranged from 1.6-2.8 cm. All patients in this group were orthodontically prepared. Using this method, the surgeon can achieve the following results: maximum narrowing of the palatopharyngeal ring, long soft palate, complete closure of the hard palate, alveolar process, and the mucous membrane of the nasal cavity is also sutured, which creates good conditions for the next stage of osteoplastic surgery. Based on the result obtained, patients have positive results of working with a speech therapist. In all patients, the dynamics were positive without complications. Conclusions: Based on our observation, tongue flap uranoplasty is one of the effective techniques for patients with wide clefts of the hard and soft palate. The use of a flap from the tongue makes it possible to reduce the number of repeated reoperations and improve the quality of social adaptation of this group of patients, which is one of the important stages of rehabilitation. Upon completion of the stages of rehabilitation, all patients had the maximum improvement in functional, anatomical and social indicators.

Keywords: congenital cleft lips and palate, bilateral cleft, child surgery, maxillofacial surgery

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34 A Strategy Therapy for Retinitis Pigmentosa Induced by Argon Laser in Rabbits by High Dose Adult Stem Cells

Authors: Hager E. Amer, Hany El Saftawy, Laila Rashed, Ahmed M. Ata, Fatma Metwally, Hesham Mettawei, Hossam E. Sayed, Tamer Adel, Kareem M. El Sawah

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Aim: The purpose of this study is to regenerate the damaged photoreceptor cells as a result of argon laser as a model of Retinitis Pigmentosa in rabbits' retina by using adult stem cells from rabbits' bone marrow. Background: Retinitis pigmentosa (RP) is a group of inherited disorders that primarily affect photoreceptor and pigment epithelium function. RP leads to loss of the rod outer segment and shorten the photoreceptor layer and expose the photoreceptor cell body to high-pressure levels in oxygen (oxidative stress) leads to apoptosis to the rod and cone cells. In particular, there is no specific treatment for retinitis pigmentosa. Materials and Methods: Forty Two Giant (Rex) rabbits were used in this experiment divided into 3 groups: Group 1: Control (6 rabbits), Group 2: Argon laser radiated as a model of retinitis pigmentosa (12 rabbits), Group 3: Laser radiated and treated by 6 million stem cells (12 rabbits). The last two groups are divided each into two subgroups each subgroup contains 6 rabbits, the ophthalmological examination was performed on rabbits, blood samples and retina samples were taken after 25 days and after 36 days from the laser radiation (10 days and 21 days after stem cells insertion in group 3) to perform the biochemical analysis. Results: Compared to control Group, a decrease of ERG wave amplitude and antioxidant substances (Glutathione) in blood and retina in group 2, and an increase of oxidative stress substances (Nitric oxide, Malonaldehyde, and carponyl protein) and apoptotic substances (Advanced glycation end product and M-metalloproteinase) in blood and retina. Compared to group 2, mostly increases of antioxidant substances and ERG wave amplitude in group 3, and mostly decreases in oxidative stress substances and apoptotic substances. Conclusion: Insertion of 6 million stem cells intravitreous gives good results in regeneration of the damaged photoreceptor cells after 21 days.

Keywords: retinitis pigmentosa, stem cells, argon laser, oxidative stress, apoptosis

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33 Evaluation of Antibody Titer Produced in Layer Chicken after Vaccination with an Experimental Ornitobacterium rhinotracheal Vaccine

Authors: Mohammad Javad Mehrabanpour, Mohammad Hosein Hosseini, Ali Shirazi, Dorsa Mehrabanpour

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Respiratory infections are the most important diseases that affect poultry. Ornithobacterium rhinotracheale is a bacterium that causes respiratory infections including alveolar inflation and pneumonia in birds. The aim of this study was to evaluated antibody titer against Ornitobacterium rhinotracheal in layer chicken sera after vaccination with an experimental ORT vaccine that produced in Razi Vaccine and Serum Research Institute. Cultured bacteria were inactivated by formalin, and controlled tests were conducted on it. The obtained antigens were formulated using Montanide oil and were homogenized using homogenizer. Eighty SPF chickens were kept until the age of 14 days under existing standards for temperature, humidity, and light. At the age of 14 days, chickens were divided into 3 groups. The first group included 50 chickens injected with prepared ORT vaccine, the second group, as control group, included 15 chickens injected with sterile PBS to get stress of infection and the third group included 15 chickens with no injection performed to them. All 3 groups were kept in separate cages at same room. Blood samples were regularly taken from the chickens every week for serum separation and evaluation of antibody titer. During the fifth week post vaccination, booster vaccine was injected into the chickens of vaccinated group. The chickens were inspected every day in terms of mortality as well as any injection site reactions. Three weeks after the booster injection, blood samples were taken from all chickens of all groups, and sera were isolated. The sera of immunized (vaccinated) SPF chickens with ORT vaccine as well as that of SPF chickens in the control groups were reviewed according to the recommendations of ELISA kit manufacturer to examine the chicken’s humeral immune response to the studied vaccine. Potency, stability and sterility tests were also performed on the above mentioned vaccine. Results obtained indicate high antibody titer in sera of chickens vaccinated with experimental ORT vaccine as compared with the control groups that emphasize the ability of experimentally prepared ORT vaccine to stimulate humoral immune response of chicken. After the second injection, antibody titer increased and remained almost stable up to 9 weeks after the injection. ORT vaccine can cause potency in chickens and can protect them against disease.

Keywords: antibody, layer chicken, Ornithobactrium rhinotracitis, vaccine

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32 Evaluation of Azo Dye Toxicity Using Some Haematological and Histopathological Alterations in Fish Catla Catla

Authors: Jagruti Barot

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The textile industry plays a major role in the economy of India and on the other side of the coin it is the major source for water pollution. As azo dyes is the largest dye class they are extensively used in many fields such as textile industry, leather tanning industry, paper production, food, colour photography, pharmaceuticals and medicine, cosmetic, hair colourings, wood staining, agricultural, biological and chemical research etc. In addition to these, they can have acute and/or chronic effects on organisms depending on their concentration and length of exposure when they discharged as effluent in the environment. The aim of this study was to assess the genotoxic and histotoxic potentials of environmentally relevant concentrations of RR 120 on Catla catla, important edible freshwater fingerlings. For this, healthy Catla catla fingerlings were procured from the Government Fish Farm and acclimatized in 100 L capacity and continuously aerated glass aquarium in laboratory for 15 days. According to APHA some physic-chemical parameters were measured and maintained such as temperature, pH, dissolve oxygen, alkalinity, total hardness. Water along with excreta had been changed every 24 hrs. All fingerlings were fed artificial food palates once a day @ body weight. After 15 days fingerlings were grouped in 5 (10 in each) and exposed to various concentrations of RR 120 (Control, 10, 20, 30 and 40 mg/L) and samples (peripheral blood and gills, kidney) were collected and analyzed at 96 hrs. of interval. All results were compared with the control. Micronuclei (MN), nuclear buds (NB), fragmented-apoptotic (FA) and bi-nucleated (BN) cells in blood cells and in tissues (gills and kidney cells) were observed. Prominent histopathological alterations were noticed in gills such as aneurism, hyperplasia, degenerated central axis, lifting of gill epithelium, curved secondary gill lamellae etc. Similarly kidney showed some detrimental changes like shrunken glomeruli with increased periglomerular space, degenerated renal tubules etc. Both haematological and histopathological changes clearly reveal the toxic potential of RR 120. This work concludes that water pollution assessment can be done by these two biomarkers which provide baseline to the further chromosomal or molecular work.

