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"New Year's Eve" is a terrible movie but a fantastic business plan.
Like its predecessor, "Valentine's Day" (also from director Garry Marshall and writer Katherine Fugate), the movie cobbles together an untenable number of vaguely connected plots surrounding a holiday celebration. The secret sauce is that almost every character, no matter how insignificant, is played by a familiar face, and almost all of these are current or former A-list stars. Robert De Niro? Check. Hilary Swank? Present. Sarah Jessica Parker? Aye-aye! Jon Bon Jovi? Why not?
While it's unlikely that "New Year's Eve" will do well at the box office, it can expect aftermarket immortality. Look for "New Year's Eve" to be trotted out for broadcast in the run-up to the holiday, first on premium cable, then on TNT or USA, then Lifetime before winding up on ABC Family until the movie's copyright expires. You probably won't see "New Year's Eve" in theaters, but it's unlikely you'll be able to avoid it indefinitely.
As long as a few million people tune in over the course of the week between Christmas and New Year's every year between now and the end of the world, the studio and the actors will make a pile of money. It's even possible that the product placement (including a conspicuous ad for the new Sherlock Holmes movie, also from Warner Bros.) could be digitally updated as new marketers buy into the movie. If accountants had critics, they'd be raving: "Breaks new ground in ancillary rights!" "A triumph of deferred compensation!"
The movie pivots around the New Year's Eve celebration in Times Square. It would be madness to try to summarize the dozen or so story lines that variously intersect or fizzle out. In broad strokes, Miss Swank plays a perfectionist planner charged with bringing the iconic ball-drop. Ashton Kutcher is particularly smarmy as a millennial curmudgeon determined to wait out the night in his pajamas — until he gets stuck in an elevator with singer Lea Michele. The comic talents of Katherine Heigl are squandered on the nothing role of a caterer who is still recovering from a broken relationship with a pop star played by Mr. Bon Jovi. The only winning performance comes from Michelle Pfeiffer as a mousy secretary who is bent on living out all her New Year's resolutions before midnight.
The film affords no time to delve into any character or story, so everything is kept on the most superficial, cliched level. Worse, New Year's itself is here positioned as a holiday of personal renewal, occasioning a couple of execrable and embarrassing speeches that play like grade school themes on "What the new year means to me."
Sadly, there's nothing redeeming about this holiday celebration.
★ (out of four)
TITLE: "New Year's Eve"
CREDITS: Directed by Garry Marshall; written by Katherine Fugate
RATING: PG-13 for swearing
RUNNING TIME: 118 minutes


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Why Did Mortgage Rates Go Up Federal Reserve Move Indicates Mortgage Rates Will Be Going. – Federal Reserve Move Indicates That From Here on Out, Mortgage Rates Will Be Going Up. By Danielle Hale | Mar 21, 2018.. Is It Last Call for Low Mortgage Rates? Why Home Buyers Should Act Now;
20-Year Mortgage Rates. A 20-year mortgage rate is attractive to borrowers who are looking for an unchanging mortgage payment for a specific period of time. A 20-year fixed-rate mortgage will generally have a lower interest rate than a 30- or 40-year mortgage. You could spend less money on interest over the life of a 20-year mortgage loan as well.
The average 15-year mortgage rate fell to 3.05% in the latest week, down from 3.20% a week ago. It was 4.05% a year earlier. Interest rates on five-year adjustable-rate home loans averaged 3.36%, the.
5 days ago · In Denmark’s $495 billion mortgage-backed covered bond market, another milestone was reached on Wednesday as Nordea Bank Abp said it will start offering 20-year fixed-rate loans that charge no.
In the US, if the down payment is less than 20% of the total property price, typically, private mortgage insurance (PMI) is mandatory until the principal is paid down to less than 80% or 78% of the total property price. The PMI rate normally ranges from 0.3%-1.5% of.
What Is Lending Rate Una Osili: payday loans won’t boost Hoosiers’ financial security – Last month, the Indiana Senate passed a bill that would expand a new class of payday loans. The bill raises the limits on interest rates to 192 percent and eliminates the 72 percent cap currently in.
. interest rate for 30-year fixed-rate mortgages with conforming loan balances ($484,350 or less) remained unchanged at.
A year ago, the rate averaged 4.60%, according to the Freddie Mac Primary Mortgage Market Survey. The 15-year FRM averaged.
Rate is fixed. The payment on a $203,500, 30-year fixed rate loan at 4.375% and 76.22% loan-to-value (LTV) is $1148.38 with 1.875 Points due at closing. Payment includes a one time upfront mortgage insurance premium (MIP) at 1.75% of the base loan amount and a monthly MIP calculated at 0.80% of the base loan amount.
Current Par Mortgage Rates Mortgage Rates, Mortgages At NerdWallet, we adhere to strict standards of editorial integrity to help you make decisions with confidence. Many or all of the products featured here are from our.
What is a 20 year fixed rate mortgage? The 20 year fixed mortgage is a simple loan program, just like it’s much more popular relative the 30 year fixed. This fixed rate mortgage is a home loan with an interest rate that remains the same throughout the 20 year term. At the end of the 20 year repayment period, the loan is fully amortized.
· View and compare urrent (updated today) 15 year fixed mortgage interest rates, home loan rates and other bank interest rates. Fixed and ARM, FHA, and VA rates.
· Mortgage rates are dropping to fresh lows. July could provide some of the lowest rates seen in over 2 years. This is the chance mortgage rate shoppers have been waiting for.

domain: finance


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GAULEY BRIDGE, WV (LOOTPRESS) – A Clay County man faces drug charges in Fayette County.
According to Sheriff Mike Fridley, a Fayette County Sheriff’s Deputy conducted a traffic stop on a vehicle in Gauley Bridge for an infraction last night. A search of the vehicle following a positive K-9 indication revealed multiple individually packaged units of heroin and methamphetamines, weight scales, and a sum of cash.
Matthew D. Evans of Lizemores is charged with Possession With Intent to Deliver Narcotics.
He will now await court proceedings.

domain: news


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About The Site
I introduced AbstractArtistGallery.org, a website designed for abstract artists, as side project in early 2011. As an abstract artist myself I saw the need to create it for other abstract artists. From the moment I launched the site, there was an interest by a number of abstract artists. The quality of artwork we have on the site is very high and I expect the site will be a wonderful resource for artists, art collectors, and students.
Abstract Artist Gallery is a juried listing of talented abstract artists from all over the world where the visual presentation of artwork takes center stage. I stress visual presentation because that is one of the unique features of the site. We do not sell original artwork, but rather serve as a portal to connect the artist with an audience via the web. The website was designed around a simple and eloquent WordPress blogging platform, that looks great and finds its way into the search engines quite effectively.
Over the years I have learned quite a lot about search engine optimization and feel I can really put it to use. The main page is an updated gallery index of artists that instantly introduces the viewer to lots of artwork they probably have never seen before. Then there is also a search engine optimized listing for each abstract artist and a main gallery with a thumbnail image of every artist’s work. The main gallery serves as an efficient way to visually browse though great abstract art. The user can also browse by artist names or simply type in a search for the type of abstract art they are looking for. We intergrated social sharing tools into the site to boost our rankings and further increase traffic to the site. Artists are also part of our Facebook Page that has thousands of fans already.
I am very proud of our artists and I continue to seek out the very best abstract artists to include in the gallery. If you are an abstract artist, or perhaps someone who is interested in abstract art please visit the Abstract Artist Gallery and let me know what you think. Currently, artists can still submit for inclusion into the site.

