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Some of the largest chocolate and candy companies across the globe plan to reduce sugar in their products to adapt to many consumers’ efforts to eat healthier.
The group of companies—Mars Chocolate, Wrigley, Nestlé USA, Ferrero, Lindt, Ghirardelli, Russell Stover, and Ferrara Candy Co.—plan to make 50% of their individually packaged products 200 calories or less within five years. The initiative also includes making these changes obvious to consumers by labeling calories on the front of packaging, as well as making packages smaller and innovating products, the National Confectioners Association (NCA) said.
There are millions of dollars being invested in research and development by the companies—collaborating together for the first time on health and wellness initiatives.
Mars has been a leader in working to meet the needs of health-conscious consumers while at the same time satisfying consumer’s colossal sweet tooth by not sacrificing taste. The company has already debuted a variety of lower-calorie candies including M&Ms, Maltesers and Snickers.
“Chocolate and candy have always been a treat, and this is a big commitment by the participating companies to keep it that way,” John Downs, president & CEO of the NCA, said. “We are proud to make this five-year commitment with The Partnership for a Healthier America, a highly regarded nonprofit organization that will help us track and verify this meaningful initiative. This is the first step on our journey to recruit other companies to join us as we work to help consumers manage their sugar intake and ensure that they feel empowered to make informed choices.” Read the article …


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“Books contain an endless source of knowledge and pleasure for children. If we teach our children to make reading a habit, we are handing down a special kind of magic to them. There are many ways to help our children become voracious, lifelong readers. We will discover some unexpected bonuses too. Our children will do better in school, improve as a reader and enjoy a richer and fuller life and be able to keep themselves from negative things. We as parents take our job seriously by making sure that our children eat healthy food and take proper rest. But what about their minds? Are we feeding that endless curiosity that causes children’s minds to grow in a healthy way? That’s where books are imperative.”
Fine PrintBlog dedicated to promoting reading, writing, and some espionage. Includes original content by SpyWriter Jack King, as well as interesting and related snippets from around the world (used in accordance with the Fair Use Policy).
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Jack SpyWriter King
"A New King of Thrillers on the Horizon" ~ Sisters in Crime Author of Thrillers: Agents of Change, WikiJustice, The Black Vault, The Fifth Internationale... www.SPYWRITER.com

domain: literature


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Over the last three seasons The Flash has undergone a radical tonal shift, and from what we seen in this year's SDCC trailer we can expect more of the same going into Season 4. Last time out we saw a focused, and ultimately selfish, side to Barry Allen as he endeavoured to save Iris from an untimely demise. Given the series' rather glum tone over the course of the year, we probably shouldn't have been surprised that the final episode [SPOILER WARNING] saw one of the shows lighter characters H.R. sacrifice himself to save the day. Of course, Barry disappeared into the Speed Force at the end of the episode so the whole Barry-Iris relationship was doomed to be split anyway.
This is where we pick things up in the trailer for Season 4. After Barry's willingness to set everything aside in a bid to save Iris, we see that Barry's disappearance has seen her shift focus to filling in his shoes to protect Central City. This is particularly in character for Ms. West who seems to always shut herself off and get stuck into a project rather than deal with her grief. Wally and Cisco too seem to have come to the same conclusion in order to protect the city, though the latter is seen to show his usual level of emotion at the loss of his friend. Caitlin Snow also makes a brief appearance in the trailer, seeming back to her old self but with the confidence of her Killer Frost side which could mark her as one of the new season's more interesting characters. Of course, all this focus on the other members of Team Flash comes at a loss of new shots of our titular hero. We know that the team are working on getting Barry back, but can we be sure that the method shown in the trailer is successful? We'll just have to wait and see when The Flash returns in October.

domain: entertainment


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Lance Sheppard, Head of Marine Division of EcoStore in New Zealand and ISS Board Member, chats about the situation in his country, which has been in lockdown since March last year, and how he has been handling his business under these very unpredictable circumstances.
How have you carried on promoting your company outside of New Zealand while your country is shut from the rest of the world? Ecostore has partnered with leading global marine companies that share the same passion that we must protect and preserve our oceans and reduce the amount of single use plastic on Superyachts. We are working closely with Burgess Yachts and Fraser Yachts supporting and sponsoring various events they have been holding for crew (Frasers Short Story Contest, Burgess Beach Clean up) These companies also have a strong company mandate to transition the boats under their management to ensuring they become more operationally sustainable in everything they do. This strong network has been very important in times when we cannot travel ourselves.
How do you ship your products during the lockdown? Is it more complicated? Most of our shipments are by seas freight so this has not been impacted. Getting samples to boats via airfreight has been more challenging. During the first few months of this year there was some reluctance to order with such uncertainty around global transport links, but as the integrity of these were maintained our clients felt more comfortable to continue to order.
Have there been production or supply problems on your end to manufacture your products? All our products and natural ingredients are manufactured in New Zealand so we have not had issues with product availability. Our only issue was getting pumps for the hand wash bottles which come out of China. Expanding the suppliers in our supply chain has mitigated the pump shortage but we have had an interesting variety of pump styles on our products.
Have you had to re-think your company strategy? Our marine strategy has had to be flexible with the uncertainty that has been created globally. Many boats are using this time to go into refit or their cruising season has extended making logistics a little tricky when the boats are on the move. Overall because the strategy we have in place has been built around a clientele which is always mobile we have had the flexability to built in to manage the curve balls this year has thrown us.
How do you envision the next few months or year? For Ecostore, growth into the marine sector will continue to strengthen. The demand for our plant and mineral based environmentally sustainable products on Superyachts is continuing to grow as more crew hear about us and the quality and performance of our products. There is a global increasing awareness for sustainability. The Americas Cup in New Zealand was certainly going to be one of our launch events, with upwards of 150 Superyachts registered to come down pre pandemic. With our borders still closed many boats have decided not to come. However we are seeing several Superyachts a week still arriving so we are very confident that momentum to help Superyachts transition operationally to more sustainable products used on board will continue to keep us busy during this time.
Ecostore has been voted New Zealand’s most trusted brand 5 years in a row.Our marine specific website www.ecostoreocean.com Also our Instagram @ecostoreocean.
Interview by Christelle Holler

domain: news


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Here are several versions of what might be one of the best songs ever written, Cole Porter's haunting (and slightly unsettling) 'Night and Day', ranging from the inspired to the downright odd. I had no idea Ringo Starr recorded it, nor do I know anything about the Japanese girl-group version except that it has a very weird video. If it weren't for a slightly messy ending, the version by Everything But The Girl would definitely get my vote, as its pared-down approach and Tracey Thorn's wonderful voice really allows the song's melody to shine. And I love the ukelele version!
Sister Q - Night and Day by ChibiLune


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Hours before the executive order was signed, Pope Francis said he stood with the USA bishops, who had condemned the family separation policy, which has led to children being held in government shelters while their parents are sent to federal prisons.
Immigration and Customs Enforcement (ICE) now separates most families when they are apprehended for illegal border crossing, Reid notes.
Die-hard supporters of Donald Trump say they remain steadfast despite an uproar over his "zero-tolerance" policy on illegal immigration that led to the separation of some children from families. But immigration advocates criticize that move as allowing a more indefinite detention of families until criminal and removal proceedings are completed, signaling legal battles ahead.
The executive order does not undo or diminish Sessions' zero tolerance policy, which has caused the spike in parent-child separations at the southern border.
Under the administration's policy, all unlawful crossings are referred for prosecution - a process that moves adults to the custody of the U.S. Marshals Service and sends many children to facilities run by the Department of Health and Human Services.
"We're going to have very strong borders, but we're going to keep families together", he said.
Video footage of children sitting in cages and an audiotape of wailing children had sparked anger as the images were broadcast worldwide.
India takes direct control in Kashmir after alliance ends
The state BJP claims to be prepared to face elections whenever it happens but the Central leadership of the party is in no hurry. Omar hit back, saying he had no doubt about his MLAs and referred to the BJP's alleged horse-trading bids in Jammu and Kashmir.
United chief executive Oscar Munoz said: "We want no part of it".
"The administration must end this gratuitous, cruel, and counterproductive policy that has brought such pain to innocent children and so much shame on this nation", Schumer said, adding that no law requires the separation of children from their families. Several White House officials advised the President to accept a stand alone bill addressing family separations or some other solution to this issue.
Last year, Ivanka Trump did offer strong words against Roy Moore, the Republican candidate for Senate in Alabama, who was accused of sexual misconduct with teenage girls during the late 1970s, when he was in his 30s.
One possible reason for Trump's about-face may very well be his wife. I feel very strongly about it.
"You have people that want absolute security and safety, and you have people that do look at the children and then you have people like me, and I think most of the people in this room, that want both", he said. For teens housed in secured facilities, the placement process became more arduous in 2017 after policy changes required Office of Refugee Resettlement Director Scott Lloyd to sign off on any placement. Instinctively combative and fond of chaos, Trump usually digs in on controversial policies, rather than backing down.
British Prime Minister Theresa May declared the United States is wrong to separate migrant children from their parents, but rejected calls to cancel Trump's visit to the UK next month. She told The Associated Press at the time: "There's a special place in hell for people who prey on children".
At the same time, the Justice Department went to court in an attempt to overturn a decades-old settlement that limits to 20 days the amount of time migrant children can be locked up with their families. Any attempt by the government to modify the agreement would likely face opposition from the lawyers who negotiated it, according to representatives of immigrant rights organizations.
New Zealand Prime Minister gives birth to healthy baby girl
Ms Ardern arrived at the country's largest public hospital at 5:50am (local time) with Gayford, her spokesman said . The first female prime minister was Sirima Bandaranaike, who was elected to lead Sri Lanka (then Ceylon) in 1960.
In addition, the order blames Congress and court decisions made in the past for putting the Trump administration in a position where it had to separate families in order to effectively enforce the law. Media reports have appeared about heart-rending scenes of children crying for their parents.
Nearly 2,000 immigrant children were forcibly separated from their parents between April 19 and May 31 following arrests involving illegal entry into the United States, according to the Department of Homeland Security (DHS).
It is not immediately clear whether, or when, detained children will be reunited with their parents.
What does the executive order mean for House Republican legislation?
Even as late as Tuesday evening, Trump adamantly insisted to staff that his hardline stance on immigration - the most potent issue of his 2016 presidential campaign - would help the Republican Party with voters in November's midterm elections.
House Republicans will postpone a vote on their immigration compromise legislation until Friday, Majority Leader Kevin McCarthy told reporters as he left a leadership meeting Thursday afternoon.
Republicans have never been able to pass a big immigration overhaul on their own, despite talking about the issue for years.
FPJ's dream XI predictions for Serbia and Switzerland; Live Blog
The result left Brazil, 2-0 winners over Costa Rica, top with four points, ahead of Switzerland on goal difference with Serbia on three.

