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1.
Antimicrob Resist Infect Control ; 13(1): 71, 2024 Jul 04.
Article in English | MEDLINE | ID: mdl-38965593

ABSTRACT

The use of antimicrobials in Thailand has been reported as one of the highest in the world in human and animal sectors. Our engagement project aimed to improve our understanding of the issue of antimicrobial use and antimicrobial resistance (AMR) among adult Thai communities, and co-create locally relevant solutions to AMR, especially those focusing on raising awareness to improve related policies in Thailand.We conducted a series of online and in-person 'conversations' according to Wellcome's 'Responsive Dialogues' engagement approach, designed to bring together different voices to understand complex AMR problems and find potential solutions. This approach enabled key AMR stakeholders and policy makers to hear directly from communities and members of the public, and vice versa. Conversations events took place between 25 November 2020 and 8 July 2022, and we engaged 179 AMR key stakeholders and members of the public across Thailand.The issues found were: there were quite a lot of misunderstandings around antimicrobials and AMR; participants felt that communications and engagement around antimicrobial resistance had limited reach and impact; asking for and taking antibiotics for self-limiting ailments is a social norm in Thailand; and there appeared to be a wide availability of cheap antimicrobials. To mitigate the spread of AMR, participants suggested that the messages around AMR should be tailored to the target audience, there should be more initiatives to increase general health literacy, there should be increased availability of AMR related information at the local level and there should be increased local leadership of AMR mitigation efforts.Trial registration Thaiclinicaltrials.org registration: TCTR20210528003 (28/05/2021).


Subject(s)
Anti-Bacterial Agents , Drug Resistance, Bacterial , Thailand , Humans , Anti-Bacterial Agents/therapeutic use , Community Participation , Health Knowledge, Attitudes, Practice , Antimicrobial Stewardship
2.
BMJ Glob Health ; 8(12)2023 12 11.
Article in English | MEDLINE | ID: mdl-38081771

ABSTRACT

INTRODUCTION: Causes of deaths often go unrecorded in lower income countries, yet this information is critical. Verbal autopsy is a questionnaire interview with a family member or caregiver to elicit the symptoms and circumstances preceding a death and assign a probable cause. The social and cultural aspects of verbal autopsy have gotten less attention than the technical aspects and have not been widely explored in South and Southeast Asia settings. METHODS: Between October 2021 and March 2023, prior to implementing a verbal autopsy study at rural sites in Bangladesh, Cambodia, Laos, Myanmar and Thailand, focus group discussions were conducted with village heads, religious leaders and community members from varied demographic backgrounds. Thematic analysis elucidated customs and traditional views surrounding death to understand local ethnocultural sensitivities. RESULTS: We found that death rituals varied greatly among religions, ethnicities and by socioeconomic status. Mourning periods were reported to last 3-100 days and related to the cause of death, age and how close the deceased person was to the family. Participants advised that interviews should happen after mourning periods to avoid emotional distress, but not long after so as to avoid recall bias. Interviewers should be introduced to respondents by a trusted local person. To provide reassurance and confidentiality, a family's residence is the preferred interview location. Interview questions require careful local language translation, and community sensitisation is important before data collection. CONCLUSION: Verbal autopsy is acceptable across a wide range of cultural settings in Southeast Asia, provided that local norms are preidentified and followed.


Subject(s)
Family , Humans , Cause of Death , Autopsy , Surveys and Questionnaires , Asia, Southeastern
3.
BMC Public Health ; 23(1): 1085, 2023 06 06.
Article in English | MEDLINE | ID: mdl-37280573

