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Representative Anika T. Omphroy Tours Hugh Taylor Birch State Park
01/07/2022 8:00 AM | Lauren Kelly-Manders (Administrator)
FORT LAUDERDALE – Representative Anika T. Omphroy, who represents District 95, visited Hugh Taylor Birch State Park in Fort Lauderdale yesterday as a guest of the Florida State Parks Foundation and the Florida Park Service.
The visit, ahead of this year's Legislative Session, was to brief Representative Omphroy about new developments at the park and to discuss issues affecting all of Florida's 175 award-winning parks and trails. Representative Omphroy is a member of the House Appropriations Committee and the Democrat Ranking Member of the Agricultural and Natural Resources Appropriations Subcommittee.
She has long been a champion of the environment with a special interest in preventing further development west into the Everglades and addressing the threat of saltwater intrusion into the South Florida aquifer.
"We appreciate Representative Omphroy spending time at Hugh Taylor Birch State Park and for her commitment to our state parks and trails," said Florida State Parks Foundation CEO Julia Gill Woodward.
Also representing the Florida Park Service were Florida Park Service Acting Director, Chuck Hatcher, District 5 Bureau Chief Kevin Jones, Foundation Board Member Jose Romano and Park Manager LeAnn Hinson.
"Our state parks form the cornerstone of biodiversity conservation and deliver invaluable cultural, economic and social benefits to our state. That is why my commitment to preserving and protecting Florida's natural heritage is unwavering," said State Representative Anika T. Omphroy.
"Our parks are not only natural treasures, but they are also economic generators as well. Our award-winning state parks attract 24 million visitors from around the world, have an economic impact to the state of $2.2 billion and support more than 31,000 jobs," Woodward said.
The 175-acre Hugh Taylor Birch State Park attracts more than 285,000 visitors a year and generates more than $25.6 million to the state's economy and supports 359 local jobs.
The Foundation, founded in 1993 as Friends of Florida State Parks and renamed in 2018, is a 501(c)(3) nonprofit corporation whose mission is to support and help sustain the Florida Park Service, its 175 award-winning parks and trails, local Friends groups and more than 20,000 park volunteers.
The volunteer Board of Directors represents private and public sectors as well as local and statewide interests.
William G. & Marie Selby Foundation Grant Allows Myakka River State Park to Purchase All-terrain Wheelchair and Trailer
A grant from the William G. & Marie Selby Foundation will help purchase an all-terrain wheelchair and trailer for use by visitors to Myakka River State Park, south of Sarasota.
"The Florida State Parks Foundation is thrilled to utilize this grant to purchase this special wheelchair that enables visitors who use mobility devices to fully experience the scenic beauty of Myakka River State Park," said Foundation President Tammy Gustafson.
"The $2,300 grant will go towards existing funds for the purchase of the all-terrain chair as well as a trailer to transport it, both of which will be available free of charge upon visitor request," she said.
Located about 20 miles from downtown Sarasota, Myakka River State Park is one of oldest and largest parks in the award-winning Florida state park system, covering more than 37,000 acres.
"The Selby Foundation is pleased to be a partner on this very important project that will increase accessibility for so many to enjoy the beauty and joy of nature", said Carol Butera, President/CEO of The William G. and Marie Selby Foundation.
The Florida State Parks Foundation, founded in 1993 as Friends of Florida State Parks and renamed in 2018, is a 501(c)(3) nonprofit corporation whose mission is to support and help sustain the Florida Park Service, its 175 award-winning parks and trails, local Friends groups and more than 20,000 park volunteers.
Florida State Parks Foundation Celebrates A Historic Year
The Florida State Parks Foundation recently celebrated a year full of tremendous achievements and looked ahead to 2022 during President Tammy Gustafson's end-of-year message to members.
"As we approach the end of the year, it is good to reflect on all the incredible accomplishments we have achieved together to make our award-winning Florida State Parks even better," Gustafson said. "We look forward, with your help, to even greater things in 2022."
The Foundation started 2021 with a huge success by introducing a bill to establish a Florida State Park specialty license to benefit park projects, and then getting overwhelming bipartisan legislative support to get it approved. Impressively, the license plate cleared its 3,000-preorder threshold in less than 34 days, has gone into production and will be available in early 2022.
"The Foundation is truly fortunate to have the support of the many thousands of folks who recognize the importance of our fabulous state parks and the need to protect and preserve them for generations to come," Gustafson said. "The impact you have cannot be overstated. Your generosity has enabled the Foundation to make significant accomplishments in its mission of protecting, preserving, sustaining and growing our Florida State Parks."
Gustafson went on to recognize some of the Foundation's most significant milestones from the last year:
"Your generosity helped us fund, build and launch the first-ever wheelchair accessible glass bottom boat at Silver Springs State Park. In keeping with the tradition of naming the glass-bottom boats at Silver Springs after Seminole tribal chiefs, the new boat is named Chief Potackee – Betty Mae Tiger Jumper, after the only woman to serve as chief of the Seminole Tribe of Florida.
"Thanks to you, we have improved accessibility in our parks through funding a new accessible playground at Edward Ball Wakulla Springs State Park, breaking ground on the first phase of an accessible fishing pier at Oscar Scherer State Park and providing beach-accessible wheelchairs to every single coastal state park.
"Your continuing largesse has improved wildlife care and conservation through expanding resources for sea turtle needs at Sebastian Inlet State Park and John D. MacArthur Beach State Park, provided habitat maintenance for the endangered red-cockaded woodpecker at St. Sebastian River Preserve State Park, expanded manatee care at Ellie Schiller Homosassa Springs Wildlife State Park and provided funding for coral reef research at John Pennekamp Coral Reef State Park.
"You have helped improve the ability to share our historical and cultural resources through the funding of the Flight to Freedom Trail at Fort Mose Historic State Park, telling the story of the first freed Black settlement in the precolonial United States and also commemorating two civil rights heroes and the park's namesakes at the Dr. Von D. Mizell – Eula Johnson State Park, who will finally have their stories told through interpretive signage.
"You have helped make our parks even greener by funding longleaf pine restoration work through our Plant a Pine program and providing electric-accessible trams at Hugh Taylor Birch State Park and Topsail Hill Preserve State Park.
"And, at the Florida State Parks Foundation, we have increased our membership to record levels. We have funded the training of dozens of park rangers this year and advocated for the most robust parks budget in recent history.
"Through the work of many, much has been accomplished. Together, we will continue to ensure that Florida's State Parks remain the best in the nation and a treasure that we can all be proud of."
Florida State Parks Foundation Welcomes Governor DeSantis' Recommendation To Fully Fund State Parks
12/10/2021 10:43 AM | Lauren Kelly-Manders (Administrator)
The Florida State Parks Foundation welcomed the Governor's recommendation to fully fund Florida's award-winning state parks.
"Florida's award-winning state parks protect our most precious natural and cultural resources as well as providing a safe haven for people to recreate and exercise," said Tammy Gustafson, Foundation President.
"We are delighted that the Governor in his proposed budget released yesterday has allocated $51.7 million for the Florida Park Service" she said.
$45 million has been allocated for state park facility improvements and $6.7 million for Billy Joe Rish State Park, which includes additional facilities, repairs and renovations to existing facilities and the removal of access barriers for persons with disabilities. In addition, $52 million has been allocated for springs restoration and $100 million from the Florida Forever program for land acquisition. Almost $1 billion has also been earmarked to continue Everglades restoration.
Florida State Parks represent one of the state's greatest success stories. They are a major contributor to the state's tourism industry, support thousands of jobs, and pour billions of dollars into the state's economy. At the same time, they preserve many of the jewels of our natural environment and Florida's significant historical and cultural sites.
"As the acknowledged premier state park system in the nation, it is vital that Florida State Parks receive full funding. When comparing the $51.7 million that has been provided in the budget with the $2.6 billion in economic impact the state parks generated last year from nearly 30 million visitors, it is difficult to think of a better return on the investment anywhere," she said.
Silver Springs State Park's Wheelchair-Accessible Glass-Bottom Boat Named as One of the Ten Most Significant Boats of 2021
11/23/2021 2:43 PM | Lauren Kelly-Manders (Administrator)
The Florida State Parks Foundation announced today that the new wheelchair accessible glass bottom boat at Silver Springs State Park has been named as one of the ten most significant boats of 2021.
The top ten list, compiled by WorkBoat magazine, will be used to select the Boat of the Year on Wednesday, Dec. 1, the first day of the 2021 International WorkBoat Show in New Orleans.
The Florida State Parks Foundation commissioned Lay, Pitman & Associates Naval Architects to design the 37-foot boat, which includes a flat deck and spacious aisles providing easy and convenient access for wheelchairs. It is also equipped with an induction-loop system that assists passengers using hearing aids to listen to the captain's narration.
This boat is significant in many ways, said Paula Russo, former Foundation president, who oversaw the vessel's design and construction. Although glass bottom boat tours have been operating at Silver Springs since 1870 none of the vessels was able to accommodate wheelchairs and it was not possible to retrofit them, so an entirely new boat was needed. The vessel was built by St. Johns Ship Building in Palatka using design principles that give universal access to all people.
The totally electric glass bottom boat was donated to the Florida Park Service in June. It can carry up to 28 passengers and two crew members, and has a 12" draft. The boat is powered by twin 20-hp Elco electric outboards with four Lithionics 7.5 kW lithium-ion batteries. Ship's service power comes from two Lithionics lithium-ion 7.5-kW batteries.
In keeping with the tradition of naming the glass-bottom boats at Silver Springs after Seminole tribal chiefs, the new boat is named Chief Potackee – Betty Mae Tiger Jumper, after the only woman to serve as chief of the Seminole Tribe of Florida.
In addition to the Foundation, a consortium of public and private partners helped fund the boat, including the Fish & Wildlife Foundation of Florida, the Felburn Foundation, the Delores Barr Weaver Legacy Fund, the Florida Park Service, Friends of Silver Springs State Park and Cape Leisure.
Florida State Parks Foundation Welcomes Chuck Hatcher as Acting Director of Division of Recreation and Parks
The Florida State Parks Foundation welcomes today's announcement that Chuck Hatcher has been named Acting Director of the Division of Recreation and Parks.
Chuck takes over from Eric Draper, who retired as Director of the Division earlier this month after four years in the position.
"The Foundation is delighted that Chuck Hatcher has been named Acting Director," said Foundation President Tammy Gustafson. "We have worked closely with Chuck and the award-winning Florida Park Service's senior management team for many years. Together we have secured many memorable successes and we look forward to achieving even greater things in the future."
Chuck joined the Department of Environmental Protection in March 2016 as Assistant Director of Field Operations for the Division of Recreation and Parks. Prior to that, he served for a decade as Director of Parks and Recreation in Jackson County, Florida, where he received numerous awards including the Excellence in Leadership Award presented by the Jackson County Chamber of Commerce.
Ivanhoe Park Brewing Company Marks Partnership with Florida State Parks Foundation with a New Beer
The Florida State Parks Foundation today announced a partnership with the Ivanhoe Park Brewing Company and a new collaboration beer, Lake Hopp'r Florida Lager.
As part of the partnership Ivanhoe Park Brewing Company, an independent craft brewery and tasting room in Orlando, is brewing a collaboration beer, Lake Hopp'r Florida Lager, with a portion of proceeds benefiting the Foundation, as well as offering corporate volunteering at the local Florida State Parks.
"Ivanhoe Park Brewing Co. continues to carry on our Commitment to Community," CEO and Founder, Glenn Closson said. "Working with Florida State Parks Foundation, we are not only excited to be able to offer additional funds, but also our time to really help their mission and volunteering to assist in projects at the Florida State Parks."
Their next corporate volunteer day will be at Lake Louisa State Park on November 21, he said.
"We are excited to enter into this partnership with Ivanhoe Park Brewing which will benefit our fabulous award-winning state parks," said Foundation CEO Julia Gill Woodward. "We are always delighted to work with corporate partners like Ivanhoe Park Brewing who share our commitment to preserving and protecting our state parks."
Located in Ivanhoe Village, one of Orlando's Main Street districts, the brewery has been open since 2018 and entered distribution later that year in Central Florida with City Beverages. That distribution partnership has grown and allowed Ivanhoe Park Brewing Co. to work with Wayne Densch, Inc and Daytona Beverages to expand their market reach. They are looking to add more distributors in the first quarter of 2022.
This new partnership with the Florida State Parks Foundation will allow Ivanhoe Park Brewing Co. to brew an amazing Florida Lager that will be easy drinking and 4% ABV to be enjoyed throughout the State of Florida.
Florida State Parks Specialty License Plate Exceeds Presale Target in One Month
The Florida State Parks Foundation has announced that presales of the Florida State Parks specialty license plate have already soared past the 3,000-sale target needed for the plate to go into production.
Those who preordered the plate will be able to receive their plate in early-spring 2022. Florida State Parks specialty license plates are still available for purchase online, and $25 from each plate sold will directly benefit the Florida State Parks Foundation.
"This is a fantastic achievement and shows just how popular our state parks are and why people want to support them in this way," said Foundation President Tammy Gustafson.
"We're pleased to have met our presale goal so quickly," said Eric Draper, Director of the Florida Park Service. "Not only are we grateful for this new fundraising opportunity, but we're also excited to see people showing off their Florida State Parks pride on their vehicles."
The Florida Department of Highway Safety and Motor Vehicles requires that all organizations wishing to introduce a new specialty license plate presell 3,000 units before they begin production. The DMW allows two years to meet this goal. The Florida State Parks specialty plate reached 3,000 presales in just 34 days.
"We not only hit our 3,000 target in just one month, we have flown past it," Gustafson said. "Funds from the sale of the Florida State Parks license plate will provide significant and ongoing funding to enable us to protect and preserve our award-winning park system."
Generous Bequest Promotes Birding Knowledge at Sebastian Inlet State Park
The Florida State Parks Foundation announced a series of interpretive birding panels at Sebastian Inlet State Park on Saturday during a dedication ceremony in honor of Bonnie Lizer, who loved both the park and birding.
"Bonnie Lizer passed away in June 2019, at the age of 82, in Indialantic, Florida, and she very generously left a quarter of her estate, more than $1.3 million, to the Foundation. Bonnie loved Sebastian Inlet State Park and birding was her favorite pastime, so this is a very fitting and lasting tribute," said Foundation President Tammy Gustafson.
"In her memory, a series of interpretive panels has been created to educate visitors about bird identification, bird habitat protection and appropriate bird observation techniques in the park," she said. "A plaque to commemorate Bonnie's generosity was also unveiled."
Sebastian Inlet State Park is one of Florida's most visited parks with over 760,000 visitors a year. Although known as a fishing and surfer's paradise, this 24-hour park is also part of the Great Florida Birding and Wildlife Trail, and offers many special wildlife programs. It is a stopover for many migratory birds and 225 different bird species have been recorded there.
The interpretive panels will assist visitors in identifying common migratory and resident bird species. Visitors will also learn about bird migratory patterns, feeding and nesting requirements, habitat, and life cycles, she said.
They will feature tactile and interactive elements for visitors of all ages and will include QR codes that will link visitors to additional information that can be viewed in the park or once they return home. Visitors will also learn about the park's estuary ecosystem and its importance to a variety of bird species.
Florida State Parks Foundation Announces 2021-2022 Board Officers
The Florida State Parks Foundation has announced its Board Officers for 2021-2022 to provide critical leadership to ensure the organization's continued growth and success.
Officers elected at yesterday's annual meeting are Tammy Gustafson, Board President; Kathleen Brennan, President Elect; Gil Ziffer, Immediate Past President; Audrine Finnerty, Treasurer; and Emily Lewis, Secretary.
"I am delighted to announce our new board officers", said Julia Gill Woodward, Foundation CEO. "These individuals bring a wealth of experience and expertise in their fields and will be tremendous assets to the Foundation as we further our mission of supporting Florida's fabulous award-winning state parks, the best in the nation."
Tammy Gustafson is a senior executive at Universal Orlando responsible for Sales Integration across the global sales divisions and working with international teams located in Latin America and the UK. She joined Universal Orlando in September of 1997 as part of the Universal Studios Marketing team and was responsible for building the Universal Studios Florida brand on a global basis- growing to three theme parks, a night-time entertainment complex and eight onsite resorts.
Appointed by the Governor's office to the Florida Commission on Tourism, Gustafson served as President of the VISIT FLORIDA Board of Directors from 2014-15 and has been on the board since 2005. She currently serves on the Florida Council of Tourism Leaders. She has also served on the board for the Kissimmee/Osceola County Chamber of Commerce/Osceola Resort Area Council and served as a Past President and Treasurer of the Florida State Parks Foundation. Gustafson brings years of experience and extensive travel industry knowledge to these organizations and has been a part of Leadership Florida, Leadership Orlando, the Greater Orlando Sports Commission and the Central Florida Chapter of the American Marketing Association.
"Throughout its history, the Florida State Parks greatest strength has been its volunteers. I am honored to be asked to serve in this capacity," said Tammy Gustafson, Foundation President. "The best and brightest minds are coming together to help guide and preserve the Real Florida. It is an exciting time for the Foundation as we work toward the Strategic Goals for the coming year."
It does this through programs that preserve and protect state parks, educate visitors about the value of state parks, encourage community engagement and active use of state parks, and advocacy. The volunteer Board of Directors represent private and public sectors as well as local and statewide interests.