domain: artistic


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“ Brand: Kallo / Type: Snacks „
I don't eat bread, and I am getting a bit bored of Ryvita, so I have turned to rice cakes for a bit of a change. I was originally buying Sainsbury's Be Good To Yourself rice cakes, but I might as well have chewed on some cardboard. They were really dry and bland - even by rice cake standards.
This brand was sitting beside the Sainsbury's rice cakes when I was doing my grocery shopping, and as they weren't much dearer, I decided to buy them instead. I was also convinced when I saw on the packaging that they were lightly salted.
They come stacked on top of one another inside the plastic packaging. The packaging is blue, white and green, depicting a blue sky and a green field with a scarecrow on it - it gives the impression that this is a very natural product (it is organic, after all). The ingredients list on the back is pretty simple - just wholegrain brown rice. The rice cakes themselves are made up of puffed rice, and are circular and flat. They are an off-white colour.
You can't really eat rice cakes plain, well not unless you're a masochist, so I usually spread a little houmous over them. But you can put anything on them - smoked salmon, cream cheese, ham, tomato, tapenade... anything you like. Sometimes I just put a tiny bit of margarine on them too.
Once bitten into, initially they are quite soft with a slight crunch to them. They don't seem like cardboard. They then become chewier and feel filling and more substantial than you would expect. Because they are very lightly salted, there is some flavour to them, and it compliments whatever topping you have chosen.
I really like these rice cakes and will be buying them from now on. The chewiness makes you feel that you are eating something filling. Rice cakes are never going to be a substitute for bread, BUT due to their consistency and slight salted flavour, they do serve as a good replacement if you are watching your weight. They feel more substantial and less cardboard-y than other rice cakes. Maybe I am an oddball but I find them really moreish!
These cost around 94p for 130g.
My children have to take healthy break to school for their morning break, sometimes they take fruit and other times they take, French toasts or Kallo rice cakes. My children have eaten rice cakes since they were small babies, I gave them small rice cakes as finger foods when they were being weaned onto solids so they have always had a taste for them. Personally I find rice cakes a bit bland on their own but my children just eat them plain with no topping.
These Kallo rice cakes , are wholegrain, they are large circular discs measuring approximately 9cm in diameter and 1 cm depth. They are basically pieces of puffed rice fused together to form a crispy circular snack. They are an off white colour with little bits of brown through the cake, the wholegrain rice. The rice cakes are not sweet they are a savoury item, they taste rather nutty and have a slightly salty tinge to them. They don't really taste of very much but they are not unpleasant and my children eat them all the time, you could add toppings to try and make them more interesting, perhaps cream cheese or some other spread, even just sliced tomatoes or cheese. I don't find them very filling and would consider them to be quite light in texture, not at all stodgy.
We have tried various makes of rice cake but Kallo id definitely my children's preferred choice. They come in a 130g cylindrical paper pack, with Kallo logo on the front, it is quite distinctive packaging. I never have any bother finding them in the supermarkets, they are sometimes in the biscuit aisle with the savoury crackers and sometimes they are in the "free from" section, but usually I can find them easily enough. The rice cakes have only 28 calories and are low in fat. These are a regular in our shopping basket, even when the children are not a school, they sometimes just have them as a snack between meals. My youngest child is nearly three and I find it very difficult to get him to eat a breakfast in the morning before he goes off to the childminder, but most mornings he will munch on a Kallo rice cake before we head off.
There are some other varieties in the Kallo range but we generally buy the wholegrain rice cakes. They are suitable for vegetarians and vegans and are gluten free. If they are left exposed to the air they can go a bit soft, so they could be stored in an airtight container, I usually have no storage problems with them because they get eaten quickly in my home. These are a relatively healthy snack that taste fine, they are versatile and can have toppings added or just eaten on their own. We recommend these rice cakes.
These rice cakes are amazing. I buy them every time I go shopping. I eat them mostly as a snack. Lovely if not a bit dry by themselves but you can spread whatever you like on them. I personally like jam or nutella. Not that they are the healthiest options, but it's all about moderation really. These are so cheap going at 97p. I tend to eat a pack a week so a week of snacks for 97p is just great. Great to accompany a healthy diet or to start one. The only downside to this product is that it is a bit dry but then again it is to be expected. Available in large food stores I usually get mine from tesco. I never eat them as part of a meal but I guess you could. Maybe having them with a spread for breakfast might be a good idea.
I think this would be a good product to buy if you are trying to keep a healthy diet or start one.
I am constantly on the look out for healthy, low fat snacks, as not only am I trying to lose weight I am increasingly conscious about the nutritional and health aspects of the food I am eating.
Despite all this I am a massive snacker and could easily snack my way through crisps and biscuits all day if I let myself, so I have to find healthy alternatives.
I have been eating rice cakes for years as my mum used to buy them when I was younger and had some bad food allergies (at the time I was not impressed at this alternative to additive fuelled sweets but hey she tried her best!)
We used to buy own brands of these but recently I saw the Kallo variety int eh shop and thinking they looked pretty nice , decided to try these instead. Ok so I have to admit I was swayed by the pretty blue packaging and the fact it just looked all healthy and organic with the picture of the grass etc. Shallow I know but hey I cant help it, packaging is important I think.
I think they were slightly more expensive than the supermarket own brand, they were approx £1.70, but they weren't extortionate so this didn't put me off. And I have to say that in this case you get what you pay for. I really believe that Kallo are a superior range of rice cakes.
Firstly they do a wholegrain variety, something that cheapo brands don't offer. This instantly made me feel healthier just by buying them.
You can tell the difference when you open the packet and look at them as well as they are not as stark white as some cheaper brands and you can see the draker flecks of wholegrain in them.
A rice cake is pretty hard to describe in terms of flavour but they are round with a crispy, slightly salty taste to them. My colleagues at work told me they smell like popcorn, which made them jealous every time I took them out at work. Although the taste is similar to that of any rice cake I noticed that Kallo had made theirs with a slightly salted taste to them which makes them tastier than some own brands which can taste bland in comparison.
Kallo rice cakes were also crisper and less prone to sogginess as some cheaper brands I found, which is particularly useful when taking them to work as you don't want to find them a soggy mess by the time you go to eat them in the afternoon.
Kallo's packaging is also ingenius in that they have extra packaging at the top tied with a plastic toggle (not the official term) which means that once open you can reseal them. This might seem simple to you, but having bought own brands that just come in a packet like normal biscuits they are prone to go staler quicker and you have to keep them in a container.
Rice Cakes are a brilliant snack by themselves , or used as a cracker or bread substitute. And even better than that each rice cake is only around 20 calories and really fills you up! Way hey!
PRICE: 97p for 130g at my local Morrisons
NUTRITIONAL INFORMATION (per rice cake):
(of which sugars): 0.3g
(of which saturates): 0.1g
Wholegrain brown rice, salt
Gluten-free, no artificial additives, suitable for vegetarians & vegans, may contain traces of sesame seeds
I am one of what seems to me the rare individuals who actually enjoys rice cakes, not easily deterred by the feeling of eating polystyrene. I've tried most brands, Kallo being last on the list, so a pack slipped into my supermarket trolley the other day.
Kallo Low Fat Wholegrain Rice Cakes come in a mostly blue sealed plastic pack, with the Kallo trademark and an image of a couple of silhouetted ducks and butterflies wandering across a rural scene. The rear of the pack shows nutritional information, ingredients, dietary/allergy advice and tips, plus Kallo's quality claim and contact details.
The seal on the pack is glued down very tightly and I had to attack it with scissors in order to get to the contents inside. The act of opening the packet broke the first rice cake, but I didn't become unduly panicked, as I feel there are worse things in the world to worry about.
Though I didn't actually count them, I estimate there were approximately 16 rice cakes inside the pack. Each rice cake is coloured a bright, almost steely white, with little pale brown flecks of wholemeal material. The circumference of each cake is I'd guess slightly larger than a standard digestive biscuit, having a thickness of approximately 1/4".
Great care has to be taken in removing the cakes from the packaging, as they break and shatter easily. If they aren't going to be eaten all at once, it's a good idea to store them in an airtight container, as they go stale very quickly.
My favourite way of eating rice cakes is topped with thin slices of Emmental cheese, but on this occasion I decided to sample them in their naked state, so as to get an accurate experience of their flavour and texture.
On biting into the first cake, a few grains of puffed rice fell back onto the plate, but the texture was pleasing - light, airy and delicately crispy. As I chewed, the rice cake didn't (as some other brands can do) go all gluey in my mouth, and the experience of crunching gently then swallowing, was a pleasant one.
As far as taste is concerned, I liked the wholemeal touch as it felt as though I was actually eating something worthwhile rather than chewing on air.
The only slight problem I personally had with the taste, was that despite the low-salt claim, these rice cakes are just a tad over-salted for my own palate. That's not to say I won't buy them again though, but in future I shall eat them with something else so as to drown out that salty flavour.
Gradually over a period of about three days, I finished off the packet - complete with slices of Emmental - and I found the whole experience rather delicious.
The difference between Kallo Low Fat Wholegrain Rice Cakes and other brands is negligible, except for them being maybe slightly saltier than other low-salt varieties - plus I'd estimate they are just a tiny, tiny bit thinner than others I've tried.
I wouldn't declare Kallo to have created a wondrously new and different rice cake experience, but they are quite nice and I'd probably buy them again.....always making sure I have something suitable to eat with them. I am awarding Kallo four stars, but if the product had contained just a little less salt or none at all, it would be a case of full house.
Rice cakes per se are very versatile and you can comfortably top them or eat them with pretty much anything, sweet or savoury, and they are much healthier than some crispbreads and most manufactured biscuits.
Thanks for reading!

