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She's cute as a bunny, so give the little one an adorable rabbit outfit. It includes a bodysuit, bib, socks, and more. The outfit is perfect for a little kid who still can't hop around, but whose bright eyes are full of life. MoreBack to School Sale! All Duffle Bags are 25% Off! Enter "Duff25" in promocode box to receive the discount. Offer expires: September 1st, 2016.
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domain: artistic


text:
The hormone named leptin, being discovered 22 years ago, and named based on the Greek word leptos has been hailed as the key regulator of leanness. However, the key experiments that probe the function of this protein and unravel the precise mechanism of its action as a guardian against obesity.
Two decades ago, the scientists discovered the identity of a mouse obesity gene, finding that it encodes a previously unknown hormone made by the fat cells. This leads to a rare genetic deficiency, in which the people born with two defective copies of the gene are extremely obese, and their obesity is reversed by restoring their levels of leptin using daily injections.
Both in mice and people without the mutation, studies have shown that leptin plays and important role in regulating metabolism. Even though the researchers speculated that this protein plays a key role in helping healthy lean people remain thin. Paradoxically, obese mice and people how don’t have the defective obesity gene almost always have high levels of leptin.
The treatment with leptin has been approved in the United States since 2014, for the use in congenital leptin deficiency, as well as in an unusual syndrome of lipodystrophy, as the protein has not been readily available for clinical experiments.

domain: health


text:
DeVos Place is a premier convention center located in downtown Grand
Rapids. Nestled beside the Grand River, the picturesque riverfront
setting is the perfect backdrop for your convention, tradeshow, meeting or any event. DeVos
Place offers a unique combination of meeting and exhibition space to
accommodate a variety of events. Downtown Grand Rapids is located 15
minutes from Gerald R. Ford International Airport and easily accessible
from all major interstates.
The convention center features a
162,000 square foot, column free exhibit hall, 40,000 square foot
ballroom and 26 individual meeting rooms (32,000 square feet). Twelve
spacious loading docks (with a wash bay and marshaling area) on Michigan Street allow for
easy access into the exhibit hall. Two loading docks and a ramp on Lyon Street provide access to the ballroom.
In addition, DeVos Place
features a 2,543 seat performing arts theater,
DeVos Performance Hall, with two independent loading
docks and one loading ramp. Featuring performances from the Grand Rapids Symphony, Grand
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address. DeVos Place is part of a vibrant downtown entertainment
district featuring over 50 dining establishments, nightclubs, museums and the 12,000+ seat Van Andel Arena, all within walking
Immediately adjacent to DeVos Place, world-class
accommodations are available including the Four-Star Amway Grand Plaza
Hotel, JW Marriott and the Courtyard by Marriott. All hold more than
1,100 guest rooms and are within walking distance of DeVos Place, most
of them accessible by a climate controlled skywalk. Grand Rapids
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DeVos Place is managed by ASM Global, part of a unique portfolio of convention centers around the world.
DeVos Place is a member of the Better Business Bureau.

domain: geography


The dilapidated Columbia Road structure adjacent to the North Dorchester Burying Ground in Uphams Corner could be reborn.
Abandoned since 1977, the approximately 1,200-square-foot building, which sits on an approximately 2,300-square-foot lot, has housed many different establishments since it was constructed in 1912, including public restrooms and office space. Located next to the Atco Supply Company, the building has sat underused since the late 1970s, with a deteriorating foundation, collapsing roof, and fire-damaged interior.
The land and structure is worth a combined $84,000, according to the city’s Assessing Department.
On Wednesday, neighbors and community advocates gathered at the Strand Theater to discuss the property’s future.
The Department of Neighborhood Development, which currently manages the space, would like to sell the property and see it put to a productive use.
Over the coming months, the department will develop a Request for Proposals for the site. The Request for Proposals, or RFP, is DND’s standard process for selling publicly owned property. In addition to providing details about the property, the RFP lays out what neighbors would like to see at the site. Potential developers are required to complete a RFP.
“I’ve driven past this site for years and didn’t know it existed and didn’t know its history,” said Donald Wright, deputy director of real estate management and sales for DND. “It’s an exciting opportunity.”
While the property offers a number advantages to potential businesses--including a highly visible storefront on the bustling Columbia Road--it has a number of challenges.
In addition to the work it will take to rehabilitate the deteriorating building, DND isn’t quite sure of its historical status. The property doesn’t appear on any historic registries, and it isn’t landmarked. It, however, does abut the North Dorchester Burying Ground, Dorchester's oldest remaining landmark, according to the city of Boston.
Historical status would limit the possibility of new doorways or windows or demolition for new construction, according to DND representatives.
Although it will be some time before the site’s historic credentials are sorted out, neighbors had plenty of ideas for what the space could be used for including a café, bike shop, visitor center, and restaurant.
“I’d think we’d rather have something there that’s active and serves the public,” said Kit Binns, an area resident. “Although there are number of businesses in Uphams Corner, the neighborhood lacks plenty.”
Other suggestions floated by residents included an ice cream shop, bakery, and chicken and waffles restaurant.
“With all the great things happening in the neighborhood, this is just another thing that can add to the momentum,” said Max MacCarthy, executive director of the Uphams Corner Main Street, a business development non-profit.
While some suggestions included demolishing the building, others advocated for it to be saved or at least preserved in spirit.
“I’d like to see in the RFP that the current look be retained,” said Nancy Conrad, an area resident. “It gives an indication of what our community was like a 100 years ago.”
Although DND is not adhering to a strict timeline for the project, an RFP is expected to be out before the summer. Once the RFP is out, potential developers will have 60 days to submit their proposals. Once the deadline has passed, the highest scored applicants will be brought before the community before any developer receives a tentative designation as the site’s new owner.