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Home / Australia Company Profiles / Warner Bros Entertainment Australia Pty Limited
Warner Bros Entertainment Australia Pty Limited - Premium Company Report Australia
Company Type: Proprietary Company
What does Warner Bros Entertainment Australia Pty Limited do?
Warner Bros Entertainment Australia Pty Limited is a foreign-owned private company, deriving revenue from the distribution of television programmes and movies, granting of licences for the Warner Bros trademarks and production of motion picture films. The company employs approximately 30 people, operates in Australia, and is administered from its head office in Neutral Bay, New South Wales.
The company's ultimate parent is AT&T Inc., a United States-based telecommunications, media and technology services provider.
IBISWorld partnered with the Australian Financial Review to publish a list of Australia's Top 500 Private Companies for 2019.
Warner Bros Entertainment Australia Pty Limited - Products & Brands
Warner Bros Entertainment Australia divides its operation into four business divisions:
Television Distribution - Distribution of international television programmes
Theatrical Distribution - Distribution of motion picture films
Consumer Product Licensing - Granting of licences for the right to use Warner Bros. trademarks and properties in consumable products
Feature Productions - Production of motion picture films and television series.
Warner Bros Entertainment Australia Pty Limited - Financial Statements
pie_chart Warner Bros Entertainment Australia Pty Limited Financial Profit & Loss Account
5,868 10,503 208,211 99,560 158,500
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Warner Bros Entertainment Australia Pty Limited - Financial Ratios
pie_chart Warner Bros Entertainment Australia Pty Limited Financial Growth
8.0 -61.0 6.2 -15.5 -38.2
pie_chart Warner Bros Entertainment Australia Pty Limited Financial Ratios
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Warner Bros Entertainment Australia Pty Limited operates in the following industries:
J5511 - Motion Picture and Video Production in Australia
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redpajama_set_name: RedPajamaCommonCrawl