domain: sociology


We often hear about all of the technology that goes into the modern time trial bike and equipment. Recently I read an article by some MIT cyclists about what equipment choices saved the most time.
For the racer who cannot afford a time trial rig and who is using clip-on aero bars, what other equipment choices will make the most difference? I have lost time trials to people with all the gear (i.e. disc wheel, aero helmet, skinsuit, TT bike, etc.) by only 10 to 20 seconds. The MIT guys suggest that not wearing gloves is better than an aero wheel. Assuming my engine is as good as it is going to get, what investments can I make that will give me the most bang for my buck?
Leave the gloves at home, and clean up the cable housings on the front of the bike. Trim and route them out of the wind. Fold electrical tape over bare cables along their length wherever they cross the wind (like along the down tube) to form an aero trailing edge. A thin wire has as much drag as a much wider aero wing section, which is why antennas sticking out of airplane fuselages are airfoil-shaped.
MIT results suggest that an aero helmet can save 1.5 minutes on a 40km TT; that will of course depend on which helmet you choose and which one it replaces, how you hold your head, and how fast you ride. MIT grad Mark Cote has a prototype water bottle that sits on the top tube behind the stem that saves almost two minutes. But the bottom line? All of these equipment choices are secondary in importance to good body position.
Attention cycling fans: Subscribe to VeloNews—the Journal of Competitive Cycling—and receive 15 huge issues filled with behind-the-scenes race coverage, news analysis, action photos, rider interviews, expert training advice, unbiased product reviews and more.


Guide for Integrated Urban Weather, Environment and Climate Services (IUWECS) and how it can best meet the needs of researchers and stakeholders in Salon 8
11:00 AM - 12:30 PM
The Integrated Urban Weather, Environment and Climate Services (IUWECS) is a new initiative from the World Meteorological Organization (WMO) that seeks to provide science-based integrated urban services supporting safe, healthy and resilient cities. As part of that initiative, a Guide for IUWECS is being developed. The CitiesIPCC conference provides an excellent opportunity to present and receive feedback from attendees on the guide because the conference falls within the period for open community review.
The intent of the IUWECS Guide is to document the best available knowledge, technologies and practices for producing and providing the relevant services that cities require to respond to the hazards posed by climate change. Such services include a combination of dense observation networks, high-resolution forecasts, multi-hazard early warning systems, and climate services. These services should assist cities in setting and implementing mitigation and adaptation strategies, which will enable the building of resilient and thriving cities that promote the Sustainable Development Goals (SDGs). The Guide also plans to include a multidisciplinary approach to better serve the social-economic needs of urban areas, and identify the required partnerships to establish and sustain urban services, including research, city governments, international organizations, and private sector stakeholders.
The session presentations will illustrate the best available knowledge/technologies and practices. Feedback will be obtained from participants on desired services, products and requirements for developing effective multidisciplinary partnerships among urban/city governments with meteorological and other key agencies.


he dimensions of this simple, handmade, acoustic drum set are unconventional: the kick drum is small and deep, and the snare is smaller both in diameter and depth than what might be considered usual. Needless to say, it’s built for sensitive playing, and whoever plays it needs, above all, to have a very fine touch.
The warmth of mahogany, highlighted here by a dark cherry finish, is the inspiring feature of this simple, classic drum set.
Even when a drum set consists of only a few pieces, it can still play like an orchestra. The sounds of this drum set are proof of that. Thanks to the shells made using the segmented technique, the warm sound and response of the mahogany and the tones of a small but deep kick drum combined with a snare that is drier and more precise.


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From the LAC Blog:
Crowdsourcing has arrived at Library and Archives Canada (LAC). You can now transcribe, add keywords and image tags, translate content from an image or document and add descriptions to digitized images using Co-Lab and the new Collection SearchBETA.
To make it even more interesting, we will launch what we call “challenges”. These challenges are content put together under a theme. For example one of our first challenges is on Rosemary Gilliat (Eaton)’s. Your challenge will be to transcribe her diary and describe her photographs from her Arctic travels. Or instead, try your hand at transcribing the love letters from Sir Wilfred Laurier to his sweetheart and future wife, Zoé – another challenge now available.
The launch of Co-Lab also introduces a new image viewer – which lets you scroll to zoom in on different parts of the image, or click and drag to move around the image itself. This is particularly useful when looking to transcribe or add keywords and image tags to describe small details!
The best thing about this new tool is that every contribution made by the public directly benefits fellow researchers and improves access. Every addition to a record becomes new metadata – which is searchable within 24 hours, helping LAC’s records become more “discoverable” day after day. Transcription of textual material that was previously just digital images also becomes accessible to those who use text-to-speech machines or screen readers, and translation of transcribed documents opens the door to unilingual Canadians.
Direct to Co-Lab

domain: library


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Boathouse setting for College of West Anglia art student's chair which hits new heights
IT S a chair but not just any old chair. And whatever you do, don t try and sit on it. Because this is a work of art created by Heather Adamson, an 18-year-old student at the College of West Anglia. It now stands on permanent display in the foyer of the B
IT'S a chair but not just any old chair. And whatever you do, don't try and sit on it.
Because this is a work of art created by Heather Adamson, an 18-year-old student at the College of West Anglia.
It now stands on permanent display in the foyer of the Boathouse, Fenland District Council's business centre in Wisbech, where it first went on show last year.
Heather is currently in the second year of her National Diploma in Art and Design at the college. She is hoping to go on to university to study Fine Art Illustration.
You may also want to watch:
The initial inspiration for her piece was a dress featured in the work of the artist/photographer Chris Anthony.
Heather's interpretation features a combination of ideas from a caged petticoat at the base to a city skyline across the top.
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Sian Thomas, the course director at the College of West Anglia, said: "The brief for the students was to design a chair or some form of seating, based on the work of a particular artist or art movement. Heather interpreted that in a really exciting and original way, creating a very ambitious, large-scale artwork."
Heather said: "We were originally told just to create a model of something but I thought this would work bigger. I wanted to do something I hadn't done before."
The chair was included in "Form and Image 2", a showcase of work by COWA students that was displayed in the Boathouse last October.
Councillor Mac Cotterell, Fenland District Council's portfolio holder responsible for special projects, said: "Heather's eye-catching piece fits in perfectly with the contemporary architecture of the building. It really grabs your attention when you come in and has become quite a talking point for visitors."
The exhibition was one of a number of recent community events held at the Boathouse.
There is space available in the gallery free of charge to community (not for profit) and charitable groups. Women in Rural Enterprise (WIRE) hold regular meetings there on the last Friday of every month.
For more details, call the Boathouse on 01945 586700.