ABSTRACT

BACKGROUND: A nationwide Movement Control Order (MCO) was enforced in Malaysia on 18 March 2020 in view of the global COVID-19 pandemic. Malaysia implemented various public health measures and later raced against time to administer COVID-19 vaccines when they became available. As a result of various public health measures to curb the spread of the virus, people in Malaysia faced unprecedented circumstances and new challenges. This study addressed the knowledge gap in our understanding the experiences, coping strategies and perspectives of the people in Malaysia about infection countermeasures by investigating their experiences during the COVID-19 pandemic. METHODS: A sequential mixed method approach was used to conduct an online survey and in-depth interviews among residents in Malaysia. A total of 827 respondents participated in the online survey from 1st May to 30th June 2020. Nineteen in-depth interviews were conducted online and by phone with key informants and members of the public, who were selected through maximum variation purposive sampling between 2nd May 2020 to 20th December 2021. The semi-structured interviews employed a phenomenological approach and transcripts were analysed using thematic analysis. The survey data were analysed using descriptive statistics in Stata 15.0. RESULTS: The survey reflected significant economic impacts of the pandemic, the maximum number of days that people could cope during the MCO, and their coping strategies, which generally entailed changes in lifestyle. The internet and social media were vital platforms to mitigate against the impact of public health measures. Thematic analysis of the interview data revealed participant experiences and perceptions of COVID-19 and public health measures in four main themes: (1) work and business; (2) emotional impact (3) coping with change and (4) the COVID-19 vaccine. CONCLUSIONS: This study provides insights into the experiences, coping strategies and perspectives of people in Malaysia living through the first-ever MCO during the COVID-19 pandemic. Such insights into COVID-19-related public health measures are pertinent for successfully planning and implementing future responses to pandemics.


Subject(s)
COVID-19 , Humans , COVID-19/epidemiology , COVID-19/prevention & control , COVID-19 Vaccines , Pandemics/prevention & control , Malaysia/epidemiology , Adaptation, Psychological
4.
J Antimicrob Chemother ; 78(6): 1317-1321, 2023 06 01.
Article in English | MEDLINE | ID: mdl-37071582

ABSTRACT

Non-academic partners can be vital in successful public engagement activities on antimicrobial resistance. With collaboration between academic and non-academic partners, we developed and launched an open-access web-based application, the 'antibiotic footprint calculator', in both Thai and English. The application focused on a good user experience, addressing antibiotic overuse and its impact, and encouraging immediate action. The application was unveiled in joint public engagement activities. From 1 Nov 2021 to 31 July 2022 (9 month period), 2554 players estimated their personal antibiotic footprint by using the application.


Subject(s)
Anti-Bacterial Agents , Drug Resistance, Bacterial , Anti-Bacterial Agents/pharmacology , Anti-Bacterial Agents/therapeutic use , Thailand , Software
5.
JMIR Res Protoc ; 12: e44875, 2023 Mar 06.
Article in English | MEDLINE | ID: mdl-36877564

ABSTRACT

BACKGROUND: The increasing emphasis to share patient data from clinical research has resulted in substantial investments in data repositories and infrastructure. However, it is unclear how shared data are used and whether anticipated benefits are being realized. OBJECTIVE: The purpose of our study is to examine the current utilization of shared clinical research data sets and assess the effects on both scientific research and public health outcomes. Additionally, the study seeks to identify the factors that hinder or facilitate the ethical and efficient use of existing data based on the perspectives of data users. METHODS: The study will utilize a mixed methods design, incorporating a cross-sectional survey and in-depth interviews. The survey will involve at least 400 clinical researchers, while the in-depth interviews will include 20 to 40 participants who have utilized data from repositories or institutional data access committees. The survey will target a global sample, while the in-depth interviews will focus on individuals who have used data collected from low- and middle-income countries. Quantitative data will be summarized by using descriptive statistics, while multivariable analyses will be used to assess the relationships between variables. Qualitative data will be analyzed through thematic analysis, and the findings will be reported in accordance with the COREQ (Consolidated Criteria for Reporting Qualitative Research) guidelines. The study received ethical approval from the Oxford Tropical Research Ethics Committee in 2020 (reference number: 568-20). RESULTS: The results of the analysis, including both quantitative data and qualitative data, will be available in 2023. CONCLUSIONS: The outcomes of our study will offer crucial understanding into the current status of data reuse in clinical research, serving as a basis for guiding future endeavors to enhance the utilization of shared data for the betterment of public health outcomes and for scientific progress. TRIAL REGISTRATION: Thai Clinical Trials Registry TCTR20210301006; https://tinyurl.com/2p9atzhr. INTERNATIONAL REGISTERED REPORT IDENTIFIER (IRRID): DERR1-10.2196/44875.