domain: food


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Delayed wound healing is commonly associated with diabetes. It may lead to amputation and death if not treated in a timely fashion. Limited treatments are available partially due to the poor understanding of the complex disease pathophysiology. Here, we investigated the role of leucine-rich α-2-glycoprotein 1 (LRG1) in normal and diabetic wound healing. First, our data showed that LRG1 was significantly increased at the inflammation stage of murine wound healing, and bone marrow–derived cells served as a major source of LRG1. LRG1 deletion causes impaired immune cell infiltration, reepithelialization, and angiogenesis. As a consequence, there is a significant delay in wound closure. On the other hand, LRG1 was markedly induced in diabetic wounds in both humans and mice. LRG1-deficient mice were resistant to diabetes-induced delay in wound repair. We further demonstrated that this could be explained by the mitigation of increased neutrophil extracellular traps (NETs) in diabetic wounds. Mechanistically, LRG1 mediates NETosis in an Akt-dependent manner through TGFβ type I receptor kinase ALK5. Taken together, our studies demonstrated that LRG1 derived from bone marrow cells is required for normal wound healing, revealing a physiological role for this glycoprotein, but that excess LRG1 expression in diabetes is pathogenic and contributes to chronic wound formation.
Wound healing is a natural reparative response to tissue injury. It proceeds through four continuous and overlapping phases: homeostasis, inflammation, proliferation, and tissue remodeling (1). Failure to progress through these phases in an orderly manner leads to impaired wound healing, which represents one of the common causes of morbidity associated with diabetes, affecting ∼25% of individuals with diabetes (2). These wounds frequently serve as portals of entry for bacterial infection that may lead to sepsis and lower-extremity amputation (3). Staggeringly, patients with lower-extremity amputation have a 5-year mortality rate of up to 50% (4). With the rising prevalence of diabetes, the incidence of wound complications is expected to increase substantially, posing a significant socioeconomic burden (5).
A plethora of factors contributes to delayed wound closure in patients with diabetes, such as excessive neutrophil infiltration and activation, impaired angiogenesis, and defective epithelial cell migration and proliferation (6). These defects lock the wound into a self-perpetuating inflammatory stage (7), which causes further tissue injury by increasing the production of inflammatory cytokines, reactive oxygen species, destructive enzymes, and cytotoxic extracellular traps in a process termed NETosis (8) (where NET is neutrophil extracellular trap). Thus, targeting inflammation serves as an attractive strategy to kick-start the proliferation phase of wound healing and promote repair. A number of anti-inflammatory agents have been developed over the last 20 years (9). Despite effectiveness in promotion of wound closure in rodent models, limited success has been achieved in clinical trials (10). This is likely due to the highly dynamic and complex interactions between different types of cell, extracellular matrix components, and soluble factors present in the wound microenvironment. A better understanding of the molecular mechanisms underlying diabetes-associated healing deficiency will guide the development of more effective therapeutics to treat wounds that do not respond sufficiently to good standard care.
Leucine-rich α-2-glycoprotein 1 (LRG1) is a secreted glycoprotein that was previously reported to regulate pathological neovascularization in the eye by switching the angiostatic TGFβ1-Smad2/3 signaling toward the proangiogenic TGFβ1-Smad1/5/8 signaling in endothelial cells (11). Besides its role in ocular angiogenesis, LRG1 is intimately associated with many inflammatory and autoimmune conditions (12–14) and tumor malignancy (15–17), which shares fundamental molecular mechanisms with chronic wound healing (18). Recently, elevated serum LRG1 levels were reported in patients with diabetes with peripheral arterial disease (19), a major risk factor for diabetic foot ulcers (DFU) (20). Paradoxically, exogenous LRG1 was reported to accelerate wound healing by promoting keratinocyte migration in animal models (21). Here, we characterized LRG1 expression level and pattern in wound tissue, investigated its contribution to wound healing under normal and diabetic condition using Lrg1-null mice, and explored its mechanism of action.
Research Design and Methods
Human Sample Analysis
This study was approved by Khoo Teck Puat Hospital Ethics Review Board (NHG Domain Specific Review Board). Adults between 21 and 90 years old with type 2 diabetes seen in Diabetes Centre of Khoo Teck Puat Hospital were enrolled into this study. Fasting blood and debrided tissues were collected from patients with ulcers during their podiatry assessment clinic. Fasting blood samples were centrifuged within 1 h after collection and kept at 4°C during this period. Thereafter, they were stored at −80°C in aliquots and used without additional freeze-thaw cycles. Devitalized tissue was obtained from desloughing and debridement performed as part of usual care. These samples were stored in liquid nitrogen until retrieval for assays described below. Control samples were obtained from patients without type 2 diabetes with venous ulcers in the same clinic. Serum level of LRG1 was measured using an ELISA kit (Immuno-Biological Laboratories, Hamburg, Germany) according to the manufacturer’s instructions.
Animals and Induction of Diabetes
C75BL/6J mice were purchased from InVivos (Singapore). Lrg1−/− mice were originally generated by the University of California, Davis, Knockout Mouse Project (KOMP) Repository Collection (https://www.komp.org) and were a generous gift from J. Greenwood and S.E. Moss (UCL Institute of Ophthalmology). Animal experiments were performed in compliance with the guidelines of the Institutional Animal Care and Use Committee (ARF-SBS/NIE-A0268/A19036) of Nanyang Technological University and the Guide for Care and Use of Laboratory Animals published by the National Institutes of Health. Diabetes was induced in 6- to 8-week-old male mice by intraperitoneal injection of 50 mg/kg streptozotocin (STZ) (50 mmol/L sodium citrate buffer, pH 4.5) for five consecutive days as previously described (22). Diabetes was confirmed when fasting blood glucose (FBG) was >200 mg/dL.
Creation of Full-Thickness Cutaneous Wounds
Six full-thickness cutaneous wounds were created on mouse dorsal skin using 4-mm Integra Miltex Standard Biopsy Punches (Thermo Fisher Scientific). Wounds were imaged daily with a digital camera. Wound size was quantified with use of ImageJ (National Institutes of Health). We used 6-mm Integra Miltex Standard Biopsy Punches (Thermo Fisher Scientific) for biopsy collection at different time points following injury. The mouse excisional wound splinting model was employed as previously described (23).
Bone Marrow Transplantation
Six-week old mice were irradiated at two doses of 5.5 Gy irradiation using a BIOBEAM GM γ irradiation device (Gamma-Service Medical, Leipzig, Germany). Bone marrow cells (BMCs) from female mice were harvested and filtrated through a 70-μm cell strainer (Falcon). BMCs (3 × 106) were intravenously injected into the irradiated recipient mice through the tail vein 24 h after the irradiation. Eight weeks after reconstitution, wounds were created at flanks of recipient mice.
Isolation and Flow Cytometry Analysis of Myeloid Cells From Wound Tissue
Wound tissues were digested in Iscove’s modified Dulbecco’s medium (Thermo Fisher Scientific) containing 2% FBS, 2 units/mL DNase I (Roche, Switzerland), and 1 mg/mL collagenase D (Roche) and passed through a 40-mm cell strainer for obtaining single-cell suspension. Red blood cells were lysed using 0.89% NH4Cl lysis buffer and removed by centrifugation. Cell pellet was resuspended and preincubated with anti–Fc receptor antibody (clone 2.4G2) followed by further incubation with anti-mouse BUV737-labeled CD45 (clone 30-F11), allophycocyanin-Cy7–labeled anti-mouse CD11b (clone M1/70), anti-mouse F4/80 (clone BM8), BUV395-labeled anti-mouse Ly6G (clone 1A8), BV605-labeled anti-mouse Ly6c (clone HK1.4), phycoerythrin-Cy7–labeled anti-mouse CD11c (clone N418), and BV421-labeled anti-mouse I-A/I-E clone (M5/114.15.2). Dead cells were labeled with fixable viability stain 510 (BD Biosciences, San Jose, CA). Cells were then fixed and permeabilized before being stained with anti–LRG-1 antibody (cat. no. 13224-1-AP; Proteintech) followed by FITC-labeled donkey anti-rabbit IgG (clone Poly4064). Cells were washed and subjected to analysis on a five-laser flow cytometer (BD LSRFortessa; BD Biosciences). Data were analyzed with FlowJo software (TreeStar, Ashland, OR).
Cells and Cell Culture
Primary mouse and human peripheral blood neutrophils (Institutional Review Board of Nanyang Technological University [IRB-2014-04/27]) were isolated, purified, and cultured as previously described (24,25). Human neutrophil-like cells (dHL-60) were derived from human promyelocytic leukemia cell line (HL-60; ATCC) by incubation with 1% DMSO (Sigma-Aldrich) for 7 days. Human dermal microvascular endothelial cells (HDMECs) (PromoCell), normal human dermal fibroblasts (PromoCell), human keratinocyte line HaCaT (ATCC), and Freestyle 293-F cells (Gibco, Thermo Fisher Scientific) were maintained according to the supplier’s instruction. Cells were treated with 20 μg mL−1 recombinant human LRG1 (rhLRG1), LDN193189 (100 nmol/L) (Sigma-Aldrich), SB431542 (10 μmol/L) (Sigma-Aldrich), and MK-2206 (10 μmol/L) (Selleck Chemicals) and as indicated.
Histology, Immunohistochemistry, and Immunofluorescence
Mice skin tissues were fixed in 4% paraformaldehyde and embedded in paraffin following a standard protocol. Paraffin sections (5 μm thick) were subjected to staining with hematoxylin-eosin or LRG1 antibody (cat. no. 13224-1-AP; Proteintech). For immunofluorescence staining, skin tissues were embedded in O.C.T. Compound (Thermo Fisher Scientific). Cryopreserved skin sections (5 μm thick) were stained with primary antibodies against LRG1 (13224-1-AP; Proteintech), F4/80 (MCA497; Bio-Rad), CD11b (130-113–235; Miltenyi Biotec), CD31 (550274; BD Biosciences), Myeloperoxidase (MPO) (ab9535; Abcam, Cambridge, U.K.), and Ki67 (ab15580; Abcam) followed by staining with Alexa 488 or Alexa 594 secondary antibodies (Thermo fisher Scientific). Images were captured using Leica DM5500 microscope (Leica Microsystems) or Carl Zeiss LSM 710 confocal microscopy (Zeiss, Berlin, Germany), processed using Adobe Photoshop CS6, and analyzed using ImageJ by investigators who were blinded to the identity of the experimental groups.
Quantitative Real-time PCR
Total RNA was extracted and purified with RNAzol RT (cat. no. 888-841-0900; Molecular Research Center) before being reverse transcribed to cDNA with qScript cDNA SuperMix (157031; Quanta Biosciences). PCR was conducted with PrecisionFAST qPCR Master Mix (FASR-LR-SY; Primerdesign Ltd, U.K.) with use of Applied Biosystems StepOnePlus Real-Time PCR System (Life Technologies). The expression levels of respective target genes were normalized to GAPDH, and relative gene expressions were calculated using standard 2−ΔΔCT method. Primers used in this study are listed in Table 1.
|Gene .||Forward primer sequence .||Reverse primer sequence .|
|VCAM 1||5′-GTCTCCAATCTGAGCAA-3′||5′-TGGGAAAAACAGAAAAGAGGTG −3′|
|Gene .||Forward primer sequence .||Reverse primer sequence .|
|VCAM 1||5′-GTCTCCAATCTGAGCAA-3′||5′-TGGGAAAAACAGAAAAGAGGTG −3′|
SDS-PAGE and Western Blotting
Cells or tissues were lysed on ice in radioimmunoprecipitation assay buffer containing 0.0037 mg/mL protease inhibitor (Roche), 1 mmol/L dithiothreitol (cat. no. D9779; Sigma-Aldrich), 1 mmol/L phenylmethylsulfonyl fluoride (P7626; Sigma-Aldrich), and 100 mmol/L phosphatase inhibitors (P0044 [for cell signaling experiments]; Sigma-Aldrich). Proteins were separated by 10% SDS-PAGE before being transferred onto an Immobilon-PSQ PVDF Membrane (ISEQ-00010; Merck Millipore). Blots were probed with LRG1 antibody (rabbit monoclonal, 13224-1-AP; Proteintech), phosphorylated (phospho-)Smad1/5 antibody (rabbit monoclonal, 9516; Cell Signaling Technology), anti-SMAD1+SMAD5 antibody (mouse monoclonal, ab75273; Abcam), histone H3 (citrulline R2 + R8 + R17) antibody (rabbit polyclonal, ab5103; Abcam), histone H3 antibody (rabbit polyclonal, ab1791; Abcam), phospho-Akt antibody (rabbit monoclonal, 4060; Cell Signaling Technology), Akt antibody (rabbit monoclonal, 9272; Cell Signaling Technology), cyclin D1 antibody (rabbit monoclonal, 2922; Cell Signaling Technology), or GAPDH antibody (rabbit polyclonal, sc-25778; Santa Cruz Biotechnology), followed by horseradish peroxidase–conjugated secondary antibodies (Santa Cruz Biotechnology). Densitometry was performed by use of ImageJ software.
Molecular Biological Methods
Human LRG1 (NM_052972) carrying a 6xHis tag expression vector, pcDNA3.1-LRG1, was generated as previously described (11). rhLRG1 protein was expressed in Freestyle 293 T cells (Invitrogen) and purified as previously described (11). siRNA oligonucleotides (cat. no. L-015179-01; Dharmacon) were used for LRG1 gene knockdown, while control siRNA (D-001810-10-20; Dharmacon) was used as a negative control. Transfection was performed using Lipofectamine 2000 (Invitrogen) for HaCaT cells and RNAiMAX (Invitrogen) for dHL-60 cells according to the manufacturer’s protocol.
Cell-surface CD11b and L-selectin were measured using flow cytometry. dHL-60 cells were treated with rhLRG1 (20 μg/mL) for 1 h, 6 h, or 24 h before being washed with flow buffer (0.1% FBS/PBS). Cells were then incubated with L-selectin antibody (mouse monoclonal, cat. no. sc-390756; Santa Cruz Biotechnology), followed by staining with Alexa Fluor 488 goat anti-mouse (IgG) secondary antibody (Thermo Fisher Scientific) and CD11b-phycoerythrin (130-113-235; Miltenyi Biotec) and fixation in 1% paraformaldehyde. Cell acquisition (10,000 cells per sample) was carried out on BD LSRFortessa X-20 (BD Biosciences) and analyzed with use of FlowJo software (BD).
Neutrophil Adhesion Assay
Lrg1-knockdown dHL-60 cells or dHL-60 cells treated with rhLRG1 (20 μg/mL) cells were labeled with CellTracker Green CMFDA Dye (Invitrogen) before being seeded onto the confluent HDMEC monolayer. In the case of Lrg1-knockdown dHL-60 cells, HDMECs were pretreated with tumor necrosis factor (TNF)α (50 ng/mL; PeproTech). Two hours later, nonadherent neutrophils were subsequently removed by washing with prewarmed PBS. Adherent neutrophils were imaged with the Eclipse Ti-E Inverted Research Microscope (Nikon Instruments, Tokyo, Japan) and quantified by measurement of the fluorescence intensity with Synergy H1 microplate reader (BioTek) at the wavelength of 492 nm/517 nm.
HDMECs were cultured in EGM-2MV media (Lonza, Basel, Switzerland) until 30% confluent and starved in EBM-2 medium (Lonza) containing 0.2% FBS for 16 h before being treated with rhLRG1 (20 μg/mL) for 48 h. Cells were then fixed and stained with Ki67 antibody (rabbit polyclonal antibody, cat. no. ab15580; Abcam) for detection of proliferating cells and DAPI (Thermo Fisher Scientific) for staining cell nuclei. Images were taken with Eclipse Ti-E Inverted Research Microscope and analyzed with ImageJ. Proliferation rate was calculated as the percentage of Ki67+ cells.
Transwell Migration Assay
HDMECs were pretreated with rhLRG1 for 24 h before being seeded onto rat tail collagen I (100 μg/mL; Corning)–coated Transwell Inserts (8 μm) (Corning). EBM2 medium containing 5% FBS served as a chemoattractant. After 5-h incubation, migrated cells were fixed and stained with DAPI (Thermo Fisher Scientific). Images were taken with Eclipse Ti-E Inverted Research Microscope and analyzed with Image J.
Matrigel Tube Formation Assay
Matrigel Growth Factor Reduced Basement Membrane Matrix (Corning) (60 μL) containing vehicle control or rhLRG1 (20 μg/mL) was added to each well of a 96-well plate and incubated for 30 min at 37°C for polymerization. HDMECs in 100 μL EBM-2 medium containing vehicle control or rhLRG1 (20 μg/mL) were seeded onto the polymerized Matrigel gel. HDMEC tube formation was imaged using phase-contrast mode on Eclipse Ti-E Inverted Research Microscope following overnight incubation, and tube formation was analyzed with ImageJ.
Trypan Blue Exclusion Assay
Transfected and rhLRG1 (2 μg/mL)–treated HaCaT cells were trypsinized and stained with Trypan blue before being counted with a hemocytometer under a phase-contrast microscope.
Scratch Wound Healing Assay
Confluent HaCaT cells were starved in DMEM medium containing 0.2% FBS (Thermo Fisher Scientific) for 24 h. A scratch was made to HaCaT cell monolayer with a sterile p200 pipette tip. The cells were washed and subsequently cultured in complete DMEM. Cells were imaged at 0 h and 24 h after scratching with Eclipse Ti-E Inverted Research Microscope 24 h after scratching. Images were analyzed with ImageJ.
SYTOX Green Assay
Primary mouse and human peripheral blood neutrophils were seeded onto each well of a 96-well black polystyrene microplate with a clear flat bottom (Corning). Cells were treated with 5 μmol/L calcium ionophore A23187 or rhLRG1 (100 μg/mL) for 4 h before incubation with the DNA dye SYTOX Green (1 μmol/L) (Invitrogen) for 15 min. NET formation was determined by measurement of the fluorescence intensity with Synergy H1 microplate reader (BioTek) at the wavelength of 504 nm/523 nm.
Induction of NETosis
Primary mouse neutrophils in RPMI-1640 medium containing 25 mmol/L HEPES were seeded onto each well of an eight-well chamber slide. Twenty minutes later, attached cells were incubated with 5 μmol/L calcium ionophore A23187 for 2 h before being fixed and stained with H3Cit antibody (1:100 dilution; Abcam) and DAPI (Thermo Fisher Scientific). Images were taken with Carl Zeiss LSM 710 confocal microscopy and analyzed with ImageJ.
Data are represented as mean ± SEM. Statistical analyses were performed using unpaired, two-tailed Student t test or one-way/two-way ANOVA followed by Tukey/Bonferroni post-test analysis using Prism 5 (GraphPad Software Inc.).
Data and Resource Availability
Complete data sets generated and analyzed during the current study are available from X.W. on request.
LRG1 Is Produced by Wound-Infiltrating Bone Marrow–Derived Cells Following Injury
To address the role of LRG1 in wound healing, we examined the expression of LRG1 in normal C56BL/6 mouse skin tissues by immunohistochemistry and revealed a very weak staining in the dermis (Fig. 1A). Western blot was used to evaluate LRG1 expression in wound tissues at various time points following injury (Fig. 1B). Our data showed that LRG1 was increased as early as 6 h post-injury and reached its highest level 24 h after wounding. LRG1 level then declined gradually and went back to basal level on day 5 following injury. It is worth noting that LRG1 expression in surrounding intact skin tissues remained low throughout the wound healing process (Supplementary Fig. 1).
Immunofluorescence staining was used to identify the source of LRG1 during wound healing. We found that Lrg1+ cells were mainly present in the provisional matrix and were colocalized with CD11b+ myeloid cells in day 1 wound tissues (Fig. 1C). Flow cytometry analysis revealed that LRG1 is expressed by Ly6G+/CD11b+ neutrophils, Ly6C+/CD11b+ monocytes, Ly6C−/F4/80+/CD11b+ macrophages, and Ly6C−/F4/80+/MHC II+/CD11c+/CD11b+ dendritic cells (Supplementary Fig. 2), all of which are bone marrow–derived cells. For confirmation of this observation, allogenic bone marrow transplantation (BMT) study was carried out in irradiated wild-type mice with use of BMCs from Lrg1−/− mice and wild-type littermate controls. Similar to what we observed in unirradiated C56BL/6 mice, quantitative real-time-PCR (qRT-PCR) showed that Lrg1 transcript was significantly higher in day 1 wound tissues of wild-type mice transplanted with wild-type BMCs, whereas Lrg1 was not induced in those that received Lrg1−/− BMCs (Fig. 1D). These data suggest that wound-infiltrating bone marrow–derived cells (BMDCs) serve as a major source of LRG1 during cutaneous wound healing.