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Schistosoma mansoni p48 eggshell protein gene: characterization, developmentally regulated expression and comparison to the p14 eggshell protein gene
Li ly Chen, David M. Rekosh, Philip T. LoVerde
Biochemistry & Structural Biology
Egg production by worm pairs is a major cause of pathogenesis in schistosomiasis. To further the understanding of female reproductive development, we have isolated and characterized a complete copy of an eggshell protein precursor gene, p48. Sequence analysis reveals that the gene has 3 open reading frames and does not contain an intron. One of the open reading frames, ORF1, encodes a polypeptide of 50 kDa which shows strong homology to insect chorion proteins. Determination of the position of the mRNA cap-site facilitated identification of putative regulatory elements in the 5′ upstream region of the gene. Some of these elements (e.g. TCACGT) have been shown to play a role in the regulation of chorion gene expression in insects. p48 mRNA is detectable only in mature female worms and the ability to detect the mRNA coincides temporally with worm pairing. Quantitative comparisons, during female reproductive development, of p48 transcripts to those from another eggshell protein precursor gene, p14, show that the p48 mRNA is significantly less abundant than p14 mRNA. In mature female worms, p48 mRNA can only be detected in vitelline cells. Antibodies made against the polypeptide sequence deduced from ORF1 of the p48 gene recognize a 50-kDa molecule in extracts from mature female worms, but not in extracts from immature females or males.
Molecular and Biochemical Parasitology
https://doi.org/10.1016/0166-6851(92)90034-H
Chorion gene
Developmentally regulated gene expression
Eggshell protein gene
Schistosome female reproductive development
10.1016/0166-6851(92)90034-H
Fingerprint Dive into the research topics of 'Schistosoma mansoni p48 eggshell protein gene: characterization, developmentally regulated expression and comparison to the p14 eggshell protein gene'. Together they form a unique fingerprint.
chorion proteins Medicine & Life Sciences
Messenger RNA Medicine & Life Sciences
Protein Precursors Medicine & Life Sciences
Genes Medicine & Life Sciences
Open Reading Frames Medicine & Life Sciences
Insect Proteins Medicine & Life Sciences
Chorion Medicine & Life Sciences
Peptides Medicine & Life Sciences
Chen, L. L., Rekosh, D. M., & LoVerde, P. T. (1992). Schistosoma mansoni p48 eggshell protein gene: characterization, developmentally regulated expression and comparison to the p14 eggshell protein gene. Molecular and Biochemical Parasitology, 52(1), 39-52. https://doi.org/10.1016/0166-6851(92)90034-H
Schistosoma mansoni p48 eggshell protein gene : characterization, developmentally regulated expression and comparison to the p14 eggshell protein gene. / Chen, Li ly; Rekosh, David M.; LoVerde, Philip T.
In: Molecular and Biochemical Parasitology, Vol. 52, No. 1, 05.1992, p. 39-52.
Chen, LL, Rekosh, DM & LoVerde, PT 1992, 'Schistosoma mansoni p48 eggshell protein gene: characterization, developmentally regulated expression and comparison to the p14 eggshell protein gene', Molecular and Biochemical Parasitology, vol. 52, no. 1, pp. 39-52. https://doi.org/10.1016/0166-6851(92)90034-H
Chen LL, Rekosh DM, LoVerde PT. Schistosoma mansoni p48 eggshell protein gene: characterization, developmentally regulated expression and comparison to the p14 eggshell protein gene. Molecular and Biochemical Parasitology. 1992 May;52(1):39-52. https://doi.org/10.1016/0166-6851(92)90034-H
Chen, Li ly ; Rekosh, David M. ; LoVerde, Philip T. / Schistosoma mansoni p48 eggshell protein gene : characterization, developmentally regulated expression and comparison to the p14 eggshell protein gene. In: Molecular and Biochemical Parasitology. 1992 ; Vol. 52, No. 1. pp. 39-52.
@article{43b7282002614e4fa87d97fe600d5920,
title = "Schistosoma mansoni p48 eggshell protein gene: characterization, developmentally regulated expression and comparison to the p14 eggshell protein gene",
abstract = "Egg production by worm pairs is a major cause of pathogenesis in schistosomiasis. To further the understanding of female reproductive development, we have isolated and characterized a complete copy of an eggshell protein precursor gene, p48. Sequence analysis reveals that the gene has 3 open reading frames and does not contain an intron. One of the open reading frames, ORF1, encodes a polypeptide of 50 kDa which shows strong homology to insect chorion proteins. Determination of the position of the mRNA cap-site facilitated identification of putative regulatory elements in the 5′ upstream region of the gene. Some of these elements (e.g. TCACGT) have been shown to play a role in the regulation of chorion gene expression in insects. p48 mRNA is detectable only in mature female worms and the ability to detect the mRNA coincides temporally with worm pairing. Quantitative comparisons, during female reproductive development, of p48 transcripts to those from another eggshell protein precursor gene, p14, show that the p48 mRNA is significantly less abundant than p14 mRNA. In mature female worms, p48 mRNA can only be detected in vitelline cells. Antibodies made against the polypeptide sequence deduced from ORF1 of the p48 gene recognize a 50-kDa molecule in extracts from mature female worms, but not in extracts from immature females or males.",
keywords = "Chorion gene, Developmentally regulated gene expression, Eggshell protein gene, Schistosoma mansoni, Schistosome female reproductive development",
author = "Chen, {Li ly} and Rekosh, {David M.} and LoVerde, {Philip T.}",
doi = "10.1016/0166-6851(92)90034-H",
journal = "Molecular and Biochemical Parasitology",
T1 - Schistosoma mansoni p48 eggshell protein gene
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AU - Chen, Li ly
AU - Rekosh, David M.
AU - LoVerde, Philip T.
N2 - Egg production by worm pairs is a major cause of pathogenesis in schistosomiasis. To further the understanding of female reproductive development, we have isolated and characterized a complete copy of an eggshell protein precursor gene, p48. Sequence analysis reveals that the gene has 3 open reading frames and does not contain an intron. One of the open reading frames, ORF1, encodes a polypeptide of 50 kDa which shows strong homology to insect chorion proteins. Determination of the position of the mRNA cap-site facilitated identification of putative regulatory elements in the 5′ upstream region of the gene. Some of these elements (e.g. TCACGT) have been shown to play a role in the regulation of chorion gene expression in insects. p48 mRNA is detectable only in mature female worms and the ability to detect the mRNA coincides temporally with worm pairing. Quantitative comparisons, during female reproductive development, of p48 transcripts to those from another eggshell protein precursor gene, p14, show that the p48 mRNA is significantly less abundant than p14 mRNA. In mature female worms, p48 mRNA can only be detected in vitelline cells. Antibodies made against the polypeptide sequence deduced from ORF1 of the p48 gene recognize a 50-kDa molecule in extracts from mature female worms, but not in extracts from immature females or males.
AB - Egg production by worm pairs is a major cause of pathogenesis in schistosomiasis. To further the understanding of female reproductive development, we have isolated and characterized a complete copy of an eggshell protein precursor gene, p48. Sequence analysis reveals that the gene has 3 open reading frames and does not contain an intron. One of the open reading frames, ORF1, encodes a polypeptide of 50 kDa which shows strong homology to insect chorion proteins. Determination of the position of the mRNA cap-site facilitated identification of putative regulatory elements in the 5′ upstream region of the gene. Some of these elements (e.g. TCACGT) have been shown to play a role in the regulation of chorion gene expression in insects. p48 mRNA is detectable only in mature female worms and the ability to detect the mRNA coincides temporally with worm pairing. Quantitative comparisons, during female reproductive development, of p48 transcripts to those from another eggshell protein precursor gene, p14, show that the p48 mRNA is significantly less abundant than p14 mRNA. In mature female worms, p48 mRNA can only be detected in vitelline cells. Antibodies made against the polypeptide sequence deduced from ORF1 of the p48 gene recognize a 50-kDa molecule in extracts from mature female worms, but not in extracts from immature females or males.
KW - Chorion gene
KW - Developmentally regulated gene expression
KW - Eggshell protein gene
KW - Schistosoma mansoni
KW - Schistosome female reproductive development
U2 - 10.1016/0166-6851(92)90034-H
DO - 10.1016/0166-6851(92)90034-H
JO - Molecular and Biochemical Parasitology
JF - Molecular and Biochemical Parasitology


