Search results for: osaka martis
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 13

Search results for: osaka martis

13 Maritime English Communication Training for Japanese VTS Operators in the Congested Area Including the Narrow Channel of Akashi Strait

Authors: Kenji Tanaka, Kazumi Sugita, Yuto Mizushima

Abstract:

This paper introduces a noteworthy form of English communication training for the officers and operators of the Osaka-Bay Marine Traffic Information Service (Osaka MARTIS) of the Japan Coast Guard working in the congested area at the Akashi Strait in Hyogo Prefecture, Japan. The authors of this paper, Marine Technical College’s (MTC) English language instructors, have been holding about forty lectures and exercises in basic and normal Maritime English (ME) for several groups of MARTIS personnel at Osaka MARTIS annually since they started the training in 2005. Trainees are expected to be qualified Maritime Third-Class Radio Operators who are responsible for providing safety information to a daily average of seven to eight hundred vessels that pass through the Akashi Strait, one of Japan’s narrowest channels. As of 2022, the instructors are conducting 55 remote lessons at MARTIS. One lesson is 90 minutes long. All 26 trainees are given oral and written assessments. The trainees need to pass the examination to become qualified operators every year, requiring them to train and maintain their linguistic levels even during the pandemic of Corona Virus Disease-19 (COVID-19). The vessel traffic information provided by Osaka MARTIS in Maritime English language is essential to the work involving the use of very high frequency (VHF) communication between MARTIS and vessels in the area. ME is the common language mainly used on board merchant, fishing, and recreational vessels, normally at sea. ME was edited and recommended by the International Maritime Organization in the 1970s, was revised in 2002, and has undergone continual revision. The vessel’s circumstances are much more serious at the strait than those at the open sea, so these vessels need ME to receive guidance from the center when passing through the narrow strait. The imminent and challenging situations at the strait necessitate that textbooks’ contents include the basics of the phrase book for seafarers as well as specific and additional navigational information, pronunciation exercises, notes on keywords and phrases, explanations about collocations, sample sentences, and explanations about the differences between synonyms especially those focusing on terminologies necessary for passing through the strait. Additionally, short Japanese-English translation quizzes about these topics, as well as prescribed readings about the maritime sector, are include in the textbook. All of these exercises have been trained in the remote education system since the outbreak of COVID-19. According to the guidelines of ME edited in 2009, the lowest level necessary for seafarers is B1 (lower individual users) of The Common European Framework of Reference for Languages: Learning, Teaching, Assessment (CEFR). Therefore, this vocational ME language training at Osaka MARTIS aims for its trainees to communicate at levels higher than B1. A noteworthy proof of improvement from this training is that most of the trainees have become qualified marine radio communication officers.

Keywords: akashi strait, B1 of CEFR, maritime english communication training, osaka martis

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12 Lung Parasites in Stone Martens (Martes foina L.) from Bulgaria

Authors: Vassilena Dakova, Mariana Panayotova-Pencheva

Abstract:

The present work focused on the study of pulmonary helminth-fauna of the stone marten in Bulgaria in terms of which the data are little. For the purpose, four stone martens were helminthologically necropsied according to the common technique. In addition, some of the injured lung parts were investigated after their boiling in lactic acid and subsequent compression. Four nematode species from different families of order Strongylida and Trichocephalida were found in the lungs. These were Crenosoma petrowi Morosov, 1939; Eucoleus aerophilus Creplin, 1839; Filaroides martis Werner, 1782 and Sobolevingylus petrowi Romanov, 1952. Some of the parasite structures with taxonomic importance were measured and described. According to our best knowledge, the species F. martis and S. petrowi are recorded for the first time as a part of the helminth-fauna of Southeast Europe and Bulgaria in particular.