6.
PLoS One ; 17(1): e0262421, 2022.
Article in English | MEDLINE | ID: mdl-35061789

ABSTRACT

This qualitative study explores the impact of non-pharmaceutical interventions (NPIs), including social distancing, travel restrictions and quarantine, on lived experiences during the first wave of the COVID-19 pandemic in Thailand (TH), Malaysia (MY), Italy (IT) and the United Kingdom (UK). A total of 86 interviews (TH: n = 28; MY: n = 18; IT: n = 20; UK: n = 20) were conducted with members of the public, including healthcare workers (n = 13). Participants across countries held strong views on government imposed NPIs, with many feeling measures lacked clarity. Most participants reported primarily negative impacts of NPIs on their lives, including through separation, isolation and grief over missed milestones; work-related challenges and income loss; and poor mental health and wellbeing. Nonetheless, many also experienced inadvertent positive consequences, including more time at home to focus on what they most valued in life; a greater sense of connectedness; and benefits to working life. Commonly employed coping strategies focused on financial coping (e.g. reducing spending); psycho-emotional coping (e.g. engaging in spiritual practices); social coping and connectedness (e.g., maintaining relationships remotely); reducing and mitigating risks (e.g., changing food shopping routines); and limiting exposure to the news (e.g., checking news only occasionally). Importantly, the extent to which participants' lived experiences were positive or negative, and their ability to cope was underpinned by individual, social and economic factors, with the analysis indicating some salient differences across countries and participants. In order to mitigate negative and unequal impacts of NPIs, COVID-19 policies will benefit from paying closer attention to the social, cultural and psychological-not just biological-vulnerabilities to, and consequences of public health measures.


Subject(s)
Adaptation, Psychological , Attitude , COVID-19 , Public Health , Adolescent , Adult , Aged , Aged, 80 and over , Communicable Disease Control , Female , Health Surveys , Humans , Italy , Malaysia , Male , Middle Aged , Thailand , United Kingdom , Young Adult
7.
PLOS Glob Public Health ; 2(7): e0000723, 2022.
Article in English | MEDLINE | ID: mdl-36962432

ABSTRACT

Following the first Thai COVID-19 case in January 2020, the Thai government introduced several non-pharmaceutical interventions (NPIs) in March 2020 (e.g., contact tracing, travel restrictions, closure of businesses, curfews, stay at home orders) to control COVID-19 transmissions. This study aimed to understand the views and experiences of a small number of Thai residents related to public health measures implemented during the first COVID-19 wave in Thailand. A total of 28 remote in-depth interviews with Thai residents (18-74 years old) were conducted between 8 May and 21 July 2020. Interviews were audio recorded, transcribed, and analysed using thematic analysis based on the Framework Method. Our results describe participants' views, challenges, and coping strategies relating to COVID-19 restrictions. Most participants expressed support for the introduction of strict public health measures, while some criticized lacking enforcement or rational of certain measures. Participants identified four major challenges, namely financial hardship; social isolation and loneliness; stigma and shaming; and fear of COVID-19 infection. Strategies adopted to address these challenges included practical coping strategies (e.g., reducing risks and fear of COVID-19 infection; mitigating financial, social, and mental health impacts), and embedded socio-cultural ways of coping (e.g., turning to religion; practicing acceptance; kindness, generosity and sharing ('Namjai'); 'making merit' ('Tham-bun')). The challenges identified from this study, in particular the role of stigma and discrimination, may be relevant to other infectious disease outbreaks beyond COVID-19. Findings from this study underscore the need for policies and interventions that mitigate the negative impacts of NPIs on the public, particularly on vulnerable groups, and highlight the importance of considering socio-cultural context to support community resilience in times of crisis. Our findings remain relevant in light of low COVID-19 vaccine availability and the potential need to implement further public health restrictions in Thailand and elsewhere against COVID-19 or future infectious disease threats.