Keywords: micronuclei, genotoxicity, RR 120, Catla catla

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31 Utilization Of Guar Gum As Functional Fat Replacer In Goshtaba, A Traditional Indian Meat Product

Authors: Sajad A. Rather, F. A. Masoodi, Rehana Akhter, S. M. Wani, Adil Gani

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Modern trend towards convenience foods has resulted in increased production and consumption of restructured meat products and are of great importance to the meat industry. In meat products fat plays an important role in cooking properties, texture & sensory scores, however, high fat contents in particular animal fats provide high amounts of saturated fatty acids and cholesterol and are associated with several types of non communicable diseases such as obesity, hypertension and coronary heart diseases. Thus, fat reduction has generally been seen as an important strategy to produce healthier meat products. This study examined the effects of reducing fat level from 20% to 10% and substituting mutton back fat with guar gum (0.5%, 1% & 1.5%) on cooking properties, proximate composition, lipid and protein oxidation, texture, microstructure and sensory characteristics of goshtaba- a traditional meat product of J & K, India were investigated and compared with high fat counterparts. Reduced- fat goshtaba samples containing guar gum had significantly (p ≤ 0.05) higher yield, less shrinkage, more moisture retention and more protein content than the control sample. TBARs and protein oxidation (carbonyl content) values of the control was significantly (p ≤ 0.05) higher than reduced fat goshtaba samples and showed a positive correlation between lipid and protein oxidation. Hardness, gumminess & chewiness of the control (20%) were significantly higher than reduced fat goshtaba samples. Microstructural differences were significant (p ≤ 0.05) between control and treated samples due to an increased moisture content in the reduced fat samples. Sensory evaluation showed significant (p ≤ 0.05) reduction in texture, flavour and overall acceptability scores of treatment products; however the scores for 0.5% and 1% treated samples were in the range of acceptability. Guar gum may also be used as a source of soluble dietary fibre in food products and a number of clinical studies have shown a reduction in postprandial glycemia and insulinemia on consumption of guar gum, with the mechanism being attributed to an increased transit time in the stomach and small intestine, which may have been due to the viscosity of the meal hindering the access of glucose to the epithelium.

Keywords: goshtaba, guar gum, traditional, fat reduction, acceptability

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30 A Comparative Study: Comparison of Two Different Fluorescent Stains -Auramine and Rhodamine- with Ehrlich-Ziehl-Neelsen, Kinyoun Staining, and Culture in the Determination of Acid Resistant Bacilli

Authors: Recep Keşli, Hayriye Tokay, Cengiz Demir, İsmail Ceyhan

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Objective: In many countries, tuberculosis (TB) is still one of the most important diseases. Tuberculosis is among top 10 causes of death worldwide. The early diagnosis of active tuberculosis still depends on the presence of acid resistant bacilli (ARB) in stained smears. In this study, we aimed to investigate the diagnostic performances of Erlich Ziehl Neelsen (EZN), Kinyoun and two different fluorescent stains. Methods: The specimens were obtained from the patients who applied to Chest Diseases Departments of Ankara Atatürk Chest Diseases and Thoracic Surgery Training and Research Hospital, and Afyon Kocatepe University, ANS Research and Practice Hospital. The study was carried out in the Medical Microbiology Laboratory, School of Medicine, Afyon Kocatepe University. All the non-sterile specimens were homogenized and decontaminated according to the EUCAST instructions. Samples were inoculated onto the Löwenstein-Jensen agars (bio-Merieux Marcy l'Etoile, France) and then incubated at 37˚C, for 40 days. Four smears were prepared from each specimen. Slides were stained with commercial EZN (BD, Sparks, USA), Kinyoun (SALUBRIS Istanbul, Turkey), Auramine (SALUBRIS Istanbul, Turkey) and Rhodamine (SALUBRIS Istanbul, Turkey) kit. While EZN and Kinyoun stainings were examined by light microscope, Auramine and Rhodamine slides were examined by fluorescence microscopy. Results: A total of 158 respiratory system samples (sputum, broncho alveolar lavage fluid…etc) were enrolled into the study. A hundred and two of the samples that processed were found as culture positive. The sensitivity, specificity, positive predictive, and negative predictive values were detected as 100%, 67.5%, 73.5%, and 100% for EZN, 100%, 70.9%, 77.4%, and 100% for Kinyoun, 100%,77.8%, 84.3%, 100% for Auramine, and 100%, 80% , 86.3%, and 100% for Rhodamine respectively. Conclusions: According to our study auramine and rhodamine staining methods showed the best diagnostic performance among the four investigated staining methods. In conclusion, the fluorochrome staining method may be accepted as the most reliable, rapid and useful method for diagnosis of the mycobacterial infections truly.

Keywords: acid resistant bacilli (ARB), auramine, Ehrlich-Ziehl-Neelsen (EZN), Kinyoun, Rhodamine

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29 Stomach Specific Delivery of Andrographolide from Floating in Situ Gelling System

Authors: Pravina Gurjar, Bothiraja Pour, Vijay Kumbhar, Ganesh Dama

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Andrographolide (AG), a bioactive phytoconstituent, has a wider range of pharmacological action. However, due to the intestinal degradation, shows low oral bioavailability. The aim of the present work was to develop Floating In-situ gelling Gastro retentive System (FISGS) for AG in order to enhance its site specific absorption and minimize pH dependent hydrolysis in alkaline environment. Further to increase its therapeutic efficacy for peptic ulcer disease caused by H. pyroli. Gellan based floating in situ gelling system of AG were prepared by using sodium citrate and calcium carbonate. The 32 factorial designs was used to study the effect of gellan and calcium carbonate concentration (independent variables) on dependent variable such as viscosity, floating lag time and drug release. Developed system was evaluated for drug content, floating lag time, viscosity, and drug release studies. Drug content, viscosity, and floating lag time was found to be 81-99%, 67-117 Cps, and 3-5 sec, respectively. The obtained system showed good in vitro floating ability for more than 12 h using 0.1 N HCl as dissolution medium with initial burst release followed by the controlled zero order drug release up to 24 hrs. In vivo testing of FISGS of AG to rats demonstrated significant antiulcer activity that were evaluated by various parameters like pH, volume, total acidity, millimole equivalent of H+ ions/30 min, and protein content of gastric content. The densities of all the formulation batches were found to be near about 0.9 and floating duration above 12 hr. It was observed that with the increase in conc. of gellan there was increase in the viscosity of formulation but all formulations were in optimum range. The drug content of optimized batch was found to be 99.23. In histopathology study of stomach, the villi at the mucosal surface, the intercellular junction, the intestinal lumen were intact; no destruction of the epithelium, and submucosal gland in formulation treated and control group animals as compared to pure drug AG and standard ranitidine. Gellan-based in situ gastro retentive floating system could be advantageous in terms of increased bioavailability of AG to maintain an effective drug conc. in gastric fluid as well as in serum for longer period of time.