To install the Vectorworks Modelo Plug-in for Mac, you first need to download the Plug-in. Download Plug-in here. When completing these steps make sure that Vectorworks is not currently running.
Please note, if you unzip and see a _MACOSX folder within, ignore it.
Step 1 Move the .vsm and .vwlibrary plug-ins to the following location: applications/Vectorworks 2016/Plug-Ins.
In your applications folder, expand the "Vectorworks 2016" folder. Right click the "Vectorworks 2016" executable file and select "Show Package Contents".
Copy-Paste the "requests" folder here.
Step 3 Open Vectorworks 2016 and go to Tools>Workspaces>Edit Current Workspace... A Workspace Editor will display with two lists: Commands list on the left and Menus list on the right.
In the Commands list, expand "Import/Export" scroll down and select "Export to Modelo"
In the menu list expand "File" scroll down and expand "Export"
In the Commands list click and drag "Export to Modelo" into "Export" under the Menus list.
The Workspace should now be updated. To use the Plug-in, open a VWX file in a 3D view.
Please note, the file must be in a 3D view in order to export.
Open the File menu, select Export and "Export to Modelo" should be at the top of the list.
Once you select it, a new dialog will display asking you for a token. Open Modelo in your web browser. At the top right there is an option called "Upload Token" select this token. You will then see two drop-down menus for Projects and Folders within those projects. Select the Project and Folder you want to upload your Vectorworks model to. Once selected a token will be generated. Copy and paste this into the Vectorworks dialog box and select Upload.
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Terrestrial geothermal fields and oceanic hydrothermal vents are considered as candidate environments for the emergence of life on Earth. Nevertheless, the ionic strength and salinity of oceans present serious limitations for the self-assembly of amphiphiles, a process that is fundamental for the formation of first protocells. Consequently, we systematically characterized the efficiency of amphiphile assembly, and vesicular stability, in terrestrial geothermal environments, both, under simulated laboratory conditions and in hot spring water samples (collected from Ladakh, India, an Astrobiologically relevant site). Combinations of prebiotically pertinent fatty acids and their derivatives were evaluated for the formation of vesicles in aforesaid scenarios. Additionally, the stability of these vesicles was characterized over multiple dehydration-rehydration cycles, at elevated temperatures. Among the combinations that were tested, mixtures of fatty acid and its glycerol derivatives were found to be the most robust, also resulting in vesicles in all of the hot spring waters that were tested. Importantly, these vesicles were stable at high temperatures, and this fatty acid system retained its vesicle forming propensity, even after multiple cycles of dehydration-rehydration. The remaining systems, however, formed vesicles only in bicine buffer. Our results suggest that certain prebiotic compartments would have had a selective advantage in terrestrial geothermal niches. Significantly, our study highlights the importance of validating results that are obtained under 'buffered' laboratory conditions, by verifying their plausibility in prebiotically analogous environments.
Potential niches that could have supported the origin of life on the early Earth have long been a topic of study, and that of even intense debate. A given environment needs to fulfill certain criteria in order to be considered as a potential site for the origin of life. To highlight some of the important ones, the environment should have a source of, both, energy and precursor molecules, mechanisms for concentrating these precursors, and the potential for catalyzing energetically uphill polymerization reactions . One such site is marine hydrothermal vents, which led to the popular hypothesis that life on the Earth might have originated and evolved in the oceans . However, it has been argued that the ionic strength and salinity of oceans would have severely limited the facilitation of fundamental processes that are considered to be important for the formation of protocellular entities. These processes mainly include the self-assembly of amphiphiles and the nonenzymatic polymerization of mononucleotides . Additionally, the pH of prebiotic oceans could have been much more acidic than that of the present day oceans because of what has been hypothesized as resulting from elevated levels of CO2 in the early Earth atmosphere . This would make it difficult for simple amphiphiles, like fatty acids, to form vesicles as they self-assemble into these higher ordered entities at a pH that is around their pKa (in a range of pH 7 to 9). Furthermore, the problem of dilution of the reactants and products, if left un-sequestered, is an important constraint for this hypothesis [5,6].
An alternative hypothesis, which alleviates the aforementioned issues, is the proposal that life would have originated in terrestrial geothermal fields, such as geysers, hot springs, and tidal pools [7,8,9]. In addition to fulfilling basic criteria for being a plausible hatchery for the beginning of cellular life, their ionic strengths are much lower than that of oceans. This would have made them more amenable for the occurrence and sustenance of prebiotically pertinent processes, including the formation of vesicular membranes by amphiphilic systems. Additionally, the dehydration-rehydration (DH-RH) conditions, which are quite common in these regimes, would have catalysed the nonenzymatic extension of monomers that were capable of encoding information [10,11], and their subsequent encapsulation into membranous compartments to result in the first protocells . Although, it is impossible to know the exact ionic composition of prebiotic oceans and terrestrial geothermal fields, one could take a somewhat top-down approach by systematically studying the aforementioned processes in prebiotically pertinent analogous environments that are found on the extant Earth. This would also provide a more realistic context to discern whether the results that we obtain in the laboratory, under "buffered" conditions, would indeed hold true in realistic natural environments. After all, life started in a natural environment and not in the laboratory.
Towards this extent, an exploration of prebiotically analogous sites was undertaken as a part of an expedition to Ladakh, which is situated in the northern most state of Jammu and Kashmir, in India. This was done under the aegis of the NASA Spaceward Bound program in the August of 2016 . Ladakh is an Astrobiologically relevant site, which is dotted with topological features that are analogous to the landscape of Mars and, hence, it is emulative of early Earth conditions. As part of the geochemistry team, water samples were collected by our group from various sites pertaining to three different hot springs in Ladakh, namely Puga (PU), Chumathang (CH), and Panamic (PA). All of these are alkaline hot springs where the water boils at 85 °C due to the high altitude (3200–4500 m above sea level; Table S1). It is interesting to note that PU is analogous in nature to the Pilbara hot spring deposits of Western Australia, where the earliest signs of terrestrial life has recently been confirmed .
These hot spring water samples provide a prebiotically pertinent backdrop for studying fundamental processes that would have been crucial for the formation of the first protocells. As mentioned earlier, one of these processes would be the self-assembly of prebiotically relevant amphiphiles, such as fatty acids, to form membranous compartments. Fatty acid vesicle formation has been shown to be affected by several parameters, such as temperature, pH, concentration of the fatty acid, ionic strength of the medium, etc. Studies have been performed to understand the effect of single or multiple ions (mostly independently) , and that of temperature , on the formation and stability of fatty acid vesicles. However, the combined effect of these two crucial factors i.e., ionic composition and temperature, on the aforementioned aspects, is yet to be elucidated. In this study, we set out to address a two-fold question. The first goal was to delineate the effect of multiple ions in varying concentrations, on vesicle formation using different fatty acid systems. We also characterized the stability of these vesicles in niches with varying ionic strengths and at a high temperature. Furthermore, we studied the effect of DH-RH conditions on these systems, as they mimic natural processes (e.g., day-night and wet-dry cycles) that would have played out in prebiotic terrestrial geothermal fields. As a part of the second goal, we undertook studies to delineate the formation and stability of fatty acid vesicles in prebiotic analog conditions, to facilitate comparison between 'buffered' versus more 'realistic' conditions. We found that the fatty acid and its glycerol derivative was the most robust system among the combinations that were tested. This binary system formed vesicles in all of the hot spring water samples, which had varying ionic strengths. These mixed fatty acid vesicles were also stable at high temperature and retained their propensity to form vesicles under multiple DH-RH cycles. Furthermore, all the combinations of different fatty acids and their derivatives formed vesicles in buffered conditions, but not in the actual hot spring samples, with the exception of the aforementioned binary system. These results clearly indicate that the outcome of experiments that are performed under laboratory conditions are highly context dependent, and, significantly, may not always hold true under natural conditions.
All fatty acids and their derivatives were purchased from Nu-Chek-Prep (Elysian, MN, USA), except for 1-Decanoyl-rac-glycerol, which was purchased from Sigma-Aldrich (Bengaluru, India). All other reagents and solvents that were used were of the highest commercially available grade.
The water samples were collected from Puga, Chumathang, and Panamic hot springs, from both origin and run-off sites. However, only the samples that were collected from the sites of origin were used for this study. This was because these sites lacked any sort of obvious microbial growth and vegetation, thus reducing concerns of biological contamination and contamination due to human interference. All of the samples were filtered at the site of collection through a 200 nm Whatman syringe filter to avoid microbial contamination, if any, and their temperature and pH were measured. The pH was reanalyzed in the laboratory for all of the water samples that were used in this study (Table S1).
Different concentrations of fatty acids and their derivatives were selected for the experiment (Table S2), which is similar to what has been previously reported [3,15]. In a typical reaction, the fatty acid alone, or with the pertinent derivative, was first melted by heating above their melting temperature. Appropriate concentrations were then dissolved in chloroform and the chloroform was subsequently evaporated under vacuum to form a fatty acid film. The desired volume of solvent (bicine buffer/hot spring water) was then added to this film to form vesicles. Importantly, the solvent was preheated above the melting temperature of the system. The resultant solution containing vesicles was mixed using a vortex mixer and then visualized under Differential Interference Contrast microscope (AxioImager Z1, Carl Zeiss, Germany), using 40X objective (NA = 0.75).
The DH-RH experimental setup was similar to the one that has been used in previous studies . Briefly, a 200 µL solution containing vesicles was taken in a glass vial and was kept on a bench-top heating block, maintained at 75 °C under constant gentle CO2 flow. The rehydration was performed using either the same starting solvent (bicine buffer/hot spring water) or with milli-Q water, as indicated appropriately. The duration of each DH-RH cycle was 1 h (Figure S1). Aliquots were taken at intermediate time points and were checked for the presence of vesicles using DIC microscopy. Unless and otherwise mentioned, all of the experiments were performed at least in triplicates.
The water samples that were collected from the three different hot springs (from their sites of origin) were analysed to discern the major ions that were present in them and to also quantify their concentrations. These measurements were carried out following standard protocols . Briefly, the alkalinity of the samples was measured using an auto-titrator Titrino plus 877 (Metrohm, Switzerland). The concentrations of the major cations (Na+, K+, Ca2+, Mg2+ and Li+) and anions (Cl−, SO42−) were measured using ion chromatography instrument Compact IC plus 882 (Metrohm, Switzerland). The accuracy and precision of these analyses were regularly monitored and were found to have average values of ±4%. The net inorganic charge balance (NICB) for these samples were within ±10% (Results Section 3.5), thus ensuring good data quality.
Analysis of PU, CH, and PA water samples showed that all of them had pH around 8.3 to 8.65 (Table S1). It is known that certain fatty acids form vesicles in this pH range, and given their importance in the formation of plausible prebiotic compartments, we set out to study whether they could also form vesicles in these hot spring waters. The experiment was performed using different combinations of oleic acid (OA) and its derivatives, oleyl alcohol (OOH), and monoolein (GMO). This mixed fatty acid system was also selected because it has been widely used to study the effect of different parameters on the formation and stability of plausible prebiotic membranes.
Out of the four combinations of oleic acid and its derivatives, only OA + GMO (6 mM; 2:1 ratio) formed vesicles in the hot spring waters (Figure 1). The number of vesicles that formed qualitatively seemed more in PU followed by CH and PA, possibly owing to the differences in their ionic make up and concentration of the various species therein. Most of the resultant vesicles were large multilamellar or multivesicular vesicles. In the case of PA, a white precipitate was observed on the walls of the reaction container, which dissolved and resulted in vesicles only after prolonged heating. Additionally, in all of the hot spring samples that were analysed, there was always a mixture of vesicles and oil droplets that were observed, with the vesicles staying in the solution phase while the oil droplets adhered to the glass surface of the slide (Please refer to Appendix A for more details about the criteria that was used to distinguish between vesicles and oil droplets). However, in case of 200 mM bicine buffer pH 8.5, which was used as a positive control, vesicles were mainly observed with very few droplets present on some occasions.
Oleic acid by itself (6 mM), or when present as a mixture of OA + OOH (6 mM; 2:1 ratio), did not form vesicles in the hot spring waters, although they could form vesicles in bicine buffer (Figures S2 and S3).
Interestingly, a tertiary system composed of OA + OOH + GMO (6 mM; 4:1:1 ratio) also was unable to form vesicles in the hot spring waters, where only oil droplets were observed (Figure 2 and Figure S4). The addition of alcohol seemed to have a destabilizing effect on vesicles that otherwise could potentially form when only the OA + GMO combination was used. Once again, this effect was observed only in the hot spring waters and not in bicine buffer, wherein the tertiary system readily formed vesicles (Figure S4).
Small chain fatty acids (C8–C12) are thought to be more plausible on the early Earth as they can be synthesized by Fischer-Tropsch-type reaction under hydrothermal conditions , and have also shown to be present in meteorites [18,19]. Therefore, the vesicle formation behaviour of a small chain system was also investigated in the hot spring water samples. 10-Undecenoic acid (C11:1) was selected for this experiment to study the effect of reducing the chain length on vesicle formation ability, while keeping the unsaturation in the carbon chain similar to oleic acid and its derivatives. It is important to note that although there are few studies on vesicle formation by 10-Undecenoic acid , to our knowledge, this study reports for the first time membrane assembly by this particular fatty acid system in the context of origins of life studies. Results that were obtained were comparable to the oleic acid system, where 10-undecenoic acid (UDA) alone (90 mM), or its combination with undecanoyl alcohol (UDOH) (90 mM; 2:1 ratio), or the tertiary system composed of UDA + UDOH + monoundecanoin (UDG) (90 mM; 4:1:1 ratio), did not form vesicles in the hot spring waters. Only oil droplets were observed in all three of the aforementioned cases in large numbers (Figures S5–S7). The combination of UDA and UDG (90 mM; 2:1 ratio) did form vesicles in PU and CH, as in the oleic acid system, though only oil droplets were initially observed in the PA system. However, prolonged heating of this binary system did induce vesicle formation in PA water (Figure S8). Even in this fatty acid system, all of the four combinations of UDA and its derivatives formed vesicles in 0.2 M bicine buffer pH 8, which served as the positive control (Figures S5–S8).
Subsequently, the focus was shifted to decanoic acid (DA) and its derivatives. This is one of the most extensively studied small chain amphiphilic system that has long been considered a plausible precursor of prebiotic membranes. Being a saturated fatty acid, this system also allowed for us to examine the effect of saturation on the vesicle formation ability in the hot spring water samples. Once again, as consistent with aforementioned results, DA alone (60 mM), or its combination with decanol (DOH) (60 mM; 2:1 ratio), did not form vesicles in the hot spring waters (Figures S9 and S10). However, DA + glycerol derivative of decanoic acid (GMD) (60 mM; 2:1 ratio) readily formed vesicles in all of the hot spring water samples that were tested (Figure S11). Interestingly, the tertiary system comprising of DA + DOH + GMD (60 mM; 4:1:1 ratio) showed peculiar structures in CH and PA, in which the vesicles were typically seen surrounding a central lipid aggregate (indicated by white arrows in Figure 3). This aggregate possibly could be rich in DOH that seems to get excluded from the vesicle forming components of the system, in the form of oil droplet. These structures were not observed in bicine or PU, which showed the presence of regular vesicular structures (top two panels of Figure 3). Table 1 summarizes the vesicle formation results in all the three fatty acid systems that were studied.
In all of the vesicle formation experiments detailed above, the solvent (bicine buffer/hot spring water) was preheated to 75 °C before adding to the lipid mixtures. This was to replicate the temperature conditions that were prevalent in the hot spring sample sites, and also to make sure that the temperature of the solvent is well above the melting temperature of the fatty acid system. In order to study the stability of mixed fatty acid vesicles at this high temperature, and in the presence of the multiple ions that are inherent in the samples, the vesicular solutions of OA and GMO (6 mM; 2:1 ratio) were prepared in the respective hot spring waters, and were heated at 75 °C for up to 7 h. The resultant vesicles were reasonably stable at this temperature in, both, the hot spring water samples, as well as in the bicine buffer control (Figure 4 and Figure S12). An interesting result was observed in case of the PA system where prolonged heating led to the dissolution of the starting white precipitate (mentioned earlier), resulting in the formation of large multilamellar vesicles that were observed after 3 h of heating (Figure S12).
DH-RH cycles, a prebiotically pertinent and prevalent feature, are quite common in terrestrial geothermal fields and have been shown to facilitate prebiotically important reactions, such as nonenzymatic oligomerization of nucleotides and the encapsulation of polymers into vesicular compartments . Furthermore, to the best of our knowledge, there are no reported studies on the stability of fatty acid vesicles under DH-RH conditions. Therefore, these conditions were simulated in the presence of constant gentle CO2 flow and the stability of mixed fatty acid vesicles was checked under these conditions (the early Earth environment has been thought to contain higher amount of CO2 ).