8.
BMJ Open ; 11(7): e046863, 2021 07 20.
Article in English | MEDLINE | ID: mdl-34285007

ABSTRACT

OBJECTIVES: To understand the impact of COVID-19 and public health measures on different social groups, we conducted a mixed-methods study in five countries ('SEBCOV-social, ethical and behavioural aspects of COVID-19'). Here, we report the results of the online survey. STUDY DESIGN AND STATISTICAL ANALYSIS: Overall, 5058 respondents from Thailand, Malaysia, the UK, Italy and Slovenia completed the self-administered survey between May and June 2020. Poststratification weighting was applied, and associations between categorical variables assessed. Frequency counts and percentages were used to summarise categorical data. Associations between categorical variables were assessed using Pearson's χ2 test. Data were analysed in Stata 15.0 RESULTS: Among the five countries, Thai respondents reported having been most, and Slovenian respondents least, affected economically. The following factors were associated with greater negative economic impacts: being 18-24 years or 65 years or older; lower education levels; larger households; having children under 18 in the household and and having flexible/no income. Regarding social impact, respondents expressed most concern about their social life, physical health, mental health and well-being.There were large differences between countries in terms of voluntary behavioural change, and in compliance and agreement with COVID-19 restrictions. Overall, self-reported compliance was higher among respondents who self-reported a high understanding of COVID-19. UK respondents felt able to cope the longest and Thai respondents the shortest with only going out for essential needs or work. Many respondents reported seeing news perceived to be fake, the proportion varying between countries, with education level and self-reported levels of understanding of COVID-19. CONCLUSIONS: Our data showed that COVID-19 and public health measures have uneven economic and social impacts on people from different countries and social groups. Understanding the factors associated with these impacts can help to inform future public health interventions and mitigate their negative consequences. TRIAL REGISTRATION NUMBER: TCTR20200401002.


Subject(s)
COVID-19 , Social Change , Child , Cross-Sectional Studies , Humans , Italy , Malaysia , SARS-CoV-2 , Slovenia , Surveys and Questionnaires , Thailand , United Kingdom
9.
Wellcome Open Res ; 6: 188, 2021.
Article in English | MEDLINE | ID: mdl-35505979

ABSTRACT

Background: The use of antimicrobials in Thailand has been reported as one of the highest in the world in both the human and animal sectors. The objectives of this project are: (1) to improve understanding of the issue of antimicrobial resistance (AMR) among adult Thai communities and (2) to drive change through the national AMR policy to include context-specific and locally-driven solutions. Methods: The project contains two components conducted in parallel: the "AMR Dialogues" public engagement project and the embedded evaluation of the project. We will bring together AMR stakeholders and members of the public through a series of conversation events to co-create an AMR stakeholder map, engagement strategy, and context-specific solutions to reduce the burden of AMR. There will be a combination of regional in-person events ('regional conversations') and national online events ('national conversations') with members of the public. The conversations will follow this sequence: introduce and explore issues related to AMR, brainstorm solutions and finally propose promising/feasible solutions to take forward. Evaluation of the project will be conducted to assess if the AMR Dialogues objectives have been achieved using feedback forms and qualitative methods. Ethics: Approval of the evaluation component of the project has been obtained from the ethics committee of the Thailand Institute for the Development of Human Subject Protection (IHRP2021059) and the Oxford University Tropical Research Ethics Committee (OxTREC 529-21). Dissemination: The results of these conversation events will inform the next Thailand National Strategic Plan on AMR. The learning and outcomes will be disseminated to AMR policy makers, academic audiences, and participants of all the conversation events. Thaiclinicaltrials.org registration: TCTR20210528003 (28/05/2021).

10.
Wellcome Open Res ; 5: 90, 2020.
Article in English | MEDLINE | ID: mdl-32704548

ABSTRACT

Introduction: Vaccines and drugs for the treatment and prevention of COVID-19 require robust evidence generated from clinical trials before they can be used. Decisions on how to apply non-pharmaceutical interventions such as quarantine, self-isolation, social distancing and travel restrictions should also be based on evidence. There are some experiential and mathematical modelling data for these interventions, but there is a lack of data on the social, ethical and behavioural aspects of these interventions in the literature. Therefore, our study aims to produce evidence to inform (non-pharmaceutical) interventions such as communications, quarantine, self-isolation, social distancing, travel restrictions and other public health measures for the COVID-19 pandemic. Methods: The study will be conducted in the United Kingdom, Italy, Malaysia and Thailand. We propose to conduct 600-1000 quantitative surveys and 25-35 qualitative interviews per country. Data collection will follow the following four themes: (1) Quarantine and self-isolation (2) social distancing and travel restrictions (3) wellbeing and mental health (4) information, misinformation and rumours. In light of limitations of travel and holding in-person meetings, we will use online/remote methods for collecting data. Study participant will be adults who have provided informed consent from different demographic, socio-economic and risk groups. Discussion: At the time of writing, United Kingdom, Italy, Malaysia and Thailand have initiated strict public health measures and varying degrees of "lockdowns" to curb the pandemic. It is anticipated that these public health measures will continue in some countries (e.g. Italy, Malaysia) or be tightened further in other countries (e.g. Thailand, UK) to control the spread of the disease in the coming weeks and months. The data generated from our study could inform these strategies in real time.