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Ha-ra Goo on Netflix
This is a regularly updated list with movies, series and documentaries with Ha-ra Goo on Netflix. The best rated item with Ha-ra Goo on Netflix is "It's Okay, That's Love" and appeared on screen in 2014.
About Ha-ra Goo
Goo Ha-ra (January 13, 1991 – November 24, 2019), better known mononymously as Hara, was a South Korean singer and actress. She was a member of the South Korean girl group Kara, and had also appeared in television dramas including City Hunter (2011). She made her debut as a soloist in July 2015 with the release of her EP Alohara (Can You Feel It?). She was found dead at her home on November 24, 2019.
Below you find an overview of all movies and series with Ha-ra Goo on Netflix. We found 1 titles with Ha-ra Goo on Netflix.

domain: entertainment


La Vieille Voie topographic map
Click on the map to display elevation.
About this map
Name: La Vieille Voie topographic map, elevation, terrain.
Location: La Vieille Voie, Corravillers, Lure, Haute-Saône, Bourgogne-Franche-Comté, Metropolitan France, 70310, France (47.87790 6.62010 47.91790 6.66010)
Average elevation: 1,946 ft
Minimum elevation: 1,388 ft
Maximum elevation: 2,602 ft
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Click on a map to view its topography, its elevation and its terrain.


text:
Sunshine Coast Credit Union has selected Open Solutions's Digital Document Systems eSign, eReceipt and eReport solutions to streamline and automate business processes. Digital Document Systems is an Open Solutions business group that provides web-based information archive and delivery systems for financial institutions.
According to Open Solutions, with eSign, eReceipt and eReport, Sunshine Coast will now have the ability to automatically store, index and retrieve receipts; eliminate paper-based processes by incorporating electronic signatures for nearly every document requiring a signature within the institution; and electronically archive data from core and item processing systems to better manage complex, secure system administration and reporting such as polling, job scheduling and archive retrieval.
An existing Open Solutions client running Open Solutions’ relational core processing platform, the credit union added the new functionality to electronically capture and deliver key data including receipts and signatures, as part of its overall strategy to move to a paperless operating environment.
Shelley McDade, CEO of Sunshine Coast Credit Union, said: “We are committed to moving to a more efficient, paperless environment. The implementation of these products is a critical step to meeting our long-term goals. We looked at several providers and Open Solutions was the only company we felt had the proven industry reputation, as well as the resources needed to meet our needs. We are looking forward to the advantages we’ll gain in terms of improved member service, increased workflow efficiencies and improved productivity.”

domain: news


text:
Matthew Fletcher, ’97, the Harry Burns Hutchins Collegiate Professor of Law, recently published a story in the graphic novel collection A Howl: An Indigenous Anthology of Wolves, Werewolves, and Rougarou (Native Realities Press, 2022). The story, “Lone Wolf,” centers on a Native American lawyer.
Fletcher, a member of the Grand Traverse Band of Ottawa and Chippewa Indians, teaches and writes in the areas of federal Indian law, American Indian tribal law, Anishinaabe legal and political philosophy, constitutional law, federal courts, and legal ethics.
He sits as the chief justice of the Pokagon Band of Potawatomi Indians and the Poarch Band of Creek Indians. He also sits as an appellate judge for the Cabazon Band of Mission Indians, the Colorado River Indian Tribes, the Grand Traverse Band of Ottawa and Chippewa Indians, the Hoopa Valley Tribe, the Lower Elwha Klallam Tribe, the Mashpee Wampanoag Tribe, the Match-E-Be-Nash-She-Wish Band of Pottawatomi Indians, the Nottawaseppi Huron Band of Potawatomi Indians, the Rincon Band of Luiseño Indians, the Santee Sioux Tribe of Nebraska, and the Tulalip Tribes.
Below, he answers five questions about his passion for comics, his burgeoning career in comics, and how he incorporates graphic novels into his teaching.
1. How did you get involved with this anthology project?
I started doing my own comics during the pandemic. I began what I called a “plague journal” every day for the first several months of lockdown using a basic comics app. I just kept going after that.
I was approached about this project because I’m a friend of Beth LaPensee, who edited the anthology. I had previously published a short graphic story called “Snapping Turtle Goes to War” in a different collection.
2. Can you talk a little bit about the story you wrote and how you developed it?
I’m interested in developing comics with specific themes, which include lawyering for Indigenous peoples, Indigenous peoples in academic spaces, Star Wars, and the state of Michigan. I try to explore the lived reality and experiences of Native people who are lawyers and law students.
“Lone Wolf” is about Indian lawyering, specifically about young-ish Native lawyers who work at large firms, and more specifically about a practice known as “window lawyering,” where Native attorneys are used by non-Native partners for marketing but not for the work. It’s partially inspired by Jim Harrison's novel Wolf, which was made into a 1994 movie starring Jack Nicholson.
I was excited to work with Dale DeForest, who did the art for the story. I'm super lucky to have worked with him because he's a really big deal.
3. What do comics mean to you, and how do they relate to your life and work?
Comics are just fun and a great diversion. I use my own amateur comics to teach my Federal Indian Law and Tribal Law classes. I use the comics to detail the principles I want the students to take away from the reading and also add in what I call “historical gossip” about the cases. For talks and presentations, I prefer comics over powerpoint-type presentations.
4. What do you hope your students take away from the comics you show them?
In my federal Indian law classes, I use the cover pages of various comics I’ve generated to give students the answers to my Socratic-type questions before class, and then I turn it back to them to ask questions. This method is known as a “reverse Socratic.” I’ve noticed that once I give them the key takeaways from the reading, the class discussion becomes far more engaging and useful.
5. What’s next for your writing projects?
A few years ago I wrote a manuscript for a book called Windigo Killers that has been adapted into a graphic novel. It is in the early stages of being shopped around to publishers.

domain: literature


What is it about?
An international multi-disciplinary group of 24 researchers met to discuss ocean acidification (OA) during the Brazilian OA Network/Surface Ocean-Lower Atmosphere Study (BrOA/SOLAS) Workshop. Fifteen members of the BrOA Network (www.broa.furg.br) authored this review. The group concluded that identifying and evaluating the regional effects of OA is impossible without understanding the natural variability of seawater carbonate systems in marine ecosystems through a series of long-term observations. Here, we show that the western South Atlantic Ocean (WSAO) lacks appropriate observations for determining regional OA effects, including the effects of OA on key sensitive Brazilian ecosystems in this area. The impacts of OA likely affect marine life in coastal and oceanic ecosystems, with further social and economic consequences for Brazil and neighboring countries. Thus, we present (i) the diversity of coastal and open ocean ecosystems in the WSAO and emphasize their roles in the marine carbon cycle and biodiversity and their vulnerabilities to OA effects; (ii) ongoing observational, experimental, and modeling efforts that investigate OA in the WSAO; and (iii) highlights of the knowledge gaps, infrastructure deficiencies, and OA-related issues in the WSAO. Finally, this review outlines long-term actions that should be taken to manage marine ecosystems in this vast and unexplored ocean region
The following have contributed to this page: Dr Ruy Kenji P Kikuchi and Dr Nadine Schubert
In partnership with:


text:
GLOBAL - Baring Asset Management has launched a new Asian fund of hedge funds as investors continue to look East for opportunities.
Baring’s Asia Hedge Select Fund will invest predominantly in equity long-short Asia Pacific hedge funds and be managed by Tom Maier, head of BAM’s alternative investment team. Fauchier Partners will advise on manager selection and strategy allocation across a 200-strong universe of funds.
Commenting on the fund, Maier said: “The investment case for the Far East markets, especially outside Japan, is strong. In China, the role of the state in the economy has declined sharply in many sectors and since joining the World Trade Organisation, key economic sectors have been opened up to free trade.
“This has acted as a catalyst for further reform, improved efficiency and long-term growth in both China and in the Asia Pacific region as a whole.”
He added: “The long manager who is contracted to outperform Asia finds that the relative risk they are prepared to take can be eclipsed by the absolute risk of the index itself. For the more risk averse, the appropriate investment strategy for these more volatile areas is one that includes absolute return investments.”
The new fund is expected to generate a return of around 7% above three month US Treasury bills over the long-term.
BAM anticipates that a growing portion of investment in Asia Pacific will be made through hedge funds to protect against the possibility of adverse returns, as seen in Asian crisis of 1998 .
By using between 10 and 20 funds with different investment styles, geographical preferences and risk profiles, BAM aims to keep manager correlation low and reduce overall volatility. Volatility is expected to be between 7% and 9% compared to 21% for the Pacific market over the past 20 years.
The minimum investment level is US$100,000 and is available to both individuals and institutions from May 1. The fund will be denominated in US dollars, euros and sterling.
BAM has significant experience in Asia and has a 3 year track record in running global funds of hedge funds in which it holds £310m assets.
The proposed cold-calling ban may be ineffective if a collaborative regulatory approach between the UK and the European Union (EU) is not maintained post-Brexit, the Pensions Management Institute (PMI) has warned.
Some 56% of defined contribution (DC) asset managers do not believe they will have transaction cost information in time for pension funds' March year-end statements, according to Lane Clark & Peacock (LCP) research.
NEST has appointed Clive Elphick, Martin Turner, Mutaz Qubbaj and Chris Hitchen as trustee members of its reshaped board.
Most people want to avoid investing in projects that contribute to climate change, and would consider moving to another less-exposed provider, according to a survey commissioned by ClientEarth.


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Excellent value, perfect location
Perfect location in downtown Santiago; close to restaurants and cafes; safe neighbourhood at night; some rooms include a sitting room and views of Santa Lucia park across the street; free internet on the ground floor (both wireless and a guest computer); staff are reliable, efficient and friendly. PRICE FOR A DOUBLE WITH PRIVATE BATH: 50$USD
Well, nothing! This hotel can't be beaten at this price.
The only advice I would give is that the restaurant breakfasts are expensive -- you'd be better off at one of the cafes nearby.
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domain: travel


text:
Luminary Bakery use baking as a tool to take women who have experienced social and economic disadvantage such as homelessness, domestic violence, prostitution/sexual exploitation or the criminal justice system on a journey to employability and entrepreneurship, equipping them with transferrable skills for the working world. Through their employability programme and enterprise course, they offer skills training, paid employment and a supportive community to help women thrive.
You can find the bakery in Stoke Newington, East London where they serve delicious cakes, fresh bread, healthy breakfast and lunch options. Luminary also bake beautiful celebration and wedding cakes that can be ordered through their cake shop and delivered in and outside of London.
Based in: UK
71-73 Allen Road, Stoke Newington
Hackney, N16 8RY

domain: food


text:
The fall in the retail index in the eurozone's third-biggest economy was the worst since 1990, when the National Institute of Statistics data collection began.
Christmas shopping failed to galvanize consumers, with non-food sales dropping in December by 0.3 percent from a month earlier and food sales down 0.5 percent according to the seasonally-adjusted data.
Consumers are suffering from increased taxes and unemployment.
Italy began shaking off its longest recession since the Second World War in the fourth quarter with a preliminary estimate showing growth of 0.1 percent, but the turn-around has done little to boost consumer spirits.
The government has said it is hoping for growth of around 1.0 percent this year and the Bank of Italy is predicting a 0.7-percent result but the International Monetary Fund is predicting only 0.6 percent growth.
The disappointing retail results came as new prime minister Matteo Renzi – who has vowed to tackle the labour market and boost growth – prepared to put his coalition to a vote in the lower house of parliament.
After an initial positive reaction among investors to the new premier, who was sworn in on Saturday, Milan's FTSE Mib index was down 0.33 percent on Tuesday.
SEE ALSO: Italy's economic recovery gets underway
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text:
Abington's Week Ahead for July 10
The Planning Board on Monday will consider a proposal to open a new retail cannabis shop at 678 Adams Street
A cannabis operation proposed for Adams Street will have a public information session on Thursday.
A proposed cannabis delivery business will host a community information meeting tonight at 7 p.m.
An Abington dental group is proposing a major overhaul for a well-known, but long empty commercial property
Abington's Week Ahead features a bunch of meetings and a high school musical
Natural Agricultural Products - which is looking to open a new cannabis retail, wholesale, cultivation, and manufacturing operation on Route 18 - is holding a community tonight at 6 p.m.

domain: economics


text:
Ive insect bite on leg. i was itching and its red around. day before yesterday i was either cold or hot (maybe it was pms). should i see a doctor? i feel okay now. how to know if i got infected with the bite?
Content on HealthTap (including answers) should not be used for medical advice, diagnosis, or treatment, and interactions on HealthTap do not create a doctor-patient relationship. Never disregard or delay professional medical advice in person because of anything on HealthTap. Call your doctor or 911 if you think you may have a medical emergency.