OA + GMO vesicles (6 mM; 2:1 ratio) were quite stable in 200 mM bicine buffer pH 8.5, even after seven cycles of DH-RH (Figure 5). In PU and CH, the number of vesicles observed decreased with an increasing number of DH-RH cycles, although few vesicles were observed, even after seven DH-RH cycles (Figure 5 and Figure S13). In the case of the PA system, the vesicle solution prepared in PA water was first heated for 3 h to maximize vesicle formation, and subsequently subjected to DH-RH cycles. Vesicles were unstable even after one DH-RH cycle, and small shiny structures were apparent in subsequent cycles (Figure S13). It is pertinent to note that the pH of the bicine buffer control and the hot spring water samples did not change significantly during the DH-RH cycles, excluding the role of pH on the effect that was observed. Furthermore, to check whether the stability of small chain fatty acid vesicles were affected in the hot spring water samples under DH-RH conditions, UDA + UDG vesicles (90 mM; 2:1 ratio) were prepared in PU and were subjected to DH-RH cycles. Vesicles were observed even after seven DH-RH cycles. Additionally, small vesicles tended to fuse together to form larger vesicles as the number of DH-RH cycles increased (Figure S14).
In some of the experiments, rehydration was carried out with 200 mM bicine buffer pH 8.5, having 73.14 mM of [Na+], to test the inhibitory effect of high monovalent cation concentrations on vesicle stability. Typically, rehydrating with this bicine buffer would cause a two-fold increase in the amount of Na+ ion in the solution, with every cycle of rehydration. Vesicle aggregation was observed after five DH-RH cycles ([Na+] = 365.7 mM), (Figure S15). However, no aggregates were observed when rehydration was performed with milli-Q water, in which case the original concentration of Na+ remained constant throughout the DH-RH experiment. These results clearly illustrate the inhibitory effect of monovalent cations on vesicle formation when present in high concentrations.
In order to understand the role of ions on the vesicle forming capability of fatty acids, the ionic composition of all the hot spring samples was measured (Table 2). Multiple ions, in varying concentrations, were detected. Overall, the ionic strength of PU was higher than that of CH, followed by PA. The chemistry of the hot spring samples analysed in this study is dominated by monovalent anions (HCO3− and Cl−) and cations (Na+ and K+). In these samples, alkalinity accounted for more than 50% of the total anions, whereas Na+ and K+ concentrations together accounted for more than 90% of the total cations. These chemical patterns are consistent with earlier data reported for hot spring water samples that were collected from these or other near-by locations . Higher concentrations of Na+ and K+ in these samples confirm the intense weathering of silicate minerals that are present within the basins. Total dissolved solids (TDS) is a measure of chemical weathering intensity in a given basin. It is estimated as the summation of all the major constituents (in mg/L units) available in spring or river water. TDS is generally measured using the chemistry of filtered aliquots of the samples, and, therefore, reflects the total amount of solutes that are present in the hot springs. In the absence of SiO2 data, we could not compute the TDS values for the samples investigated herein. However, earlier studies have shown that the average TDS values for thermal springs in these locations vary widely from 550 ± 29 mg/L to 2116 ± 230 mg/L, with the highest concentrations being observed for the samples from Puga [22,23]. This may be because the Puga spring water that is situated close to the Indus Tsangpo Suture Zone has a deeper (5–7 km) source of fluids, which may promote intense chemical weathering .
The concentration of major divalent (Mg2+ and Ca2+) and monovalent (Na+ and K+) cations, however, were lower (even in PU) than what has been reported to disrupt vesicle formation . In this previous study, these ions had been individually tested for their vesicle disruption ability. However, not much is known about their combined effect on vesicle stability. Therefore, it is possible that these multiple ions (and their varying concentrations) might be causing a combinatorial effect on both vesicle formation and their stability in the hot spring water samples. Interestingly, this effect was seen across all of the fatty acid systems that were tested. Additionally, the possibility of the presence of organic compounds in these hot spring waters cannot be excluded, which could have also affected the vesicle formation process.
Results obtained in this study showed that the mixture of fatty acid and its glycerol derivative was the most resilient system among all of the combinations that were tested. This binary system formed vesicles in all of the hot spring water samples with different ionic strengths. Also, these vesicles were stable at high temperature and under DH-RH conditions. The aforementioned results pertaining to vesicle formation was consistent across all the fatty acid systems that were tested; independent of varying chain length and saturation. The results pertaining to vesicle stability was consistent in both the C18:1 and C11:1 systems, irrespective of the chain length. Furthermore, it is important to note that all of the combinations of different fatty acids and their derivatives formed vesicles in bicine buffer, but only few of them could form vesicles in the actual hot spring water samples.
This study confirms previous observations that the fatty acid and its glycerol derivative seems to be the most robust system among all of the combinations that were tested . Importantly, this binary system also showed a vesicle forming capability under the formidable 'realistic' conditions of the hot spring samples that had multiple ions in varying concentrations. Furthermore, they were resilient when subjected to high temperatures and multiple DH-RH cycles. Additionally, these results corroborate the hypothesis that mixed fatty acid vesicles, with a stabilizing counterpart, like glycerol derivative, could have withstood the tough conditions that are encountered in prebiotic environments. The stabilizing effect provided by the glycerol derivative of fatty acid was consistent across all of the fatty acid systems that were tested including the short chained fatty acid, decanoic acid. This fatty acid is long enough to easily form vesicles , but also short enough to be found in relatively large abundance under prebiotic conditions, when compared with C18 or C16 systems [18,19]. Therefore, the formation of mixed fatty acid vesicles of DA and its glycerol derivative, under hot spring conditions, indicates that the C10 system could have readily served as membrane components of the first protocells in a terrestrial geothermal field.
Interestingly, when the fatty alcohol counterpart of a particular fatty acid was included in the relevant mixture, there was an inhibitory effect on the vesicle forming capability in certain fatty acid systems. It is important to note that this inhibitory effect from the addition of the alcohol derivative, on the vesicle formation by a tertiary system, was only observed in the hot spring samples (discussed in detail in the Results section). This was not observed in the control reactions where bicine buffer was used to maintain the pH. This result was initially surprising given that previous studies involving the alcohol derivative have demonstrated the formation of stable vesicles with mixed composition [15,25]. Also, vesicle formation by a heterogeneous mixture of fatty acids and their derivatives has been thought to be more prebiotically 'realistic' as the soup would have been a composite mixture of diverse types of amphiphiles. However, the significance of our aforementioned result from the fatty alcohol experiments highlights the fact that the stabilizing effect of individual surfactants need not always be additive in all scenarios. The only exception was the DA tertiary system that formed vesicles, even in the hot spring water samples, further strengthening the pertinence of this fatty acid system as plausible prebiotic membranes.
One other significant highlight of this study was to test the stability of mixed fatty acid vesicles at high temperature (prolonged heating) and under DH-RH cycles. It has been reported previously that vesicles made up of fatty acid and their glycerol derivative are thermostable under buffered conditions . In this study, we show that this holds true even under actual hot spring conditions, which further affirms the robustness of this system. Importantly, the stability of amphiphilic systems under DH-RH conditions has not been studied in great detail. There are very few reports on the stability of phospholipid vesicles under DH-RH cycles, in buffered conditions [10,11]. However, given the importance of fatty acids as plausible prebiotic compartments, it is pertinent to study their stability under DH-RH conditions. For the first time, we report that, just like in the case of phospholipid vesicles, the vesicles that are made up of fatty acid and its glycerol derivative continue to retain their vesicle forming capability for up to seven cycles of DH-RH in bicine buffer. However, this ability decreases under hot spring conditions, and also seems to vary across the different hot spring samples.
It is important to understand that the variation in aforementioned stability will be context dependent and will also be affected by the ionic composition (and strength) of the relevant scenario. This is in addition to the temperature of the niche and other crucial parameters that could potentially lead to inhibitory outcomes in certain instances. Towards this, preliminary geochemical analyses of the three hot spring water samples was undertaken to better understand the role of their ionic makeup on, both, the feasibility of vesicle formation and their subsequent stability in these 'analog' niches. The chemistry of a hot spring is mainly governed by the relative supply of solutes that result from the weathering of different rocks that are present in the basin. The high Cl− and Na+ concentrations in the hot spring water samples that were used in this study, indicate to a supply of these solutes from basinal brines and/or buried evaporite . The remaining Na+ in excess of Cl− (i.e., Na* [= (Na+) − (Cl−)]) in these samples, varies between 6500 µM and 14,310 µM. On an average, the Na* of these samples is about 77 ± 21% of their total Na+ concentration. The main source for Na*, K+ and Li+ in these hot springs is the chemical weathering of silicate minerals. Therefore, a major part of the measured alkalinity must have a silicate origin. Furthermore, the absence of prominent amounts of Mg2+ in these samples confirms the insignificant supply of solutes that come from the weathering of dolomites and Mg-rich silicates. The spring water chemistry seems to be, by and large, regulated by weathering of silicate rocks (garnetiferous micaschist, dark phyllites, light quartzite, para gneisses, etc. ) that are present in the area. However, it is important to note that the ionic content of a hot spring depends on several factors, like geographical location, type of rock, site of collection, etc. Therefore, the ionic content of hot springs that were analyzed in this study could be different from that of other well studied hot springs, like those in Yellowstone National Park .
In conclusion, this work highlights the importance of testing laboratory 'buffered' reactions under more 'realistic' conditions. Our results clearly indicate how things could dramatically change when one moves from laboratory conditions to more prebiotically pertinent scenarios. All four combinations of fatty acids and their derivatives formed vesicles in bicine buffer controls. However, only the combination of fatty acid and its glycerol derivative could significantly form vesicles in the hot spring water samples. This indicates that terrestrial geothermal fields might have allowed for 'selection mechanisms' that would have allowed for the sustenance of certain types of reactions, and mixtures, over others, depending on the physicochemical constraints that are at play in that specific niche. These observations also underline the fact that much work remains to be done to systematically characterize various planetary analogs that might have allowed for (and sustained) chemistries that eventually would have led to the transition from chemistry to biology. Importantly, some of the observations from our stability experiments seem to have implications for discerning what biochemical signatures might survive under a certain set of environmental constraints. This, we believe, should be factored in while evaluating the habitability aspect of any planetary analog. Additionally, a logical extension of this work would be to systematically study the nonenzymatic polymerization of mononucleotides under 'realistic' hot spring conditions. Furthermore, it would be interesting and important to characterize how various prebiotically relevant co-solutes, including nucleic acids, amino acids, and other pertinent polymers, might have affected the stability of mixed fatty acid vesicles, under these early Earth and planetary analogous conditions.
The following are available online at www.mdpi.com/2075-1729/7/4/51/s1, Figure S1: Schematic of DH-RH experiment, Figure S2: Formation of vesicles by OA, Figure S3: Formation of vesicles by OA and OOH binary system, Figure S4: Formation of mixed fatty acid vesicles from the tertiary system comprising of OA, OOH and GMO, Figure S5: Formation of vesicles by UDA, Figure S6: Formation of vesicles by UDA and UDOH, Figure S7: Testing the formation of mixed fatty acid vesicles by UDA, UDOH and UDG, Figure S8: Formation of vesicles by UDA and UDG, Figure S9: Formation of vesicles by DA, Figure S10: Formation of vesicles by DA and DOH, Figure S11: Formation of vesicles by DA and GMD, Figure S12: Temperature stability of OA and GMO mixed vesicles in CH and PA, Figure S13: Stability of OA and GMO mixed vesicles in CH and PA under DH-RH conditions, Figure S14: Stability of UDA and UDG mixed vesicles in PU under DH-RH conditions, Figure S15: Effect of rehydration with bicine buffer on vesicle stability, Table S1: Collection of water samples from hot springs at Ladakh, Table S2: Fatty acids and their derivatives used in the experiment.
The authors M.P.J. and S.R. wish to thank DST-SERB, Government of India, for financial support in the form of a grant (EMR/2015/000434). G.R.T. and A.S. wish to acknowledge DST-INSPIRE Faculty award (DST/INSPIRE/04/2013/000050). M.P.J. and A.S. are grateful for graduate fellowship (JRF) support from UGC, Government of India. The authors also wish to thank IISER Pune Microscopy Facility and Vijay Vitthal for their support with imaging. Special thanks to David Deamer, Niraja Bapat and Chaitanya Mungi for their critical comments. Importantly, the authors wish to convey their sincere gratitude to the whole team that made Spaceward Bound India expedition to Ladakh a reality. It was crucial to the conception and execution of this project. Finally, thanks to the Department of Biology at IISER Pune for their financial and other forms of support.
M.P.J. and S.R. conceived and designed the experiments; M.P.J. performed most of the experiments in the manuscript; G.R.T. and A.S. performed the geochemical analysis experiments; M.P.J., G.R.T. and S.R. analyzed the data; M.P.J. and S.R. wrote the paper with crucial inputs from G.R.T. for sections pertaining to the geochemical analysis and related aspects.
There are a few different methods of making fatty acid vesicles. The most commonly used method is that of vesicle formation by titration. In this method, the pH of the solution containing the fatty acid is made alkaline, at which point all the fatty acids are present in the form of micelles. Subsequently, the pH is slowly reduced to match the pKa of the fatty acid, at which point formation of vesicles is facilitated. Further reduction of the pH to below the pKa of the fatty acid results in the formation of disordered aggregates of fatty acid, referred to as oil droplets. This titration method of vesicle formation is of particular interest because it is assumed to maximize vesicle formation. However, in this study, we did not want to alter the original pH of the hot spring water samples. Therefore, we used a second method of vesicle formation wherein an aqueous solution is added to a dry film of fatty acid molecules, which results in the spontaneous formation of vesicles. It is pertinent to note that during this process all kinds of higher order structures can form simultaneously, ranging from micelles to vesicles to droplets. Micelles and small unilamellar vesicles (0.1–0.2 µm in diameter) are below the resolving power of light microscope. Therefore, we cannot observe them using DIC microscopy at 40× objective (which was used in this study). Large vesicles (diameter of 1 µm and above) appear mostly as donut shaped structures that vary in size and shape. Also, there is not much contrast between the vesicle and its surrounding medium. Additionally, there could also be a mixture of large unilamellar vesicles, multilamellar vesicles and multivesicular vesicles in these solutions. Oil droplets, on the other hand, can be seen as shiny structures with a relatively narrow distribution of size and shape. They appear as convex shaped circles with a two phase appearance within the same structure; one is extremely shiny and other is dark. Figure A1 shown below will enable readers to distinguish between oil droplets and vesicles.
Figure A1. Distinction between vesicles and oil droplets. (A) Vesicles appear as donut shaped structures having very less contrast when compared with the surrounding medium. (B) Oil droplets typically appear as shiny convex circles. (C) One can, therefore, distinguish between vesicles (indicated by white arrows) and droplets (indicated by black arrows) in the same mixture based on their appearance.
Thompson, J.M.; DeMonge, J.M. Chemical Analyses of Hot Springs, Pools, and Geysers from Yellowstone National Park, Wyoming, and Vicinity; Open-File Report 96-68, 1-67; U.S. Department of the Interior; U.S. Geological Survey: Denver, CO, USA, 1996.
Figure 1. Oleic acid (OA) + monoolein (GMO) system (6 mM; 2:1 ratio): Vesicles formed in 200 mM bicine buffer as well as in all the hot spring water samples i.e., in Puga (PU, top right), Chumathang (CH, bottom left) and Panamic (PA, bottom right), respectively, as shown in the above figure.
Figure 2. Combination of OA, oleyl alcohol (OOH) and GMO (6 mM; 4:1:1 ratio) formed vesicles in 200 mM bicine buffer pH 8.5 but not in Puga (PU).
Figure 3. Combination of decanoic acid (DA), decanol (DOH) and glycerol derivative of decanoic acid (GMD) (60 mM; 4:1:1 ratio) formed vesicles in 200 mM bicine buffer pH 8 and also in Puga (PU), as indicated in the top two panels. Interesting structures were observed in the Chumathang (CH) and Panamic (PA) systems (bottom two panels), in which vesicles were present surrounding a central lipid aggregate (indicated by arrows in the respective panels).
Figure 4. Temperature stability of OA and GMO (6 mM; 2:1 ratio) mixed vesicles in 200 mM bicine buffer pH 8.5 (top two panels) and in Puga (PU, bottom two panels). Vesicles were observed in both the systems, albeit to varying extent, even after heating the reaction samples at 75 °C for 7 h.
Figure 5. Stability of OA and GMO mixed vesicles (6 mM; 2:1 ratio) under DH-RH conditions in 200 mM bicine buffer pH 8.5 (top two panels) and in Puga (PU, bottom two panels).
Table 1. Vesicle formation experiment results in 0.2 M bicine buffer and Puga (A), Chumathang and Panamic (B) water samples that were tested using three different fatty acid systems, as indicated. Presence of vesicles is denoted by (✓) while (✗) denote their absence. The pH of bicine buffer control was 8.5 for oleic acid (OA) system and 8 for undecenoic acid (UDA) and decanoic acid (DA) systems.
# Vesicles formed after prolonged heating.
Table 2. Geochemical analysis of hot spring water samples under study. The overall ionic strength decreases from Puga to Panamic. The net inorganic charge balance (TZ+/TZ−) around 1 indicates the high quality of analysis.