domain: artistic


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Congenital obstructive nephropathy is a common cause of chronic kidney disease and a leading indication for renal transplant in children. The cellular and molecular responses of the kidney to congenital obstruction are incompletely characterized. In this study, we evaluated global transcription in kidneys with graded hydronephrosis in the megabladder (mgb−/−) mouse to better understand the pathophysiology of congenital obstructive nephropathy. Three primary pathways associated with kidney remodeling/repair were induced in mgb−/− kidneys independent of the degree of hydronephrosis. These pathways included retinoid signaling, steroid hormone metabolism, and renal response to injury. Urothelial proliferation and the expression of genes with roles in the integrity and maintenance of the renal urothelium were selectively increased in mgb−/− kidneys. Ngal/Lcn2, a marker of acute kidney injury, was elevated in 36% of kidneys with higher grades of hydronephrosis. Evaluation of Ngalhigh versus Ngallow kidneys identified the expression of several novel candidate markers of renal injury. This study indicates that the development of progressive hydronephrosis in mgb−/− mice results in renal adaptation that includes significant changes in the morphology and potential functionality of the renal urothelium. These observations will permit the development of novel biomarkers and therapeutic approaches to progressive renal injury in the context of congenital obstruction.
Citation: Becknell B, Carpenter AR, Allen JL, Wilhide ME, Ingraham SE, Hains DS, et al. (2013) Molecular Basis of Renal Adaptation in a Murine Model of Congenital Obstructive Nephropathy. PLoS ONE 8(9): e72762. https://doi.org/10.1371/journal.pone.0072762
Editor: David Long, UCL Institute of Child Health, United Kingdom
Received: April 12, 2013; Accepted: July 15, 2013; Published: September 4, 2013
Copyright: © 2013 Becknell et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Funding: This work was supported in part by R01-DK085242, K08-DK09234 and F32-DK089733 from the NIDDK of the NIH. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
Competing interests: The authors have declared that no competing interests exist.
Congenital obstructive nephropathy (CON) is a leading cause of chronic kidney disease (CKD) and end stage renal disease (ESRD) in children, particularly in boys with prune belly syndrome and posterior urethral valves . The best available biomarker of residual renal function in children with CON is serum creatinine, even though it does not permit physicians to reliably predict which children with obstruction will develop significant CKD , . Clearly additional studies are needed to clarify the cellular and molecular responses to CON in order to develop reliable diagnostic/prognostic markers of renal injury and therapeutic strategies to reduce the impact of CKD.
A significant body of work describes the renal response to acute obstruction principally through the use of partial or complete ureteral obstruction in large animal and rodent models . In contrast, far less is known about the renal response to graded degrees of congenital hydronephrosis, a fact that is critical in understanding the pathophysiology of CON. To this end, we have developed the megabladder (mgb−/−) mouse model of CON. Mgb−/− embryos develop an atonic bladder and exhibit hydronephrosis in utero, which persists postnatally and is exacerbated in males at sexual maturation . Male mgb−/− animals eventually develop severe bilateral hydronephrosis resulting in death around six weeks of age from renal failure and severe dehydration .
Hydronephrosis in mgb−/− mice progresses at variable rates, such that by 3–4 weeks of age, a spectrum of obstruction occurs associated with mild to severe reductions in renal parenchyma and function . This variability parallels that seen in humans with obstructive uropathy . In this study, we utilized this variability to evaluate the global transcriptomes of mgb−/− versus control kidneys at varying grades of hydronephrosis. We hypothesized that distinct cellular responses would be observed in mild, moderate and severe hydronephrotic kidneys providing insight into the key molecular pathways responsible for the pathophysiology of CON as well as the identification of novel biomarkers for the staging of renal injury.
Materials and Methods
The following studies were conducted at The Research Institute at Nationwide Children’s Hospital under the Public Health Services approved Animal Welfare Assurance number A3544-01. All studies were approved by the Institutional Animal Care and Use Committee under protocol 02105AR.
Mice were maintained according to the NIH Guide for the Care and Use of Laboratory Animals as previously described . Age-matched, 23–30 day old, control and mgb−/− male mice were subject to renal ultrasound to establish the degree of hydronephrosis as previously published . Of note, our method of grading hydronephrosis is based on the degree of parenchymal preservation, and does not directly correlate with clinical grading of hydronephrosis, such as the Society of Fetal Urology Grading System (http://www.sfu-urology.org/sfu_hydrone_grading.cfm). Kidneys with ≥67% parenchyma were considered mildly, 34–66% moderately and ≤33% severely affected. Mice were euthanized, kidneys extracted, snap frozen and stored at −80°C.
RNA Extraction and Microarray Hybridization
Total RNA was extracted using mirVana™ kit (Life Technologies, Carlsbad, CA). RNA integrity was analyzed using Agilent 2100 Bioanalyzer Lab-On-A-Chip 6000 Series II chip (Agilent Technologies, Santa Clara, CA). Samples were hybridized to Agilent SurePrint G3 Mouse GE 8×60 K Microarray and scanned using Agilent G2505C Microarray Scanner. Raw data were quality-assessed, filtered for outliers and normalized to remove non-biological variation. Differential gene expression was defined using a 10% false discovery rate (FDR) and adjusted p-value <0.05 (Bioconductor LIMMA package). Data have been deposited in Gene Expression Omnibus (GEO accession: GSE48041).
Ingenuity Pathway Analysis (IPA; Ingenuity Systems, Redwood City, CA. http://www.ingenuitypathway.org; version 14197757) was used to evaluate the biological significance of co-regulated transcripts. mRNA with ≥2 fold change in expression, adjusted p<0.05, and FDR <10 were considered significant.
cDNA was prepared from kidney total RNA using random hexamers and reverse transcriptase. Triplicate PCR reactions were performed using 25 ng of cDNA as a template in 20 µL reactions containing 2X master mix and gene-specific primers/probes (Applied Biosystems, Foster City, CA). Simultaneous triplicate reactions included Gapdh as an endogenous control. Results were expressed using the 2−ΔΔCT method by normalizing to a common pool of control kidney cDNA . The average fold change ± standard error was graphed, and p-values were calculated based on comparison of individual fold change values in an unpaired two-tailed Student’s t test.
Formalin fixed, paraffin embedded kidneys were sectioned at 4 µm. Deparaffinized sections were rehydrated, subjected to antigen retrieval, peroxidase block, biotin block, and Superblock (Scytek, Logan, UT). Primary antibodies were incubated for 1 hour at the following dilutions: anti-Upk3a (Research Diagnostics Inc., Flanders, NJ.) 1∶500; anti-Krt14 (Covance, Princeton, NJ) 1∶1600; anti-Ki-67 (Abcam, Cambridge, MA) 1∶800. Biotinylated secondary antibody and HRP-conjugated streptavidin were implemented (Scytek). Slides were developed using diaminobenzamide (MP Biomedicals, Santa Ana, CA), counterstained with hematoxylin and visualized using an Olympus BX-51 microscope (Olympus, America, Center Valley, PA).
Ki-67 positive and Ki-67 negative nuclei within the renal urothelium were identified using anti-Ki-67 labeled and hematoxylin counterstained tissue sections. Longitudinally oriented, four-micron thick kidney sections were collected near the hilum. Briefly, contours were traced onto morphometric tissues of interest; kidney (red), renal urothelium (not shown), renal lumen (not shown) and large vasculature (blue). Individual Ki-67 positive (yellow crosshair) and Ki-67 negative (white dot) nuclei within the entirety of the renal urothelium contour were digitally marked. Graphic representations were rendered using StereoInvestigator software and quantitative analysis was obtained using Neurolucida Explorer software (MBF Bioscience, Williston, VT). Representative images from each experimental group are shown.
Global transcriptome analysis of 18 male mgb−/− kidneys with varying degrees of hydronephrosis and six age-matched male control kidneys identified 202 and 207 genes with ≥2-fold higher or lower expression in mgb−/− kidneys, respectively (Figure 1, Table S1). Differential expression of 70 target genes was validated by qPCR (Figures S1–S9).
Left panel: percent parenchyma of mgb−/− kidneys undergoing transcriptome analysis. Black horizontal lines indicate individual kidney measurements. Right panel: representative sonograms of mild, moderate and severely hydronephrotic mgb−/− kidneys. Dotted white line indicates longitudinal renal length. Solid white lines from right to left indicate transverse renal width, renal pelvis diameter and renal papillae width with no measureable renal papillae width in the severe kidney.
The top five toxicological functions identified by IPA comparing mgb−/− versus control kidneys included renal necrosis/cell death, kidney failure, renal inflammation, renal nephritis and renal damage (Table S2). The top twenty canonical pathways identified by IPA encompassed three primary cellular processes: 1) response to renal injury, 2) steroid hormone metabolism and 3) retinoic acid (RA) metabolism and signaling (Figure 2A). Using qPCR we validated differential expression of 22 of 23 genes associated with retinoic acid metabolism, 17 of 21 genes associated with steroid hormone metabolism, and 21 of 43 genes associated with the response to renal injury in mgb−/− versus control kidneys (Figures S2–S4). The overall distribution of these cellular processes was relatively consistent at 20% RA, 25% steroid hormone and 55% renal injury pathway whether mgb−/− kidneys were compared to controls collectively or as stratified groups (Figure 2B). The top ten up and down-regulated transcripts were also associated with these same three pathways, with nine of the genes possessing overlapping functional roles in two or more (Table 1). While the overall percentage involvement of these three canonical pathways did not appear to vary between mild, moderate and severe mgb−/− kidneys, numerous individual gene targets showed progressive increases/decreases that correlated with the degree of hydronephrosis observed in these animals. In addition, the relative positions of the individual top twenty canonical pathways varied between mild, moderate and severe mgb−/− kidneys suggesting preferential utilization based upon the degree of hydronephrosis observed.
(a) Blue bars represent the -log of p-value with bar height corresponding to level of significance. The red line represents a p-value cutoff of 0.05. The green squares/line represent the ratio of the number of genes represented within each pathway that meet the cutoff criteria to the total number of genes in a given pathway. The three main cellular processes identified by IPA include retinoic acid metabolism (blue), response to renal injury (red) and steroid hormone metabolism (green). (b) Canonical pathway distribution of retinoic acid metabolism (blue), response to renal injury (red) and steroid hormone metabolism (green) in control kidneys versus all mutant, mild, moderate and severe mgb−/− kidneys.
The top ten activated upstream regulators included three growth factors, three intracellular signaling molecules, and four pro-inflammatory cytokines (Table 2). Transforming growth factor-beta 1 (TGFβ1) was the top activated upstream regulator in mgb−/− kidneys, with expression changes in 72 canonical TGFβ1 target genes between mgb−/− and control kidneys (Table S3). In addition, TGFβ3, Smad3 and Smad4 were activated while Smad7 was inhibited (Table S3). Of the 72 canonical TGFβ1 genes, 36% represented direct targets of Smad3/4, while inhibitory Smad7 was the most downregulated transcription factor identified in mgb−/− kidneys. We validated differential regulation of 13 transcriptional targets of TGFβ1 signaling between mgb−/− and control kidneys by qPCR: Egr1, Fos, Nr4a1 (Figure S1); Dusp1, Foxo3, Ptgs2 (Figure S3); and Apoe, Cd36, Cdkn1a, Gadd45b, Nfkbia, Serpine1, and Stat3 (Figure S4).
The top ten inhibited upstream regulators had roles in a wide range of cellular processes including transcription, cell death, extracellular matrix regulation, fatty acid metabolism, and immune response (Table 3). Histone deacetylase (Hdac) was the top inhibited upstream regulator identified in mgb−/− kidneys. Hdac inhibition was associated with up-regulation of 11 known target genes including six transcription factors (Table 4). We confirmed upregulation of 5 of 11 “de-repressed” Hdac target mRNAs in mgb−/− kidneys by qPCR ().
The mRNA most differentially expressed between mgb−/− and control kidneys was Serpina6/corticosteroid-binding globulin (Cbg), a gene that displays sexually dimorphic expression , . Eighteen additional sexually dimorphic mRNAs were identified in mgb−/− kidneys including 12 male and 6 female transcripts that were expressed at lower or higher levels, respectively (Table 5). We confirmed differential expression of 14 sexually dimorphic mRNAs by qPCR (Figure S6).
No significant differentially expressed transcripts were identified when the individual transcriptomes of mild and moderate or moderate and severe kidneys were directly compared. In contrast, when mild and severe kidneys were compared, a single gene was expressed at ≥2-fold higher levels in mild kidneys (Cyp2d22) and 80 genes were expressed ≥2-fold higher levels in severe kidneys (Table S4). Interestingly, 13/80 genes including 6/10 of the most differentially expressed genes between mild and severe kidneys were expressed by renal urothelium. These included genes with roles in maintaining urothelial integrity such as uroplakins and cytokeratins, transcription factors regulating urothelial cell fate, and genes with currently unknown urothelial functions (Table 6) –. We confirmed expression differences in 11 urothelial mRNAs between mgb−/− kidneys with mild and severe hydronephrosis by qPCR (Figure S7).
Since urothelial genes appeared preferentially upregulated in mgb−/− kidneys, we compared urothelial morphology in severe kidneys versus controls (Figures 3–5). Control renal urothelium was principally comprised of a single layer of cuboidal cells with intermittent multilayered regions of 2–3 cells. In contrast, the renal urothelium of severe kidneys contained numerous multilayered regions of 3–5 cells throughout the renal pelvis and showed frequent cellular blebbing.
