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Article ; Online: Imaging Membrane Proteins Using Total Internal Reflection Fluorescence Microscopy (TIRFM) in Mammalian Cells.

Gada, Kirin D / Kamuene, Jordie M / Kawano, Takeharu / Plant, Leigh D

2023 Volume 13, Issue 4, Page(s) e4614

Abstract: ... for micromirror TIRFM analysis of optogenetically activated protein kinase C-ε in HEK293-T cells, as well ...

Abstract	The cell surfaceome is of vital importance across physiology, developmental biology, and disease states alike. The precise identification of proteins and their regulatory mechanisms at the cell membrane has been challenging and is typically determined using confocal microscopy, two-photon microscopy, or total internal reflection fluorescence microscopy (TIRFM). Of these, TIRFM is the most precise, as it harnesses the generation of a spatially delimited evanescent wave at the interface of two surfaces with distinct refractive indices. The limited penetration of the evanescent wave illuminates a narrow specimen field, which facilitates the localization of fluorescently tagged proteins at the cell membrane but not inside of the cell. In addition to constraining the depth of the image, TIRFM also significantly enhances the signal-to-noise ratio, which is particularly valuable in the study of live cells. Here, we detail a protocol for micromirror TIRFM analysis of optogenetically activated protein kinase C-ε in HEK293-T cells, as well as data analysis to demonstrate the translocation of this construct to the cell-surface following optogenetic activation. Graphic abstract.
Language	English
Publishing date	2023-02-20
Publishing country	United States
Document type	Journal Article
ZDB-ID	2833269-6
ISSN	2331-8325 ; 2331-8325
ISSN (online)	2331-8325
ISSN	2331-8325
DOI	10.21769/BioProtoc.4614
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Article ; Online: Optogenetic dephosphorylation of phosphatidylinositol 4,5 bisphosphate in Xenopus laevis oocytes.

Gada, Kirin D / Xu, Yu / Winn, Brenda T / Masotti, Meghan / Kawano, Takeharu / Vaananen, Heikki / Plant, Leigh D

STAR protocols

2023 Volume 4, Issue 1, Page(s) 102003

Abstract: Here, we present a protocol for optogenetic dephosphorylation of the phosphoinositide PI(4,5) ... ...

Abstract	Here, we present a protocol for optogenetic dephosphorylation of the phosphoinositide PI(4,5)P
MeSH term(s)	Animals ; Phosphatidylinositols/metabolism ; Phosphatidylinositol 4,5-Diphosphate/metabolism ; Xenopus laevis/metabolism ; Optogenetics ; Oocytes/metabolism ; Potassium Channels/metabolism
Chemical Substances	Phosphatidylinositols ; Phosphatidylinositol 4,5-Diphosphate ; Potassium Channels
Language	English
Publishing date	2023-01-11
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Extramural
ISSN	2666-1667
ISSN (online)	2666-1667
DOI	10.1016/j.xpro.2022.102003
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Article: Evaluating urban methane emissions from space using TROPOMI methane and carbon monoxide observations

Plant, Genevieve / Kort, Eric A. / Murray, Lee T. / Maasakkers, Joannes D. / Aben, Ilse

Remote sensing of environment. 2022 Jan., v. 268

2022

Abstract: Anthropogenic methane emissions from urban centers are important and addressable, yet remain poorly characterized, and the representativeness of studies from individual cities is unknown. A satellite-based approach provides a pathway to tackle this ... ...

Abstract	Anthropogenic methane emissions from urban centers are important and addressable, yet remain poorly characterized, and the representativeness of studies from individual cities is unknown. A satellite-based approach provides a pathway to tackle this challenge on a national or global scale. Here we present a space-based method that uses the simultaneous, daily observation of methane and carbon monoxide from the TROPOMI satellite to estimate urban methane emissions. We assess and validate the method and demonstrate that using these simultaneous observations enables robust assessment of methane emissions from urban centers without relying on atmospheric transport models. Initial assessments with this approach in eight United States cities suggest emission inventory underestimates previously discovered in older East Coast cities are more broadly representative, with aggregated emissions totaling 1.47 (0.56, 3.19, 95% confidence interval) Tg CH₄/year, compared to the Environmental Protection Agency estimate of 0.52 Tg CH₄/year. We show this data driven approach provides a pathway to study urban methane emissions across the globe and track how emissions respond if urban mitigation measures are implemented by investigating three additional megacities outside the United States.
Keywords	carbon monoxide ; coasts ; confidence interval ; environment ; environmental protection ; inventories ; methane ; satellites
Language	English
Dates of publication	2022-01
Publishing place	Elsevier Inc.
Document type	Article
ZDB-ID	431483-9
ISSN	0034-4257
ISSN	0034-4257
DOI	10.1016/j.rse.2021.112756
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Article ; Online: An optogenetic tool to recruit individual PKC isozymes to the cell surface and promote specific phosphorylation of membrane proteins.

