text:
Q: Explorer.exe: No such interface supported Lately Windows 7 has been throwing these weird errors at me and refusing to open certain applications. From what I've managed to figure out, the problem might be a lost dll.
Errors:
While trying to open up "Screen resolution", "Personalize",  from the desktop, it throws back an error with "Explorer.EXE" in the title saying "No such interface supported". All I can do is to agree with "Ok".
When programs such as Microsoft Visual studio tries to open up Explorer.exe to open a file it will fail due to unknown reasons. "The operation could not be completed".
Trying to open up certain windows ("Join a Personal Area Network" and "Show bluetooth devices") for Bluetooth activities, the window just won't open up, while others open up in an instance.
Several activities in the Control panel won't work either, such as "Uninstall program", "Adjust screen resolution" (wont work from here either :p)

I'm really clueless where to look or what to do right now, except for re-installing the whole thing, which I rather not spend my weekend doing. Help? :)

A: Some ideas:

*

*Have you installed some product just before this started happening?
If yes, it might be the cause of the problem. An uninstallation is the best way to verify.

*Try to boot in Safe mode (F8 during Windows startup). In this mode only Windows components are allowed to startup. If the problem disappears, then it's some installed product that's causing it. Download Autoruns, save a system restore point, and then use it to serially block products from starting with Windows, until you find the guilty one.

*Based on the premise that something went wrong with your own account that prevents you from executing some programs, the solution would be to create a new user profile and abandon your old one.

*Based on the premise that some dll went wrong, and if you have a real Windows installation DVD (meaning not just a restore one), you might try to do an upgrade installation. This would do an upgrade of your system to itself, meaning nothing, but will re-establish any wrong dlls.

*The last-ditch effort would be to re-install Windows. But wait with this until some more ideas for solutions come in from others on this forum.

A: Not virus  it is permissions problem you need to go into registry and correct the permissions for hkeyclasses root . Add the current user to the permissions of CLSID and give at least read access. For correct instructions go search for no such interface supported on sevenforums.com

A: I would personally see if a system restore works, and failing that, do a re install of Windows.
This is not the sort of error that happens every day, it must of been caused by some sort of failure and even though there are methods that may be able to fix it, if you do not know what caused it - it could lead to problems later on.

A: there is a corrupted or missing system file, so SFC /SCANNOW may help you recover them.

A: We have had two such issues. Same exact problem. Doing a system restore back to a time before the problem existed fixed the problem. It seems a Microsoft update caused the issues.

A: Having a similar issue on Windows 2012, where it suddenly stopped opening Explorer.exe on Windows logon, and displayed a black desktop.
Any attempt to load Explorer.exe failed with error "No such interface supported".
I found a solution in http://www.w7forums.com/threads/no-such-interface-supported.17543/ :
From Task Manager (CTRL + SHIFT + ESC), open CMD and run:
FOR /R C:\Windows\ %G IN (*.dll) DO "%systemroot%\system32\regsvr32.exe" /s "%G"

It will take some time and will open many Runtime errors messages, you can close them all.
Once completed, restart windows ( shutdown -r -t 0 ), and hold fingers...

A: In reference to all of the above, thanks.Great ideas, but I've found the only way you can really fix this issue is with a Restore to a working restore Point.Pain in the butt but it works perfectly for me.As I Purposely create restore points once a week.This problem was caused FYI by a "Scam" software I bought called, Advanced System Repair. BEWARE of this software,it really destroys a lot of things.Running Win7, i7 CPU,32ram etc.Hope this helps someone,and I recommend creating restore points on a schedule base for sure.
Jim