Keywords: andrographolide, floating drug delivery, in situ gelling system, gastroretentive system

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28 Antigen Stasis can Predispose Primary Ciliary Dyskinesia (PCD) Patients to Asthma

Authors: Nadzeya Marozkina, Joe Zein, Benjamin Gaston

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Introduction: We have observed that many patients with Primary Ciliary Dyskinesia (PCD) benefit from asthma medications. In healthy airways, the ciliary function is normal. Antigens and irritants are rapidly cleared, and NO enters the gas phase normally to be exhaled. In the PCD airways, however, antigens, such as Dermatophagoides, are not as well cleared. This defect leads to oxidative stress, marked by increased DUOX1 expression and decreased superoxide dismutase [SOD] activity (manuscript under revision). H₂O₂, in high concentrations in the PCD airway, injures the airway. NO is oxidized rather than being exhaled, forming cytotoxic peroxynitrous acid. Thus, antigen stasis on PCD airway epithelium leads to airway injury and may predispose PCD patients to asthma. Indeed, recent population genetics suggest that PCD genes may be associated with asthma. We therefore hypothesized that PCD patients would be predisposed to having asthma. Methods. We analyzed our database of 18 million individual electronic medical records (EMRs) in the Indiana Network for Patient Care research database (INPCR). There is not an ICD10 code for PCD itself; code Q34.8 is most commonly used clinically. To validate analysis of this code, we queried patients who had an ICD10 code for both bronchiectasis and situs inversus totalis in INPCR. We also studied a validation cohort using the IBM Explorys® database (over 80 million individuals). Analyses were adjusted for age, sex and race using a 1 PCD: 3 controls matching method in INPCR and multivariable logistic regression in the IBM Explorys® database. Results. The prevalence of asthma ICD10 codes in subjects with a code Q34.8 was 67% vs 19% in controls (P < 0.0001) (Regenstrief Institute). Similarly, in IBM*Explorys, the OR [95% CI] for having asthma if a patient also had ICD10 code 34.8, relative to controls, was =4.04 [3.99; 4.09]. For situs inversus alone the OR [95% CI] was 4.42 [4.14; 4.71]; and bronchiectasis alone the OR [95% CI] =10.68 (10.56; 10.79). For both bronchiectasis and situs inversus together, the OR [95% CI] =28.80 (23.17; 35.81). Conclusions: PCD causes antigen stasis in the human airway (under review), likely predisposing to asthma in addition to oxidative and nitrosative stress and to airway injury. Here, we show that, by several different population-based metrics, and using two large databases, patients with PCD appear to have between a three- and 28-fold increased risk of having asthma. These data suggest that additional studies should be undertaken to understand the role of ciliary dysfunction in the pathogenesis and genetics of asthma. Decreased antigen clearance caused by ciliary dysfunction may be a risk factor for asthma development.

Keywords: antigen, PCD, asthma, nitric oxide

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27 Effect of Cigarette Smoke on Micro-Architecture of Respiratory Organs with and without Dietary Probiotics

Authors: Komal Khan, Hafsa Zaneb, Saima Masood, Muhammad Younus, Sanan Raza

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Cigarette smoke induces many physiological and pathological changes in respiratory tract like goblet cell hyperplasia and regional distention of airspaces. It is also associated with elevation of inflammatory profiles in different airway compartments. As probiotics are generally known to promote mucosal tolerance, it was postulated that prophylactic use of probiotics can be helpful in reduction of respiratory damage induced by cigarette smoke exposure. Twenty-four adult mice were randomly divided into three groups (cigarette-smoke (CS) group, cigarette-smoke+ Lactobacillus (CS+ P) group, control (Cn) group), each having 8 mice. They were exposed to cigarette smoke for 28 days (6 cigarettes/ day for 6 days/week). Wright-Giemsa staining of bronchoalveolar lavage fluid (BALF) was performed in three mice per group. Tissue samples of trachea and lungs of 7 mice from each group were processed by paraffin embedding technique for haematoxylin & eosin (H & E) and alcian blue- periodic acid-Schiff (AB-PAS) staining. Then trachea (goblet cell number, ratio and loss of cilia) and lungs (airspace distention) were studied. The results showed that the number of goblet cells was increased in CS group as a result of defensive mechanism of the respiratory system against irritating substances. This study also revealed that the cells of CS group having acidic glycoprotein were found to be higher in quantity as compared to those containing neutral glycoprotein. However, CS + P group showed a decrease in goblet cell index due to enhanced immunity by prophylactically used probiotics. Moreover, H & E stained tracheas showed significant loss of cilia in CS group due to propelling of mucous but little loss in CS + P group because of having good protective tracheal epithelium. In lungs, protection of airspaces was also much more evident in CS+ P group as compared to CS group having distended airspaces, especially at 150um distance from terminal bronchiole. In addition, a comprehensive analysis of inflammatory cells population of BALF showed neutrophilia and eosinophilia was significantly reduced in CS+ P group. This study proved that probiotics are found to be useful for reduction of changes in micro-architecture of the respiratory system. Thus, dietary supplementation of probiotic as prophylactic measure can be useful in achieving immunomodulatory effects.

Keywords: cigarette smoke, probiotics, goblet cells, airspace enlargement, BALF

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26 Dinoflagellate Thecal Plates as a Green Cellulose Source

Authors: Alvin Chun Man Kwok, Wai Sun Chan, Wei Yuan, Joseph Tin Yum Wong

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Cellulose, the most abundant biopolymer, is the major constituent of plant and dinoflagellate cell walls. Thecate dinoflagellates, in particular, are renowned for their remarkable capacity to synthesize intricate cellulosic thecal plates (CTPs). Unlike the extracellular two-dimensional structure of plant cell walls, these CTPs are three-dimensional and reside within the cellular structure itself. The deposition of CTPs occurs with remarkable precision, and their arrangement serves as crucial taxonomic markers. It is noteworthy that these plates possess the hardness of wood, despite the absence of lignin. Partial and prolonged hydrolysis of CTPs results in the formation of uniform long bundles and lowdimensional, modular crystalline whiskers. This observation aligns with the consistent nanomechanical properties, suggesting a CTPboard structure. The unique composition and structural characteristics of CTPs distinguish them from other cellulose-based materials in the natural world. Spectroscopic studies using Raman and FTIR methods indicate a clear low crystallinity index, with the OH shift becoming more distinct following SDS treatment. Birefringence imaging confirms the highly organized structure of CTPs, demonstrating varying degrees of anisotropy in different regions, including both seaward and cytosolic passages. The knockdown of a cellulose synthase enzyme in dinoflagellates resulted in severe malformation of CTPs and hindered the life-cycle transition. Unlike certain other microalgal groups, these unique circum-spherical depositions of CTPs were not pre-fabricated and transported "to site," but synthesized within alveolar sacs at the specific site. Our research is particularly focused on unraveling the mechanisms underlying the biodeposition of CTPs and exploring their potential biotechnological applications. Understanding the processes involved in CTP formation can pave the way for harnessing their unique properties for various practical applications. Dinoflagellates play a crucial role as major agents of algal blooms and are also known for producing anti-greenhouse sulfur compounds such as DMS/DMSP, highlighting the significance of CTPs as a carbon-neutral source of cellulose. Grant acknowledgement: Research in the laboratory are supported by GRF16104523 from Research Grant Council to JTYW.

Keywords: cellulosic thecal plates, dinoflagellates, cellulose, cell wall

Procedia PDF Downloads 44
25 Prospects of Regenerative Medicine with Human Allogeneic Adipose Tissue-Derived Mesenchymal Stem Cell Sheets: Achievements and Future Outlook in Clinical Trials for Myopic Chorioretinal Atrophy

Authors: Norimichi Nagano, Yoshio Hirano, Tsutomu Yasukawa

Abstract:

Mesenchymal stem cells are thought to confer neuroprotection, facilitate tissue regeneration and exert their effects on retinal degenerative diseases, however, adverse events such as proliferative vitreoretinopathy and preretinal membrane disease associated with cell suspension transplantation have also been reported. We have recently developed human (allogeneic) adipose tissue-derived mesenchymal stem cell (adMSC) sheets through our proprietary sheet transformation technique, which could potentially mitigate these adverse events. To clarify the properties of our adMSC sheets named PAL-222, we performed in vitro studies such as viability testing, cytokine secretions by ELISA, immunohistochemical study, and migration assay. The viability of the cells exceeded 70%. Vascular Endothelial Growth Factor (VEGF) and Pigment Epithelium-Derived Factor (PEDF), which are quite important cytokines for the retinal area, were observed. PAL-222 expressed type I collagen, a strength marker, type IV collagen, a marker of the basement membrane, and elastin, an elasticity marker. Finally, the migration assay was performed and showed negative, which means that PAL-222 is stably kept in the topical area and does not come to pieces. Next, to evaluate the efficacy in vivo, we transplanted PAL-222 into the subretinal space of the eye of Royal College of Surgeons rats with congenital retinal degeneration and assessed it for three weeks after transplantation. We confirmed that PAL-222 suppressed the decrease in the thickness of the outer nuclear layer, which means that the photoreceptor protective effect treated with PAL-222 was significantly higher than that in the sham group. (p < 0.01). This finding demonstrates that PAL-222 showed their retinoprotective effect in a model of congenital retinal degeneration. As the study suggested the efficacy of PAL-222 in both in vitro and in vivo studies, we are presently engaged in clinical trials of PAL-222 for myopic chorioretinal atrophy, which is one of the retinal degenerative diseases, for the purpose of regenerative medicine.