11.
Wellcome Open Res ; 5: 245, 2020.
Article in English | MEDLINE | ID: mdl-34345714

ABSTRACT

Background: Any government needs to react quickly to a pandemic and make decisions on healthcare interventions locally and internationally with little information regarding the perceptions of people and the reactions they may receive during the implementation of restrictions. Methods: We report an anonymous online survey in Thailand conducted in May 2020 to assess public perceptions of three interventions in the Thai context: isolation, quarantine and social distancing. A total of 1,020 participants, of whom 52% were women, responded to the survey. Results: Loss of income was the main concern among respondents (>80% for all provinces in Thailand). Traditional media and social media were important channels for communication during the pandemic. A total of 92% of respondents reported that they changed their social behaviour even before the implementation of government policy with 94% reporting they performed social distancing, 97% reported using personal protective equipment such as masks and 95% reported using sanitizer products. Conclusions: This study showed a high level of compliance from individuals with government enforced or voluntarily controls such as quarantine, isolation and social distancing in Thailand. The findings from this study can be used to inform future government measures to control the pandemic and to shape communication strategies.

12.
J Exp Bot ; 68(13): 3339-3350, 2017 06 15.
Article in English | MEDLINE | ID: mdl-28605454

ABSTRACT

The plant Golgi apparatus modifies and sorts incoming proteins from the endoplasmic reticulum (ER) and synthesizes cell wall matrix material. Plant cells possess numerous motile Golgi bodies, which are connected to the ER by yet to be identified tethering factors. Previous studies indicated a role for cis-Golgi plant golgins, which are long coiled-coil domain proteins anchored to Golgi membranes, in Golgi biogenesis. Here we show a tethering role for the golgin AtCASP at the ER-Golgi interface. Using live-cell imaging, Golgi body dynamics were compared in Arabidopsis thaliana leaf epidermal cells expressing fluorescently tagged AtCASP, a truncated AtCASP-ΔCC lacking the coiled-coil domains, and the Golgi marker STtmd. Golgi body speed and displacement were significantly reduced in AtCASP-ΔCC lines. Using a dual-colour optical trapping system and a TIRF-tweezer system, individual Golgi bodies were captured in planta. Golgi bodies in AtCASP-ΔCC lines were easier to trap and the ER-Golgi connection was more easily disrupted. Occasionally, the ER tubule followed a trapped Golgi body with a gap, indicating the presence of other tethering factors. Our work confirms that the intimate ER-Golgi association can be disrupted or weakened by expression of truncated AtCASP-ΔCC and suggests that this connection is most likely maintained by a golgin-mediated tethering complex.


Subject(s)
Arabidopsis Proteins/genetics , Arabidopsis/genetics , Endoplasmic Reticulum/metabolism , Golgi Apparatus/metabolism , Membrane Proteins/genetics , Arabidopsis/metabolism , Arabidopsis Proteins/metabolism , Golgi Matrix Proteins , Membrane Proteins/metabolism , Plant Leaves/metabolism
13.
Redox Biol ; 12: 806-813, 2017 08.
Article in English | MEDLINE | ID: mdl-28441629

ABSTRACT

Adipose tissue is a complex and multi-faceted organ. It responds dynamically to internal and external stimuli, depending on the developmental stage and activity of the organism. The most common functional subunits of adipose tissue, white and brown adipocytes, regulate and respond to endocrine processes, which then determine metabolic rate as well as adipose tissue functions. While the molecular aspects of white and brown adipose biology have become clearer in the recent past, much less is known about sex-specific differences in regulation and deposition of adipose tissue, and the specific role of the so-called pink adipocytes during lactation in females. This review summarises the current understanding of adipose tissue dynamics with a focus on sex-specific differences in adipose tissue energy metabolism and endocrine functions, focussing on mammalian model organisms as well as human-derived data. In females, pink adipocytes trans-differentiate during pregnancy from subcutaneous white adipocytes and are responsible for milk-secretion in mammary glands. Overlooking biological sex variation may ultimately hamper clinical treatments of many aspects of metabolic disorders.