LRG1 Is Critical for Timely Wound Closure
For elucidation of the functional role of LRG1 in cutaneous would healing, 4-mm full-thickness wounds were created on the dorsal skin of wild-type and Lrg1−/− mice. Lrg1−/− mice demonstrated a significant delay in wound closure as compared with wild-type controls in both excisional wound model and excisional wound splinting model (Fig. 2A and Supplementary Fig. 3). As LRG1 is markedly induced at the inflammatory phase of wound healing, the number of wound-infiltrating immune cells was analyzed in the wound bed of wild-type and Lrg1−/− mice. Neutrophils are the first inflammatory cells to be recruited to the wound bed (26). After performing their functions, apoptotic neutrophils and tissue debris are cleared by macrophages, which eventually leads to the resolution of inflammation (27). Immunofluorescence staining showed a significant reduction in the number of wound-infiltrating MPO+ neutrophils (Fig. 2B) and F4/80+ macrophages (Fig. 2C) 1 day and 5 days post-injury, respectively. As BMDCs are major LRG1-producing cells, we next performed BMT between Lrg1−/− mice and wild-type controls. Mice subjected to irradiation were previously reported to show delayed wound repair (28). In this study, irradiated Lrg1−/− mice and wild-type control mice were intravenously transplanted with wild-type or Lrg1−/− BMCs, respectively (Fig. 2D). Wild-type mice transplanted with wild-type BMCs and Lrg1−/− mice transplanted with Lrg1−/− BMCs served as controls to exclude the impact of irradiation on wound healing. Consistent with what was observed in unirradiated control mice, wound closure was significantly delayed in Lrg1−/− recipients transplanted with Lrg1−/− BMCs as compared with that in wild-type recipients transplanted with wild-type BMCs. On the other hand, wound closure in wild-type recipients transplanted with Lrg1−/− BMCs was delayed substantially, whereas wild-type BMCs fully rescued the delayed wound healing in Lrg1−/− mice. Together, these data provide compelling evidence that LRG1-producing BMDCs are critical for timely wound closure.
LRG1 Promotes Neutrophil Adhesion via Inducing the Expression of L-Selectin
The ability of neutrophils to adhere to the endothelium is critical for their recruitment to the wound bed (29). To understand LRG1’s role in neutrophil infiltration, we investigated the ability of dHL-60 cells to adhere to a HDMEC monolayer in the presence and absence of rhLRG1. Our study showed that rhLRG1 significantly promoted dHL-60 cell adhesion to the HDMEC monolayer (Fig. 3A). As dHL-60 cells express higher levels of LRG1 compared with other skin cells (Supplementary Fig. 4), we went on investigating the consequence of siRNA-mediated LRG1 knockdown in neutrophil function and demonstrated reduced responsiveness to TNFα-induced adhesion to HDMECs (Fig. 3B). L-selectin, a cell adhesion molecule expressed on neutrophils, serves as a master regulator of neutrophil adhesion (30). We next examined whether LRG1 exerts its function through mediating the expression of L-selectin on neutrophil-like dHL-60 cells. Indeed, immunoblots showed a significant increase in L-selectin expression in dHL-60 cells subjected to 24-h treatment with rhLRG1 (Fig. 3C). Consistent with this, flow cytometry revealed a marked increase of the median of fluorescence intensity in dHL-60 following rhLRG1 treatment (Fig. 3D and Supplementary Fig. 5). However, rhLRG1 did not affect the expression of endothelial adhesion molecules including ICAM-1, VCAM-1, P-selectin, and E-selectin (Supplementary Fig. 6). Together, these data show that LRG1 promotes neutrophil adhesion, at least partially, by regulating the expression of L-selectin on neutrophils.
LRG1 Promotes Epithelial Cell Proliferation and Epithelial-to-Mesenchymal Transition
Besides inflammation, reepithelialization is essential to prompt wound repair. It is achieved by orchestrated migration and proliferation of epithelial cells adjacent to the wound (31). H-E analysis revealed delayed reepithelialization in day 4 wounds of Lrg1−/− mice (Fig. 4A). Although the denuded surface is completely covered by newly formed epithelium 5 days following injury, the reconstituted epidermis is significantly thinner in Lrg1−/− mice as compared with that in wild-type controls (Fig. 4B). It was previously reported that exogenous LRG1 promotes keratinocyte migration (21). Similarly, we showed that LRG1-overexpressing HaCaT cells migrated much faster as compared with control plasmid transfected cells, whereas the migration ability of the LRG1 siRNA-treated HaCaT was significantly compromised (Fig. 4C). Activation of the partial epithelial-to-mesenchymal transition (EMT) has been reported to drive keratinocyte migration (32). Our study demonstrated that rhLRG1 was able to cause a significant induction of key EMT markers, such as fibronectin (FN1) and N-cadherin (N-Cad) (Fig. 4D). To define LRG1’s role in reepithelialization further, we examined the keratinocyte proliferation as indicated by immunofluorescence staining with Ki67 in day 3 wounds of Lrg1−/− and wild-type control mice. Our study revealed a substantial reduction in the percentage of Ki67+ keratinocytes at the wound edge of Lrg1−/− mice (Fig. 4E). Consistent with this observation, the number of viable LRG1-overexpressing HaCaT cells was significantly higher than control cells, and HaCaT cells subjected to siRNA-mediated LRG1 knockdown showed reduced viability compared with control siRNA-treated HaCaT (Fig. 4F). This observation was supported by a marked increase in cell proliferation marker cyclin D1 in rhLRG1-treated HaCaT (Fig. 4G). Together, these data suggest that LRG1 facilitates reepithelialization by promoting keratinocyte proliferation and migration.
LRG1 Modulates Dermal Angiogenesis
Our previous study demonstrated an essential role of LRG1 in pathological neovascularization in the eye (11), and angiogenesis is required for the formation of granulation tissue during wound healing (1). To understand LRG1’s role in dermal angiogenesis during wound healing, day 7 wound tissues were subjected to immunofluorescence staining with an endothelial cell (EC)-specific marker, CD31. Although the total vessel area in the distal part of the skin remained unchanged (Supplementary Fig. 7), there was a significant reduction in total vessel area in the wound bed of Lrg1−/− mice (Fig. 5A). In line with the observations in macrovascular human umbilical vein endothelial cells (HUVECs) (11), rhLRG1 was able to induce HDMEC proliferation as visualized by Ki67 staining (Fig. 5B) and the ability of HDMECs to form tube-like structure in Matrigel (Fig. 5C). We also showed increased motility of rhLRG1-treated HDMECs (Fig. 5D). Mechanistically, we found that rhLRG1 significantly stimulated the phosphorylation of proangiogenic Smad1/5 in HDMECs, and blocking of TGFβ type I receptor activin-like kinase 1 (ALK1) and activin-like kinase 5 (ALK5) completely abrogated this activation (Fig. 5E). These data demonstrate an ALK1/5-dependent proangiogenic role of LRG1 in wound healing.
LRG1 Is Highly Induced in Diabetic Mice and in Humans With Diabetes
Having established an important role for LRG1 in physiological wound healing, we next examined the association between LRG1 and chronic wound healing in mice and humans with diabetes. ELISA analysis revealed a significantly higher LRG1 level in the serum of DFU patients as compared with that in venous ulcer patients (Fig. 6A). Consistent with this, we showed that LRG1 expression in ulcer tissues of DFU patients was also significantly higher than that from venous ulcer patients (Fig. 6B). To support this observation, we analyzed wound tissues collected from C57BL/6 mice subjected to STZ-induced diabetes for the expression of LRG1. As observed in DFU patients, LRG1 levels were significantly higher in wounds of diabetic mice (Fig. 6C). Consistently, qRT-PCR analysis revealed a sustained high expression of Lrg1 transcript in the wound tissue of diabetic mice throughout the wound healing process (Fig. 6D). Wound closure was significantly impaired in diabetic mice as compared with that in nondiabetic control (Fig. 6E). These data show that in the skin, LRG1 expression is further increased in both humans and mice with diabetes.
Deletion of the Lrg1 Gene Was Beneficial to Impaired Wound Healing in Diabetes
As our data thus far have demonstrated an elevated Lrg1 transcript level in wounds of both mice and humans with diabetes, we investigated whether wound closure in mice with STZ-induced diabetes is affected in the absence of Lrg1. Although STZ treatment led to a significant weight loss, there was no difference in body weight between STZ-treated wild-type and Lrg1−/− mice (Supplementary Fig. 8). Unlike what was observed in normoglycemic mice, mice with genetic deletion of Lrg1 were protected from the diabetes-induced delay in would closure (Fig. 7A). Recent studies highlighted the influence of diabetes on NET formation (8). Considering the role of LRG1 in neutrophil functions and its upregulation at the inflammatory phase of wound healing, we next studied whether LRG1 affects NETosis in mice subjected to STZ-induced diabetes. Western blot analysis showed a significant reduction in the expression of a NET marker, H3Cit, in day 3 wounds of diabetic Lrg1−/− mice (Fig. 7B). For confirmation of this observation, bone marrow–derived neutrophils were isolated from wild-type and Lrg1−/− mice and subjected to calcium ionophore–induced NETosis. Consistent with the earlier observation, Lrg1−/− neutrophils were resistant to calcium ionophore–induced expression of H3Cit (Fig. 7C). Similarly, immunofluorescence staining showed that calcium ionophore–induced NETs, as indicated by the presence of H3Cit+ neutrophils, were significantly reduced in Lrg1-deficient neutrophils (Fig. 7D). We also showed that Lrg1-deficient neutrophils formed fewer NETs in comparison with their wild-type counterparts upon calcium ionophore treatments (Fig. 7E). Complementing this observation, SYTOX Green assay showed that LRG1 supplementation significantly induced the formations of NETs in human peripheral blood neutrophils (Fig. 7F). Consistently, immunoblots demonstrated that rhLRG1 significantly induced citrullination of histone H3 in dHL-60 cells (Fig. 7G). Activation of Akt pathway was reported to mediate calcium ionophore–induced NETosis (33). We further showed that LRG1 was able to induce the phosphorylation of Akt in dHL-60 cells and the LRG1-induced expression of H3Cit and Akt phosphorylation were completely abolished in the presence of an allosteric Akt inhibitor, MK2206 (Fig. 7G). LRG1 was previously reported to signal through TGFβ type I receptor activin-like kinase 5 (ALK5) in non-ECs (34). For elucidation of whether LRG1-induced NETosis and Akt activation are dependent on ALK5, ALK5 was inhibited by SB431542, resulting in a complete abrogation of LRG1-induced phosphorylation of Akt and H3Cit (Fig. 7H). Together, our data demonstrate an important role of LRG1 in diabetic wounds and that LRG1 exerts its function through mediating NETosis in a TGFβ/ALK5/Αkt-dependent manner.
Impaired wound healing and subsequent formation of foot ulcers is one of the most common complications found in patients with diabetes (2). Considering the important role of inflammation in DFU pathophysiology, multiple anti-inflammatory drugs have been developed but have shown limited success (10). LRG1 is a multifunctional protein that was previously linked to neutrophil activation (35), EMT (16), and angiogenesis (11), all of which are essential for effective wound closure. Here, we investigated the role of LRG1 in both physiological and pathological cutaneous wound healing.
Overwhelming evidence indicated the association between LRG1 and various inflammatory and autoimmune conditions (12–14). Infiltrating myeloid cells have been reported to act as the key source of LRG1 in psoriatic skin lesions (13) and remodeling myocardium following infarction (36). In line with these observations, our study revealed that LRG1 is predominantly produced by the wound-infiltrating CD11b+ myeloid cells. Neutrophils are among the first inflammatory cells to reach the wound bed following injury. Both impaired neutrophil function and hyperactive neutrophils have been reported to compromise wound healing (26). Despite being induced during early neutrophil differentiation (37), the role of LRG1 in neutrophil function remains to be elucidated. Our study showed that LRG1 promotes neutrophil adhesion, likely by inducing the expression of neutrophil adhesion molecule, L-selectin. Consistent with this observation, the number of wound-infiltrating neutrophils was significantly reduced in the wound bed of Lrg1−/− mice. We further showed that wild-type recipients transplanted with Lrg1−/− BMCs showed a significant delay in wound closure as compared with wild-type recipients receiving wild-type BMCs. On the other hand, wild-type BMCs were sufficient to rescue the delayed wound closure in Lrg1−/− recipients. These data support the important role of BMDC-derived LRG1 in wound healing.
Reepithelialization plays an indispensable role in wound healing, and it is driven by the proliferation and migration of keratinocytes at the wound edge (31). A previous study using exogenous LRG1 showed that LRG1 does not affect keratinocyte proliferation as demonstrated by EdU+ staining (21). Here, we showed reduced number of proliferating keratinocytes as indicated by Ki67 staining at the wound edge of Lrg1-deficient mice. The discrepancy between the two studies is likely due to the use of different cellular markers for proliferating cell detection. Ki67 is a broad cell proliferation marker that is expressed throughout the active cell cycle (G1, S, G2, and M phases) (38), whereas EdU is only incorporated in nascent DNA during the S phase (39). Therefore, EdU may provide partial information regarding the extent of cell proliferation. Supporting LRG1’s role in keratinocyte proliferation, plasmid-mediated LRG1 overexpression and siRNA-mediated LRG1 knockdown treatment significantly affect the viability of keratinocytes in vitro. We also demonstrated a promoting role of LRG1 in the expression of cell-cycle marker cyclin D1. These results are in line with previous studies that LRG1 increases proliferation of different epithelial-derived cancer cells, such as colorectal cancer cells (40), pancreatic ductal adenocarcinoma (17), non-small-cell lung cancer cells (41), and gastric cancer cells (15). Consistent with the previous report (21), we showed that LRG1 overexpression and knockdown affect keratinocyte migration. To acquire migratory capacity, quiescent epithelial cells undergo phenotypic changes to gain mesenchymal characteristics (42). Our study discovered that rhLRG1 induces the expression of EMT markers, which is consistent with the promoting effect of LRG1 in EMT and colorectal cancer metastasis (16).
The increased metabolic demand of repairing triggers angiogenesis, and failure in forming functional new vessel leads to delayed wound closure (43). Although LRG1 has previously been implicated in ocular (11) and tumor (16) angiogenesis, its role in normal blood vessel formation during wound healing was largely unknown. In this study, we observed reduced blood vessel density in the wound bed of Lrg1−/− mice. We further showed that LRG1 promotes angiogenesis by mediating HDMEC proliferation, migration, and the ability to form tube-like structures. Unlike what was observed in HUVECs (11), both ALK1 and ALK5 are required for LRG1-induced Smad1/5/8 phosphorylation in HDMECs, which is not surprising, as ALK5 kinase activity is necessary for optimal TGFβ/ALK1 action (44).
Our study showed elevated LRG1 levels in serum and wound tissues of human patients with DFU and diabetic mice, which could be explained, at least partially, by the increased infiltration of immune cells, including neutrophils and macrophages, in diabetic wounds (45). While neutrophils are beneficial to normal wound repair, excessive neutrophil infiltration and NET formation are critical culprits in chronic inflammation and delayed wound closure in diabetes (8). Mechanistically, NETosis could be triggered in a NADPH oxidase (NOX)-dependent and -independent manner (33). Our study showed that there was a reduced NETosis in Lrg1−/− mice and Lrg1−/− neutrophils are resistant to calcium ionophore–induced NOX-independent NETosis. Akt is essential for NOX-independent NETosis (33). Furthermore, we demonstrated that LRG1-mediated NET formation is dependent on activation of the Akt pathway through TGFβ type I receptor ALK5, which is in agreement with LRG1-mediated TGFβ signaling in ECs (11), fibroblasts (34), glioma cells (46), and T-helper 17 cells (47). We further demonstrated that Lrg1−/− mice are resistant to diabetes-induced delay in wound closure, especially during the inflammatory phase, which is likely due to its role in NETosis. It is worth highlighting that global Lrg1−/− mice were used in this study, whereas prompt wound healing is achieved by collaborative interactions between multiple types of cells present in the wound microenvironment (1). Our BMT and in vitro experiments provided a direct evidence of BMDC-derived LRG1 on the behavior of other types of skin cells, However, this would not exclude the possible impacts of EC and keratinocyte-derived LRG1 on inflammation, reepithelialization, and angiogenesis during wound closure. To address these questions, we are now generating cell-specific knockout mice.
In conclusion, we define here a complex but critical role of LRG1 in normal and diabetic wound healing. Lrg1 deficiency leads to a significant delay in normal wound healing as a consequence of impaired inflammation, reepithelialization, and angiogenesis. On the other hand, there is a reduced NETosis in diabetic mice with ablation of Lrg1, which protects Lrg1−/− from the diabetes-induced delay in wound healing. Targeting LRG1 may represent an attractive strategy to suppress excessive NETosis, therefore accelerating wound closure in patients with diabetes.
This article contains supplementary material online at https://doi.org/10.2337/figshare.12860753.
Acknowledgments. The authors thank David Becker, Lee Kong Chian School of Medicine, for advice on histology analysis.
Funding. This work was supported by Singapore Biomedical Research Council SPF grant (SIPRAD) to X.W. and W.H. and Singapore National Medical Research Council DYNAMO NMRC/OFLCG/001/2017 and TAPP NMRC/OFLCG/004/2018 to X.W. Singapore Study of Macroangiopathy and Micro-vascular Reactivity in Type 2 Diabetes (SMART2D) is supported by the Singapore Ministry of Health’s National Medical Research Council under its Clinician Scientist Individual Research Grant (CS-IRG) (MOH-000066).
Duality of Interest. No potential conflicts of interest relevant to this article were reported.
Author Contributions. C.L. designed the study, performed experiments, analyzed data, and wrote the manuscript. M.H.Y.T. and S.L.T.P. designed and performed experiments and discussed data. M.L.L. and H.M.T. performed experiments. H.W.H., C.R., S.E.M., J.G., and S.T. contributed to discussion and reviewed and edited the manuscript. W.H. secured funding, conceived the project, contributed to discussion, and reviewed and edited the manuscript. X.W. secured funding, conceived the project, designed the study, and wrote the manuscript. X.W. is the guarantor of this work and, as such, had full access to all the data in the study and takes responsibility for the integrity of the data and the accuracy of the data analysis.
Prior Presentation. Parts of this study were presented in the abstract form at the 3rd Singapore International Conference on Skin Research, Singapore, 21 March 2018.