domain: medical


text:
Time-correlated single photon counting (TCSPC) enables acquisition of fluorescence lifetime decays with high temporal resolution within the fluorescence decay. However, many thousands of photons per pixel are required for accurate lifetime decay curve representation, instrument response deconvolution, and lifetime estimation, particularly for two-component lifetimes. TCSPC imaging speed is inherently limited due to the single photon per laser pulse nature and low fluorescence event efficiencies (<10%) required to reduce bias towards short lifetimes. Here, simulated fluorescence lifetime decays are analyzed by SPCImage and SLIM Curve software to determine the limiting lifetime parameters and photon requirements of fluorescence lifetime decays that can be accurately fit. Data analysis techniques to improve fitting accuracy for low photon count data were evaluated. Temporal binning of the decays from 256 time bins to 42 time bins significantly (p<0.0001) improved fit accuracy in SPCImage and enabled accurate fits with low photon counts (as low as 700 photons/decay), a 6-fold reduction in required photons and therefore improvement in imaging speed. Additionally, reducing the number of free parameters in the fitting algorithm by fixing the lifetimes to known values significantly reduced the lifetime component error from 27.3% to 3.2% in SPCImage (p<0.0001) and from 50.6% to 4.2% in SLIM Curve (p<0.0001). Analysis of nicotinamide adenine dinucleotide–lactate dehydrogenase (NADH-LDH) solutions confirmed temporal binning of TCSPC data and a reduced number of free parameters improves exponential decay fit accuracy in SPCImage. Altogether, temporal binning (in SPCImage) and reduced free parameters are data analysis techniques that enable accurate lifetime estimation from low photon count data and enable TCSPC imaging speeds up to 6x and 300x faster, respectively, than traditional TCSPC analysis.
© 2016 Optical Society of America
Fluorescence lifetime is the time a fluorophore remains in the excited state upon absorption of an incident photon, before emission of a subsequent photon and return to ground state. Fluorescence lifetimes range from several hundred picoseconds to several nanoseconds, and are dependent on the microenvironment of the molecule, such as temperature, pH, and proximity to quenchers . Additionally, the fluorescence lifetime is dependent upon molecule conformation and binding, with short and long lifetimes depending on quenched or unquenched fluorophore configuration [1,2]. In this way, fluorescence lifetime can be used to evaluate protein interactions.
The fluorescence lifetime can provide information about fluorophore behavior and environment and has been used to detect intracellular pH and ion concentration [3–5], differentiate cancerous tissue from normal tissue [6–8], and detect early response to cancer treatment [8,9]. However, fluorescence lifetime decays are difficult to measure due to the required sub-nanosecond temporal resolution. Fluorescence lifetime imaging (FLIM) can be performed in the time domain by directly detecting the fluorescence decay following an excitation pulse. Several methods have been developed for detecting fluorescence lifetime, including the use of gated cameras [10–12], fast oscilloscopes , and time-correlated single photon counting . While gated cameras and fast oscilloscopes are advantageous for fast imaging of bright samples, both methods are limited in decay sampling (temporal resolution) to ~0.5-1 ns which does not provide sufficient sampling of the decay curve to accurately resolve multiexponential decays.
Time correlated single photon counting (TCSPC) provides the highest temporal resolution within a decay curve for accurate analysis of multi-exponential fluorescence decays. In TCSPC, a fast, ultra-sensitive detector detects individual photons. Electronics time the arrival of the photon with respect to the laser pulse and build a histogram of fluorescence events across time. With repeated observations of fluorescence events, a fluorescence decay curve is assembled. TCSPC systems typically use a very short excitatory laser pulse, <200 fs, which approximates an impulse function. TCSPC electronics can detect the arrival photons with approximately 50 ps time resolution (12.5 ns/256 time bins) .
For all time domain approaches, the fluorescence decay can be described by an exponential decay function Eq. (1) with multiple components corresponding to the fractional lifetimes present in the sample [1,15],Eq. (2) includes the fluorescence decay convolved with the instrument response function, IRF(t).
In TCSPC data, I(t) is the histogram of detected photons. F(t) contains the lifetime endpoints, τ and α. The IRF(t) is the response of the system to a laser pulse. The shape of the IRF(t) is dependent on the laser excitation pulse, cable length, and TCSPC electronics. While the IRF(t) can be measured experimentally, either by direct detection of a laser pulse or detection of an instantaneous event such as second-harmonic generation, subsequent data deconvolution lacks an analytical solution. Therefore, deconvolution to extract fluorescence lifetime parameters is achieved though iterative reconvolution of a predicted model function (1, 2 or 3 component exponential decay) with the measured IRF until least squares residuals are minimized .
The fluorescence decay of a fluorophore which can exist in quenched and unquenched states is described by two lifetimes Eq. (3),2,16], ion concentration fluorescence dyes , and Förster Resonance Energy Transfer (FRET) donors .
While TCSPC provides the highest temporal resolution of fluorescence lifetime decays, TCSPC image acquisition and data analysis are inherently slow. TCSPC FLIM images typically require seconds to minutes to acquire for adequate SNR in each pixel. Shot noise in TCSPC data follows a Poisson distribution, where the SNR is proportional to the number of photons detected . Detector dark current and after pulsing can also contribute to TCSPC noise; however, these noise sources are effectively reduced to negligible levels with the use of hybrid GaAsP detectors . Thousands to tens of thousands of photons per pixel are required for accurate deconvolution and exponential decay fitting of multi-exponential decays. Furthermore, TCSPC can only resolve one photon per laser pulse, yet photon yield must be much less, around 10%, in order to avoid bias towards early arrival photons . Therefore, approximately 82 seconds are required for a 256x256 pixel image, with an average of 10,000 photons per pixel, imaged on a typical TCSPC FLIM system with an 80 MHz laser source and one detector. Unfortunately, this imaging speed is too slow to allow investigation of dynamic physiological processes, such as calcium fluxes in neurons, which occur much faster, within ~100ms. Multifocal mulitphoton fluorescence lifetime systems have been shown to reduce image acquisition times with multiple detectors operating in parallel [19,20]; however, these systems still rely on typical TCSPC data analysis.
The purpose of this study is to investigate the inherent limitations of photon count/SNR for accurate TCSPC data analysis and identify strategies to reduce the number of required photons and image acquisition times. Simulated fluorescence lifetime decays were generated with varying lifetime parameters and SNR. These lifetime decays were convoluted with a system response and analyzed to determine the accuracy of the extracted fluorescence decay parameters. Two techniques were investigated to increase the accuracy of fits for low photon count/SNR, binning the data temporally and reducing the unknown fit parameters. Simulated decay curves were analyzed by two software packages, SPCImage and SLIM Curve. SPCImage is a commercial fluorescence lifetime analysis program, which uses a least squares method with iterative re-convolution to determine the fluorescence decay parameters . SLIM Curve is an open-source software for analyzing fluorescence lifetime data, which uses a triple integral to estimate the single exponential lifetime value and then applies a Levenberg-Marquardt algorithm with a least squares minimization approach to compute double or triple exponential decays. These simulation experiments define the minimum number of photons required, and therefore the imaging speed, for a given fluorescence decay. Finally, the analysis techniques were tested on fluorescence lifetime data of fluorescent cofactor molecule and cofactor receptor protein, nicotinamide adenine dinucleotide – lactate dehydrogenase (NADH-LDH) solutions.