Subject(s)
Adipose Tissue/metabolism , Energy Metabolism , Metabolic Diseases/metabolism , Adipose Tissue/cytology , Animals , Cell Differentiation , Female , Humans , Male , Obesity , Sex Characteristics
14.
Methods Cell Biol ; 118: 69-83, 2013.
Article in English | MEDLINE | ID: mdl-24295301

ABSTRACT

Leaf epidermal cells make ideal specimens for the investigation of the plant secretory pathway in that it is relatively easy to tag with fluorescent proteins and visualize in vivo the various organelles of the pathway. A number of techniques can be employed to identify and study proteins within the endomembrane organelles and to study their dynamics and interactions. Here, we discuss the most commonly used approaches to express proteins within arabidopsis and tobacco leaves, the use of mutant screens to identify trafficking proteins, and the use of two in vivo techniques, Fluorescence recovery after photobleaching and Förster resonance energy transfer, to study protein dynamics in plant cells.


Subject(s)
Arabidopsis/metabolism , Golgi Apparatus/metabolism , Intracellular Membranes/metabolism , Nicotiana/metabolism , Plant Leaves/metabolism , Arabidopsis/cytology , Arabidopsis/genetics , Arabidopsis Proteins/genetics , Arabidopsis Proteins/metabolism , Endoplasmic Reticulum/metabolism , Fluorescence Resonance Energy Transfer , Green Fluorescent Proteins/biosynthesis , Green Fluorescent Proteins/genetics , Microscopy, Confocal , Microscopy, Fluorescence , Plant Leaves/cytology , Protein Transport , Secretory Pathway , Nicotiana/cytology , Nicotiana/genetics , Transformation, Genetic
15.
Plant Methods ; 9(1): 26, 2013 Jul 11.
Article in English | MEDLINE | ID: mdl-23845168

ABSTRACT

Social media now complements many parts of our lives. Facebook, Twitter, YouTube and many other social networking sites allow users to share and interact with online content and to connect with like-minded people. Its strengths - rapid dissemination and amplification of content and the ability to lead informal conversations - make it a powerful tool to use in a professional context. This commentary explains the overall concept of social media and offers suggestions on usage and possible types of scientific content. It advises researchers on the potential benefits and how to take a strategic approach towards building a social media presence. It also presents examples of effective social media use within the plant science community. Common reasons for scientists to not engage with social media include the fear of appearing unprofessional, posting something wrong or being misunderstood, or a lack of confidence in their computer skills. With the rapid changes in academic publishing, dissemination and science communication, as well as the rise of 'altmetrics' to track online engagement with scientific content, digital literacy will become an essential skill in a scientist's tool kit.

17.
Plant J ; 66(4): 613-28, 2011 May.
Article in English | MEDLINE | ID: mdl-21294794

ABSTRACT

We have identified two endoplasmic reticulum (ER)-associated Arabidopsis proteins, KMS1 and KMS2, which are conserved among most species. Fluorescent protein fusions of KMS1 localised to the ER in plant cells, and over-expression induced the formation of a membrane structure, identified as ER whorls by electron microscopy. Hydrophobicity analysis suggested that KMS1 and KMS2 are integral membrane proteins bearing six transmembrane domains. Membrane protein topology was assessed by a redox-based topology assay (ReTA) with redox-sensitive GFP and confirmed by a protease protection assay. A major loop domain between transmembrane domains 2 and 3, plus the N- and C-termini were found on the cytosolic side of the ER. A C-terminal di(tri)-lysine motif is involved in retrieval of KMS1 and deletion led to a reduction of the GFP-KMS1 signal in the ER. Over-expression of KMS1/KMS2 truncations perturbed ER and Golgi morphology and similar effects were also seen when KMS1/KMS2 were knocked-down by RNA interference. Microscopy and biochemical experiments suggested that expression of KMS1/KMS2 truncations inhibited ER to Golgi protein transport.