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August 03, 2018 @ 13:00 +03:00
Republican and Democratic U.S. senators introduced legislation on Thursday to impose stiff new sanctions on Russia and combat cyber crime, the latest effort by lawmakers to punish Moscow over interference in U.S. elections and its activities in Syria and Ukraine.
The bill includes restrictions on new Russian sovereign debt transactions, energy and oil projects and Russian uranium imports, and new sanctions on Russian political figures and oligarchs. It also expresses strong support for NATO and would require that two-thirds of the Senate to vote in favor of any effort to leave the alliance.
Russian markets reacted quickly to the measure, with the rouble slumping toward two-week lows. “The current sanctions regime has failed to deter Russia from meddling in the upcoming 2018 midterm elections,” said Republican Senator Lindsey Graham, one of the measure’s lead sponsors. Earlier this week, Graham had told reporters he planned a “sanctions bill from hell” to punish Russia. Congress passed a Russia sanctions bill last summer but some lawmakers chafed at what they saw as President Donald Trump’s reluctance to implement it; he signed it only after Congress passed it with huge majorities.
Republicans and Democrats united last month in repudiating Trump’s failure to publicly condemn Russian President Vladimir Putin for interfering in the 2016 U.S. elections. Still, Congress failed to pass anything before lawmakers left Washington for their weeks-long summer recess. Democratic Senator Bob Menendez said the administration had not fully complied with those sanctions. “This bill is the next step in tightening the screws on the Kremlin and will bring to bear the full condemnation of the United States Congress so that Putin finally understands that the U.S. will not tolerate his behavior any longer,” Menendez said.