Immunohistochemical localization (brown, arrows) of Upk3a shows relatively uniform expression in urothelial cells in cortical (a, d) and cortical-medullary (b, e) regions of control (a, b) and severe (d, e) kidneys. The intensity of Upk3a staining is greater within urothelial cells of severe kidneys compared to controls (compare inset b to e). Increased Upk3a expression was observed in urothelial cells of the medullary papillae in severe (f) versus control (c) kidneys. Scale bars are 100 microns (a, c, d, f), 200 microns (b, e) and 40 microns (inset b, e).
Immunohistochemical localization (brown, arrows) of Krt14 shows relatively uniform expression in urothelial cells in cortical (a, d) and cortical-medullary (b, e) regions of control (a, b) and severe (d, e) kidneys. Increased Krt14 expression was observed in urothelial cells of the medullary papillae in mutant (f) versus control (c) kidneys. Scale bars are 100 microns (a, c, d, f), 200 microns (b, e).
Immunohistochemical localization (brown, arrows) of Ki-67 expression is limited to sparse individual cells within control kidney urothelium (a). Severe kidney urothelium shows an increase in the number of Ki-67 positive cells (b) including long stretches of positive cells in mutant cortical urothelium (c) and in mutant ureteropelvic urothelium (d). Scale bar is 100 microns.
Uroplakin 3a (Upk3a), a transmembrane protein that forms apical complexes on urothelial cells, was uniformly expressed along control urothelium but showed decreased expression ascending the renal papillae resulting in only scarce individually stained cells found on the upper 2/3 of the papillae (Figure 3A–C). A similar pattern of expression was observed in the urothelium of severe kidneys except that the intensity of corticomedullary Upk3a staining appeared greater and the number of positive cells on the renal papillae appeared to increase (Figure 3D–F).
Cytokeratin 14 (Krt14), a progenitor cell marker in renal urothelium, was expressed throughout control urothelium with decreased expression as urothelial cells ascended the renal papillae (Figure 4A–C) . In contrast, Krt14 expression in severe kidneys was uniformly expressed throughout the renal urothelium including the papillae where expression was significantly increased versus controls (Figure 4D–F).
Cellular proliferation within the renal urothelium was assessed using Ki-67 (Figure 5) . Minimal Ki-67 staining was observed in control renal urothelium with only rare individual positive cells identified. In contrast, severe kidneys showed a 20-fold increase (p≤0.02) in the percentage of Ki-67-positive cells within the renal urothelium versus controls (Figure 6). Long clusters of positive cells were observed along the corticomedullary (Figure 5C) and ureteropelvic (Figure 5D) junctions.
(a) Schematic reconstruction of representative control and mild, moderate and severe kidneys showing kidney capsule (red), renal vasculature (blue), Ki-67 negative renal urothelial cells (white dots) and Ki-67 positive renal urothelial cells (yellow crosshairs). Scale bar is 1000 microns. (b) Quantitation of the number of Ki-67 positive (black) versus negative (white) urothelial cells with percentage of Ki-67 positive urothelial cells indicated above each bar.
Biomarker analysis identified a subset of three moderate and one severe hydronephrotic kidneys expressing high levels of Ngal/Lcn2, a well-characterized marker of tubular injury –. qPCR analysis validated Ngal expression in all 24 kidneys used in this study confirming that these four kidneys had Ngal mRNA expression levels at least three standard deviations higher than the mean (Figure S8). We next grouped mgb−/− transcriptomes with elevated Ngal expression (Ngalhigh) and analyzed for global differences in expression compared to mgb−/− kidneys with low Ngal expression (Ngallow). This analysis identified nine potential biomarkers of renal injury including several genes previously associated with tubular injury (Havcr1, Il1f6, Fgg, Timp1) as well as a set of novel genes not previously implicated as biomarkers of renal injury including Sprr2f, Sprr2g, Serpina10, Itih4 and Saa2 (Figure 7). We confirmed differential mRNA expression of 8 of these known or candidate tubular injury markers between Ngalhigh and Ngallow mgb−/− kidneys by qPCR (Figure S9).
Microarray results showing fold-change in expression of potential biomarkers of renal injury comparing Ngalhigh versus Ngallow kidneys. (Sprr2f) small proline-rich protein 2F, Sprr2g, (Timp1) tissue inhibitor metalloproteinase 1, (Serpina10) serpin peptidase inhibitor, clade A, member 10, (Fgg) fibrinogen gamma chain, (Itih4) inter-alpha-trypsin inhibitor heavy chain family, member 4, (Il1f6) interleukin-1 family member 6, (Saa2) serum amyloid A2 and (Havcr1) hepatitis A virus cellular receptor 1.
The mgb−/− mouse represents a unique model for the study of CON and progressive renal injury. Prior studies show that mgb−/− kidneys display an adaptive response to the development of progressive hydronephrosis . To better understand the molecular mechanisms controlling this adaptive response, whole genome expression arrays were compared between mild, moderate and severe kidneys versus controls. This analysis suggested that the progressive development of hydronephrosis in mgb−/− kidneys principally involves three canonical pathways including: 1) renal response to injury, 2) RA pathway and 3) steroid hormone metabolism.
Over half of the top twenty canonical pathways identified in this study involved renal response to injury. This finding is consistent with prior studies showing that the progression of renal injury in mgb−/− kidneys correlates with the severity of hydronephrosis . Affected kidneys showed an adaptive response that involves the appearance of α-smooth muscle actin (SMA) positive myofibroblasts, expanded TGFβ1 and CTGF expression, and the development of limited renal fibrosis . This study confirms and expands those original observations at the molecular level by demonstrating that the most activated upstream regulator in mgb−/− kidneys is the TGFβ pathway. This observation is consistent with current literature and confirms the key role that TGFβ plays in modulating progressive renal injury and fibrosis following obstruction –.
Although our analysis identified a group of genes that have been previously shown to be upregulated in newborn rats with UUO, it did not identify any of the key inflammatory genes previously found to be dysregulated in UUO such as interleukin-1 beta, interferon gamma regulating factor 1, and monocyte chemoattractant protein 1 . Activation of the host inflammatory response is an early event in the UUO model, attributed to apoptosis of tubular epithelial cells, resulting in the release of TGFβ and reactive oxygen species which recruit and activate macrophages . In contrast, apoptosis is a rare event in mgb−/− tubular epithelium, and macrophages are only detected in a subset of mgb−/− kidneys with advanced hydronephrosis . We propose that the mgb−/− kidney is a low-pressure system with a more gradual onset of increased tubular hydrostatic pressure, rendering it less susceptible to tubular injury and macrophage infiltration than kidneys following UUO. The acute inflammatory response observed in certain mgb−/− kidneys with advanced hydronephrosis most likely represents a key “second hit” that worsens tubular injury resulting in expanded myofibroblast transformation/activation leading to the development of severe interstitial fibrosis. A similar “second hit” model in children with CON may be found in the form of recurrent urinary tract infections/pyelonephritis or increased back pressure resulting in ischemic injury, both of which have been associated with the subsequent development of severe interstitial fibrosis and progression to ESRD , .
Comparison of transcriptomes with varying degrees of hydronephrosis identified a urothelial gene expression signature associated with severe obstruction. Many of the urothelial genes upregulated in severe kidneys have been implicated in urothelial integrity/function, and prior studies have shown that urothelial permeability is compromised in mice deficient in Upk2, Upk3a, and Grainyhead-like 3 –. The changes in Upk3a and Krt14 expression and distribution observed in severe kidneys would therefore be predicted to alter the structural integrity, permeability and functionality of the renal urothelium.
The renal urothelium also showed alterations in proliferative index and organization in severe kidneys. The corticomedullary region of proliferation identified in our study appears similar to the previously identified renal fornix, which has been proposed as a site of urothelial progenitor cells , . These same studies showed an Fgf7/Fgfr2 mediated increase in urothelial proliferation near the renal fornix in response to ischemia-reperfusion injury and UUO. Fgf2 activation was predicted in our current analysis (p≥5.3×10−10) suggesting that this same signaling pathway may play a key role in renal urothelial expansion in CON.
These results suggest that urothelial cells may actively participate in the renal response to obstruction. We postulate that during the development of progressive hydronephrosis, distension of the renal pelvis results in increased proliferation of the renal urothelium and altered expression of key structural genes. Although these changes may have short-term deleterious effects on membrane permeability and function, they may be critical in initiating an adaptive, organ-protective response during pathogenic insult to the kidney. This hypothesis is consistent with our previous observation that renal fibrosis first initiates immediately beneath the renal urothelium of mgb−/− kidneys . In addition to distending pelvic urothelium, obstruction leads to tubular dilatation and exposure of the entire nephron to increased hydrostatic pressure which may activate additional signaling pathways that participate in renal injury and account for some of the transcriptional changes observed in mgb−/− kidneys.
RA has been implicated in kidney and urinary tract development , . We hypothesize that these same developmental functions of RA may be recapitulated during renal pathogenesis as a transient repair mechanism. Prior studies in our lab have found evidence of delayed maturation in mgb−/− kidneys including prolonged expression of early glomerular markers as well as expanded medullary expression of α-SMA . RA has been shown to promote cell survival and antagonize renal fibrosis in models of glomerular injury, while administration of isotretinoin (13-cis-RA) or all-trans RA attenuates interstitial fibrosis and reduces TGFβ1 expression in UUO , . In addition, administration of 13-cis RA to mice with ischemia-reperfusion injury results in decreased expression of the proapoptotic molecule, Nur77, as well as improved renal function, decreased renal injury, and reduced levels of pro-inflammatory cytokines . Nur77/Nr4a1 was among the top induced transcripts in mgb−/− kidneys suggesting a similar role for this molecule in our model system.
RA has also been shown to play a role in differentiation of urothelium . This observation provides a mechanism whereby RA signaling may represent an important early urothelial response to impending renal injury. As discussed above, this hypothesis is consistent with our prior observations that the first site of collagen deposition in mgb−/− kidneys is immediately underlying the renal urothelium. In addition, the results of this study indicate that one of the earliest detectable changes in mgb−/− kidneys is an increase in renal urothelial proliferation in the face of limited pelvic distension. We postulate that these initial changes in urothelial morphology represent an early adaptive response to progressive renal injury that is designed to generate a localized and reversible repair mechanism. In the face of continued renal insult, these early remodeling/repair responses may become exacerbated and irreversible by additional contributors to renal injury resulting in permanent kidney damage.
The most significantly upregulated mRNA in mgb−/− kidneys was Cbg, encoding the major transport protein for glucocorticoids and progestins in the blood. Cbg expression in the developing and postnatal kidney is highly regulated at the mRNA and protein level . While the significance of increased Cbg expression by mgb−/− kidneys remains to be determined, it seems plausible that it functions to increase the local concentration of glucocorticoid/progestin thereby downregulating the local inflammatory response. This hypothesis provides a molecular mechanism for the previous observation that mgb−/− kidneys display limited inflammatory infiltrates, suggesting that increased Cbg expression may be critical in promoting remodeling/repair following congenital obstruction by attenuating the acute inflammatory response.
In addition to altered Cbg levels, our study identified differential expression of genes with roles in sex hormone metabolism between mgb−/− and control kidneys. A variety of studies have suggested gender-based differences in the progression of renal injury –. Among the hypothetical mechanisms proposed for renal protection are gender-specific gene expression, gender-specific synthesis and metabolism of sex hormones, and/or modification of epigenetic factors controlling gene activation/inactivation. Interestingly, our study identified the skewed expression of a group of sexually dimorphic genes in male mgb−/− kidneys. Although the underlying mechanism(s) and significance of this observation remain to be determined, we postulate that male mgb−/− kidneys transiently alter expression of mRNAs to adopt a more “female” transcriptome under the influence of estrogen and/or other hormones. Epidemiologic data supports the concept that female gender is protective for non-diabetic renal disease progression, with higher rates of CKD progression and ESRD in males , . Studies in animal models support a specific role for estrogen in limiting glomerulosclerosis and inflammation . Tamoxifen, an estrogen partial agonist, has been shown to have anti-fibrotic effects in rats, while testosterone administration has been shown to exacerbate renal injury in the mouse UUO model , . These observations suggest that the expression of a more “female” transcriptome in male mgb−/− kidneys may initiate a cytoprotective genetic program that transiently supports the remodeling/repair response observed in mgb−/− kidneys.
The most inhibited upstream pathway observed in mgb−/− kidneys involved Hdacs, a class of enzymes that remove acetyl groups from histones permitting tight DNA packaging that often results in the downregulation/inactivation of gene transcription . Prior studies have shown that androgen, estrogen and glucocorticoid receptors are substrates/binding partners for various members of the Hdac family and that steroid hormone expression can influence perinatal programing . These observations suggest that mgb−/− kidneys may reactivate Hdac-regulated genes under the modulation of local steroid hormone activity providing a potential dual role for steroids in renal pathogenesis that includes suppression of an acute inflammatory response as well as modulation of the genetic programs controlling cellular differentiation.