Keywords: Bulgaria, Crenosoma petrowi, Eucoleus aerophilus, Filaroides martis, lung parasites, Sobolevingylus petrowi, stone martens

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11 Improvements of the Difficulty in Hospital Acceptance at the Scene by the Introduction of Smartphone Application for Emergency-Medical-Service System: A Population-Based Before-And-After Observation Study in Osaka City, Japan

Authors: Yusuke Katayama, Tetsuhisa Kitamura, Kosuke Kiyohara, Sumito Hayashida, Taku Iwami, Takashi Kawamura, Takeshi Shimazu

Abstract:

Background: Recently, the number of ambulance dispatches has been increasing in Japan and it is, therefore, difficult to accept emergency patients to hospitals smoothly and appropriately because of the limited hospital capacity. To facilitate the request for patient transport by ambulances and hospital acceptance, the emergency information system using information technology has been built up and introduced in various communities. However, its effectiveness has not been insufficiently revealed in Japan. In 2013, we developed a smartphone application system that enables the emergency-medical-service (EMS) personnel to share information about on-scene ambulance and hospital situation. The aim of this study was to assess the introduction effect of this application for EMS system in Osaka City, Japan. Methods: This study was a retrospective study with population-based ambulance records of Osaka Municipal Fire Department. This study period was six years from January 1, 2010 to December 31, 2015. In this study, we enrolled emergency patients that on-scene EMS personnel conducted the hospital selection for them. The main endpoint was difficulty in hospital acceptance at the scene. The definition of difficulty in hospital acceptance at the scene was to make >=5 phone calls by EMS personnel at the scene to each hospital until a decision to transport was determined. The definition of the smartphone application group was emergency patients transported in the period of 2013-2015 after the introduction of this application, and we assessed the introduction effect of smartphone application with multivariable logistic regression model. Results: A total of 600,526 emergency patients for whom EMS personnel selected hospitals were eligible for our analysis. There were 300,131 smartphone application group (50.0%) in 2010-2012 and 300,395 non-smartphone application group (50.0%) in 2013-2015. The proportion of the difficulty in hospital acceptance was 14.2% (42,585/300,131) in the smartphone application group and 10.9% (32,819/300,395) in the non-smartphone application group, and the difficulty in hospital acceptance significantly decreased by the introduction of the smartphone application (adjusted odds ration; 0.730, 95% confidence interval; 0.718-0.741, P<0.001). Conclusions: Sharing information between ambulance and hospital by introducing smartphone application at the scene was associated with decreasing the difficulty in hospital acceptance. Our findings may be considerable useful for developing emergency medical information system with using IT in other areas of the world.

Keywords: difficulty in hospital acceptance, emergency medical service, infomation technology, smartphone application

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10 Comparison of the Curvizigzag Incision with Transverse Stewart Incision in Women Undergoing Modified Radical Mastectomy for Carcinoma Breast

Authors: John Joseph S. Martis, Rohanchandra R. Gatty, Aaron Jose Fernandes, Rahul P. Nambiar

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Introduction: Surgery for breast cancer is either mastectomy or breast conservation surgery. The most commonly used incision for modified radical mastectomy is the transverse Stewart incision. But this incision may have the disadvantage of causing disparity between the closure lines of superior and inferior skin flaps in mastectomy and can cause overhanging of soft tissue below and behind the axilla. The curvizigzag incision, on principle, may help in this regard and can prevent scar migration beyond the anterior axillary line. This study aims to compare the two incisions in this regard. Methods: 100 patients with cancer of breast were included in the study after satisfying inclusion and exclusion criteria. They underwent surgery at Father Muller Medical College, Mangalore, India, between November 2019 to September 2021. The patients were divided into two groups. Group A patients were subjected to modified radical mastectomy with curvizigzag incision and group B patients with transverse Stewart incision. Results: Seroma on postoperative day1, day 2 was 0% in both the groups. Seroma on postoperative day 30 was present in 14% of patients in group B. 60% of patients in group B had sag of soft tissue below and behind the axilla, and none of the patients in group A had this problem. In 64% of the patients in group B, the incision crossed the anterior axillary fold, 64% of the patients in group B had tension in the incision site while approximation of the skin flaps. Conclusion: Curvizigzag incision is statistically better with lesser complications when compared to the transverse Stewart incision for modified radical mastectomy for carcinoma breast.

Keywords: breast cancer, curvizigzag incision, transverse Stewart incision, seroma, modified radical mastectomy

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9 The Concentration of Formaldehyde in Rainwater and Typhoon Rainwater at Sakai City, Japan

Authors: Chinh Nguyen Nhu Bao, Hien To Thi, Norimichi Takenaka

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Formaldehyde (HCHO) concentrations in rainwater including in tropical storms in Sakai City, Osaka, Japan have been measured continuously during rain event by developed chemiluminescence method. The level of formaldehyde was ranged from 15 µg/L to 500 µg/L. The high concentration of HCHO in rainwater is related to the wind direction from the south and west sides of Sakai City where manufactures related to chemicals, oil-refinery, and steel. The in-situ irradiated experiment on rainwater sample was conducted to prove the aqueous phase photo-production of HCHO and the degradation of HCHO. In the daytime, the aqueous phase photolysis is the source of HCHO in rainwater (4.52 ± 5.74 µg/L/h for UV light source in-situ condition, 2.84-8.96 µg/L/h under sunlight). However, in the night time, the degradation is the function of microorganism.