Keywords: cell sheet, clinical trial, mesenchymal stem cell, myopic chorioretinal atrophy

Procedia PDF Downloads 54
24 Morphological and Molecular Evaluation of Dengue Virus Serotype 3 Infection in BALB/c Mice Lungs

Authors: Gabriela C. Caldas, Fernanda C. Jacome, Arthur da C. Rasinhas, Ortrud M. Barth, Flavia B. dos Santos, Priscila C. G. Nunes, Yuli R. M. de Souza, Pedro Paulo de A. Manso, Marcelo P. Machado, Debora F. Barreto-Vieira

Abstract:

The establishment of animal models for studies of DENV infections has been challenging, since circulating epidemic viruses do not naturally infect nonhuman species. Such studies are of great relevance to the various areas of dengue research, including immunopathogenesis, drug development and vaccines. In this scenario, the main objective of this study is to verify possible morphological changes, as well as the presence of antigens and viral RNA in lung samples from BALB/c mice experimentally infected with an epidemic and non-neuroadapted DENV-3 strain. Male BALB/c mice, 2 months old, were inoculated with DENV-3 by intravenous route. After 72 hours of infection, the animals were euthanized and the lungs were collected. Part of the samples was processed by standard technique for analysis by light and transmission electronic microscopies and another part was processed for real-time PCR analysis. Morphological analyzes of lungs from uninfected mice showed preserved tissue areas. In mice infected with DENV-3, the analyzes revealed interalveolar septum thickening with presence of inflammatory infiltrate, foci of alveolar atelectasis and hyperventilation, bleeding foci in the interalveolar septum and bronchioles, peripheral capillary congestion, accumulation of fluid in the blood capillary, signs of interstitial cell necrosis presence of platelets and mononuclear inflammatory cells circulating in the capillaries and/or adhered to the endothelium. In addition, activation of endothelial cells, platelets, mononuclear inflammatory cell and neutrophil-type polymorphonuclear inflammatory cell evidenced by the emission of cytoplasmic membrane prolongation was observed. DEN-like particles were seen in the cytoplasm of endothelial cells. The viral genome was recovered from 3 in 12 lung samples. These results demonstrate that the BALB / c mouse represents a suitable model for the study of the histopathological changes induced by DENV infection in the lung, with tissue alterations similar to those observed in human cases of DEN.

Keywords: BALB/c mice, dengue, histopathology, lung, ultrastructure

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23 Steps of the Pancreatic Differentiation in the Grass Snake (Natrix natrix) Embryos

Authors: Magdalena Kowalska, Weronika Rupik

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The pancreas is an important organ present in all vertebrate species. It contains two different tissues, exocrine and endocrine, that act as two glands in one. The development and differentiation of the pancreas in reptiles is poorly known in comparison to other vertebrates. Therefore, the aim of this study was to investigate the particular steps concerning the differentiation of the pancreas in the grass snake (Natrix natrix) embryos. For this, histological methods (including hematoxylin and eosin, and Heidenhain's AZAN staining), transmission electron microscopy and three-dimensional (3D) reconstructions from serial paraffin sections were used. The results of this study indicated that the first step of pancreas development in Natrix was the connection of the two pancreatic buds: dorsal and ventral one. Then, duct walls in both buds started to be remodeled from the multilayered to single-layered epithelium. This remodeling started in the dorsal bud and was simultaneously with the differentiation of the duct lumens which occurred by the cavition. During this process, the cells that had no contact with the mesenchyme underwent cell death named anoikis. These findings indicated that the walls of ducts in the embryonic pancreas of the grass snake were initially formed by the abundant principal and single endocrine cells. Later the basal and goblet cells differentiated. Among the endocrine cells, as the first the B and A cells differentiated, then the D and PP cells. The next step of the pancreatic development was the withdrawing of the endocrine cells from the duct walls to form the pancreatic islets. The endocrine cells and islets were found only in the dorsal part of the pancreas in Natrix embryos what is different than in other vertebrate species. The islets were formed mainly by the A cells. Simultaneously, with the differentiation of the endocrine pancreas, the acinar tissue started to differentiate. The source of the acinar cells were pancreatic ducts similar as in other vertebrates. The acini formation began at the proximal part of the pancreas and went towards the caudal direction. Differentiating pancreatic ducts developed into the branched system that can be divided into extralobular, intralobular, and intercalated ducts, similarly as in other vertebrate species. However, the pattern of branching was different. In conclusions, particular steps of the pancreas differentiation in the grass snake were different than in other vertebrates. It can be supposed that these differences are related to the specific topography of the snake’s internal organs and their taxonomy position. All specimens used in the study were captured according to the Polish regulations concerning the protection of wild species. Permission was granted by the Local Ethics Commission in Katowice (41/2010; 87/2015) and the Regional Directorate for Environmental Protection in Katowice (WPN.6401.257.2015.DC).

Keywords: embryogenesis, organogenesis, pancreas, Squamata

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22 Effects of Bone Marrow Derived Mesenchymal Stem Cells (MSC) in Acute Respiratory Distress Syndrome (ARDS) Lung Remodeling

Authors: Diana Islam, Juan Fang, Vito Fanelli, Bing Han, Julie Khang, Jianfeng Wu, Arthur S. Slutsky, Haibo Zhang

Abstract:

Introduction: MSC delivery in preclinical models of ARDS has demonstrated significant improvements in lung function and recovery from acute injury. However, the role of MSC delivery in ARDS associated pulmonary fibrosis is not well understood. Some animal studies using bleomycin, asbestos, and silica-induced pulmonary fibrosis show that MSC delivery can suppress fibrosis. While other animal studies using radiation induced pulmonary fibrosis, liver, and kidney fibrosis models show that MSC delivery can contribute to fibrosis. Hypothesis: The beneficial and deleterious effects of MSC in ARDS are modulated by the lung microenvironment at the time of MSC delivery. Methods: To induce ARDS a two-hit mouse model of Hydrochloric acid (HCl) aspiration (day 0) and mechanical ventilation (MV) (day 2) was used. HCl and injurious MV generated fibrosis within 14-28 days. 0.5x106 mouse MSCs were delivered (via both intratracheal and intravenous routes) either in the active inflammatory phase (day 2) or during the remodeling phase (day 14) of ARDS (mouse fibroblasts or PBS used as a control). Lung injury accessed using inflammation score and elastance measurement. Pulmonary fibrosis was accessed using histological score, tissue collagen level, and collagen expression. In addition alveolar epithelial (E) and mesenchymal (M) marker expression profile was also measured. All measurements were taken at day 2, 14, and 28. Results: MSC delivery 2 days after HCl exacerbated lung injury and fibrosis compared to HCl alone, while the day 14 delivery showed protective effects. However in the absence of HCl, MSC significantly reduced the injurious MV-induced fibrosis. HCl injury suppressed E markers and up-regulated M markers. MSC delivery 2 days after HCl further amplified M marker expression, indicating their role in myofibroblast proliferation/activation. While with 14-day delivery E marker up-regulation was observed indicating their role in epithelial restoration. Conclusions: Early MSC delivery can be protective of injurious MV. Late MSC delivery during repair phase may also aid in recovery. However, early MSC delivery during the exudative inflammatory phase of HCl-induced ARDS can result in pro-fibrotic profiles. It is critical to understand the interaction between MSC and the lung microenvironment before MSC-based therapies are utilized for ARDS.