Biomarkers of CON
The identification of novel biomarkers of renal injury that provide prognostic value for progression to ESRD remains a major goal in the study of kidney disease. The mgb−/− mouse model provides a unique opportunity to search for stage-specific diagnostic and prognostic markers of renal injury. Unfortunately, in this current study, we were unable to detect a single individual gene that directly correlated with our current ultrasound stratification criteria. This suggests that the degree of hydronephrosis may not always directly correlate with the molecular severity of disease progression in affected kidneys, and is consistent with clinical observations in children with CON . The basis for this discordance is most likely complex, involving both genetic and epigenetic factors that may include individual genetic background, understanding of the precise chronicity of hydronephrosis/renal injury, cellular heterogeneity of whole kidney and potential loss of individual cellular phenotypes as disease severity increases.
Therefore, we sought to further stratify mgb−/− kidneys at the molecular level based upon Ngal expression. While Ngal expression was heterogeneous and did not accurately predict the degree of obstruction, a subset of moderate and severe kidneys did express significantly higher levels of Ngal. Assuming that this subset of Ngalhigh kidneys represented a consistent molecular signature associated with acute renal injury, we compared this group to the remainder of affected kidneys and controls. The detection of three previously identified markers of tubular injury validated this approach and studies are currently underway to determine the potential efficacy of the remaining five genes as stage-specific and/or prognostic markers of obstructive renal injury. Finally, identification of the renal urothelium as an early responder in kidney remodeling/repair may provide a novel target for the development of stage-specific markers of CON.
The results of this study indicate that the development of progressive hydronephrosis in mgb−/− mice results in a highly orchestrated adaptive response in the chronically affected kidney (Figure 8). We propose that renal adaptation involves a balance between RA-initiated remodeling/repair and TGFβ directed pathogenesis. In this model, the renal urothelium plays a key role in sensing and transmitting the early signals responsible for initiating the remodeling/repair mechanisms observed in mgb−/− kidneys. We postulate that increasing back-pressure within the renal pelvis causes the gradual expansion of the renal urothelium from morphologically well-defined regions of proliferation. Cellular proliferation results in significant phenotypic alterations within the renal urothelium changing the composition of apical plaques and altering the functionality and permeability of the urothelial membrane. These urothelial changes most likely initiate the activation of submucosal myofibroblasts resulting in collagen deposition immediately underlying the renal urothelium – the first major histopathological finding observed in mgb−/− kidneys (6). Steroid hormones may play a dual role in the remodeling/repair response by directly suppressing acute inflammation and modulating the transcriptional control of gene expression thereby providing a potential mechanism for gender-based differences in renal pathogenesis and progression.
Renal adaptation in mgb−/− kidneys involves a balance between tissue remodeling promoted by retinoic acid (RA, left side of dotted line) and renal pathogenesis promoted by transforming growth factor beta (TGFβ, right side of dotted line) in context of the specific steroid hormone background. Normal renal urothelium (light yellow cells) and submucosal fibroblasts (light red cells) maintain a controlled permeability barrier (brown apical plaques). As renal pelvic pressure increases during development of congenital obstruction (black arrows), the urothelium (gold cells) responds by dedifferentiating, increasing urothelial cellular proliferation (mitotic nuclei) and altering apical plaque production and composition (green). These changes result in urothelial blebbing and loss to the renal space, breakdown of membrane permeability, activation of submucosal myofibroblast (red cells) and increased deposition of extracellular matrix including collagen (blue lines). Neurovascular beds are indicated by: A (artery), V (vein), and blue dots (neural bundles).
The distribution of pathways associated with RA signaling, steroid hormone metabolism, and renal injury did not vary between mildly and severely hydronephrotic mgb−/− kidneys. Nonetheless, we propose that these pathways modulate proliferation and remodeling of the renal urothelium and activation of submucosal myofibroblasts, processes that occur in all mgb−/− kidneys (Figure 6) . We further postulate that these pathways may act both in a reparative fashion and to render the kidney susceptible to future pathologic changes including interstitial fibrosis and tubular injury under the influence of TGFβ, and that steroid hormones may serve a role in modulating this process (Figure 8).
Progression of renal injury in our model may be exacerbated by a “second hit” in the affected kidney. In mgb−/− mice, this “second hit” may occur through several processes including the development of urolithiasis, ascending infection and pyelonephritis, and/or acute high-pressure obstruction. Each of these events drives the molecular balance away from renal remodeling/repair towards expanded renal pathogenesis. Under the influence of TGFβ, increased activation of myofibroblasts leads to severe interstitial fibrosis that results in further loss of renal tubules and glomeruli and reduced renal function. It is intriguing to postulate that the structural changes induced during RA or steroid hormone mediated remodeling/repair alter the functionality of the renal urothelium increasing the kidney’s susceptibility to a “second hit” and the subsequent development of ESRD. Future studies will focus on the refinement of our current model including evaluation of the relationship between urothelial proliferation/integrity and renal injury pathways. Elucidation of these pathogenic mechanisms will lead to identification of biomarkers for risk stratification and development of therapeutic strategies designed to reduce the impact of CON.
qPCR validation of a subset of differentially expressed genes in all mutant versus control kidneys. (a) mRNA expression of the six most increased and (b) three most decreased genes in mgb−/− kidneys versus control.
qPCR validation of a subset of differentially expressed genes implicated in retinoic acid metabolism. Fold change of mRNA expression of 22 genes in mgb−/− (mutant) kidneys versus controls. *p = <0.05, **p = <0.005, ***p = 0.0005.
qPCR validation of a subset of differentially expressed genes implicated in steroid hormone metabolism. Fold change of mRNA expression of 17 genes in mgb−/− (mutant) kidneys versus controls. *p = <0.05, **p = <0.005, ***p = 0.0005.
qPCR validation of a subset of differentially expressed genes implicated in the response to renal injury. Fold change of mRNA expression of 21 genes in mgb−/− (mutant) kidneys versus controls. *p = <0.05, **p = <0.005, ***p = 0.0005.
qPCR validation of a subset of differentially expressed Hdac target genes. Fold change of mRNA expression of 5 genes in mgb−/− (mutant) kidneys versus controls. *p = <0.05, ***p = 0.0005.
qPCR validation of a subset of differentially expressed sexually dimorphic genes. Fold change of mRNA expression of 14 genes in mgb−/− (mutant) kidneys versus controls. *p = <0.05, **p = <0.005, ***p = 0.0005.
qPCR validation of a subset of urothelium specific genes between mild and severe kidneys. Fold change of mRNA expression of 11 genes in severe mgb−/− kidneys versus mild mgb−/−. *p = <0.05, **p = <0.005, ***p = 0.0005.
qPCR validation of Ngal expression in mutant and control kidneys. The average fold change of the four Ngalhigh mutant kidneys, all Ngallow (Ngallow control+Ngallow mutant kidneys), mutant Ngallow and control Ngallow kidneys compared to a control pool of kidneys. *p = 0.018, **p = 0.019 and ***p = 0.017.
qPCR validation of potential biomarkers of renal injury. Fold change of mRNA expression of 8 genes in Ngalhigh kidneys versus Ngallow mgb−/− kidneys. *p = <0.05, **p = <0.005, ***p = 0.0005.
Genes with ≥2-fold differential expression between all mgb−/− and wildtype kidneys.
Top Toxicological Functions Identified by IPA of All Mutant Kidneys to Controls.
Differentially Expressed TGF-β/Smad Target Genes Comparing All Mutant and Control Kidneys.
We thank David E. Newsom and Dr. Peter White and of the Biomedical Genomics Core of the Research Institute at Nationwide Children’s Hospital, Columbus, Ohio for their help with the Agilent Microarray and pathway analysis.
Conceived and designed the experiments: KM BB ARC DH SEI. Performed the experiments: ARC JA BB MW. Analyzed the data: KM BB ARC SEI. Contributed reagents/materials/analysis tools: DH SEI. Wrote the paper: KM ARC BB.
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A new battle is looming over access to antiviral medicines in developing countries - this time for treating hepatitis C - more than a decade after a global showdown over the price of AIDS drugs in Africa.
Modern pills being launched in western markets could cure the liver-destroying infection in tens of millions of people from China to Congo, or even eradicate the disease entirely. But that will only happen if the cost falls dramatically.
Drugmakers like Gilead Sciences, whose product Sovaldi won U.S. approval this month with a $1,000 a day price tag, are under mounting pressure to strike deals to avoid a rerun of the disputes that stalled early access to HIV therapy.
"Affordability is an urgent and pressing issue," World Health Organization (WHO) Director General Margaret Chan told Reuters during a visit to London.
"These drugs are very expensive. How can we address this? I hope we can learn from the lesson of HIV and find solutions without confrontations."
In the 1990s, HIV/AIDS drugs costing more than $10,000 per patient a year were simply out of reach for millions of people in the developing world. Today, thanks to cheap generics from India, the cost for the poor has been slashed to around $100.
Like HIV, hepatitis C (HCV) can be spread through blood, often via contaminated needles. The WHO estimates that 150 million people worldwide are chronically infected, putting them at risk of cirrhosis and liver cancer.
But whereas the burden of HIV is largely in sub-Saharan Africa, most cases of HCV are in middle-income countries like China, India and Russia, where drug companies are more reluctant to accept rock-bottom prices.
Chan said options for maximizing use of the drugs could include granting licenses to low-cost generic drug manufacturers in India and other countries, as has happened with HIV drugs and also Roche's flu pill Tamiflu.
Gregg Alton, Gilead's head of corporate and medical affairs, said his company was working on plans to help ensure access to Sovaldi in resource-limited countries and aimed to set out details early in 2014.
Other companies developing all-oral treatment regimens for HCV - such as Johnson & Johnson, AbbVie, Bristol-Myers Squibb and Merck & Co - also recognize they need to tackle the issue.
"We are going to be responsible players to make sure that people get access," said Paul Stoffels, head of pharmaceuticals at J&J. "There is a lot of pressure on us to make this available."
Like Gilead, J&J is not yet ready to disclose its access plans and Stoffels said in an interview it would take a couple of years before really simple all-tablet regimens suitable for use in poorer nations were widely available.
While Gilead's Sovaldi has broken new ground as the first pill to treat some HCV patients without injections of interferon, which can cause severe flu-like symptoms, it is still supposed to be given alongside interferon in others.
There is little doubt the new drugs will transform HCV treatment - and prove hugely profitable. In the developed world, they are tipped to become major blockbusters, with consensus sales forecasts for Sovaldi alone standing at $6.8 billion in 2018, according to Thomson Reuters Pharma.
Healthcare campaigners expect corporate schemes for poorer countries will focus on price discounts in middle- and low-income markets - but they argue this as a poor substitute for full generic competition, which could send prices tumbling.
In a bid to force the issue and allow Indian generic manufacturers to make cheap copies of the drug for distribution at home and in some other poor countries, a legal group has filed a motion opposing the patenting of Sovaldi in India.
The New York-based Initiative for Medicines, Access and Knowledge, or I-MAK, which claims the science behind Gilead's drug is not new, says it is taking the action to push prices down massively from the $84,000 that 12 weeks of treatment will cost in the United States.
"We think it has to be below $500," said Tahir Amin, the group's co-founder. "We have seen with HIV that only when you really bring the price down do people really get access."
Medecins Sans Frontieres (MSF), a campaigning group which provides humanitarian care, has thrown its weight behind the I-MAK patent challenge and Leena Menghaney, MSF campaign manager for India, said a sub-$500 price was a prerequisite for many governments to start considering use of the new drugs.
Looking simply at the cost of production, Andrew Hill, a pharmacologist at the University of Liverpool, thinks such a price target is certainly achievable. In fact, he calculates that large-scale production is possible for just $1 a day - or one thousandth of the Sovaldi price tag.
For a three-drug combination, he estimates a target price of between $100 and $250 for 12 weeks of treatment is realistic, including a margin for profits.
Of course, drug prices take account of more than just the cost of goods. They also reflect the value that medicines can offer society and Gilead argues its price is fair based on the significant benefits it brings.
The total cost of the 12-week regimen is also not so different to current treatment, which requires 24 or 48 weeks of interferon-based therapy and is less effective. And Gilead has ploughed an awful lot of money into this new medicine after paying $11 billion in 2012 for Pharmasset, the company that initially developed Sovaldi.
Still, the $1,000-a-day price tag has raised concerns even among some healthcare providers in the United States.
"Obviously, drug companies need to make a return on investment," said Hill. "The question is how much?"