We all basically know what we need to do to become healthier. A new study offers some insights into how to get us to do it.
Get up off the couch and go buy some fruit and vegetables. According to a recently published study, that's a road to better health that's easiest for most people to take. A little financial incentive doesn't hurt either.
It's hardly news that Americans eat too much saturated fat, too little fruit and vegetables, sit too much and don't exercise nearly enough. But it would be news if someone found out to get them to change these habits for the better. And researchers at Northwestern University may have found the way.
As an incentive, the participants could earn $180 for meeting their goals for the two habits they were trying to improve during the first three (target) weeks of the study.
The researchers looked at people with all four of these unhealthy habits who wanted to change them. And they tested which pair of behaviors--one dietary and one physical--was easiest for people to improve. The pairs were increase fruit/vegetables and sit less, increase fruit/vegetables and increase exercise, lower saturated fat and sit less, and lower saturated fat and increase exercise.
By far, people found it easier to decrease their couch time and increase their fruit and vegetable intake, making major changes within three weeks. These changes persisted, though not as strongly, for a full 20 weeks, the length of the study. As a bonus, people also lowered their saturated fat intake slightly.
It might just be time to change the formula for better health from "eat less and exercise more" to "get up off the couch and start eating more fruit and vegetables."
As an incentive, the participants could earn $180 for meeting their goals for the two habits they were trying to improve during the first three (target) weeks of the study: eating five or more servings of fruit and vegetables daily, lowering saturated fats to below 8% of their daily calorie total, decreasing sedentary leisure time to under 90 minutes per day and/or increasing moderate or vigorous physical activity to at least 60 minutes per day. All information was self-reported.
After the first three weeks, participants could earn an additional $30-$80 by continuing to upload data to the researchers for the next 17 weeks (follow-up), though there was no longer any need to actually meet any dietary or activity goals to receive this payment.
For the decrease sitting/increase fruit and vegetable group, their fruit and vegetable consumption started at 1.9 servings per day, rose to 5.5 during the three week target phase and dropped off to 2.9 during the 17-week follow-up phase. Their sedentary leisure time went down from an initial 219.2 minutes to day to 89.3 during the target phase and 125.7 during follow-up. And their saturated fat consumption as a percent of daily calories went from 12% to 9.4% to 9.9%.
In other words, people were able to make significant healthy changes during their first three weeks. And while these improvements slacked off a bit afterwards, they still remained 17 weeks later, five months after the start of the trial.
Improvements were also seen in the other three test groups, but they weren't as large.
Every hour you're not sitting on the couch watching TV is an hour that you're not eating chocolate chip cookies, chips or the other tempting snacks that are a standard part of TV time. And while the researchers didn't actually test this, it seems reasonable that less couch time is the first step towards an overall healthier lifestyle and probably the easiest step to take.
So get up off the couch and go invest in some fruit and vegetables.
An article on the study appears in Archives of Internal Medicine.