2.1 One-component exponential decay simulations
One-component exponential decay simulation experiments were performed to evaluate the accuracy of fluorescence lifetime deconvolution and parameter fitting in SPCImage (Becker &Hickl) and SLIM Curve (http://fiji.sc/SLIM_Curve). Simulated 256-time channel TCSPC fluorescence decay curves (256 per input conditions) were generated in MATLAB® with Poisson noise and lifetime values from 0.1 ns to 6 ns and total photons per curve from 100 to 10000. These decay curves were convoluted with an IRF measured from the SHG of a urea crystal on a TCSPC system, as previously described [8,21]. The resulting I(t) decay was analyzed by SPCImage and SLIM Curve to extract the lifetime value. The percent errors of the extracted lifetime values were computed, error = [|true-computed|/true]*100, and analyzed.
2.2 Two-component exponential decay simulations
Two-component TCSPC exponential decays (Eq. (4)) with Poisson noise and 256-time channels were simulated in MATLAB.
2.3 Temporal binning
The temporal resolution of the simulated decay curves created in 2.2 was reduced from 256 time bins to 128, 85, 64, 51, 42, 36, and 25 by summing the number of photons in 2, 3, 4, 5, 6, 7, and 10 consecutive time bins, respectively. These new fluorescence decay curves were analyzed by SPCImage and SLIM Curve to extract τ1, τ2, α1, and α2 values and the errors of the lifetime parameters analyzed. The shift value of the multiexponential decay (in SPCImage) was set to a median value obtained by randomly sampling points in a representative image and was uniform across a data set with the same number of time bins.
2.4 Reduced parameters
The decay curves simulated in 2.2 and 2.3 were analyzed in SPCImage and SLIM Curve with the τ1 and τ2 parameters fixed to the appropriate value. The error in the resulting α1 value was analyzed.
2.5 NADH-LDH experiments
Tris-buffered saline at pH 7.6 was used as the solvent for all solutions. Varying concentrations of lactate dehydrogenase (LDH) from porcine heart (Sigma) were mixed with 100 μM NADH (Sigma) in order to generate solutions with desired free-to-bound ratios of NADH (Table 1). Required concentrations were calculated based on a published value for the dissociation constant of NADH to LDH (4.6 μM), and the fact that each LDH molecule has four identical binding sites .
Following mixing, a 100 μl droplet of each solution was placed in a separate 35 mm glass-bottom imaging dish (MatTek Corp.), and a coverslip was placed over each droplet to reduce evaporation.
Fluorescence lifetime images were acquired using a custom-built multiphoton fluorescence microscope system (Bruker) using a 40x oil-immersion objective (1.3 NA) and an inverted microscope (Nikon). A titanium:sapphire laser (Chameleon Ultra II, Coherent) was tuned to 750 nm for excitation of NADH, and a 440/80 nm bandpass filter was used to collect fluorescence emission. A pixel dwell time of 4.8 μs was used to collect images that were 256x256 pixels, and a GaAsP PMT (H7422P-40, Hamamatsu) detected emitted photons. Imaging was performed in the solutions at a depth of 30 μm. Time-correlated single photon counting electronics (SPC-150, Becker and Hickl) were used to acquire fluorescence decay curves. Laser power and integration time were varied in order to generate fluorescence decay curves with an average of 50, 100, 200, 400, 600, 800, 1000, 2500, 5000, 7000, and 10000 photons per pixel. A single image with 256x256 pixels was obtained for each evaluation yielding 65,536 decay curves. The second harmonic generated signal from urea crystals at 900 nm excitation was used to measure the instrument response function, which was found to have a full width at half maximum of 220 ps.
NADH-LDH experimental data was analyzed in SPCImage. The scatter value was set to 0 as there was no evidence of SHG or laser light being detected. The offset and shift values were set to a median value obtained by randomly sampling points within an image, but kept uniform among data with consistent photon counts (offset) and time bins (shift). Data were first analyzed as collected with 256 time bins with unknown lifetimes. Next, data were binned temporally as described in 2.3 and the resulting decays analyzed in SPCImage. Finally, repeat analysis was performed with the τ1 and τ2 parameters fixed to 0.52 ns and 1.25 ns, respectively. These values for free and bound NADH were determined experimentally and agree with published values of free and protein-bound NADH [2,8,21].
3.1 Accuracy for single exponential decays
The SPCImage and SLIM Curve calculated fluorescence lifetimes closely match the true values of the single exponential decays. The mean lifetime error for SPCImage is less than 5% for lifetime values between 0.1 and 6 ns (Fig. 1(a)). The error for lifetimes fit by SLIM Curve was less than 5% for lifetimes above 0.5 ns (Fig. 1(a)). Below 0.5 ns, the error for lifetimes computed by SLIM Curve increased exponentially (Fig. 1(a)). The error of the estimated lifetime for decays with τ = 0.6 ns, increased as the number of photons/SNR decreased (Fig. 1(b)). If a lifetime error of 5% is tolerated, fewer than 500 photons per pixel are required to resolve lifetimes greater than 0.5 ns in SLIM Curve (Fig. 1(c)). If a 1% τ error is tolerated, additional photons are required, ~3000 for SLIM Curve to resolve lifetimes greater than 1 ns (Fig. 1(d)).
3.2 Lifetime analysis accuracy for double exponential decays
Both SPCImage and SLIM Curve fail to accurately fit some two component fluorescence lifetimes. At 10,000 photons per decay curve (SNR = 100), SPCImage fails to accurately fit (component error > 20%) fluorescence lifetimes with α1 values < 0.5 (Fig. 2(a)), τ1 values < 0.3 ns (Fig. 2(b)), and τ2 values < 1 ns (Fig. 2(c)). Likewise, SLIM Curve fails to accurately fit fluorescence lifetimes with α1 values > 0.5 (Fig. 2(d)) and τ1 values < 0.3 ns (Fig. 2(e)). SLIM Curve accurately fits all τ2 values, provided that α1 is < 0.8 (Fig. 2(f)).
For the two component lifetimes that can be accurately fit, the minimum number of photons required for certain error thresholds, α1 error < 10%, τ1 error < 25%, and τ2 error < 25%, was determined (Fig. 3). For SPCImage, the fewest photons are required for decay curves of high α1 values (Fig. 3(a)-3(c)) and (τ2 - τ1)> ~2 ns (Fig. 3(c)). For SLIM Curve, the number of photons required for good fits is minimized for α1 values = 0.2 (Fig. 3(d)) and τ2~ = τ1 (Fig. 3(d), 3(f)). For the fluorescence lifetimes of free and bound NADH, τ1 = 0.5 ns and τ2 = 1.2 ns, respectively, lifetime component error is minimized with increasing photons, when analyzed with both SPCImage and SLIM Curve (Fig. 4).
3.3 Improved accuracy with temporal binning