Subject(s)
Arabidopsis Proteins/metabolism , Arabidopsis/metabolism , Endoplasmic Reticulum/ultrastructure , Nicotiana/genetics , SNARE Proteins/metabolism , Secretory Pathway , Amino Acid Sequence , Arabidopsis/genetics , Arabidopsis Proteins/genetics , Cloning, Molecular , Endoplasmic Reticulum/metabolism , Gene Knockdown Techniques , Green Fluorescent Proteins/metabolism , Hydrophobic and Hydrophilic Interactions , Molecular Sequence Data , Protein Transport , RNA Interference , Recombinant Fusion Proteins/metabolism , Sequence Alignment , Sequence Analysis, Protein , Nicotiana/metabolism , Nicotiana/ultrastructure
18.
Curr Opin Plant Biol ; 13(6): 731-5, 2010 Dec.
Article in English | MEDLINE | ID: mdl-21093352

ABSTRACT

Laser trapping of micron-sized particles can be achieved utilizing the radiation pressure generated by a focused infrared laser beam. Thus, it is theoretically possible to trap and manipulate organelles within the cytoplasm and remodel the architecture of the cytoplasm and membrane systems. Here we describe recent progress, using this under utilized technology, in the manipulation of cytoplasmic strands and organelles in plant cells.


Subject(s)
Cytoplasm , Micromanipulation/methods , Optical Tweezers , Organelles , Plant Cells
19.
Traffic ; 11(11): 1429-44, 2010 Nov.
Article in English | MEDLINE | ID: mdl-20716110

ABSTRACT

Herein, we report the stepwise transport of multiple plant Golgi membrane markers during disassembly of the Golgi apparatus in tobacco leaf epidermal cells in response to the induced expression of the GTP-locked Sar1p or Brefeldin A (BFA), and reassembly on BFA washout. The distribution of fluorescent Golgi-resident N-glycan processing enzymes and matrix proteins (golgins) with specific cis-trans-Golgi sub-locations was followed by confocal microscopy during disassembly and reassembly. The first event during Golgi disassembly was the loss of trans-Golgi enzymes and golgins from Golgi membranes, followed by a sequential redistribution of medial and cis-Golgi enzymes into the endoplasmic reticulum (ER), whilst golgins were relocated to the ER or cytoplasm. This event was confirmed by fractionation and immuno-blotting. The sequential redistribution of Golgi components in a trans-cis sequence may highlight a novel retrograde trafficking pathway between the trans-Golgi and the ER in plants. Release of Golgi markers from the ER upon BFA washout occurred in the opposite sequence, with cis-matrix proteins labelling Golgi-like structures before cis/medial enzymes. Trans-enzyme location was preceded by trans-matrix proteins being recruited back to Golgi membranes. Our results show that Golgi disassembly and reassembly occur in a highly ordered fashion in plants.


Subject(s)
Biomarkers/metabolism , Golgi Apparatus/physiology , Nicotiana/metabolism , Brefeldin A/pharmacology , Epidermal Cells , Golgi Apparatus/drug effects , Microscopy, Confocal , Organelle Biogenesis , Plant Leaves/metabolism , Protein Synthesis Inhibitors/pharmacology
20.
Biochem Soc Trans ; 38(3): 761-7, 2010 Jun.
Article in English | MEDLINE | ID: mdl-20491662

ABSTRACT

It has long been assumed that the individual cisternal stacks that comprise the plant Golgi apparatus multiply by some kind of fission process. However, more recently, it has been demonstrated that the Golgi apparatus can be experimentally disassembled and the reformation process from the ER (endoplasmic reticulum) monitored sequentially using confocal fluorescence and electron microscopy. Some other evidence suggests that Golgi stacks may arise de novo in cells. In the present paper, we review some of the more recent findings on plant Golgi stack biogenesis and propose a new model for their growth de novo from ER exit sites.


Subject(s)
Endoplasmic Reticulum/metabolism , Golgi Apparatus/metabolism , Golgi Apparatus/ultrastructure , Plants/ultrastructure , Animals , Endoplasmic Reticulum/ultrastructure , Microscopy, Confocal , Microscopy, Electron , Models, Biological , Plant Proteins/metabolism , Plants/metabolism
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