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Sustained Virological Response to PegIntron Plus Ribavirin in Asian and Hispanic Patients
Retrospective examination of available data strongly suggests that Asian patients experience a higher rate of sustained virological response (SVR) to interferon plus ribavirin therapy for chronic hepatitis C compared with Caucasian patients, while Hispanic patients have a lower SVR than Caucasians. It is well established that African-American patients have poorer response than other racial/ethnic groups.
In the current study, presented at the 57th AASLD annual meeting taking place this week in Boston, researchers at 17 U.S. medical centers* evaluated SVR rates in Asian and Hispanic patients with chronic hepatitis C enrolled in the WIN-R trial, a large U.S. multicenter study comparing fixed dose (FD) vs weight based dosing (WBD) of ribavirin combined with pegylated interferon alfa-2b (PegIntron).
Patients were randomized to receive PegIntron 1.5 mcg/kg once weekly combined with ribavirin 800 mg (FD) or 800-1400 mg/day (WBD). Treatment was for 48 weeks, with 24 weeks follow-up.
The SVR rate was based on intent-to-treat (ITT) analysis and defined as undetectable HCV RNA by central quantitative TaqMan assay (lower limit of detection < 29 IU/mL or 100 copies/mL) 24 weeks after the completion of treatment.
Dose reductions of ribavirin were required for hemoglobin (Hgb) levels < 10 g/dL and discontinuation for Hgb < 8.5. Erythropoietin (EPO) was permitted concomitant with ribavirin dose reduction for Hgb < 10.
Approximately 5000 patients were randomized in this trial.
The present report concerns 65 Asian patients (genotype 1=37; genotypes 2/3=25) and 251 Hispanic patients (genotype 1=166; genotype 2/3=77) who received at least 1 dose of ribavirin and were ? 65 kg.
Of the 65 Asians, 34 (52.3%) achieved SVR vs 45.7% of Caucasians (P = 0.29).
" SVR rate for Asians with genotype 1 was 15 of 37 (40.5%) vs 34.9% of Caucasians (P = 0.48).
SVR rate for Asians with genotype 2/ 3 was 18 of 25 (72%) vs 61.6% of Caucasians (P = 0.29).
SVR rate for Asians receiving ribavirin WBD was 21 of 32 (66%) vs 13 of 33 (39%) who received ribavirin FD (P = 0.04).
Of the 251 Hispanic patients, 84 (33.5%) achieved SVR vs 45.7% of Caucasians (P = 0.0002)
Hispanics were 1.7 times less likely to achieve SVR than Caucasians.
SVR rate for 159 Hispanics who finished a full course of treatment was 46.5% vs 55.3% for Caucasians (P = 0.0006).
SVR for Hispanics with genotype 1 was 44 of 166 (24.1%) vs 34.9% for Caucasians (P = 0.005).
SVR for Hispanics with genotype 2/3 was 41 of 77 (53.6%) vs 61.6% for Caucasians (P = 0.14).
SVR for Hispanics receiving ribavirin WBD was 39 of 122 (32%) vs 45 of 129 (35%) receiving ribavirin FD (P = 0.6).
Based on these findings, the authors concluded, "Compared to Caucasian patients, [PegIntron] 1.5 mcg/kg results in a significantly lower SVR for Hispanic patients by ITT analysis and for those who completed therapy."
They added that, "SVR for Asian patients vs Caucasian patients trended higher, but was not statistically significant."
This study was supported by Schering-Plough Corp.
*Baptist Medical Center, Kansas City, MO; University of California Irvine, Orange, CA; Weill Cornell Medical College, New York, NY; Columbia Presbyterian Medical Center, New York, NY; Beth Israel Deaconess Medical Center, Boston, MA; Indiana University School of Medicine, Indianapolis, IN, USA; Atlantic Gastroenterology Associates, PA, Egg Harbor Township, NJ; Austin Gastroenterology, Austin, TX; East Bay Liver Clinic, San Francisco, CA; Indianapolis Gastrointestinal Research Foundation, Indianapolis, IN; Bach and Godofsky, MD PA, Bradenton, FL; Northwest Georgia Gastroenterology Associates, PC, Marietta, GA; North Shore Long Island Jewish Health System, Manhasset, NY; Northwestern University, Chicago, IL; Gastroenterology Consultants, PA, Houston, TX; Albert Einstein Medical Center, Philadelphia, PA; Schering-Plough Research Institute, Kenilworth, NJ;
B Freilich, K Hu, I Jacobson, and others. Prospective Analysis of Sustained Virologic Response (SVR) to Peg-interferon (PEG IFN) Alfa-2b and Ribavirin Treatment in Asian and Hispanic Patients with Chronic Hepatitis C: Results from the WIN-R Trial. 57th AASLD. October 27-31, 2006. Boston, MA. Abstract 349.
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Richard G. Frank
Economics and the Transformation of the Mental Health System
Sherry Glied, Richard G. Frank
Measuring the Economics of Health
Journal of Health Politics, Policy and Law (1 June 1996) 21 (3): 617–624.
Regulatory Policy and Information Deficiencies in the Market for Mental Health Services
Per Case Prospective Payment for Psychiatric Inpatients: An Assessment and Alternatives
Richard G. Frank, Judith R. Lave
Journal of Health Politics, Policy and Law (1 February 1986) 11 (1): 83–96.
DOI: https://doi.org/10.1215/03616878-11-1-83
Mental Health Care and National Health Insurance
Journal of Health Politics, Policy and Law (1 February 1985) 10 (1): 195–197.
Medical Costs, Moral Choices
Journal of Health Politics, Policy and Law (1 June 1984) 9 (3): 529–532.
Professionalism and the Public Interest: Price and Quality in Optometry