Gada, Kirin D / Kawano, Takeharu / Plant, Leigh D / Logothetis, Diomedes E

The Journal of biological chemistry

2022 Volume 298, Issue 5, Page(s) 101893

Abstract: ... molecules. Here, we developed an optogenetic blue light-activated PKC isozyme that harnesses a plant-based ...

Abstract	The PKC family consists of several closely related kinases. These enzymes regulate the function of proteins through the phosphorylation of hydroxyl groups on serines and/or threonines. The selective activation of individual PKC isozymes has proven challenging because of a lack of specific activator molecules. Here, we developed an optogenetic blue light-activated PKC isozyme that harnesses a plant-based dimerization system between the photosensitive cryptochrome-2 (CRY2) and the N terminus of the transcription factor calcium and integrin-binding protein 1 (CIB1) (N-terminal region of the CRY2-binding domain of CIB1). We show that tagging CRY2 with the catalytic domain of PKC isozymes can efficiently promote its translocation to the cell surface upon blue light exposure. We demonstrate this system using PKCε and show that this leads to robust activation of a K
MeSH term(s)	Cell Membrane/metabolism ; Isoenzymes/genetics ; Isoenzymes/metabolism ; Membrane Proteins/genetics ; Membrane Proteins/metabolism ; Optogenetics ; Phosphorylation ; Transcription Factors/metabolism
Chemical Substances	Isoenzymes ; Membrane Proteins ; Transcription Factors
Language	English
Publishing date	2022-03-31
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Extramural
ZDB-ID	2997-x
ISSN	1083-351X ; 0021-9258
ISSN (online)	1083-351X
ISSN	0021-9258
DOI	10.1016/j.jbc.2022.101893
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Article ; Online: Underestimated Dry Season Methane Emissions from Wetlands in the Pantanal.

Li, Mengze / Kort, Eric A / Bloom, A Anthony / Wu, Dien / Plant, Genevieve / Gerlein-Safdi, Cynthia / Pu, Tianjiao

Environmental science & technology

2024

Abstract: Tropical wetlands contribute ∼30% of the global methane ( ... ...

Abstract	Tropical wetlands contribute ∼30% of the global methane (CH
Language	English
Publishing date	2024-02-07
Publishing country	United States
Document type	Journal Article
ISSN	1520-5851
ISSN (online)	1520-5851
DOI	10.1021/acs.est.3c09250
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Article ; Online: Internuclear ophthalmoplegia.

Virgo, Jonathan D / Plant, Gordon T

Practical neurology

2017 Volume 17, Issue 2, Page(s) 149–153

Abstract: A brainstem lesion of any type that involves the medial longitudinal fasciculus (MLF) can cause internuclear ophthalmoplegia (INO). This primarily affects conjugate horizontal gaze and classically manifests as impaired adduction ipsilateral to the lesion ...

Abstract	A brainstem lesion of any type that involves the medial longitudinal fasciculus (MLF) can cause internuclear ophthalmoplegia (INO). This primarily affects conjugate horizontal gaze and classically manifests as impaired adduction ipsilateral to the lesion and abduction nystagmus contralateral to the lesion. Here, we describe the anatomy of the MLF and review the clinical features of INO. We also describe conjugate horizontal gaze palsy and some of the 'INO-plus' syndromes.
MeSH term(s)	Afferent Pathways/pathology ; Afferent Pathways/physiopathology ; Brain Stem Infarctions/complications ; Brain Stem Infarctions/diagnostic imaging ; Eye Movements ; Female ; Functional Laterality ; Humans ; Magnetic Resonance Imaging ; Male ; Nystagmus, Pathologic/etiology ; Ocular Motility Disorders/diagnosis ; Ocular Motility Disorders/etiology
Language	English
Publishing date	2017-04
Publishing country	England
Document type	Journal Article ; Video-Audio Media
ZDB-ID	2170881-2
ISSN	1474-7766 ; 1474-7758
ISSN (online)	1474-7766
ISSN	1474-7758
DOI	10.1136/practneurol-2016-001428
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Article ; Online: Custom-engineered hydrogels for delivery of human iPSC-derived neurons into the injured cervical spinal cord.