Temporal binning of fluorescence decay curves was evaluated as a method to improve fitting accuracy. Typical TCSPC decay curves contain 256 time bins over 12.5 ns. Here, sequential time bins were summed to reduce the total number of time bins to 128, 85, 64, 51, 42, 36, and 25. A significant improvement in α1 error (red, Fig. 5(a)) and τ1 error (blue, Fig. 5(a)) was observed for decays with 85, 64, 51, 42, and 36 time bins analyzed in SPCImage. Likewise, τ2 error (green, Fig. 5(a)) improved for decays with 51, 42, and 36 time bins. This improvement in fitting accuracy was dependent on the number of photons per curve, with the greatest improvement observed for curves with 400-1000 photons (Fig. 5(b)). For fluorescence lifetimes which can be resolved with a cumulative error in all endpoints < 25% (decay curves with high α1 values and τ1 and τ2 separations > 2 ns), the average minimum number of photons required decreased from 4185 photons to a minimum average of 681 photons at 42 time bins (Fig. 5(c)). Additionally, the number of simulated decay curves that could be accurately fit increased with temporal binning to 42 bins (Table 2). Temporally binned fluorescence decay curves were evaluated in SLIM Curve; however, no improvement in fitting accuracy was observed (Fig. 5(d), Table 2).
3.4 Improved accuracy with fixed lifetime components
The values of the short and long lifetime components may be known in many cases, or determined experimentally, as is the case for FLIM dyes and FRET pairs. If the fluorescence lifetimes are known values, the values can be fixed in the decay models of SPCImage and SLIM Curve to improve the accuracy of the estimated α parameters. To evaluate the α parameter estimation, the same two-component simulated decay curves evaluated in 3.2 and 3.3 were analyzed with the lifetimes fixed in the analysis. The accuracy of the calculated α1 parameter increased when the τ1 and τ2 values were fixed in the analysis model. For SPCImage, the average α1 error decreased from 26.7% to 3.2% (p<0.0001; Fig. 6(a)). For SLIM Curve, the average α1 error decreased from 50.6% to 4.2% (p<0.0001; Fig. 6(b)). While the α1 error can be improved by temporal binning of data analyzed by SPCImage with free τ1 and τ2 values (Fig. 7(a)), no significant improvement in α1 error is achieved by fixed-parameter analysis of temporally binned data (Fig. 7(b)). However, an overall more accurate calculation of α1 can be achieved and at a fewer required number of photons when the lifetime parameters are fixed in SPCImage (Fig. 7(a)-7(b)). Likewise, for SLIM Curve, an overall more accurate estimate of α1 at a fewer required number of photons is achieved when the lifetime parameters are fixed, and there is no improvement with temporal binning (Fig. 7(c)-7(d)).
3.5 Improvement in photon requirement and imaging speed
As shown in Sections 3.3 and 3.4, temporal binning and fixed lifetime parameters in decay analysis reduces the required number of photons for an accurate decay fit. The minimum photons required for total error (τ1 error + τ2 error + α error) to be less than 10% was evaluated using the simulated NADH fluorescence decays where τ1 = 0.5, τ2 = 1.2 and α1 = 0.8, for the original data and the data with processing techniques (Table 3). With no SNR improvement techniques, 30,000 photons are required for accurate fitting (total error < 10%; Fig. 4), which would take approximately 246 s to acquire a 256x256 pixel image. With temporal binning in SPCImage, the required number of photons is reduced to 4000 (data not shown), and the image capture time reduced to 32.8 s. By fixing the lifetime values, the required number of photons is reduced to 100 (Fig. 7(b)), and imaging time reduced to 0.819 s. By combining fixed parameter analysis and spatially binning 9 pixels, accurate fits can be achieved with as few as 11 photons/pixel (108 total for the binned area), and an imaging time of 90 ms.
3.6 Validation in NADH-LDH solution experiments
The simulated fluorescence decay curve experiments demonstrate improvement in SPCImage analysis of fluorescence lifetime decays with temporal binning and fixed lifetime parameters. To ensure these analysis techniques improve SPCImage fitting of real data, fluorescence lifetime images of solutions of NADH-LDH with α1 values of 0.3 and 0.5 were acquired and analyzed. Representative images demonstrate the improvement in α1 values between the orginal data (α1 = 0.5) with 800 photons and 256 time bins evaluated with free components (Fig. 8(a)), fixed components (Fig. 8(b)), and temporally binned to 64 time bins (Fig. 8(c)). As a representative example, temporal binning from 256 to 51 time bins of fluorescence decay curves with 800 photons improved the total error from 108% to 55% (p<0.0001) for α1 = 0.3 (Fig. 8(d)). Likewise, temporal binning from 256 time bins to 64 time bins of fluorescence decay curves with 800 photons improved the total error from 76% to 44.0% (p<0.0001) for α1 = 0.5 (Fig. 8(d)). This reduction in error is comparable to that obtained by spatially binning 9 pixels of the original 256 time-bin image, 44.0% vs. 44.3%, respectively. In addition to improving the lifetime fitting by temporal binning, fixing the lifetime values in SPCImage greatly improves the estimation of α1 (Fig. 8(e)) and allows accurate estimations of α1 at reduced photon numbers. For NADH, error in the mean lifetime is reduced with large photon counts (Fig. 8(f)) and a reduction in error with temporal binning in SPCImage is observed (Fig. 8(g)).
With a 90% error threshold tolerance, temporal binning improves the required number of photons to fit NADH decays with α1 = 0.3 from 10,000 photons to 800 photons, which corresponds to an imaging time improvement of a 256x256 pixel image from 82s to 6.55s (Table 3). Likewise, temporal binning reduces the number of photons and imaging time required from 7000 photons, 58s, to 400 photons, 3.3s, for NADH decays with α1 = 0.5. With the lifetimes fixed in SPCImage analysis, α1 errors less than 10% are achieved with as few as 400 photons for α1 = 0.3 and 200 photons for α1 = 0.5 (Table 3, Fig. 8(e)), equating to imaging times of 3.3 s and 1.64 s. When combined with a 9-pixel moving spatial bin, further reductions in imaging time can be achieved, down to 0.37 s for α1 = 0.3 and 0.188 s for α1 = 0.5, although with reduction in spatial resolution.
TCSPC remains the optimal method to obtain fluorescence lifetime decay data with high temporal resolution within a decay curve. However, conventional imaging and analysis protocols require a large number of photons per pixel, 10,000 photons or more, for accurate deconvolution of the instrument response function and lifetime parameter estimation. This high photon count number for image resolution requires lengthy image acquisition times (>1 min) that are not conducive to imaging fast biological phenomena. Therefore, this study applied simulation experiments to define the limitations of conventional FLIM analysis algorithms and determine the photon count requirements for accurate analysis of fluorescence decay curves. Additionally, two methods to improve fitting accuracy are demonstrated on simulated fluorescence decay curves and NADH-LDH experimental data.
The deconvolution of the measured signal into the system response and the fluorescence decay curve does not have an analytical solution. Therefore, the fluorescence decay parameters are approximated by iterative analysis of the convolution of the instrument response with a 'best guess' fluorescence decay curve. This method assumes an exponential shape of the fluorescence decay with a set number of components. Commercial software, SPCImage uses a least squares method with iterative re-convolution to determine the fluorescence decay parameters . An open-source alternative, SLIM Curve, uses a triple integral to estimate the single exponential lifetime value and then applies a Levenberg-Marquardt algorithm with a least squares minimization approach.
Simulation experiments demonstrate the fluorescence decays which can accurately be evaluated by TCSPC and conventional analysis. The single-component exponential decay simulation results show that both SPCImage and SLIM Curve can accurately evaluate single-component fluorescence decays for lifetimes greater than 0.5 ns (Fig. 1). Furthermore, both SPCImage and SLIM Curve accurately describe fluorescence decays of two components with large lifetime separation and either high or low α1 values, respectively (Fig. 2). However, even at high photon counts, both analysis programs fail to accurately fit certain decay curves (Fig. 3). SPCImage fails to fit fluorescence lifetimes with α1 values < 0.4, τ1 values < 0.3 ns, and τ2 values < 1 ns. SLIM Curve fails to accurately fit fluorescence lifetimes with α1 values > 0.5 and τ1 values < 0.3 ns. Fortunately, the short and long lifetimes of most two-component fluorescence decay systems are less than 0.4 ns and greater than 1 ns, respectively, and have a separation greater than 0.5 ns [2,21].