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Mortgage buyer Freddie Mac said Thursday the average rate on the 30-year, fixed-rate mortgage held steady from last week at 3.82 percent, its lowest point since September 2017. By contrast, a year ago.
Mortgage rates valid as of 19 Jul 2019 08:28 am CDT and assume borrower has excellent credit (including a credit score of 740 or higher). Estimated monthly payments shown include principal, interest and (if applicable) any required mortgage insurance. ARM interest rates and payments are subject to increase after the initial fixed-rate period (5 years for a 5/1 ARM, 7 years for a 7/1 ARM and 10.
Mortgage rates could change daily.. 30-Year Fixed-Rate Mortgage: The payment on a $200,000 30-year Fixed-Rate Loan at 3.99% and 75.00% loan-to-value (LTV) is $953.68 with 2 points due at closing. The Annual Percentage Rate (APR) is 4.253%. Payment does not include taxes and insurance premiums.
FHA loan rates can be lower than conventional loan rates like the 30-year fixed, but they can end up being more expensive due to mortgage insurance costs. mortgage loans with less than 20 percent down generally have to carry mortgage insurance, but the insurance on FHA loans is more expensive than insurance on conventional loans.
All rates are based upon a 30-year term, owner occupied, and up to 60% of home value. Rates and Fees disclosed are for loans that meet Secondary Mortgage Market underwriting standards; additional rate and fees may apply for loans outside of those guidelines.
Difference Between Fha And Conventional Loans fha 30 year fixed rates current Mortgage Rates For Second Home Second Mortgage Rates – Mortgage Calculator – A second mortgage is an additional loan that can be acquired after the first. The same assets that were used to secure the first, must be used to secure the second. Generally, the interest rate on a second mortgage is higher than that of a first. equity determines the quantity and type of second mortgage an individual qualifies for.Use annual percentage rate APR, which includes fees and costs, to compare rates across lenders.Rates and APR below may include up to .50 in discount points as an upfront cost to borrowers. Select product to see detail. Use our Compare Home Mortgage Loans Calculator for rates customized to your specific home financing need.Home Loans for Veterans: Everything You Need to Know – But the 3.3 percent funding fee can be cost-prohibitive for veterans refinancing from an FHA or conventional. use a jumbo VA loan and buy a more expensive property, you just have to make a down.
· The most popular form of fixed-interest home financing is the 30-year fixed mortgage, which spreads the principal repayment over a long period of.
Fixed mortgage rates didn’t go down much, but they did go down for the fourth week in a row. According to the latest data released Thursday by Freddie Mac, the 30-year fixed-rate average slipped to.
When you're looking at different upfront charges, interest rates and. Total 30- Year Cost, The total cost of an FHA loan, including down.
Mortgage buyer Freddie Mac said Thursday the average rate on the 30-year, fixed-rate mortgage dipped to 3.82% from 3.99% last week. By contrast, a year ago the benchmark rate stood at 4.54%. The.
Fha Or Conventional Mortgage FHA.com Reviews. FHA.com is a one-stop resource for homebuyers who want to make the best decisions when it comes to their mortgage. With our detailed, mobile-friendly site, individuals can access information about different FHA products, the latest loan limits, and numerous other resources to make their homebuying experience easier.