Keywords: acute respiratory distress syndrome (ARDS), mesenchymal stem cells (MSC), hydrochloric acid (HCl), mechanical ventilation (MV)

Procedia PDF Downloads 649
21 [Keynote Talk]: Monitoring of Ultrafine Particle Number and Size Distribution at One Urban Background Site in Leicester

Authors: Sarkawt M. Hama, Paul S. Monks, Rebecca L. Cordell

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Within the Joaquin project, ultrafine particles (UFP) are continuously measured at one urban background site in Leicester. The main aims are to examine the temporal and seasonal variations in UFP number concentration and size distribution in an urban environment, and to try to assess the added value of continuous UFP measurements. In addition, relations of UFP with more commonly monitored pollutants such as black carbon (BC), nitrogen oxides (NOX), particulate matter (PM2.5), and the lung deposited surface area(LDSA) were evaluated. The effects of meteorological conditions, particularly wind speed and direction, and also temperature on the observed distribution of ultrafine particles will be detailed. The study presents the results from an experimental investigation into the particle number concentration size distribution of UFP, BC, and NOX with measurements taken at the Automatic Urban and Rural Network (AURN) monitoring site in Leicester. The monitoring was performed as part of the EU project JOAQUIN (Joint Air Quality Initiative) supported by the INTERREG IVB NWE program. The total number concentrations (TNC) were measured by a water-based condensation particle counter (W-CPC) (TSI model 3783), the particle number concentrations (PNC) and size distributions were measured by an ultrafine particle monitor (UFP TSI model 3031), the BC by MAAP (Thermo-5012), the NOX by NO-NO2-NOx monitor (Thermos Scientific 42i), and a Nanoparticle Surface Area Monitor (NSAM, TSI 3550) was used to measure the LDSA (reported as μm2 cm−3) corresponding to the alveolar region of the lung between November 2013 and November 2015. The average concentrations of particle number concentrations were observed in summer with lower absolute values of PNC than in winter might be related mainly to particles directly emitted by traffic and to the more favorable conditions of atmospheric dispersion. Results showed a traffic-related diurnal variation of UFP, BC, NOX and LDSA with clear morning and evening rush hour peaks on weekdays, only an evening peak at the weekends. Correlation coefficients were calculated between UFP and other pollutants (BC and NOX). The highest correlation between them was found in winter months. Overall, the results support the notion that local traffic emissions were a major contributor of the atmospheric particles pollution and a clear seasonal pattern was found, with higher values during the cold season.

Keywords: size distribution, traffic emissions, UFP, urban area

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20 Adjustment of the Level of Vibrational Force on Targeted Teeth

Authors: Amin Akbari, Dongcai Wang, Huiru Li, Xiaoping Du, Jie Chen

Abstract:

The effect of vibrational force (VF) on accelerating orthodontic tooth movement depends on the level of delivered stimulation to the tooth in terms of peak load (PL), which requires contacts between the tooth and the VF device. A personalized device ensures the contacts, but the resulting PL distribution on the teeth is unknown. Furthermore, it is unclear whether the PL on particular teeth can be adjusted to the prescribed values. The objective of this study was to investigate the efficacy of apersonalized VF device in controlling the level of stimulation on two teeth, the mandibular canines and 2nd molars. A 3-D finite element (FE) model of human dentition, including teeth, PDL, and alveolar bone, was created from the cone beam computed tomography images of an anonymous subject. The VF was applied to the teeth through a VFdevice consisting of a mouthpiece with engraved tooth profile of the subject and a VF source that applied 0.3 N force with the frequency of 30 Hz. The dentition and mouthpiece were meshed using 10-node tetrahedral elements. Interface elements were created at the interfaces between the teeth and the mouthpiece. The upper and lower teeth bite on the mouthpiece to receive the vibration. The depth of engraved individual tooth profile could be adjusted, which was accomplished by adding a layer of material as an interference or removing a layer of material as a clearance to change the PL on the tooth. The interference increases the PL while the clearance decreases it. Fivemouthpiece design cases were simulated, which included a mouthpiece without interference/clearance; the mouthpieces with bilateral interferences on both mandibular canines and 2nd molars with magnitudes of 0.1, 0.15, and 0.2-mm, respectively; and mouthpiece with bilateral 0.3-mm clearances on the four teeth. Then, the force distributions on the entire dentition were compared corresponding to these adjustments. The PL distribution on the teeth is uneven when there is no interference or clearance. Among all teeth, the anterior segment receives the highest level of PL. Adding 0.1, 0.15, and 0.2-mm interferences to the canines and 2nd molars bilaterally leads to increase of the PL on the canines by 10, 62, and 73 percent and on the 2nd molar by 14, 55, and 87 percent, respectively. Adding clearances to the canines and 2nd molars by removing the contactsbetween these teeth and the mouthpiece results in zero PL on them. Moreover, introducing interference to mandibular canines and 2nd molarsredistributes the PL on the entireteeth. The share of the PL on the anterior teeth are reduced. The use of the personalized mouthpiece ensures contactsof the teeth to the mouthpiece so that all teeth can be stimulated. However, the PL distribution is uneven. Adding interference between a tooth and the mouthpiece increases the PL while introducing clearance decreases the PL. As a result, the PL is redistributed. This study confirms that the level of VF stimulation on the individual tooth can be adjusted to a prescribed value.

Keywords: finite element method, orthodontic treatment, stress analysis, tooth movement, vibrational force

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19 Mechanisms of Atiulcerogenic Activity of Costus speciosus Rhizome Extract in Ethanol-Induced Gastric Mucosal Injury in Rats

Authors: Somayeh Fani, Mahmood Ameen Abdulla

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Costus speciosus is an important Malaysian medicinal plant commonly used traditionally in the treatment of many aliments. The present investigation is designed to elucidate preventive effects of ethanolic extracts of C. speciosus rhizome against absolute ethanol-induced gastric mucosal injury in Sprague-Dawley rats. Five groups of rats were orally pre-treated with vehicle, carboxymethylcellulose (CMC) as normal control group (Group 1), ethanol as ulcer control group (Group 2), omeprazole 20 mg/kg (reference group) (Group 3), and 250 and 500 mg/kg of C. speciosus extract (experimental groups) (Group 4 and 5), respectively. An hour later, CMC was given orally to Group 1 rats and absolute ethanol was given orally to Group 2-5 rats to generate gastric mucosal injury. After an additional hour, the rats were sacrificed. Grossly, ulcer control group exhibited severe of gastric mucosal hemorrhagic injury and increased in ulcer area, whereas groups pre-treated with omeprazole or plant’s rhizomes exhibited the significant reduction of gastric mucosal injury. Significant increase in the pH and mucous of gastric content was observed in rats re-treated with C. speciosus rhizome. Histology, ulcer control rats, demonstrated remarkable disruption of gastric mucosa, increased in edema and inflammatory cells infiltration of submucosal layer compared to rats pre-treated with rhizomes extract. Periodic acid Schiff staining for glycoprotein, rats pre-fed with C. speciosus C. displayed remarkably intense uptake of magenta color by glandular gastric mucosa compared with ulcer control rats. Immunostaining of gastric epithelium, rats pre-treatment with rhizome extract provide evidence of up-regulation of HSP70 and down-regulation of Bax proteins compared to ulcer control animals. Gastric tissue homogenate, C. speciosus significantly increased the activity of superoxide dismutase (SOD), and catalase (CAT), increased the level of non-protein sulfhydryl (NP-SH) and decreased the level of lipid peroxidation after ethanol administration. Acute toxicity test did not show any signs of toxicity. The mechanisms implicated the gasrtoprotective property of C. speciosus depend upon the antisecretory activity, increased in gastric mucus glycoprotein, up-regulation of HSP70 protein and down-regulation of Bax proteins, reduction in the lipid peroxidation and increase in the level of NP-SH and antioxidant enzymes activity in gastic homogenate.