TCSPC noise follows a Poisson distribution where , where N is the number of photons . Therefore, the SNR of fluorescence decays is improved by increasing the number of photons. Post-imaging, this can be achieved by binning spatial pixels. However, spatial binning degrades the resolution of the image and assumes congruent fluorophore states within the binned area. As an alternative to spatial binning, temporal binning was evaluated to increase SNR. Temporal binning was achieved by reducing the number of decay time bins from 256 to 128, 85, 64, 51, 42, 36, and 25 time bins by summing sequential time bins of the original 256 bin decay curve. By combining the photons in consecutive time bins within a pixel, the error due to shot noise in the time bin is reduced by the averaging effect of the multiple bins combined. Temporal binning greatly improved the fitting by SPCImage with the greatest improvements in fluorescence parameters observed for the curves with 42 time bins (Fig. 4). Evaluated by SPCImage, temporal binning resulted in a greater number of fluorescence decays that could be accurately resolved and reduced the average number of photons required for accurate fitting from 4500 photons for decay curves with 256 time bins to about 500 photons for decay curves with 42 time bins, a 9x reduction in the required number of photons (Fig. 4). The improved results for SPCImage evaluation of temporally binned data suggests that for many two-component exponential decay curves, the time axis is oversampled, at the expense of added noise into the decay curve. Temporal binning reduces the noise in each time bin and yields a more accurate decay approximation. Temporal binning also improved SPCImage fitting of NADH-LDH data (Fig. 7(a)). While temporal binning to 42 bins improved fits in SPCImage, further binning to 36 or 25 bins increased error, indicating that with too low temporal resolution, the shape of the decay is degraded and the two-component fit accuracy is reduced.
SLIM Curve did not show improvements with temporal binning, which suggests SLIM Curve is less influenced by noise in the decay curveor the improvement in noise is offset by the degradation of temporal resolution. SPCImage uses a least squares method with iterative re-convolutionwhile SLIM curve uses a triple integral to estimate the single exponential lifetime value and then applies a Levenberg-Marquardt algorithm with a least squares minimization approach to determine the multi-component lifetime parameters. These subtle differences in algorithms lead to different lifetime approximations, lifetime parameter accuracies, and behaviors when data is binned temporally to improve SNR.
Additional instrumentation configurations exist for fast FLIM and fast data processing. Camera-gated methods can be used to image whole-field fluorescence; however, these are typically limited to 2-20 gates/time bins [10–12] which, as shown in Fig. 5(a), is insufficient for accurate analysis of two component lifetime decays. Rapid lifetime determination (RLD)  and Laguerre estimation methods [24–26] exist for improved speed of lifetime approximation, yet linear least squares remains the most accurate for lifetime determination. Additionally, single-photon avalanche diode (SPAD) arrays can be used to rapidly resolve fluorescence lifetime decays in a sensitive manner not limited by the single photon aspect of TCSPC [27–29]. SPAD acquisition can be combined with center-of-mass method processors for hardware-based temporal binning for single-component lifetime estimation . Furthermore, hardware-based temporal binning to 4 (16 time bins) enables faster transfer of data to the computer, demonstrating improved imaging speeds with in-pixel histogramming to fewer time bins . This method supports the advantages of temporal binning for improved FLIM imaging. For TCSPC, the data analysis method of temporal bining described here improves exponential decay fits in SPCImage analysis and may enable robust results with fewer collected photons.
In addition to increasing SNR, lifetime fitting can be improved by reducing the number of unknown parameters. By fixing the lifetime values in SPCImage and SLIM Curve, the average α1 error decreased significantly from 26.7% to 3.2% and from 50.6% to 4.2%, respectively. While the fluorescence lifetime of the sample may not always be known or may be influenced by the environment, there are many experiments where the lifetime components are known. For example, the fluorescence lifetime of dyes designed to report ion concentrations exist in one state when unquenched by the ion and a second when quenched [4,5]. These lifetime values can be measured independently and then these values used to evaluate mixed samples. Furthermore, often the fluorescence lifetime of a FRET donor is known to be a certain value determined by the quenched or unquenched state of the molecule . As demonstrated by the NADH-LDH experimental data (Fig. 7), fixing the lifetime parameters improves the estimation of α1, and the substantial improvement suggests lifetime values should be fixed when known. However, for in vivo experiments, fluorescence lifetimes may exist in a continuum of values due to local changes in temperature, pH, viscosity, proximity to quenchers. In this case, when fluorophore lifetime is largely variant, fixing the lifetimes to set values may introduce error.
Improvements in image acquisition time and fluorescence lifetime decay estimation benefit all TCSPC FLIM studies. In particular, more accurate estimation of the lifetime weight components, α, will improve FRET experiments and yield more accurate approximations of FRET donor and acceptor concentrations. Additionally, clinical and in vivo FLIM studies have been limited by the lengthy image acquisition times which introduce motion error and other artifacts into the images. Faster image acquisition may enable additional clinical FLIM studies, previously hindered by image acquisition time. Furthermore, increasing evidence supports characteristic changes in intrinsic fluorophore lifetimes with diseases such as cancer and precancers [7,8,31]. Therefore, increased accuracy in lifetime estimation will increase the sensitivity and specificity of FLIM-based screens.
Altogether, these simulation and NADH-LDH experiments demonstrate that fluorescence lifetime fitting accuracy is highly dependent on the lifetime parameters and number of photons/SNR. Temporal binning in SPCImageand fixed lifetime parameters can achieve high fidelity fits of noisy or low photon count data, enabling accurate lifetime component estimation with reduced number of photons and faster imaging. Combining analysis techniques amplifies the effect with theoretical improvements in imaging speed from 246 s for a 256x256 pixel image to 0.82 s, a 300X improvement. Image acquisition times can be reduced further, to as low as 90 ms with a 9-pixel moving spatial bin; however, this approach reduces spatial resolution. These improvements in photon count and imaging time will allow improved imaging of weakly fluorescent signals, such as autofluorescence, or fast phenomena.
Funding sources include the National Research Council Research Associateship Program (AW), AFOSR LRIR#14RH02COR, R01 CA185747, and the NSF GRFP (JTS).
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30. N. Krstajić, S. Poland, J. Levitt, R. Walker, A. Erdogan, S. Ameer-Beg, and R. K. Henderson, “0.5 billion events per second time correlated single photon counting using CMOS SPAD arrays,” Opt. Lett. 40(18), 4305–4308 (2015). [CrossRef] [PubMed]