Keywords: chemiluminescence, formaldehyde, rainwater, typhoon

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8 Producing Outdoor Design Conditions based on the Dependency between Meteorological Elements: Copula Approach

Authors: Zhichao Jiao, Craig Farnham, Jihui Yuan, Kazuo Emura

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It is common to use the outdoor design weather data to select the air-conditioning capacity in the building design stage. The outdoor design weather data are usually comprised of multiple meteorological elements for a 24-hour period separately, but the dependency between the elements is not well considered, which may cause an overestimation of selecting air-conditioning capacity. Considering the dependency between the air temperature and global solar radiation, we used the copula approach to model the joint distributions of those two weather elements and suggest a new method of selecting more credible outdoor design conditions based on the specific simultaneous occurrence probability of air temperature and global solar radiation. In this paper, the 10-year period hourly weather data from 2001 to 2010 in Osaka, Japan, was used to analyze the dependency structure and joint distribution, the result shows that the Joe-Frank copula fit for almost all hourly data. According to calculating the simultaneous occurrence probability and the common exceeding probability of air temperature and global solar radiation, the results have shown that the maximum difference in design air temperature and global solar radiation of the day is about 2 degrees Celsius and 30W/m2, respectively.

Keywords: energy conservation, design weather database, HVAC, copula approach

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7 Proposal of Analytical Model for the Seismic Performance Evaluation of Reinforced Concrete Frames with Coupled Cross-laminated Timber Infill Panels

Authors: Velázquez Alejandro, Pradhan Sujan, Yoon Rokhyun, Sanada Yasushi

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The utilization of new materials as an alternative solution to decrease the environmental impact of the construction industry has been gaining more relevance in the architectural design and construction industry. One such material is cross-laminated timber (CLT), an engineered timber solution that excels for its faster construction times, workability, lightweight, and capacity for carbon storage. This material is usually used alone for the entire structure or combined with steel frames, but a hybrid with reinforced concrete (RC) is rarer. Since RC is one of the most used materials worldwide, a hybrid with CLT would allow further utilization of the latter, and in the process, it would help reduce the environmental impact of RC construction to achieve a sustainable society, but first, the structural performance of such hybrids must be understood. This paper focuses on proposing a model to predict the seismic performance of RC frames with CLT panels as infills. A series of static horizontal cyclic loading experiments were conducted on two 40% scale specimens of reinforced concrete frames with and without CLT panels at Osaka University, Japan. An analytical model was created to simulate the seismic performance of the RC frame with CLT infill based on the experimental results. The proposed model was verified by comparing the experimental and analytical results, showing that the load-deformation relationship and the failure mechanism agreed well with limited error. Hence, the proposed analytical model can be implemented for the seismic performance evaluation of the RC frames with CLT infill.

Keywords: analytical model, multi spring, performance evaluation, reinforced concrete, rocking mechanism, wooden wall

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6 The Effect of Ultrasound as Pre-Treatment for Drying of Red Delicious and Golden Delicious Apples

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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5 The Influence of Apple Pomace on Colour and Chemical Composition of Extruded Corn Snack Product

Authors: Jovana Petrovic, Biljana Pajin, Ivana Loncarevic, Aleksandar Fistes, Antun Jozinivic, Durdica Ackar, Drago Subaric