redpajama_set_name: RedPajamaStackExchange


Do you want to get rid of that belly fat hanging loose around your tummy? This HIIT workout to target abs may be the solution to that problem!
Amy Roberts, CPT says these HIIT moves can burn your upper body and build up muscle. The duration is only 10 minutes but this can be pretty intense and long. These moves lets you pay close attention to your abs, with fat-torching and muscle-toning moves, you'll be burning quite a good amount of your calories.
Mark Beier, co-director at Shred415 in Chicago, is the one who came up with the workout. He says that this routine uses multi-plane movements that target each area of your core muscles for ten minutes.
Just take note, when you start, aim for quality not quantity. You'd want this to be effective so make sure your execution is correct.
Nora Tobin from shape.com says that this is the best workout for tight and toned abs. Your stomach muscles just need to be revealed and we all can do that if we follow this intense training that she uses for her beach volleyball season. She says that her go-to exercise is a combination of jumping rope and six core exercises.
This is the right workout for you if you want to sculpt that stomach. You can turn this into your main workout three to five days a week or insert these moves into your regular routine.
Ben Boudro from muscleandfitness.com says that this tri-set routine is your ticket to a ripped form. If you're repeatedly asking yourself how to get rid of that stubborn belly fat, commit to this routine and say bye-bye to your bulging tummy.
Mind you, this is fast and extreme so prepare your body well. This will shred those abs and you'll definitely end up looking fantastic!
Boudro's tri-set routine requires you to do exercises A, B, and C without resting. This is Round 1; the target is three rounds. Perform as many reps as possible!
Since these are extreme routines, you may want to think about your hydration needs. MusclePharm Amino1 might do the trick for you. This can totally compensate your lost energy and help your body recover.
The post HIIT Workout to Target Abs: Get Your Body Ready With Amino1 appeared first on Muscle Pharm Reviews.
You want to turn your arms into big guns? We've got it all right here for you. Check out our gun training tips to turn your skinny arms into cannons.
Anthony Bevilacqua, CPT from Muscle shares his own techniques how to turn your upper extremities into cannons. Muscles are indeed magnificent and most guys would usually work themselves out just to achieve glorious arms.
The author mentioned blood flow restriction training, increasing volume, and utilizing all angles. He also included workout tips for your arms. These are designed to help you reach those goals. Follow this guide and get ready to be noticed! Your muscle definition will reach its peak!
Author Edward Lane from Men's Health UK compiled 16 best exercises for bigger arms. If you still feel like you can still fill out that t-shirt, you can trust that these exercises will make things work for you. Each exercise outlined here is capable of pushing your muscles into performing at an optimum level.
You have the potential to make yourself stronger and better. This has detailed instructions how to perform each exercise so you'll know what to do in each rep.
Frank McGrath for Flexonline.com shares his advice how to train your arms and stimulate muscle growth. He gives us tips how he does it on his own, and he reveals how he builds his arms through this workout. He has a variety of exercises focusing on arm work to make his arm muscles body bigger.
For increasing muscle protein synthesis, you have to take the right supplement for your muscles. MusclePharm's Combat Protein is a pure protein source that is capable of supporting muscle building.
The post Gun Training Tips: Improve Your Upper Body With MusclePharm appeared first on Muscle Pharm Reviews.
Put some spice into your routine using these unique bodyweight moves. These are bound o get you toned in no time. You won't even have to use any equipment. Just these exercises!
Lara McGlashan MFA CPT from muscleandperformance.com says that these 5 moves can tone your legs in no time. You don't even need to go to the gym anymore. These exercises target different muscle groups in your body. In addition to that, it burns fat and improves endurance.
You can incorporate these exercises into your daily leg routine in order to give your muscles a good workout. This workout is also doable outdoors. You'll stir things up a bit, and that's enough to have an effect on your metabolism. You'll get those shapely legs you've been dreaming of!
Craig Ballantyne says these awesome and unique bodyweight exercises are even more effective than pushups and crunches. Why? You don't see these on a daily basis.
The author collaborated with his two friends Adam and Ryan to come up with these unique moves. His two friends are innovative: they're masters in reinventing exercises. These exercises can shake things up, and can add variety to your normal routine. Spice things up by incorporating your current training plan with these moves!
Mitchell Graves is a personal trainer and author. He shares his thoughts about bodyweight moves and why it's becoming a new trend in the fitness world; programs such as P90X and Insanity are just a few examples. The author points out why enthusiasts are getting hooked in this traditional form of exercise.
These programs emphasize that you don't need equipment to stay fit. If you've got enough motivation and the right knowledge, you can stay ripped. He shares a combination of workouts that you can perform at the comfort of your home. In addition to that, these can be done within four minutes!
It must be tough to finish all these workouts. You should have something that encourages body recovery. MusclePharm's BCAA is the best example. It has Leucine, which has the capacity slow muscle damage after intense training so it's right for your workout!
The post Best Unique Bodyweight Moves: Tone With These Exercises appeared first on Muscle Pharm Reviews.
A lot of people are struggling with their weight especially those who are in their mid-20s and beyond. It can get quite frustrating for some who didn't even have this problem before. When you reach a certain age, deposited fat can be your greatest enemy.
Losing weight is the easy part. Keeping it off is what's really hard.
To enhance your fat-burning regimen, MusclePharm's Amino1 has L-Carnitine that promotes fat oxidation. It's just the right drink for someone who wants to lose more fat. You can take Amino1 before, after, or during your workout.
The post Ways To Keep Fat Off: Maintaining Weight With MusclePharm appeared first on Muscle Pharm Reviews.


Control4 has announced their partnership with MainStreet America (MSA), a home product theme park and interactive shopping destination.
The 14-acre site, which is located 30 minutes from downtown Houston, on Interstate 45 in Spring, TX, will showcase 12 fully-furnished showcase homes equipped with the latest in home technology, appliances and interior décor. Control4 smart home solutions will be featured in two of the homes: a 2,600 square foot, 3-story, contemporary, LEED Platinum Certified Mid-town home and an expansive 6,000 square foot Villa Lago Mediterranean-inspired home. Control4 is working with local integrator, Randal Duncan of Data Smart, to design a unique suite of automated solutions that fits the distinct needs of each home.
Main Street America is designed to be a year-round interactive home show and shopping destination. It will be outfitted with a two-story 44,000 square foot guest center, cooking demonstration studio, a custom home building office, a home remodeling center, mortgage financing services, an interior design studio, specialty café and childcare area. Visitors will also be encouraged to engage with products that interest them in the homes using high-tech handheld devices and RFID technology, and to participate in home product demonstrations, how-to workshops and home buying seminars.