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Kaljo Raid (4 March 1921 - 21 January 2005) was an Estonian composer, cellist and pastor.
He was born in Tallinn. One of three children, he had an older brother and a twin sister. He studied composition at Tallinn Conservatory under Heino Eller. His Symphony No. 1 was performed in 1944, the year of his graduation. He studied theology in Stockholm from 1945 to 1946 and then at the Andover Newton Theological School in Massachusetts from 1946 to 1949. He taught music at Bethel College in St. Paul, Minnesota, meeting Jacques Ibert and Darius Milhaud.
In 1954 he moved to Canada and became the pastor of the Estonian Baptist Church in Toronto; he continued in this capacity for 35 years. As a result of a late marriage in 1982 he became stepfather to five children. After retiring in 1989 he devoted himself full-time to composition. He died at Richmond Hill, Ontario in 2005.
Among his works are four symphonies and an opera on the life of Polycarp of Smyrna, Fiery Chariots (1993). He also completed the first movement of Eduard Tubin's unfinished Symphony No. 11.
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Ensemble, Madame Morrible U/S
Training: The Musical Theatre Academy.
Theatre includes: understudy ‘Carrie’ in Carousel (English National Opera, London Coliseum); ‘Fortune Teller’ in Side Show (Southwark Playhouse); ‘Frenchie’ in Cool Rider Live (Lyric Theatre) and West End Women (UK tour).
Workshops include: ‘Tara’ in Crash! (Theatre Delicatessen).
Concerts include: Of Thee I Sing (Royal Festival Hall); World Aids Day (St Paul's, Covent Garden) and Julie Atherton: Rush of Life (album launch).


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2017 Summer Reading Program registration
KENNEWICK, Wash. — The Mid-Columbia Libraries is hoping you’ll pick up a book with your child and read this summer as part of the 2017 Summer Reading Program
Registration begins Monday, May 15 and the program starts June 1.
Multiple activities and prizes are up for grabs and you can find that full list, here.
If you register before the deadline, you will be entered to win a LEGO Batman set.


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Lycoming County, the largest county in Pennsylvania, is positively the place for the outdoor enthusiasts, offering a wide range of year-round recreational opportunities. The most picturesque portions of the northeast United States are here in Lycoming County. There are year round sporting events, campgrounds, trails for hiking, biking and ATV riding, areas for skiing, hunting, fishing, boating and more. Enjoy the Susquehanna River and all of its many tributaries like the Pine, Loyalsock or Muncy Creeks. Lycoming County is home to several of Pennsylvania’s most beautiful State Parks & forests. There are lakes, waterfalls, creeks, trails, camping, boating, fishing, great views and so much more.
- Riverfront Park/Greevy Boat Launch Montoursville. There are bike paths and a boat dock.
- Little Pine State Park in Waterville PA is inTiadaghton State Forest. Little Pine Lake, hiking trails and the campground are prime attractions to the park. The lake and connecting stream provide fishermen with the perfect spot to cast their lines.
- Susquehanna State Park is a 20-acre recreational area situated along the Susquehanna River. The city of Williamsport operates this park in cooperation with the Pennsylvania Bureau of State Parks.
- Tiadaghton State Forest, Waterville, the home of Worlds End State Park, a narrow S-shaped valley of the Loyalsock Creek south of Forksville, Sullivan County. Loyalsock Canyon Vista, reached via Mineral Spring and Cold Run roads, provides breathtaking views, especially during fall foliage.
- Loyalsock State Forest Dushore is named for the Loyalsock Creek, a scenic freestone stream that winds through the heart of its 114,494 acres. The forest spans across the northern tier’s “Endless Mountains” and features flaming fall foliage and cascading waterfalls.
- Susquehanna River Basin Water Trails: Here are the trails in the area West Branch Susquehanna River Water Trail, Middle Susquehanna River Water Trail, PIne Creek Trail and Lehigh River Water Trail.
Whether you canoe, Kayak, Ski or Bike, Country Ski and Sports is the place to shop. We are a full service shop offering bicycle repair and tune up, minor repairs on canoes and kayaks including all accessories. We rent kayaks, canoes, cross country skis and snowboards. We offer kayak classes for the beginner or the experienced kayaker.
Thank you for visiting our website.

domain: geography


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The Rhoads-Cande profile photographic camera and its updated version, the video REMOTS (Remote Ecological Monitoring of the Seafloor) system, are described for in situ remote monitoring of the seafloor. These systems allow one to identify temporal and spatial changes in both the benthic fauna and physical-chemical aspects of the sedimentary environment. Image interpretation is based on a model which allows one to deduce the stage of infaunal succession from observed organism-sediment relations. The REMOTS system was developed because existing methods of obtaining ecological information are labor intensive, expensive, and data return is slow. The REMOTS system may be used for efficient monitoring of dredge-spoil activities and as a reconnaissance mapping tool for detecting change in benthic habitats in the region of outer continental shelf drilling fields.