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Recovery of food wastes and their conversion to economically viable products will play a vital role for the management strategies in the years to come. Apple pomace may be considered as wastes, but they contain considerable amounts of high value reusable materials. Apple pomace, the by-product of apple juice and cider production, is a good source of fibre, particularly insoluble one. The remaining apple pulp contains 12% dry residue, which is half dietary fibre. Another remarkable aspect is its richness in polyphenols, components with antioxidant activity. Apple pomace could be an interesting alternative source for fibre and polyphenols in extruded corn meals. The extruded corn meals with the addition of finely ground apple pomace were prepared (the ratio of corn meal: apple pomace was 85:15 and 70:30). Characterization of the extrudates in terms of determining the chemical composition and colour was performed. The color of samples was measured by MINOLTA Chroma Meter CR-400 (Minolta Co., Ltd., Osaka, Japan) using D 65 lighting, a 2º standard observer angle and an 8-mm aperture in the measuring head. The following CIELab color coordinates were determined: L* – lightness, a* – redness to greenness and b* – yellowness to blueness. Protein content decreased significantly from 7.91% to 5.19% with increase in pomace from 0% to 30%, while total fibre content increase from 3.39% to 16.62%. The apple pomace addition produced extrudates with a significantly lower L* value and significantly higher a* value. This study has been fully supported by the Provincial Secretariat for High Education and Scientific Research of the Government of Autonomous Province of Vojvodina, Republic of Serbia, project 142-451-2483/2017 and the Ministry of Science and Technological Development of the Republic of Serbia (Project no. 31014).

Keywords: apple pomace, chemical composition, colour, extruded corn snack products, food waste recovery

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4 Stigmatization of Individuals Who Receive Mental Health Treatment and the Role of Social Media: A Cross-Generational Cohort Design and Extension

Authors: Denise Ben-Porath, Tracy Masterson

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In the past, individuals who struggled with and sought treatment for mental health difficulties were stigmatized. However, the current generation holds more open attitudes around mental health issues. Indeed, public figures such as Demi Lovato, Naomi Osaka, and Simone Biles have taken to social media to break the silence around mental health, discussing their own struggles and the benefits of treatment. Thus, there is considerable reason to believe that this generation would hold fewer stigmatizing attitudes toward mental health difficulties and treatment compared to previous ones. In this study, we explored possible changes in stigma on mental health diagnosis and treatment seeking behavior between two generations: Gen Z, the current generation, and Gen X, those born between 1965-1980. It was hypothesized that Gen Z would hold less stigmatizing views on mental illness than Gen X. To examine possible changes in stigma attitudes between these two generations, we conducted a cross-generational cohort design by using the same methodology employed 20 years ago from the Ben-Porath (2002) study. Thus, participants were randomly assigned to read one of the following four case vignettes employed in the Ben-Porath (2002) study: (a) “Tom” who has received psychotherapy due to depression (b) “Tom” who has been depressed but received no psychological help, (c) “Tom” who has received medical treatment due to a back pain, or (d) “Tom” who had a back pain but did not receive medical attention. After reading the vignette, participants rated “Tom” on various personality dimensions using the IFQ Questionnaire and answered questions about their frequency of social media use and willingness to seek mental health treatment on a scale from 1-10. Identical to the results 20 years prior, a significant main effect was found for diagnosis with “Tom” being viewed in more negative terms when he was described as having depression vs. a medical condition (back pain) [F (1, 376) = 126.53, p < .001]. However, in the study conducted 20 years earlier, a significant interaction was found between diagnosis and help-seeking behavior [F (1, 376) = 8.28, p < .005]. Specifically, “Tom” was viewed in the most negative terms when described as depressed and seeking treatment. Alternatively, the current study failed to find a significant interaction between depression and help seeking behavior. These findings suggest that while individuals who hold a mental health diagnosis may still be stigmatized as they were 20 years prior, seeking treatment for mental health issues may be less so. Findings are discussed in the context of social media use and its impact on destigmatization.

Keywords: stigma, mental illness, help-seeking, social media

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3 The Impact of Encapsulated Raspberry Juice on the Surface Colour of Enriched White Chocolate

Authors: Ivana Loncarevic, Biljana Pajin, Jovana Petrovic, Aleksandar Fistes, Vesna Tumbas Saponjac, Danica Zaric