domain: finance


text:
Added to shopping list. Go to shopping list.
Ingredients10 m servings 96 cals
Original recipe yields 12 servings
- Place Brie into individual ramekins or small bowls. Drizzle honey over the wedges; surround with blueberries and raspberries. Garnish with mint.
Per Serving: 96 calories; 4.4 g fat; 11.7 g carbohydrates; 3.5 g protein; 16 mg cholesterol; 99 mg sodium. Full nutrition

domain: food


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On June 21 and 23, 2021, CRRC and DPP co-sponsored a virtual event entitled “NOS Hurricane Preparedness Summit.” The purpose of the summit was to put NOAA in a better posture for the 2021 hurricane season by identifying current best practices, lessons learned, and future challenges. Participants represented academia, federal, state, and local agencies. The two-day summit included plenary presentations from academics, federal and state agency representatives outlining: preparedness, response, and virtual and in-person incident command lessons learned; emergency support function (ESF) 3/10 response gaps; the NOAA/FEMA perspective to hurricane preparedness and response; managing pandemic fatigue; unreliability and/or loss of utilities; coordinating deployment logistics; and anticipated challenges and mitigation methods for the 2021 hurricane season.
Coastal Response Research Center, "NOS Hurricane Preparedness Summit 2021 Report" (2021). Coastal Response Research Center. 28.


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From Blizzard Entertainment:
Requires an Internet connection and iPhone 4S, iPod touch 5th Generation, iPad 2 or newer.DECEPTIVELY SIMPLE. INSANELY FUN.Pick up your cards and throw down the gauntlet! In Hearthstone, you play the hero in a fast-paced, whimsical card game of cunning strategy. In minutes, youll be unleashing powerful cards to sling spells, summon minions, and seize control of an ever-shifting battlefield. Whether its your first card game or youre an experienced pro, the depth and charm of Hearthstone will draw you in. JUMP RIGHT IN: Fun introductory missions bring you into the world of Hearthstones intuitive gameplay. BUILD YOUR DECK: With hundreds of additional cards to win and craft - your collection grows with you.HONE YOUR SKILLS: Play in practice matches against computer-controlled heroes of the Warcraft universe. Thrall, Uther, Guldan - theyre all here! COLLECTION TRAVELS WITH YOU: Your card collection is linked to your Battle.net account - enabling you to switch your play between tablet and desktop with ease. AND FIGHT FOR GLORY: When youre ready, step into the Arena and duel other players for the chance to win awesome prizes!Post feedback about the game in our forums: http://us.battle.net/hearthstone/en/forum/13619661/Languages Supported: * English * Franais * Deutsch * Espaol (Latinoamrica) * Espaol (Europa) * Italiano * Portugus * Polski* (Russian)* (Korean) * (Simplified Chinese) * (Traditional Chinese) * (Japanese)* (Thai) 2016 Blizzard Entertainment, Inc. All rights reserved. Hearthstone and Heroes of Warcraft are trademarks, and Battle.net and Blizzard Entertainment are trademarks or registered trademarks of Blizzard Entertainment, Inc., in the U.S., and/or other countries.
What's new in this version:
This Hearthstone patch prepares the Tavern for the arrival of Whispers of the Old Gods! This patch also paves the way for a new way to play Hearthstone and introduces adjustments to key Basic and Classic cards. Along with bug fixes, we've also added new card backs and Tavern Brawls.Read on for details!* Do you hear them? Whispers of the Old Gods expansion will arrive on April 26! You can purchase Whispers of the Old Gods card packs from the in-game shop and play them beginning April 26.* As of A...
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Carol Hills: I’m Carol Hills in for Marco Werman and this is The World, a co-production of the BBC World Service, PRI and WGBH here in Boston. There are no more Ebola cases in the US, but that doesn’t mean the fear is gone. And immigrants to the US from Africa are still facing harassment and discrimination because of Ebola. Now a Bronx-based nonprofit organization for African immigrants has launched a phone and web hotline to help. Amaha Kassa is the executive director for African Communities Together. Amaha, what kind of stories are you hearing from the immigrants you’re working with?
Amaha Kassa: We’ve heard a range of stories, both from the immigrants we’re working with here in New York, and then as we’ve been collecting stories happening all across the country. The incidents have ranged from small -- street harassments that people have experienced, insults and jokes, all the way up to more serious issues, to children in the Bronx who were essentially assaulted by their classmates while they yelled â€œEbola!â€; people who have been kept off of work and who are not able to return to work, have been essentially suspended because --
Hills: Where are the kids from at the school you mentioned?
Kassa: There were two little boys from Senegal, and they were recently in the country. But as with a lot of these cases, it was unrelated to any actual contact with anyone infected with Ebola, or have even traveled to regions that have been infected by Ebola. We’ve seen cases where it’s about travel to Rwanda, Kenya, Nigeria, places that are often thousands of miles from Ebola-affected areas, and it’s just because people are African.
Hills: How are you trying to help? What are you offering to do for these kind of people who are facing this stigma?
Kassa: We think the first step is to make this problem, this crisis, more visible. There’s no good numbers about how many people have experienced this, but if you talk to people in the communities, if you talk to the organizations serving African immigrants, everyone has stories of this happening. So, the first thing we want to do is just collect and document and publicize the stories so that people are aware of the scope of the problem. Then it’s going to depend on what kind of experiences people are having, what kinds of problems people are facing. In some cases, it might be a criminal matter -- people have been physically assaulted. In some cases, there may be violations of anti-discrimination law. In other cases, it’s not going to be a legal matter at all, but it could be a matter of connecting people with support services. One problem that we’ve been hearing a lot about is African businesses, such as hair braiders, that have lost business due to fear and misinformation about Ebola. That might be a case where we can help those businesses with health education for their customers or support services.
Hills: In the case of businesses that are asking or telling employees, who they fear are somehow linked to Ebola, not to come to work -- that’s illegal. You can’t force somebody to not come to work.
Kassa: In some cases, it definitely is illegal. What’s particularly surprising or shocking about some of these cases is they often involve healthcare providers -- hospitals, hospices, those kinds of businesses that should have better information about dealing with infectious diseases. In many cases, people are receiving pay during the time that they’re not working, but frankly they would just rather be on the job. We’ve also seen cases where children have been kept from registering for school because of panic among parents; several cases, actually, around the country. In all those cases, there was no actual risk of Ebola. They were often involving countries thousands of miles away from the region where infections have been reported. But again, it’s a sort of guilt by association.
Hills: It’s interesting, because just in preparing for this interview, I sort of looked up â€œEbolaâ€ and â€œhotline,â€ and you also see Ebola hotlines for information about Ebola issued by states. It’s almost like you’re getting two versions when you search for hotlines. You’re getting â€œHere’s how to not get Ebola, not be exposed,â€ and then there’s hotlines for â€œTell us how you’ve been discriminated against.â€
Kassa: The two things are actually related. Our message is â€œTarget Ebola, not Africans,â€ as African immigrants and many of our members are from the countries that have been touched by Ebola in West Africa. We want to see the Ebola virus eradicated in Africa. We don’t want to see an outbreak in our communities here. But the thing is that profiling, harassment -- that doesn’t make anybody safer or healthier. In fact, it creates greater risks. People are now afraid sometimes to get routine medical treatment for things like the flu or asthma because they’re afraid they’re going to be profiled, quarantined, that they’re going to experience harassment, and those aren’t baseless fears because those incidents have happened. So, we think part of the best way to actually stem the flow of the disease is to engage African communities as part of the solution and not treat them as part of the problem.
Hills: Amaha Kassa is the executive director for African Communities Together in New York. Thanks for your time, Amaha.
Kassa: Thanks very much Carol.