The US Patent & Trademark Office has supplied Impact Cryotherapy with a broad patent that covers the fundamental features of its cryotherapy chambers.
Under the patent Impact Cryotherapy will receive protection for its technology related to the delivery of nitrogen gas that allows for rapid pre-cooling of the cryotherapy chamber.
The patent claims cover several aspects of the cryo chamber’s unique safety features. These features allow for safe and efficient operations during a cryotherapy session.
“We entered the market four years ago by first developing a product that set new standards in terms of its innovative design and safety,” said Richard E Otto, President and CEO at Impact Cryotherapy.
“We became the US market leader in large part because of the quality of our engineering, which is now protected by this patent, which we believe will be the first of many.”
“Over 500 Impact Cryotherapy cryo chambers have been sold in the US and internationally. The issuance of this patent will help assure the protection of our proprietary design and technology,” Otto concluded.
Source: Impact Cryotherapy
Impact Cryotherapy also has additional patents covering other aspects of the company’s innovative and leading cryotherapy chambers and related processes.
Month on month, the gasworld website is the market-leading news portal for the global industrial gas sector, growing at an unprecedented rate and keeping its readers at the forefront of breaking news, insightful analysis and must-see features across the industry. Launched in 2004 and continually evolving, it is the only independent online news, views and intelligence portal for the global industrial gas community and the larger end-user markets – and home to the ever-increasing range of gasworld platforms.
To find out more about gasworld's advertising opportunities click here.


text:
$9.00 Add to Cart
Rapala proBass Fishing
Only 1 available
Shipping: US-Mainland: $3.60 (more destinations)
Returns: 10 days, buyer pays return shipping (more)
Rapala proBass Fishing, Wii Video Game. Activision 2010. plays perfect, no subscription needed. comes in original case with instruction booklet, both mint condition. Dominate the bass tournament. Used game like new.

domain: entertainment


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We’re here to tell you what we know, but don’t take it as medical advice. Talk to your medical provider about your specific health concerns.
Everything you need to know about the dermatologist-designed formula that’s gentle on acne-prone skin yet tough on UV rays.
What to do about annoying cysts on your back.
Get to the bottom of what’s inside your pimple and how to get rid of pus-filled blemishes.
Get to the bottom of what’s causing acne and blemishes so you can help stop them for good.
Everything you need to know about clearing up infected pimples—and steering clear of new ones.
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Say goodbye to pesky pimples and learn how to zap those zits.
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Say goodbye to painful, swollen acne with these tips to help reduce and prevent it.
A little bacteria can have a big role in forming pimples.
Acne vulgaris is an extremely common skin condition—here’s how to treat it.
Say goodbye to moderate acne with the help of these treatments that can help stop it in its tracks.


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Nestled beneath a pine forest and situated right on the waterfront Camp Wellington: Campervan Park is in a wonderful location. Just 15 minutes to the city and 7 minutes to the airport means it's the perfect place to stay and discover Wellington or just recharge your batteries before you continue your journey through New Zealand.
This place has it all.... unlimited powerful hot showers, clean plentiful toilets, cosy lounge, games room and that all important FAST, UNLIMITED WIFI.
Each of the Campervan sites has a sea view and because there are only 6 spaces it feel spacious despite being a very small park. 2 of the sites have 2 power , the other 4 are unpowered. Each site has it's own picnic table and there is also an outdoor games & picnic area as well as a small garden area directly adjacent to the sea for everyone to enjoy.
- Large cosy lounge
- Games room with pool table and vinyl player
- Large kitchen with all the kitchen appliances you could ever need
- Unlimited hot, powerful showers
- Clean, plentiful UNISEX toilets
- UNLIMITED FAST FIBRE WIFI
- Close to lots of walking and cycling tracks
- Safe swimming across the road
- Little Blue Penguin (Korora) nest underneath the lodge (you can sometimes see them cross the road at night)
- Cafe within 200 metres , open from 8am to 5pm daily (try their amazing seafood sandwiches)
- 5 minutes to Weta Cave and Weta Workshop
- Miramar village just 5 minutes drive away (Supermarket, bars, pharmacy, cinema, take-aways)
- "Bandit" the managers sweet, fur-baby (Huntaway cross dog)
Te Āti Awa settled on the bay before European arrival, Maru-Kai-Kuru was the name of the village sited at the northern end of the bay.
In 1839 the bay was sold to the New Zealand Company along with most of Wellington. In 1907 the land was transferred to the Royal New Zealand Navy, and in 1946 it was transferred again to the Royal New Zealand Air Force to accommodate up to 300 staff. The New Zealand Defence Force owned land on Shelly Bay until 1995 when the then Air Force Base was closed.
On 14 February 2009 the land was handed over to Taranaki Whanui ki Te Upoko o Te Ika as part of a Treaty of Waitangi settlement.
For more information or to make a reservation go to www.campwellington.co.nz
or call Simon on +64 27 233 8269
Camp Wellington is run and managed by Si (Simon Craggs) who also runs and owns BEDVANZ ;)

domain: travel


text: Our pomegranates also come from Selcuk/Izmir in Turkey. The trees that can reach up to five metres in height belong to the loosestrife family. The fruit resembles to an apple and firstly, it is green as it only gets its characteristic red colour at a later stage. Pomegranates can be eaten raw. Between September and December, the ripened fruits are picked from the trees and prepared for sale.


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When i have fears that i may cease to be ( 1818 1848) this sonnet, is possibly john keats first use of the shakespearean form, is usually. The poem `ode on a grecian urn' is a poem written by john keats in the form of an ode in its original (greek) form, an ode is an elaborately structured poem. John keats was an english romantic poet he was one of the main figures of the second it was lockhart at blackwoods who coined the defamatory term the cockney school for hunt and his the largest collection of the letters, manuscripts, and other papers of keats is in the houghton library at harvard university. The subject of this letter from keats to his publishers john taylor and keats' expression “manufactured rags” was a slang term for paper. Im writing a term paper for my honors brit lit class on to autumn and im a lil stuck on clearly defining 'the theme' of the poem if an yone at all.
John keats research papers seek to trace the development of the romantic elements get help with your research paper on keats and have one of paper masters' gothic revival - gothic revival term papers analyze the architectural style. Full-text paper (pdf): john keats and john clare: the ivory tower poets of been changing constantly, this term has undergone the same.
Free john keats papers, essays, and research papers piece of literature conveys in terms of motif and attempts to portray that aesthetic value onto the screen. According to the romantic english poet john keats (1795-1821), artists of a progressive and anti-monarchal newspaper,” says wolfson. Read the odes of john keats (paper) (belknap press) book reviews & author here's how (terms and conditions apply) go cashless: pay with bhim upi, get.
Revise and learn about john keat's poem, to autumn with bbc bitesize gcse english literature poetry resources. James najarian delivered a paper for the roundtable on keats's discusses thomas hood embracing the shadow of john keats on the poets' relationship, sequels in poetry, and the often derided term “minor poet. This paper seeks to understand keats's early poems as an attempt to keywords: john keats, 'sleep and poetry', 'i stood tip-toe upon a little.
Nature and beauty in keats great odes - paola bertolino - seminar paper publish your bachelor's or master's thesis, dissertation, term paper or essay this writing focuses itself on john keats, who lived a short time between the 18th and. John keats and his siblings were quite popular at school also if you need a custom essay, dissertation, thesis, term paper or research paper.
In this paper i should like to raise questions about the kind all use subject to 4adonais: a life of john keats (london 1938), p. The consecrated urn: an interpretation of keats in terms of growth and form london: john keats: his life and poetry, his friends, critics and after-fame. John keats was born in london on 31 october 1795, the eldest of thomas and frances and taking first prizes in essay contests his last two or three terms.