Keywords: antioxidant, Costus speciosus, gastric ulcer, histology, omeprazole

Procedia PDF Downloads 281
18 Association between TNF-α and Its Receptor TNFRSF1B Polymorphism with Pulmonary Tuberculosis in Tomsk, Russia Federation

Authors: K. A. Gladkova, N. P. Babushkina, E. Y. Bragina

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Purpose: Tuberculosis (TB), caused by Mycobacterium tuberculosis, is one of the major public health problems worldwide. It is clear that the immune response to M. tuberculosis infection is a relationship between inflammatory and anti-inflammatory responses in which Tumour Necrosis Factor-α (TNF-α) plays key roles as a pro-inflammatory cytokine. TNF-α involved in various cell immune responses via binding to its two types of membrane-bound receptors, TNFRSF1A and TNFRSF1B. Importantly, some variants of the TNFRSF1B gene have been considered as possible markers of host susceptibility to TB. However, the possible impact of such TNF-α and its receptor genes polymorphism on TB cases in Tomsk is missing. Thus, the purpose of our study was to investigate polymorphism of TNF-α (rs1800629) and its receptor TNFRSF1B (rs652625 and rs525891) genes in population of Tomsk and to evaluate their possible association with the development of pulmonary TB. Materials and Methods: The population distribution features of genes polymorphisms were investigated and made case-control study based on group of people from Tomsk. Human blood was collected during routine patients examination at Tomsk Regional TB Dispensary. Altogether, 234 TB-positive patients (80 women, 154 men, average age is 28 years old) and 205 health-controls (153 women, 52 men, average age is 47 years old) were investigated. DNA was extracted from blood plasma by phenol-chloroform method. Genotyping was carried out by a single-nucleotide-specific real-time PCR assay. Results: First, interpopulational comparison was carried out between healthy individuals from Tomsk and available data from the 1000 Genomes project. It was found that polymorphism rs1800629 region demonstrated that Tomsk population was significantly different from Japanese (P = 0.0007), but it was similar with the following Europeans subpopulations: Italians (P = 0.052), Finns (P = 0.124) and British (P = 0.910). Polymorphism rs525891 clear demonstrated that group from Tomsk was significantly different from population of South Africa (P = 0.019). However, rs652625 demonstrated significant differences from Asian population: Chinese (P = 0.03) and Japanese (P = 0.004). Next, we have compared healthy individuals versus patients with TB. It was detected that no association between rs1800629, rs652625 polymorphisms, and positive TB cases. Importantly, AT genotype of polymorphism rs525891 was significantly associated with resistance to TB (odds ratio (OR) = 0.61; 95% confidence interval (CI): 0.41-0.9; P < 0.05). Conclusion: To the best of our knowledge, the polymorphism of TNFRSF1B (rs525891) was associated with TB, while genotype AT is protective [OR = 0.61] in Tomsk population. In contrast, no significant correlation was detected between polymorphism TNF-α (rs1800629) and TNFRSF1B (rs652625) genes and alveolar TB cases among population of Tomsk. In conclusion, our data expands the molecular particularities associated with TB. The study was supported by the grant of the Russia for Basic Research #15-04-05852.

Keywords: polymorphism, tuberculosis, TNF-α, TNFRSF1B gene

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17 Biophysical Analysis of the Interaction of Polymeric Nanoparticles with Biomimetic Models of the Lung Surfactant

Authors: Weiam Daear, Patrick Lai, Elmar Prenner

Abstract:

The human body offers many avenues that could be used for drug delivery. The pulmonary route, which is delivered through the lungs, presents many advantages that have sparked interested in the field. These advantages include; 1) direct access to the lungs and the large surface area it provides, and 2) close proximity to the blood circulation. The air-blood barrier of the alveoli is about 500 nm thick. The air-blood barrier consist of a monolayer of lipids and few proteins called the lung surfactant and cells. This monolayer consists of ~90% lipids and ~10% proteins that are produced by the alveolar epithelial cells. The two major lipid classes constitutes of various saturation and chain length of phosphatidylcholine (PC) and phosphatidylglycerol (PG) representing 80% of total lipid component. The major role of the lung surfactant monolayer is to reduce surface tension experienced during breathing cycles in order to prevent lung collapse. In terms of the pulmonary drug delivery route, drugs pass through various parts of the respiratory system before reaching the alveoli. It is at this location that the lung surfactant functions as the air-blood barrier for drugs. As the field of nanomedicine advances, the use of nanoparticles (NPs) as drug delivery vehicles is becoming very important. This is due to the advantages NPs provide with their large surface area and potential specific targeting. Therefore, studying the interaction of NPs with lung surfactant and whether they affect its stability becomes very essential. The aim of this research is to develop a biomimetic model of the human lung surfactant followed by a biophysical analysis of the interaction of polymeric NPs. This biomimetic model will function as a fast initial mode of testing for whether NPs affect the stability of the human lung surfactant. The model developed thus far is an 8-component lipid system that contains major PC and PG lipids. Recently, a custom made 16:0/16:1 PC and PG lipids were added to the model system. In the human lung surfactant, these lipids constitute 16% of the total lipid component. According to the author’s knowledge, there is not much monolayer data on the biophysical analysis of the 16:0/16:1 lipids, therefore more analysis will be discussed here. Biophysical techniques such as the Langmuir Trough is used for stability measurements which monitors changes to a monolayer's surface pressure upon NP interaction. Furthermore, Brewster Angle Microscopy (BAM) employed to visualize changes to the lateral domain organization. Results show preferential interactions of NPs with different lipid groups that is also dependent on the monolayer fluidity. Furthermore, results show that the film stability upon compression is unaffected, but there are significant changes in the lateral domain organization of the lung surfactant upon NP addition. This research is significant in the field of pulmonary drug delivery. It is shown that NPs within a certain size range are safe for the pulmonary route, but little is known about the mode of interaction of those polymeric NPs. Moreover, this work will provide additional information about the nanotoxicology of NPs tested.

Keywords: Brewster angle microscopy, lipids, lung surfactant, nanoparticles

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16 Histological and Ultrastructural Study on the Effect

Authors: Olfat Mohamed Hussien Yousef

Abstract:

Tamoxifen (TM) is a synthetic non-steroidal antiestrogen. It is one of the most effective drugs for treatment of estrogen-dependent cancer by binding to estrogen receptors, suppressing of epithelial proliferation and as a chemotherapeutic agent. Recently, more attention has been paid to the protective effects of natural antioxidants against toxicities induced by anti-cancer drugs involving free radical-mediated oxidative stress and tissue injury. Vitamin C is a potent antioxidant that has the ability to scavenge factors causing free radical formation in animals receiving tamoxifen. The present study aims at pinpointing the TM-induced histopathological and ultrastructural changes in the kidneys and to assess the possible chemoprotective role of vitamin C against such TM-induced microscopic changes. Thirty adult male CD-1 mice, 25-30 g in weight and 3 months old, were divided into three groups. The first group served as control. The second group received the therapeutic dose of TM at daily oral dose of 40 mg/kg body weight for 28 days. The third group received the therapeutic dose of vitamin C at a daily dose of 500 mg/kg body weight simultaneously with the therapeutic dose of TM used in group two for 28 days. Animals were sacrificed and kidney samples were obtained and processed for histological and ultrastructural examination. Histological changes induced by TM included damage of the renal corpuscles including obliteration of the subcapsular space, congestion of the glomerular blood capillaries, segmental mesangial cell proliferation with matrix expansion, capsular adhesions with the glomerular tuft especially at the urinary pole of the corpuscles. Moreover, some proximal and distal tubules suffered various degrees of degeneration in some lining cells. Haemorrhage and inflammatory cell infiltration were also observed in the intertubular spaces. Ultrastructural observations revealed damage of the parietal epithelium of Bowman’s capsule, fusion and destruction of the foot processes of podocytes and great increase of mesangial cells and mesangial matrix. The cells of the proximal convoluted tubules displayed marked destruction of the microvilli constituting the brush borders and degeneration of the mitochondria; besides, abundant lysosomes, numerous vacuoles and pyknotic nuclei were observed. The distal convoluted tubules displayed marked distruction of both the basal infolding and the mitochondria in some areas. Histological and ultrastructural results revealed that treatment of male mice with TM simultaneously with vitamin C led to apparent repair of the injured renal tissue. This might suggest that vitamin C (an antioxidant agent) can minimize the toxic effects of TM (an antiestrogen).