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Younger Witnesses less fanatical or fundamental
It would seem to me that there are a lot of lazy people in it today! Sitting by some trolley with books and magazines on it, no one really stopping to look and the three people guarding it would more than likely be annoyed when someone stops to look, as they have to pause their conversations.
JW's as a whole have gotten soft! Yes no one could explain most of the Franz doctrines, most didn't care to even try. Yet some of these younger ones,are still very haughty when it comes to those who have left.
And, by the way, regardless of how moderate it gets, it is still ALL BS!
geevee - "...JWs as a whole have gotten soft..."
Outlaw once said that back in the day, JWs were "Watchtower Warrors".
These days, though?
Not so much.
Thus as it ever was.
Likewise, I find that younger ones in the organization, say 40 and under, also seem to be more rational and reasonable. I've known a lot of "true believers" in my time but I find I rarely meet any around my age. From what I've seen as well many have a more tolerant attitude towards alcohol as well as entertainment. Although no one really advertises this
Not only is this true, but we can go even further. Most young Jdrones barely know anything about the troof aside from the basic precepts. For them, being JW is more a social identity, and being part of a group. Going to the meetings and standing next to a cart for a few hours looking at their phones, is just the price they pay for keeping their friends and having fun at gatherings and outings.
They don't give two shiites about doctrine, and in reality this is helpful for the Org.
If by "less fanatical" you mean less factual and not giving a f*** then, yes, I agree, they are. I doubt they could debate that water is wet, because they would be too afraid of getting wet to do so.
But, to be fair the "less fanatical" 😉 thing isn't confined to JW young people it's everywhere. There seems to be an epidemic of mentally lazy and super-sensitive these days. Not a good combination I might add.


Ever since I was 12 back in 1996, and remember sitting behind a drum kit for the first time, I knew exactly what career I wanted to pursue and I have never looked back. For me drumming isn’t and has never ever been just a hobby, it’s been a passion and I love to share that passion through my students and team activities.
I teach to all ages and music tastes, so maybe you just want to learn your favourite Rock tune or maybe Hip Hop’s your thing, whatever your music preference is I will teach you. You may want to learn something more serious with the aim of working towards grades, I will get you there!
Drumming isn’t about being noisy and hitting hard, it involves what I describe as the “three ation’s”…… concentration, co-ordination and dedication, teamed with lots of fun thrown in for good measure. Anyone can hit a drum, but it takes a certain person to make it look and sound cool.
If you are interested in a Team Building Activity event or ice breaker, we cover the whole of the United Kingdom, please feel free to get in touch for more information or visit the information page at the top of this page.
Thank you for taking the time to look at my page, enjoy the site and please contact me for all questions/enquiries.


text:
NOM president Brian Brown has penned a lengthy tribute to late Justice Antonin Scalia’s attempts to keep homosexuality criminalized in the United States. An excerpt:
When the majority of his colleagues, led by Justice Anthony Kennedy, ruled in Romer v Evans that the voters of Colorado could not prohibit the granting of special rights to homosexuals, Scalia took them to task for the abject lack of constitutional basis for the ruling.
He reminded the Court that just ten years earlier in Bowers v. Hardwick they had ruled that there was no fundamental constitutional right for someone to engage in homosexual acts and that the constitution had not changed during the intervening decade. Despite this, the liberal Kennedy wing decreed that it is unconstitutional for voters to prevent special rights to be granted to gays and lesbians because doing so supposedly amounted to animus and discrimination, and that expressing moral disapproval of homosexual conduct was akin to racial and religious bigotry.
To this, Justice Scalia chastised the majority for its utter lack of judicial basis: “The Court today… employs a constitutional theory heretofore unknown to frustrate Colorado’s reasonable effort to preserve traditional American moral values…But the Court today has done so, not only by inventing a novel and extravagant constitutional doctrine to take the victory away from traditional forces, but even by verbally disparaging as bigotry adherence to traditional attitudes.”
The constitutional battle over marriage and traditional American values came more clearly into focus in the case of Lawrence v Kansas, which revisited whether states could penalize people for engaging in homosexual conduct such as sodomy (which was at issue in the Bowers case years before). Once again, Justice Anthony Kennedy led the majority to strike down state laws penalizing sodomy, and once again Justice Scalia dissented — not because he personally favored penalties for homosexual conduct but because the US Constitution simply does not contain a fundamental right to homosexuality.
In his dissent in Lawrence, Justice Scalia presciently warned that the ruling would inevitably lead to gay marriage and the striking down of laws against polygamy, bigamy, adult incest and bestiality because the Court was effectively banning states from considering the morality of sexual practices as an intrusion into the personal and private life of the individual.


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The Red Lion Public House Thurleigh
The Red Lion in 1960 [Z53/122/2]
The Bedfordshire Historic Environment Record [HER] contains information on the county’s historic buildings and landscapes and summaries of each entry can now be found online as part of the Heritage Gateway website. The HER tells us [HER 5995] that the building containing the former Red Lion used to be listed but, due to extensive modernisations, was de-listed in 1997. The core of the building dates from the 17th century and is timber-framed with colour-washed roughcast render and a thatched roof. It is built in an L-shape and has a 20th century block. It has one storey with attics for north and central sections and two storeys to the south-east.
The inn is first mentioned in the countywide licensing register of 1825 [CLP13]. As it is not mentioned in the previous registers of 1822 to 1824 it seems as if this is when it was first licensed. However, this is not the first reference to the building. In 1713 John Harvey devised a cottage with a pightle (a small close) and cow pasture near the church to his wife for her life and, after her death, ordering that profits be given at Christmas to the poorest families in the village [P97/25/2/1].
The east end of 2 High Street January 2015
The countywide licensing register of 1876 confirms that the Red Lion was owned by Rev Benjamin Trapp, Vicar of Thurleigh, presumably as trustee of John Harvey’s Charity. The countywide licensing register of 1891 confirms the owners as Harvey’s Charity and the pub as a free house. In 1903, however, the trustees leased the Red Lion to Bedford brewers Newland and Nash Limited [P97/25/2/2].
The Rating and Valuation Act 1925 specified that every building and piece of land in the country was to be assessed to determine its rateable value. The valuer visiting the Red Lion [DV1/C73/124] noted that the lease was now held by Biggleswade brewers Wells and Winchwho had bought Newland and Nash in 1924. The landlord (“only been here 3 months”) paid rent of £12 per annum for a tap room, a bar, a parlour, a club room, a cellar and three bedrooms. Outside were three loose boxes, a cart shed and two more loose boxes. The valuer commented: "plaster and thatched, artistic looking, roomy place". He also noted: "Does not do 18 [gallons?] a week. No spirits, does not do a bottle of whisky a week, small tobacco trade". The public house seems to have closed around 1960.
By 1965 the place was a private house. In May that year it was gutted by fire. The Bedfordshire Times reported that the owners had purchased it three years before “and worked every moment of their spare time to make it into a beautiful showpiece home. On Saturday, their task completed, a house-warming party was in progress when fire destroyed the house. Customers from the Jackal public house opposite gave all the help they could and saved much of the furniture. Several small children who were in the house were taken to safety and no one was hurt”.
The former Red Lion from the churchyard January 2015
- P97/25/2/1: will of John Harvey: 1713;
- CLP13: Register of Alehouse Recognizances: 1825-1828;
- PSS3/1: Register of Alehouse Licences - Sharnbrook Petty Sessional Division: c.1901;
- PSS3/2: Register of Alehouse Licences - Sharnbrook Petty Sessional Division: c.1903;
- P97/25/2/2: counterpart lease: 1903;
- PSS3/3: Register of Alehouse Licences - Sharnbrook Petty Sessional Division: 1904-1930;
- P97/25/2/3: payment of rent: 1907-1919;
- P97/25/2/5: insurance policy: 1930;
- GK297/3: conveyance from Newland and Nash to Wells and Winch: 1938;
- Z53/122/2: photograph: 1960;
- Z50/122/1-7: interior photographs showing construction: 1962;
- Z1091/6/10g: three snapshots: c. 1962;
- FSD/PC7: newspaper report of the cottage following destruction by fire: 1965;
List of Licensees: note that this is not a complete list. Italics indicate licensees whose beginning and/or end dates are not known:
1825-1828: James Packwood;
1847-1876: Edward Long;
1885: James Hart;
1890-1898: John Thomas Russell;
1903: John Zanker;
1906: George Edward Philpott;
1910-1913: Jane Philpot;
1913-1921: William Asque;
1921-1925: Sidney Stratford;
1925-1926: Thomas Frederick Bond;
1926-1928: Charles Thomas Deacon;
1928-1929: Arthur Thomas Thorne;
1929-1931: Frederick Henry Phelps;
1931-1932: John Edward Herton;
1932: Henry Gardener;
1940: William H Coleman.
Closed about 1960.

domain: history