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Chocolate is a complex rheological system usually defined as a suspension consisting of non-fat particles dispersed in cocoa butter as a continuous fat phase. Dark chocolate possesses polyphenols as major constituents whose dietary consumption has been associated with beneficial effects. Milk chocolate is formulated with a lower percentage of cocoa bean liquor than dark chocolate and it often contains lower amounts of polyphenols, while in white chocolate the fat-free cocoa solids are left out completely. Following the current trend of development of functional foods, there is an idea to create enriched white chocolate with the addition of encapsulated bioactive compounds from berry fruits. The aim of this study was to examine the surface colour of enriched white chocolate with the addition of 6, 8, and 10% of raspberry juice encapsulated in maltodextrins, in order to preserve the stability, bioactivity, and bioavailability of the active ingredients. The surface color of samples was measured by MINOLTA Chroma Meter CR-400 (Minolta Co., Ltd., Osaka, Japan) using D 65 lighting, a 2º standard observer angle and an 8-mm aperture in the measuring head. The following CIELab color coordinates were determined: L* – lightness, a* – redness to greenness and b* – yellowness to blueness. The addition of raspberry encapsulates led to the creation of new type of enriched chocolate. Raspberry encapsulate changed the values of the lightness (L*), a* (red tone) and b* (yellow tone) measured on the surface of enriched chocolate in accordance with applied concentrations. White chocolate has significantly (p < 0.05) highest L* (74.6) and b* (20.31) values of all samples indicating the bright surface of the white chocolate, as well as a high share of a yellow tone. At the same time, white chocolate has the negative a* value (-1.00) on its surface which includes green tones. Raspberry juice encapsulate has the darkest surface with significantly (p < 0.05) lowest value of L* (42.75), where increasing of its concentration in enriched chocolates decreases their L* values. Chocolate with 6% of encapsulate has significantly (p < 0.05) highest value of L* (60.56) in relation to enriched chocolate with 8% of encapsulate (53.57), and 10% of encapsulate (51.01). a* value measured on the surface of white chocolate is negative (-1.00) tending towards green tones. Raspberry juice encapsulates increases red tone in enriched chocolates in accordance with the added amounts (23.22, 30.85, and 33.32 in enriched chocolates with 6, 8, and 10% encapsulated raspberry juice, respectively). The presence of yellow tones in enriched chocolates significantly (p < 0.05) decreases with the addition of E (with b* value 5.21), from 10.01 in enriched chocolate with a minimal amount of raspberry juice encapsulates to 8.91 in chocolate with a maximum concentration of raspberry juice encapsulate. The addition of encapsulated raspberry juice to white chocolate led to the creation of new type of enriched chocolate with attractive color. The research in this paper was conducted within the project titled ‘Development of innovative chocolate products fortified with bioactive compounds’ (Innovation Fund Project ID 50051).

Keywords: color, encapsulated raspberry juice, polyphenols, white chocolate

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2 Hegemonic Salaryman Masculinity: Case Study of Transitional Male Gender Roles in Today's Japan

Authors: D. Norton

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This qualitative study focuses on the lived experience and displacement of young white-collar masculinities in Japan. In recent years, the salaryman lifestyle has undergone significant disruption - increased competition for regular employment, rise in non-regular structurings of labour across public/private sectors, and shifting role expectations within the home. Despite this, related scholarship hints at a continued reinforcement of the traditional male gender role - that the salaryman remains a key benchmark of Japanese masculine identity. For those in structural proximity to these more ‘normative’ performativities, interest lies their engagement with such narratives - how they make sense of their masculinity in response to stated changes. In light of the historical emphasis on labour and breadwinning logics, notions of respective security or precarity generated as a result remain unclear. Similarly, concern extends to developments within the private sphere - by what means young white-collar men construct ideas of singlehood and companionship according to traditional gender ideologies or more contemporary, flexible readings. The influence of these still-emergent status distinctions on the logics of the social group in question is yet to be explored in depth by gender scholars. This project, therefore, focuses on a salaryman archetype as hegemonic - its transformation amidst these changes and socialising mechanisms that continue to legitimate unequal gender hierarchies. For data collection, a series of ethnographic interviews were held over a period of 12 months with university-educated, white-collar male employees from both Osaka and the Greater Tokyo Area. Findings suggest a modern salaryman ideal reflecting both continuities and shifts within white-collar employment. Whilst receptive to more contemporary workplace practices, the narratives of those interviewed remain imbued with logics supporting patterns of internal hegemony. Regular/non-regular distinction emerged as the foremost variable for both material and discursive patterns of white-collar stratification, with variants of displacement for each social group. Despite the heightened valorisation of stable employment, regular workers articulated various concerns over a model of corporate masculinity seen to be incompatible with recent socioeconomic developments. Likewise, non-regular employees face detachment owing to a still-inflexible perception of their working masculinity as marginalized amidst economic precarity. In seeking to negotiate respective challenges, those interviewed demonstrated an engagement with various concurrent social changes that would often either accommodate, reinforce, or expand upon traditional role behaviours. Few of these narratives offered any notable transgression of said ideal, however, suggesting that within the spectre of white-collar employment in Japan for the near future, any substantive transformation of corporate masculinity remains dependant upon economic developments, less so the agency of those involved.