Keywords: tamoxifen, vitamin c, mammalian kidney, histology, ultrastructure

Procedia PDF Downloads 351
15 Mycotoxin Bioavailability in Sparus Aurata Muscle After Human Digestion and Intestinal Transport (Caco-2/HT-29 Cells) Simulation

Authors: Cheila Pereira, Sara C. Cunha, Miguel A. Faria, José O. Fernandes

Abstract:

The increasing world population brings several concerns, one of which is food security and sustainability. To meet this challenge, aquaculture, the farming of aquatic animals and plants, including fish, mollusks, bivalves, and algae, has experienced sustained growth and development in recent years. Recent advances in this industry have focused on reducing its economic and environmental costs, for example, the substitution of protein sources in fish feed. Plant-based proteins are now a common approach, and while it is a greener alternative to animal-based proteins, there are some disadvantages, such as their putative content and intoxicants such as mycotoxins. These are naturally occurring plant contaminants, and their exposure in fish can cause health problems, stunted growth or even death, resulting in economic losses for the producers and health concerns for the consumers. Different works have demonstrated the presence of both AFB1 (aflatoxin B1) and ENNB1 (enniatin B1) in fish feed and their capacity to be absorbed and bioaccumulate in the fish organism after digestion, further reaching humans through fish ingestion. The aim of this work was to evaluate the bioaccessibility of both mycotoxins in samples of Sparus aurata muscle using a static digestion model based on the INFOGEST protocol. The samples were subjected to different cooking procedures – raw, grilled and fried – and different seasonings – none, thyme and ginger – in order to evaluate their potential reduction effect on mycotoxins bioaccessibility, followed by the evaluation of the intestinal transport of both compounds with an in vitro cell model composed of Caco-2/HT-29 co-culture monolayers, simulating the human intestinal epithelium. The bioaccessible fractions obtained in the digestion studies were used in the transport studies for a more realistic approach to bioavailability evaluation. Results demonstrated the effect of the use of different cooking procedures and seasoning on the toxin's bioavailability. Sparus aurata was chosen in this study for its large production in aquaculture and high consumption in Europe. Also, with the continued evolution of fish farming practices and more common usage of novel feed ingredients based on plants, there is a growing concern about less studied contaminants in aquaculture and their consequences for human health. In pair with greener advances in this industry, there is a convergence towards alternative research methods, such as in vitro applications. In the case of bioavailability studies, both in vitro digestion protocols and intestinal transport assessment are excellent alternatives to in vivo studies. These methods provide fast, reliable and comparable results without ethical restraints.

Keywords: AFB1, aquaculture, bioaccessibility, ENNB1, intestinal transport.

Procedia PDF Downloads 27
14 A Preliminary Randomized Controlled Trial of Pure L-Ascorbic Acid with Using a Needle-Free and Micro-Needle Mesotherapy in Treatment of Anti-Aging Procedure

Authors: M. Zasada, A. Markiewicz, A. Erkiert-Polguj, E. Budzisz

Abstract:

The epidermis is a keratinized stratified squamous epithelium covered by the hydro-lipid barrier. Therefore, active substances should be able to penetrate through this hydro-lipid coating. L-ascorbic acid is one of the vitamins which plays an important role in stimulation fibroblast to produce collagen type I and in hyperpigmentation lightening. Vitamin C is a water-soluble antioxidant, which protects skin from oxidation damage and rejuvenates photoaged skin. No-needle mesotherapy is a non-invasive rejuvenation technique depending on electric pulses, electroporation, and ultrasounds. These physicals factors result in deeper penetration of cosmetics. It is important to increase the penetration of L-ascorbic acid, thereby increasing the spectrum of its activity. The aim of the work was to assess the effectiveness of pure L-ascorbic acid activity in anti-aging therapy using a needle-free and micro-needling mesotherapy. The study was performed on a group of 35 healthy volunteers in accordance with the Declaration of Helsinki of 1964 and agreement of the Ethics Commissions no RNN/281/16/KE 2017. Women were randomized to mesotherapy or control group. Control group applied topically 2,5 ml serum containing 20% L-ascorbic acid with hydrate from strawberries, every 10 days for a period of 9 weeks. No-needle mesotherapy, on the left half of the face and micro-needling on the right with the same serum, was done in mesotherapy group. The pH of serum was 3.5-4, and the serum was prepared directly prior to the facial treatment. The skin parameters were measured at the beginning and before each treatment. The measurement of the forehead skin was done using Cutometer® (measurement of skin elasticity and firmness), Corneometer® (skin hydration measurement), Mexameter® (skin tone measurement). Also, the photographs were taken by Fotomedicus system. Additionally, the volunteers fulfilled the questionnaire. Serum was tested for microbiological purity and stability after the opening of the cosmetic. During the study, all of the volunteers were taken care of a dermatologist. The regular application of the serum has caused improvement of the skin parameters. Respectively, after 4 and 8 weeks improvement in hydration and elasticity has been seen (Corneometer®, Cutometer® results). Moreover, the number of hyper-pigmentated spots has decreased (Mexameter®). After 8 weeks the volunteers has claimed that the tested product has smoothing and moisturizing features. Subjective opinions indicted significant improvement of skin color and elasticity. The product containing the L-ascorbic acid used with intercellular penetration promoters demonstrates higher anti-aging efficiency than control. In vivo studies confirmed the effectiveness of serum and the impact of the active substance on skin firmness and elasticity, the degree of hydration and skin tone. Mesotherapy with pure L-ascorbic acid provides better diffusion of active substances through the skin.

Keywords: anti-aging, l-ascorbic acid, mesotherapy, promoters

Procedia PDF Downloads 243
13 Alternate Optical Coherence Tomography Technologies in Use for Corneal Diseases Diagnosis in Dogs and Cats

Authors: U. E. Mochalova, A. V. Demeneva, Shilkin A. G., J. Yu. Artiushina

Abstract:

Objective. In medical ophthalmology OCT has been actively used in the last decade. It is a modern non-invasive method of high-precision hardware examination, which gives a detailed cross-sectional image of eye tissues structure with a high level of resolution, which provides in vivo morphological information at the microscopic level about corneal tissue, structures of the anterior segment, retina and optic nerve. The purpose of this study was to explore the possibility of using the OCT technology in complex ophthalmological examination in dogs and cats, to characterize the revealed pathological structural changes in corneal tissue in cats and dogs with some of the most common corneal diseases. Procedures. Optical coherence tomography of the cornea was performed in 112 animals: 68 dogs and 44 cats. In total, 224 eyes were examined. Pathologies of the organ of vision included: dystrophy and degeneration of the cornea, endothelial corneal dystrophy, dry eye syndrome, chronic superficial vascular keratitis, pigmented keratitis, corneal erosion, ulcerative stromal keratitis, corneal sequestration, chronic glaucoma and also postoperative period after performed keratoplasty. When performing OCT, we used certified medical devices: "Huvitz HOCT-1/1F», «Optovue iVue 80» and "SOCT Copernicus Revo (60)". Results. The results of a clinical study on the use of optical coherence tomography (OCT)of the cornea in cats and dogs, performed by the authors of the article in the complex diagnosis of keratopathies of variousorigins: endothelial corneal dystrophy, pigmented keratitis, chronic keratoconjunctivitis, chronic herpetic keratitis, ulcerative keratitis, traumatic corneal damage, sequestration of the cornea of cats, chronic keratitis, complicating the course of glaucoma. The characteristics of the OCT scans are givencorneas of cats and dogs that do not have corneal pathologies. OCT scans of various corneal pathologies in dogs and cats with a description of the revealed pathological changes are presented. Of great clinical interest are the data obtained during OCT of the cornea of animals undergoing keratoplasty operations using various forms of grafts. Conclusions. OCT makes it possible to assess the thickness and pathological structural changes of the corneal surface epithelium, corneal stroma and descemet membrane. We can measure them, determine the exact localization, and record pathological changes. Clinical observation of the dynamics of the pathological process in the cornea using OCT makes it possible to evaluate the effectiveness of drug treatment. In case of negative dynamics of corneal disease, it is necessary to determine the indications for surgical treatment (to assess the thickness of the cornea, the localization of its thinning zones, to characterize the depth and area of pathological changes). According to the OCT of the cornea, it is possible to choose the optimal surgical treatment for the patient, the technique and depth of optically constructive surgery (penetrating or anterior lamellar keratoplasty).; determine the depth and diameter of the planned microsurgical trepanation of corneal tissue, which will ensure good adaptation of the edges of the donor material.

Keywords: optical coherence tomography, corneal sequestration, optical coherence tomography of the cornea, corneal transplantation, cat, dog

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12 Sublethal Effects of Industrial Effluents on Fish Fingerlings (Clarias gariepinus) from Ologe Lagoon Environs, Lagos, Nigeria

Authors: Akintade O. Adeboyejo, Edwin O. Clarke, Oluwatoyin Aderinola

Abstract:

The present study is on the sub-lethal toxicity of industrial effluents (IE) from the environment of Ologe Lagoon, Lagos, Nigeria on the African catfish fingerlings Clarias gariepinus. The fish were cultured in varying concentrations of industrial effluents: 0% (control), 5%, 15%, 25%, and 35%. Trials were carried out in triplicates for twelve (12) weeks. The culture system was a static renewable bioassay and was carried out in the fisheries laboratory of the Lagos State University, Ojo-Lagos. Weekly physico-chemical parameters: Temperature (0C), pH, Conductivity (ppm) and Dissolved Oxygen (DO in mg/l) were measured in each treatment tank. Length (cm) and weight (g) data were obtained weekly and used to calculate various growth parameters: mean weight gain (MWG), percentage weight gain (PWG), daily weight gain (DWG), specific growth rate (SGR) and survival. Haematological (Packed Cell Volume (PCV), Red blood cells (RBC), White Blood Cell (WBC), Neutrophil and Lymphocytes etc) and histological alterations were measured after 12 weeks. The physico-chemical parameters showed that the pH ranged from 7.82±0.25–8.07±0.02. DO range from 1.92±0.66-4.43±1.24 mg/l. The conductivity values increased with increase in concentration of I.E. While the temperature remained stable with mean value range between 26.08±2.14–26.38±2.28. The DO showed significant differences at P<0.05. There was progressive increase in length and weight of fish during the culture period. The fish placed in the control had highest increase in both weight and length while fish in 35% had the least. MWG ranged from 16.59–35.96, DWG is from 0.3–0.48, SGR varied from 1.0–1.86 and survival was 100%. Haematological results showed that C. gariepinus had PCV ranging from 13.0±1.7-27.7±0.6, RBC ranged from 4.7±0.6–9.1±0.1, and Neutrophil ranged from 26.7±4.6–61.0±1.0 amongst others. The highest values of these parameters were obtained in the control and lowest at 35%. While the reverse effects were observed for WBC and lymphocytes. This study has shown that effluents may affect the health status of the test organism and impair vital processes if exposure continues for a long period of time. The histological examination revealed several lesions as expressed by the gills and livers. The histopathology of the gills in the control tanks had normal tissues with no visible lesion, but at higher concentrations, there were: lifting of epithelium, swollen lamellae and gill arch infiltration, necrosis and gill arch destruction. While in the liver: control (0%) show normal liver cells, at higher toxic level, there were: vacoulation, destruction of the hepatic parenchyma, tissue becoming eosinophilic (i.e. tending towards Carcinogenicity) and severe disruption of the hepatic cord architecture. The study has shown that industrial effluents from the study area may affect fish health status and impair vital processes if exposure continues for a long period of time even at lower concentrations (Sublethal).

Keywords: sublethal toxicity, industrial effluents, clarias gariepinus, ologe lagoon

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11 Gastroprotective Effect of Copper Complex On Indomethacin-Induced Gastric Ulcer In Rats. Histological and Immunohistochemical Study

Authors: Heba M. Saad Eldien, Ola Abdel-Tawab Hussein, Ahmed Yassein Nassar

Abstract:

Background: Indomethacin is a non-steroidal anti inflammatory drug. Indomethacin induces an injury to gastrointestinal mucosa in experimental animals and humans and their use is associated with a significant risk of hemorrhage, erosions and perforation of both gastric and intestinal ulcers. The anti-inflammatory action of copper complexes is an important activity of their anti-ulcer effect achieved by their intermediary role as a transport form of copper that allow activation of the several copper-dependent enzymes. Therefore, several copper complexes were synthesized and investigated as promising alternative anti-ulcer therapy. Aim of the work: The purpose of this study was to evaluate a copper chelating complex consisting of egg albumin and copper as one of the copper peptides that can be used as anti-inflammatory agent and effective in ameliorates the hazards of the indomethacin on the histological structure of the fundus of the stomach that could be added to raise the efficacy of the currently used simple and cheap gastric anti-inflammatory drug mucogel. Material &methods: This study was carried out on 40 adult male albino rats,divided equally into 4 groups;Group I(control group) received distilled water,Group II(indomethacin treated group) received (25 mg/kg body weight, oral intubation) once, Group III (mucogel treated group)2 mL/rat once daily, oral incubation, Group IV(copper complex group) 1 mL /rat of 30 gm of copper albumin complex was mixed uniformly with mucogel to 100 mL. Treatment has been started six hour after Induction of Ulcers and continued till the 3rd day. The animals sacrificed and was processed for light, transmission electron microscopy(TEM) and immunostaining for inducible nitric oxide synthase(iNOS). Results: Fundic mucosa of group II, showed exfoliation of epithelial cells lining the gland, discontinuity of surface epithelial cells (ulcer formation), vacuolation and detachment of cells, eosinophilic infiltration and congestion of blood vessels in the lamina propria and submucosa. There was thickening and disarrangement of mucosa, weak positive reaction for PAS and marked increase in the collagen fibers lamina propria and the submucosa of the fundus. TEM revealed degeneration of cheif and parietal cells.Marked increase positive reactive of iNOS in all cells of the fundic gland. Group III showed reconstruction of gastric gland with cystic dilatation and vacuolation, moderate decrease of collagen fibers, reduced the intensity of iNOS while in Group IV healthy mucosa with normal surface lining epithelium and fundic glands, strong positive reaction for PAS, marked decrease of collagen fibers and positive reaction for iNOS. TEM revealed regeneration of cheif and parietal cells. Conclusion: Co treatment of copper-albumin complex seems to be useful for gastric ulcer treatment and ameliorates most of hazards of indomethacin.

Keywords: copper complex, gastric ulcer, indomethacin, rat

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