For the last generation or two, modern society has been pulling away from acknowledging the reality of sin. Yet, when people believe that God's law is no longer valid, they deceive themselves. Martin Collins surveys scriptures that urge Christians to admit. . .
Martin Collins, noting that the foundational way of life as outlined by Jesus Christ is not much followed in mainstream Christianity, and observing that the five foolish Virgins also belonged to the visible church, reminds us that we are only Christ's if w. . .
John Ritenbaugh insists that we must be aware of our awesome status as a unique, called-out, chosen, royal priesthood—teachers of a way of life and builders of bridges between people and God. Because God owns us, we differ from the rest of the people. . .
In this sermon focusing on meekness and forgiveness, John Ritenbaugh indicates that when we are sinned against, our ego gets extremely strong and our emotions get muddled, making it difficult to give forgiveness. Because God is the Creator of everything, o. . .


The planned move, however, is not without opposition. Justin Trudeau, Liberal Party leader, along with Tom Mulcair, head of the New Democratic Party (NDP), have publicly voiced their stand against the tax credit. Lindsay Tedds, an economist with the University of Victoria, says that it may not actually help northern mining. "It does not increase investment to the point where you're going to have increased exploration," she reiterates.
Introduced in 2006, the 15 percent Mineral Exploration Tax Credit was meant to help companies raise capital by providing investors with an incentive to finance mineral exploration. The credit is an appendage to the deduction provided to the investor for the exploration expenses "flowed through" by the company that issues the shares. According to the Conservative Party website, the tax credit has helped mining companies raise over $5.5 billion for exploration since its inception.
Whether the 15 percent tax credit is an effective policy or not, the idea to encourage more mining exploration is a sound one. When mineral deposits that exhibit great potential are discovered, mine development soon follows. In turn, new mines require underground equipment such as, skips, cages, loading systems, rail haulage cars, mine car wheels, etc. which is good for the mining supply and services sector.
Top mining equipment manufacturers, like Wabi Iron & Steel Corp., can supply superior conveyance systems for your mining operation. Companies such as Wabi have the engineering capabilities and fabrication facilities that enable them to produce quality equipment, from wheels for mining cars to loading and dumping systems, skips and cages.


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La bataille de Turbigo est un épisode de la seconde guerre d'indépendance italienne. Elle se déroule le 3 juin 1859 à Turbigo.

Le 2 juin, la division de voltigeurs de la Garde du général Camou atteint Turbigo, suivi du  corps d'armée du général Mac Mahon le jour suivant. Le reste de l'armée se concentre autour du Novara. Le village de Turbigo est inoccupé, mais dénué de pont. Les sapeurs du général Camou travaillent toute la nuit pour construire un pont sur le Tessin qui sera finalement effectif le 3 juin à l'aube, couvert par cinq batteries de six canons. Le  corps d'armée de Mac Mahon traverse le ponton de Turbigo pour consolider la tête de pont. Il continue en direction du village de Robecchetto, où le  de tirailleurs algériens s'illustre en pourchassant une colonne autrichienne explorant tardivement le secteur au nord de Magenta. Robecchetto est aux mains des Français à 14h30. Les trois divisions de Mac Mahon bivouaquent autour de Turbigo, prêtes à s'élancer le long de la rive gauche du Tessin le lendemain matin.

Bibliographie
 .

Article connexe
 Pont de Turbigo

Notes et références

Histoire de la Sardaigne
Histoire de la Lombardie
Bataille de la campagne d'Italie (1859)
Turbigo
Risorgimento
1859 en Italie
Bataille de 1859
Juin 1859
Turbigo

redpajama_set_name: RedPajamaWikipedia


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She Started an Initiative to Help Women in Domestic Violence Shelters — It Grew Into A Sustainable Apparel Business
Hayley Santell is the founder of MADI Apparel, a social enterprise fashion line that donates to a woman in need for every purchase made.
By Victoria Flexner
When Hayley Santell learned that someone close to her had been a victim of domestic violence, she knew instinctively that she wanted to help. But Santell also wanted to find a way to help other women who were suffering or had suffered from domestic violence. She did research and found that underwear tops the most urgent needs list of nearly every domestic violence and homeless shelter and rape crisis center. What has emerged since is MADI Apparel, a line of staple basics for women and men made from bamboo. With each purchase, MADI Apparel donates a new pair of underwear to women in need. Today the Kansas City, Missouri entrepreneur is taking pride in the 7,5000 pairs of underwear they have donated so far and continuing to grow her eco-friendly social enterprise.
Santell's story, as told to The Story Exchange 1,000+ Stories Project:
A few years back I leared that a close family member had been a victim of domestic violence. She was a strong, independent women, and it was difficult to believe that she had been a victim for three years. I wanted to find a way to make a difference in the lives of women who suffer the ultimate abuse, abuse from someone they love. After researching the needs of domestic violence shelters, my path and my passion became clear. I began with an Indiegogo fundraiser to raise money for the first production run of six styles of MADI underwear in one color. Since then, I've added new colors, and a full line of basic staples for women and men from underwear, sleep tops and bralettes, to skirts, joggers dresses and long underwear, and men's jockeys.
My definition of success is at its core helping others and inspiring others to do the same. Close your eyes and picture a world where businesses all did something to benefit others or the environment, even if that "something" was small. What if every person in this dream world made a commitment to help others and beam positivity on a daily basis? We can get there if we all do our part.
There are so many success stories to share, especially the financial success of the men's wear launch during the pandemic. But one story in particular is very close to my heart. A few months ago, I received an email from a Kenyan woman named Beth who is the founder of the non-profit Smart Child Kenya. She helps to rescue girls from FGM and early childhood marriages and fights for their education by looking for scholarships for their primary and secondary schools. She also told be about how many girls miss and drop out of school because of a lack of menstrual products. MADI Apparel has since started donating panties to Beth's organization.
The postal system in Kenya, however, is rather difficult. Smart Child Kenya is in a remote area a few hours outside of Nairobi. We had a tough time getting the underwear there. But in the end we found a way. A few weeks later, Beth sent me a photo of the women opening their packages. They were elated. This really made my year.
[Related: Meet the Fashion 'Trash Nerd' Who's Won Money and Love from Vogue]
To be honest, I knew absolutely nothing about everything involved in starting a fashion business from the ground up. At 24 years old, I was fresh out of college and fresh out of money. I had zero experience in business management, money management, sourcing fabrics, manufacturing, designing products, opening a store, launching a website, managing employees, reporting taxes or scaling and driving product sales. Literally, name an area, and I promise I hadn't done it before.
My first big challenge was fundraising $20,000 to pay for the first manufacturing run of products and launch online sales. I used my circumstances and my gifts to my advantage. I networked through building genuine relationships with locals, travelers, business owners and even a few investors. I told everyone I met about what I was starting. I launched an Indiegogo crowdsourcing campaign with a $14,000 goal, and I worked by butt off to spread the word. The restaurant where I worked even let me host a local fundraiser. Between the Indiegogo campaign, the local fundraiser and help from close family, I started my business debt-free with over $25,000.
For perspective, ethical fashion and sustainable fashion brands weren't really a thing back in 2013. TOMS Shoes was really the first leader in the social impact business world. Then, other giving brands started sprouting up. Hardly any clothing brands were making their products in the U.S. anymore; the bigger brands had years prior moved their production to mostly Asian countries where labor was cheaper and less ethical. So, it wasn't easy finding a U.S. based cut-and-sew team, and we had to do a ton of homework. No one was really working with organic or biodegradable fabrics at the time either, so we were on our own a bit here, too. But that's what you sign up for when you commit to entrepreneurship — a lifetime of facing a steep learning curve and tackling it over and over! You fall on your face, you get back up, you climb the mountain and then you do it all over again.
There weren't (and still aren't that we know of), any brands doing what we are — making a difference in every element of the business model. This is similar for many new businesses; you may very well find yourself comparing your business to others — feeling like a sole turtle in a sea of jellyfish. And that's okay. That's actually great. DO YOU and bring your gifts to the table as a leader. For everything else, find a team full of people who make you greater and stronger and lean on them.
[Related: Why This Nonprofit Cares About Periods. And How Meghan Markle Might Help]
I have had too many mentors and role models to name, but here are a few. My parents have led the way as my biggest cheerleaders since day one. They're both very intelligent, well-educated, business-savvy and have always encouraged me to follow my gut and heart. They have been my biggest investors — not only financially using personal funds to help push MADI Apparel along, but also investing in the day-in and day-out operations. My husband and I met a few years after I launched MADI Apparel, but he goes so above and beyond to help in any way. He's a musician and videographer, which comes in handy. He's a great creative influence in my life. He's always dreaming big and introducing new ideas to our business — like building a stage in our shop backyard to host community events.
The rest of my role models and supporters are made up of my extended family, my best friends, our board members, my team, investors, neighbors, and more. Other close friends and business owners in New Smyrna Beach and Orlando, Florida contributed to our grassroots humble beginnings. They volunteered their energy and time to help me fundraise, brainstorm, build a business plan, pop up merchandise at concerts and count inventory in my guest room closet. I met one of my early mentors, donors and board members, Roy Johnson, while bartending in Florida. He passed away in a bicycle accident a few years ago, and I think about how he lived his life to the fullest and put his all into everything he touched. It's because of his encouragement that we launched our non-profit subsidiary. It's also because of him that I look at sunsets a little differently. I'll always remember the time he took me to watch manatees in a kayak spot on the intercoastal that we both loved at sunset. He asked me to never forget the real reason I started MADI Apparel.
It takes a whole tribe to start a small business from scratch and to keep it moving. All of this is not because of me, it's because of them. I'm just a little ant in the equation.
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