Doulames, V M / Marquardt, L M / Hefferon, M E / Baugh, N J / Suhar, R A / Wang, A T / Dubbin, K R / Weimann, J M / Palmer, T D / Plant, G W / Heilshorn, S C

Biomaterials

2023 Volume 305, Page(s) 122400

Abstract: Cervical damage is the most prevalent type of spinal cord injury clinically, although few preclinical research studies focus on this anatomical region of injury. Here we present a combinatorial therapy composed of a custom-engineered, injectable hydrogel ...

Abstract	Cervical damage is the most prevalent type of spinal cord injury clinically, although few preclinical research studies focus on this anatomical region of injury. Here we present a combinatorial therapy composed of a custom-engineered, injectable hydrogel and human induced pluripotent stem cell (iPSC)-derived deep cortical neurons. The biomimetic hydrogel has a modular design that includes a protein-engineered component to allow customization of the cell-adhesive peptide sequence and a synthetic polymer component to allow customization of the gel mechanical properties. In vitro studies with encapsulated iPSC-neurons were used to select a bespoke hydrogel formulation that maintains cell viability and promotes neurite extension. Following injection into the injured cervical spinal cord in a rat contusion model, the hydrogel biodegraded over six weeks without causing any adverse reaction. Compared to cell delivery using saline, the hydrogel significantly improved the reproducibility of cell transplantation and integration into the host tissue. Across three metrics of animal behavior, this combinatorial therapy significantly improved sensorimotor function by six weeks post transplantation. Taken together, these findings demonstrate that design of a combinatorial therapy that includes a gel customized for a specific fate-restricted cell type can induce regeneration in the injured cervical spinal cord.
MeSH term(s)	Rats ; Humans ; Animals ; Cervical Cord ; Induced Pluripotent Stem Cells ; Hydrogels/chemistry ; Reproducibility of Results ; Spinal Cord ; Neurons ; Spinal Cord Injuries
Chemical Substances	Hydrogels
Language	English
Publishing date	2023-11-17
Publishing country	Netherlands
Document type	Journal Article
ZDB-ID	603079-8
ISSN	1878-5905 ; 0142-9612
ISSN (online)	1878-5905
ISSN	0142-9612
DOI	10.1016/j.biomaterials.2023.122400
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Article ; Online: Teaching Video NeuroImages: Cerebellar esotropia: A pitfall in ophthalmology and neurology.

Chapelle, Anne-Catherine / Plant, Gordon T / Kaski, Diego

Neurology

2019 Volume 93, Issue 1, Page(s) e114–e115

MeSH term(s)	Adolescent ; Arnold-Chiari Malformation/diagnosis ; Brain/diagnostic imaging ; Diagnosis, Differential ; Esotropia/diagnosis ; Female ; Humans
Language	English
Publishing date	2019-07-01
Publishing country	United States
Document type	Case Reports ; Journal Article ; Video-Audio Media
ZDB-ID	207147-2
ISSN	1526-632X ; 0028-3878
ISSN (online)	1526-632X
ISSN	0028-3878
DOI	10.1212/WNL.0000000000007729
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Article: Retention and the Intersection of Structural Inequities in a Breastfeeding Intervention Study.

LaPlant, Helen Wilde / Francis-Edoziuno, Confidence / Guan, Zhe / Aderibigbe, Tumilara / Chang, Xiaolin / Alhabodal, Ashwag Saad / Delaney, Kristen / Scott, Dana / Marshall-Crim, Mary / Freytes, Idelisa / Henderson, Wendy A / Walsh, Stephen / Lucas, Ruth F

medRxiv : the preprint server for health sciences

2024

Abstract: Introduction: Women below the poverty threshold have lower representation and retention in breastfeeding studies.: Methods: A secondary analysis of a longitudinal randomized controlled self-management for breast and nipple pain during breastfeeding ... ...