Keywords: gender ideologies, hegemonic masculinity, Japan, white-collar employment

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1 Oncolytic Efficacy of Thymidine Kinase-Deleted Vaccinia Virus Strain Tiantan (oncoVV-TT) in Glioma

Authors: Seyedeh Nasim Mirbahari, Taha Azad, Mehdi Totonchi

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Oncolytic viruses, which only replicate in tumor cells, are being extensively studied for their use in cancer therapy. A particular virus known as the vaccinia virus, a member of the poxvirus family, has demonstrated oncolytic abilities glioma. Treating Glioma with traditional methods such as chemotherapy and radiotherapy is quite challenging. Even though oncolytic viruses have shown immense potential in cancer treatment, their effectiveness in glioblastoma treatment is still low. Therefore, there is a need to improve and optimize immunotherapies for better results. In this study, we have designed oncoVV-TT, which can more effectively target tumor cells while minimizing replication in normal cells by replacing the thymidine kinase gene with a luc-p2a-GFP gene expression cassette. Human glioblastoma cell line U251 MG, rat glioblastoma cell line C6, and non-tumor cell line HFF were plated at 105 cells in a 12-well plates in 2 mL of DMEM-F2 medium with 10% FBS added to each well. Then incubated at 37°C. After 16 hours, the cells were treated with oncoVV-TT at an MOI of 0.01, 0.1 and left in the incubator for a further 24, 48, 72 and 96 hours. Viral replication assay, fluorescence imaging and viability tests, including trypan blue and crystal violet, were conducted to evaluate the cytotoxic effect of oncoVV-TT. The finding shows that oncoVV-TT had significantly higher cytotoxic activity and proliferation rates in tumor cells in a dose and time-dependent manner, with the strongest effect observed in U251 MG. To conclude, oncoVV-TT has the potential to be a promising oncolytic virus for cancer treatment, with a more cytotoxic effect in human glioblastoma cells versus rat glioma cells. To assess the effectiveness of vaccinia virus-mediated viral therapy, we have tested U251mg and C6 tumor cell lines taken from human and rat gliomas, respectively. The study evaluated oncoVV-TT's ability to replicate and lyse cells and analyzed the survival rates of the tested cell lines when treated with different doses of oncoVV-TT. Additionally, we compared the sensitivity of human and mouse glioma cell lines to the oncolytic vaccinia virus. All experiments regarding viruses were conducted under biosafety level 2. We engineered a Vaccinia-based oncolytic virus called oncoVV-TT to replicate specifically in tumor cells. To propagate the oncoVV-TT virus, HeLa cells (5 × 104/well) were plated in 24-well plates and incubated overnight to attach to the bottom of the wells. Subsequently, 10 MOI virus was added. After 48 h, cells were harvested by scraping, and viruses were collected by 3 sequential freezing and thawing cycles followed by removal of cell debris by centrifugation (1500 rpm, 5 min). The supernatant was stored at −80 ◦C for the following experiments. To measure the replication of the virus in Hela, cells (5 × 104/well) were plated in 24-well plates and incubated overnight to attach to the bottom of the wells. Subsequently, 5 MOI virus or equal dilution of PBS was added. At the treatment time of 0 h, 24 h, 48 h, 72 h and 96 h, the viral titers were determined under the fluorescence microscope (BZ-X700; Keyence, Osaka, Japan). Fluorescence intensity was quantified using the imagej software according to the manufacturer’s protocol. For the isolation of single-virus clones, HeLa cells seeded in six-well plates (5×105 cells/well). After 24 h (100% confluent), the cells were infected with a 10-fold dilution series of TianTan green fluorescent protein (GFP)virus and incubated for 4 h. To examine the cytotoxic effect of oncoVV-TT virus ofn U251mg and C6 cell, trypan blue and crystal violet assay was used.

Keywords: oncolytic virus, immune therapy, glioma, vaccinia virus

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