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A Whiting Landmark
By John Hmurovic
You can call it Chase Bank. Or, you might still call it Bank One, or Bank of Indiana. If you are older, it may be the State Bank of Whiting to you. The oldest among us may still remember it as the First National Bank. But most of us would agree that the building on the northwest corner of 119th Street and New York Avenue should be called a Whiting Landmark.
The Merriam-Webster dictionary says a landmark is “a structure (such as a building) of unusual historical and usually aesthetic interest.” For such a small community, Whiting-Robertsdale is blessed with several landmarks, including the Whiting Public Library, the Community Center, and the Hoosier Theatre. Those buildings add to the beauty of the community and give Whiting-Robertsdale a personality that makes it different from other cities. Landmarks are buildings which have been a part of our lives from our first days here. No matter how different we might be from our neighbors, we are bound together by living in the same community. Landmarks are a part of our common identity.
The bank building on the northwest corner of 119th and New York fits into the landmark category. It certainly holds “aesthetic interest.” More than any other bank in Whiting-Robertsdale, it looks like a bank. Or at least, it looks like a bank looked in 1914. That’s the year it was built. Its stately appearance adds to the attractiveness of downtown Whiting, much more than a modern building would at that busy intersection. That’s because it fits in with its neighbors: the 1930s-era Post Office across New York Avenue; the early 20th century buildings on the corners across 119th Street; and the Whiting City Hall building next door. Of all these old structures, the bank on the northwest corner is, arguably, the most interesting.
But besides aesthetic interest, it has historical importance. The building was constructed in 1914. It was built to be the home of the First National Bank of Whiting. “It will be one of the most beautiful and impressive buildings in Whiting,” wrote Louise Mattern and Marguerite Schaub in the 1914 edition of the Reflector, the yearbook of Whiting High School. “The people of Whiting can look forward with pride to its completion,” said the Whiting Call newspaper in 1913 when plans for the new building were announced.
The bank was established late in 1902, with Gallus J. Bader and Fred J. Smith as its principal creators. Its original home was just a block away, on the northwest corner of 119th and Oliver. It was in Dr. G.S. Hilliard’s brick building, a structure which still stands and is currently the home of the Comfort Roast coffee shop. Dr. Hilliard was a dentist who came to Whiting in 1897. His office was on the second floor of his building, while the bank shared the lower floor with a doctor’s office. The First National Bank grew rapidly, starting with $40,000 in deposits in 1903, to half a million dollars by 1914. More cashiers and clerks had to be hired. The Hilliard building was too small.
The new building was ground-breaking for Whiting in several ways. It was, for instance, the first major building in the city which reflected the fact that women were increasingly moving into the workforce. “An interesting illustration of the feminine invasion of the business world,” Miss Mattern and Miss Schaub wrote in the Reflector, “is in the waiting, or rest room, provided for the ladies.”
Another unique feature, which all city residents and visitors were able to enjoy, was the clock attached to the front of the building. It was nine feet high, three feet wide, with thirty-nine-inch dials. The name of the bank was above the face of the clock. The clock could be seen from a distance, in either direction, on 119th Street, even at night, because it was illuminated by electricity, another uncommon feature for signs in 1914. The clock could also be heard. Every quarter of an hour it had a different sound as it chimed. Then, at the top of each hour, the full Westminster chimes would play, followed by the strike of a chime for each hour, to let listeners know the time of day.
When the bank was built, the Congregational Church was just to its west, where Whiting City Hall is now located. The space between the church building and the bank was intentionally kept open. The idea was to create a space that was green and cheerful. It remains open to this day, and since 1970 it has gone by the name of Binhammer Park, named in honor of the bank’s long-time president, Carl Binhammer.
Another feature of the building, in its early years, was the placement of flower boxes on the ledges of the second-floor windows. “The First National Bank has blossomed out with window gardens,” the newspaper said in 1915, “greatly to the satisfaction and pleasure of all concerned.”
Architect M. Clifford Wiley of East Chicago designed the building. It cost between fifty and sixty thousand dollars to build in 1914. Its grand opening on March 27, 1915, was a big event in Whiting. Over four thousand people came to look. It was too early in the season for the outdoor flower boxes to be in bloom, but indoors there were flowers everywhere, and their fragrance filled the air. Most of the flowers came from other businesses, sent as a grand opening gift. The bank itself, greeted all women who visited that day with a large, American Beauty rose, while all girls received a carnation. The fresh smell of flowers was probably mixed with cigar smoke, because every adult male who visited received a cigar. The cigars were called “Jim’s Best,” and were made in Whiting.
But the bank building itself was the main attraction. Visitors were told to notice the floor of Tennessee marble; the fixtures of Skyros marble from Greece; and the African mahogany in the director’s office. The cages of the tellers were on the first floor. So were two offices which were important in a town filled with immigrants: A foreign exchange for money brought from overseas, and a steamship booking department to arrange journeys between Whiting and the home country of the immigrants. “Two immense vaults with time lock combinations on their massive doors,” were also available for viewing, according to an article in the Whiting Call.
Open house visitors could also go to the second floor, which had its own entrance on New York Avenue. There were five office suites upstairs, consisting of four rooms in each suite. The first occupants were attorneys (John H. Fetterhoff and George Michaely), and doctors (A.J. Lauer, B.W. Avery, and G.H. Hoskins).
The First National Bank building was constructed in a time of prosperity in Whiting. The refinery was booming, thanks in part to the huge demand for automobiles, and the need for gasoline to fuel those newfangled vehicles. That growth created more jobs, and more jobs led to a doubling of the city’s population between the bank’s founding in 1902, to the time its new building was constructed in 1914. So, it was no wonder that even though the building was two stories tall, it was built with expansion in mind. “The massive foundations and walls will permit its being enlarged to five stories, whenever the need shall require.”
But the good times did not last. In 1929, the national economy almost collapsed. Banks across the country shut down, never to reopen, as they ran out of money during the Great Depression. In 1932, every bank in Hammond shut its doors. In Whiting, the Central State Bank, which had just constructed Whiting’s tallest building a few years earlier, had to close forever. Two other banks were very close to collapse. But, then, Standard Oil stepped in. It was one of the best examples in Whiting history of the close relationship that existed between the refinery and the city.
“Officials of the Standard Oil Company realized that the banks…would close if something wasn’t done quickly,” said Walter E. Schrage, head of the Bank of Whiting. Standard, one of the nation’s largest companies, did not want to see the banks fail in the hometown of its largest refinery. The company stepped in to save the two that were closest to failure: First Trust and Savings, and the First National Bank. “A conference was held,” Schrage said, “and a new institution, the State Bank of Whiting, was organized.” Standard Oil used its resources to back up the new bank. The knowledge that Standard Oil was there, and would not let the new bank fail, kept depositors from panicking and withdrawing their money from the bank. The president of the new State Bank of Whiting was Edward G. Seubert, the president of the Standard Oil Company of Indiana.
Over the years, the State Bank of Whiting changed owners and names several times. The building, however, still stands on the corner of 119th and New York Avenue. Shortly after it was constructed, the building was hailed as “one of the great landmarks in Whiting history.” It was unique for its time, but it was probably premature to call it a landmark in 1914. But in the century-plus that has gone by, the building’s stately appearance has become something which every living generation of Whiting-Robertsdale residents recognizes. It has become a part of what gives downtown Whiting its character. Combine that with its rich history, and there can be no doubt that no matter what it is called, the bank on the northwest corner of 119th and New York is a Whiting Landmark.


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Jason Daniels, Ph.D.
Early America, Colonial Caribbean
Dr. Jason Daniels earned a Ph.D.from the University of Warwick (U.K.) in 2013, a M.A (2008), and a B.A. (2006) from the University of Florida. Prior to joining CSU-East Bay, Daniels held positions at Black Hills State University in Spearfish, South Dakota (2015-2020), Knox College in Galesburg, Illinois (2014-15) and Indian River State College in Fort Pierce, Florida (2008-2014). His research focuses primarily on the eighteenth-century, Anglo Francophone Atlantic world. He is particularly interested in the relationship between the Caribbean and North American colonies. He has published articles in the The Journal of Caribbean History, The Florida Historical Quarterly, and Pennsylvania History: A Journal of Mid-Atlantic Studies. He teaches classes in early America, the Caribbean, and the Atlantic World.


Leicester, Northampton dominate 2022/23 Premiership Dream Team XV
Leicester and Northampton have led the way in the 2022/23 Gallagher Premiership Dream Team, the East Midlands rivals getting three players each selected on the XV chosen by a BT Sport selection panel. Nine of the 11 clubs that finished the financially-hit tournament’s regular season – Wasps and Worcester didn’t make it past the first month – had representatives picked, with only Heineken Champions Cup semi-finalists Exeter and ninth-place Bristol missing out.
Tigers forwards Julian Montoya, George Martin and Jasper Wiese made the cut, as did the Saints trio of Fraser Dingwall, Alex Mitchell and David Ribbans. Leicester, who are on their travels this Sunday in the Premiership semi-final, will come up against a Sale side that had two players named in the Dream Team – Joe Carpenter and Rob du Preez.
The other semi-finalists Saracens, who host Northampton on Saturday, also had two players selected, tighthead Marco Riccioni and back-rower Ben Earl.
A statement read: “The Gallagher Premiership Rugby season has brought no shortage of impressive individual showings and the BT Sport selection panel have been hard at work selecting their 2022/23 Dream Team.
“Their final selections, revealed at the Premiership Rugby Awards presented by Gallagher, saw no less than nine teams represented. East Midlands duo Leicester Tigers and Northampton Saints lead the way with three players apiece making up part of the coveted XV.
“All three Tigers stars are in the pack, with Julian Montoya, George Martin and Jasper Wiese all selected. Montoya packs down between props Val Rapava-Ruskin, Gloucester Rugby’s sole representative, and Saracens’ Marco Riccioni.
“Martin is joined in the second row by Saints ace David Ribbans, while Earl is joined by two players who were nominated for individual awards in the back row – Tom Pearson, who won the breakthrough player of the season award, and Gallagher player of the season nominee Jasper Wiese.
“Robert du Preez joined Wiese on that shortlist and is selected at fly-half as one of two Sale Sharks players in the back line, with full-back Joe Carpenter also integral to leading Alex Sanderson’s side to their first home semi-final since 2006.
“Ollie Lawrence, who was voted Gallagher player of the season nominee, joins Fraser Dingwall in midfield while the top two try scorers in 2022/23, Harlequins’ Cadan Murley and Newcastle Falcons flyer Mateo Carreras, are on the wings.
“Both players walked away with individual awards from the evening, Murley picking up the top try scorer prize after crossing 15 times across the season – two clear of Carerras, who picked up the try of the season prize for his superb solo effort against Northampton in round six.
“Saints’ Alex Mitchell, meanwhile, is selected at scrum-half following a string of excellent displays in the number nine jersey.
BT Sport Dream Team 2022/23
15. Joe Carpenter (Sale Sharks)
14. Cadan Murley (Harlequins)
13. Ollie Lawrence (Bath Rugby)
12. Fraser Dingwall (Northampton Saints)
11. Mateo Carreras (Newcastle Falcons)
10. Robert du Preez (Sale Sharks)
9. Alex Mitchell (Northampton Saints)
1. Val Rapava-Ruskin (Gloucester Rugby)
2. Julian Montoya (Leicester Tigers)
3. Marco Riccioni (Saracens)
4. George Martin (Leicester Tigers)
5. David Ribbans (Northampton Saints)
6. Ben Earl (Saracens)
7. Tom Pearson (London Irish)
8. Jasper Wiese (Leicester Tigers)
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Gavin Hastings, was playing was he? 🤔Go to comments
Let Moana go and setup Timi Samoa and a Timi Tonga. I guarantee you that the support will be even stronger than that for the Fiji Drua. You only have to remember the support from The respective communities for Mate ma Tonga and Toa Samoa in Rugby League and also the national teams in Rugby Union. Both teams would be able to field teams equal to The Fiji drua or even better.Go to comments


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Starter Houston Oilers Satin Full-Snap Blue Jacket
The Houston Oilers Blue Satin Jacket is made from satin material. The satin material used to fabricate this jacket is of the best quality. It is sure to fell super warm, cozy and comfortable on your skin. The color of this jacket is blue. It has a logo on its chest which makes it a perfect Houston Oilers jacket. It will make you look dapper and sturdy. This jacket can be worn by both the genders because of its neutral look. The inner side of Blue Houston Oilers Retro Jacket is of viscose lining which makes it very warm and cozy. The material is also very much breathable. The Houston Oilers Blue Satin Jacket has buttoned closure along with a stand up collar. The standup collar gives it a classy look. This jacket has full length sleeves along with rib knitted cuffs. It also has two waist pockets to keep your hands warm.

domain: design


text:
From delayed reactive medical services to evidence-based Predictive, Preventive & Personalised Medicine.
March 2019: "Innovative Predictive, Preventive and Personalised Medical Approaches in Dental Care, Suboptimal Health Conditions and Stroke"
Issue Editors: Babak Baban, Vincenzo Costigliola, Mahmood Mozaffari
June 2019: “Predictive, Preventive and Personalised Medical Approach – Multiprofessional Expertise”
Issue Editors: Rostyslav Bubnov, Vincenzo Costigliola, Romano Danesi, Josef Flammer, Marko Kapalla, Mahmood Mozaffari, Halina Podbielska, Byong Chul Yoo
September 2019: “Predictive, Preventive and Personalised Medical Approach in Pregnancy, Diabetes mellitus, Cancer and Visual Impairment”
Issue Editors: Marko Kapalla, Mahmood Mozaffari, Niva Shapira, Byong Chul Yoo, Xianquan Zhan