Abstract	Introduction: Women below the poverty threshold have lower representation and retention in breastfeeding studies. Methods: A secondary analysis of a longitudinal randomized controlled self-management for breast and nipple pain during breastfeeding study. Participants completed online surveys at discharge, weeks 1, 2, 3, 6, 9, 12, 18, and 24, with face-to-face interviews at 6 and 24 weeks. Text messages were sent to participants when modules and surveys were due. Retention was assessed in R with descriptive statistics, Mann-Whitney, Pearson's chi-square, and Cox Proportional Hazard Regression. Results: Two hundred and forty-four women (89 ≤$50,000 and 155 >$50,000) were recruited. Retention rates at 1 (93%), 2 (87%), 6 (82%), 9 (77%) and 24 (72%) weeks. For women of low income compared to those of high income there was a hazard ratio (HR) of 2.5 (p=0.0001) for retention. For non-Hispanic Black and Hispanic women compared to the combined non-Hispanic White and Other group, HRs for retention were 3.3 and 2.6 respectively (p=0.0001). Adjustment for age in the final hazard regression model of income, age, race and ethnicity decreased the HR for women of low income to 1.6 and HRs for non-Hispanic Black and Hispanic women to 2.1 and 1.9, respectively (p=.0001). However, none of the individual factors in the model achieved statistical significance. Discussion: Retention in breastfeeding studies impacts breastfeeding duration, a key lifelong preventative health behavior. Despite accessible study design, retention of women desiring to breastfeed was adversely affected by the intersection of income, race and ethnicity, and age.
Language	English
Publishing date	2024-03-16
Publishing country	United States
Document type	Preprint
DOI	10.1101/2024.03.15.24304355
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Article ; Online: Telehealth and virtual health monitoring in cystic fibrosis.

Vagg, Tamara / Shanthikumar, Shivanthan / Morrissy, David / Chapman, Wendy W / Plant, Barry J / Ranganathan, Sarath

Current opinion in pulmonary medicine

2021 Volume 27, Issue 6, Page(s) 544–553

Abstract: Purpose of review: At many institutions, the Covid-19 pandemic made it necessary to rapidly change the way services are provided to patients, including those with cystic fibrosis (CF). The purpose of this review is to explore the past, present and ... ...

Abstract	Purpose of review: At many institutions, the Covid-19 pandemic made it necessary to rapidly change the way services are provided to patients, including those with cystic fibrosis (CF). The purpose of this review is to explore the past, present and future of telehealth and virtual monitoring in CF and to highlight certain challenges/considerations in developing such services. Recent findings: The Covid-19 pandemic has proven that telehealth and virtual monitoring are a feasible means for safely providing services to CF patients when traditional care is not possible. However, both telehealth and virtual monitoring can also provide further support in the future in a post-covid era through a hybrid-model incorporating traditional care, remote data collection and sophisticated platforms to manage and share data with CF teams. Summary: We provide a detailed overview of telehealth and virtual monitoring including examples of how paediatric and adult CF services adapted to the need for rapid change. Such services have proven popular with people with CF meaning that co-design with stakeholders will likely improve systems further. In the future, telehealth and virtual monitoring will become more sophisticated by harnessing increasingly powerful technologies such as artificial intelligence, connected monitoring devices and wearables. In this review, we harmonise definitions and terminologies before highlighting considerations and limitations for the future of telehealth and virtual monitoring in CF.
MeSH term(s)	Adult ; Artificial Intelligence ; COVID-19 ; Child ; Cystic Fibrosis/therapy ; Humans ; Pandemics ; SARS-CoV-2 ; Telemedicine
Language	English
Publishing date	2021-08-02
Publishing country	United States
Document type	Journal Article ; Review
ZDB-ID	1285505-4
ISSN	1531-6971 ; 1070-5287 ; 1078-1641
ISSN (online)	1531-6971
ISSN	1070-5287 ; 1078-1641
DOI	10.1097/MCP.0000000000000821
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