text:
A new species of Leptomyxida, named Rhizamoeba neglecta was found during studies of the amoeba fauna of the inner Lake Leshevoe located at Valamo archipelago (The Lake Ladoga, North-Western Russia). Light-microscopical and ultrastructural studies indicated that it represents a new species of Leptomyxida. The partial 18S rDNA sequence of this amoeba is very similar to that of Leptomyxa reticulata.. These organisms, however, are very different in LM morphology and biology. Organisms assigned to the genus Rhizamoeba do not form a single clade in the 18S rDNA tree. This may indicate that the genus is an artificial grouping or that a number of studied strains were misidentified. The phylogeny and the systematics of leptomyxids require further investigation.
see on Pubmed

domain: biology


text:
Relatives refuse to receive body of raped, murdered girl
- Kanchanpur rape case
Jul 29, 2018-
Body of a 13-year-old girl, who was raped and murdered at Nimbukheda of Bhimdatta Municipality-18 in Kanchanpur district on Thursday, remained at the Mahakali Zonal Hospital for the third day on Sunday after her family and locals refused to receive the body until the culprits were booked.
Repeated talks between the administration and the victim's kin failed to reach an agreement after the relatives demanded strong action against the guilty, compensation for the bereaved family and a police beat in the Nimbukheda area. The irate relatives said they will receive the body only if the administration addresses their demand to establish a police beat in Nimbukheda area.
It has been learnt that the parents of the deceased girl had agreed to receive the body for final rites but were stopped by the locals. Chief District Officer Kumar Bahadur Khadka said family relatives had agreed to perform the final rites after the local administration agreed to book the culprits and establish the police beat as demanded. But a large crowd of locals came and stopped them.
Nirmala Panta, of Bhimdatta-2, was found dead at a roadside sugarcane field along Mahandranagar-Ultakham road on Thursday. She had left her home early that Thursday to meet her friend.
Police investigation showed the girl was murdered after rape.
Published: 29-07-2018 19:02

domain: law


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text:
When Roy writes, ‘The youth, in preparation to an attack, marked each venue by reading from their prayer books in an act most of us are familiar with as a precursor to a holy war or fight’, he comes close to demonising a community.
Watching her from our elephant-back vantage, there was a peace that was starting to descend on that wretched Dhikala chaur. The chital went back to their grass. The elephant breathed slowly, then slower, and then no more. The tigers of Corbett had claimed their largest victim yet


text:
Calligraphy and Crest Design Styles
The following calligraphy and crest styles are all hand-written and custom to each bride and groom. They are not fonts. These styles can be applied to any custom wedding invitation design, logo design, digital invitation design, or package invitation suite.
The Seattle is a slanted script, with more space between letters to give the script a romantic yet modern feel. This script is for the bride that wants an updated invitation that still invokes romantic calligraphy.
The Whimsical is a tight script that merges the modern and the Seattle and Portland scripts. For the bride that wants an updated invitation that isn't too stiff or sharp.
The Modern is a loopy, rounded script that is best for the traditional bride that doesn't want stuffy or too formal invitations. A lot of loops make the Modern a wonderful addition to a formal suite with lots of pretty details.
The Portland is a variation on the Seattle with more even line heights and letters that all start at the same point on the line. It's an updated
Crest and Monogram Styles
Crests are available in shield shapes, as simple laurels, as shields decorated in laurels, and with banners below.
Monograms are available in multiple formats:
- one letter (the future last name of the couple)
- a full, formal monogram (first name of the groom, future last name of the couple, and first name of the bride)
- the first initials of the bride and groom
Add-ons can include an "established" line, a wedding date, a location


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Houston Community Newspapers sold to Nevada company
Houston Community Newspapers has been sold to Nevada-based 1013 Star Communications, an operator of suburban newspapers.
The sale is the final divestiture for ASP Westward, which had owned the Houston group since 2002, HCN reports. Earlier this year, ASP Westward sold newspaper operations in east Texas and suburban Denver.
“1013 brings a considerable knowledge of community newspaper publishing to the operation,” ASP Westward CEO Jim Hopson told HCN.
Miller told HCN the Houston group is a “great fit for us.”
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text:
Escape. Bed N Beach. Breakaway Suite.Escape Bed n Beach is away from the madding crowd. We have an almost deserted beach with a view of Redang and Bidong Islands. Turtles come up on our beach to lay eggs from March till September. We welcome guests who love the sound of Nature and silence. Day trips to the islands can be arranged for you, as well as home cooked dinners by us, ordered in advance. Please let us know in your inquiry the exact number of guests, including children and infants.Thank you.
Teratak SekuchiTeratak Sekuchi is a semi-traditional timber beach-house facing the South China Sea. Originally built in downtown Kuala Terengganu, the house was relocated to a typical fishing village in Mengabang Telipot. Furnished mainly with old wooden furnitures from Terengganu and other parts of Malaysia, it offers a taste of coastal village living with basic modern comforts. There’s no wifi, TV or air-cond. Have a look at this video: https://www.facebook.com/mengabang.telipot/videos/235672950364234/
The ResthouseA wooden house, on its own, by the sea... A large portion of proceeds from The Resthouse are channeled into our conservation project - Lang Tengah Turtle Watch - on a neighbouring island. We encourage all our guests to spend a day visiting and enjoying this project, to see how your stay is contributing to the conservation of these majestic creatures.


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