لا يجتمع فيها دينان (no non-Muslim can become its citizen till the Day of Judgement).Earlier, the same status was enjoyed by Palestine for centuries. The addressees of Islam and the Islamic shari'ah here too are individuals in their various capacities. However, if for such a state it is said that its religion is Islam and only Islam shall reign in it, then this statement is very comprehensible on all grounds. It cannot be objected to.
Second, the states whose boundaries would be determined by their conquerors. They would govern them by making their inhabitants subservient to them. In such states, the religion of the royal family or of the ruler would be considered as the religion of the state. Disregarding whether their birth was legitimate or illegitimate, if these states are dubbed as Muslim, Christian or Communist states, then this too cannot be regarded as incomprehensible.
Third, the nation-states of the modern era whose boundaries are ascertained by international treaties and which become a source of nationhood themselves for their citizens as soon as they come into existence. Thus in spite of having commonality or diversity in colour, ancestry, language and culture they call themselves Egyptians, Americans, Afghans and Pakistanis and express their nationhood in this respect. No one is superior or subservient here. All, in fact, are regarded as equal citizens in all respects and in this capacity participate in the affairs of the state.
You are free; you are free to go to your temples, you are free to go to your mosques or to any other place of worship in this State of Pakistan. You may belong to any religion or caste or creed that has nothing to do with the business of the State. As you know, history shows that in England, conditions, some time ago, were much worse than those prevailing in India today. The Roman Catholics and the Protestants persecuted each other. Even now there are some States in existence where there are discriminations made and bars imposed against a particular class. Thank God, we are not starting in those days. We are starting in the days where there is no discrimination, no distinction between one community and another, no discrimination between one caste or creed and another. We are starting with this fundamental principle that we are all citizens and equal citizens of one State. The people of England in course of time had to face the realities of the situation and had to discharge the responsibilities and burdens placed upon them by the government of their country and they went through that fire step by step. Today, you might say with justice that Roman Catholics and Protestants do not exist; what exists now is that every man is a citizen, an equal citizen of Great Britain and they are all members of the Nation.
A question that can be raised on this is: does Islam acknowledge such a state? I had tried to answer this question by saying that Islam addresses the rulers of a society.If they are Muslims, then they are the real addressees of Islam's directives related to the collectivity. Islam has not given any directive to Islamize a state. Thus its followers can live as citizens in such nation-states and as a nation on the basis of nationhood the way they currently live in many states. Nothing in this is against Islam and the Islamic shari'ah.
This is my stance regarding the nation-states of these times.
Consider next the government. Only two statements endorsed by knowledge and reason can be given about it: first, the rulers of a state will be chosen by the Almighty, and second, they will be chosen by the inhabitants. After the termination of the institution of prophethood the first of the above options is not possible. Only the second one remains. Its essential consequence is that the majority will have the right to rule. If Muslims have this majority and if on its basis they become the rulers of a state, then it is their democratic and human right that if their religion has given them a directive regarding the collectivity, then they should follow it and also decide all matters of the followers of this religion in accordance with the shari'ah revealed by God through His last prophet. It is precisely this what the Quaid e Azam implied when he would speak of Islam, Islamic civilization and the Islamic shari'ah.
The relationship of the shari'ah with the nation-states of today is of the nature just described. I had endeavoured to put across this very understanding Thus, in accordance with this understanding, I had also presented a comprehensive list of directives of the shari'ah that relate to the collectivity. I had also written that Muslims have been given these directives with the warning that those who do not accept the verdicts of the Book of God after acknowledging it will be regarded as wrongdoers (zalim), defiant (fasiq) and disbelievers (kafir) on the Day of Judgedment. Men of learning can differ with the list I have compiled and also alter it. However, if even after understanding this difference between state and government and after viewing this list they claim that I have confined the shari'ah to an individual or negated all directives that relate to the spheres of politics, economics and the social set-up and they are also not feigning ignorance, then I dare say that they have not understood my views on this topic at all.
.Abu 'AbdullahMalik ibn Anas, Mu'atta', vol. 2 (Cairo: Dar ihya' al-turath al-'arabi, n.d.), 892, (no. 1584).
.Reference is to the article: "Islam and the State: A Counter Narrative."


Sir Arthur Foulkes writes To The Point on Tuesdays in the Nassau Tribune. He was a reporter and editor at the Tribune from 1948 to 1962 when he became founding editor of the Bahamian Times. His column first appeared in the Tribune in the early 1970s. Elected to parliament in 1967, he was a cabinet minister in the first PLP government and a founder of the Free National Movement in 1971. As a delegate to the Bahamas Independence Conference in London in 1972, he drafted the opposition memorandum for the conference. In 1992 he was appointed high commissioner to the United Kingdom and ambassador to the European Union. Since 1999 he has been ambassador to China. Email: sirarthurfoulkes@hotmail.com.
Craig Butler studied law at Wolverhampton University in England and at the Norman Manley Law School in Jamaica. He also has a degree in economics from Rollins College in Florida. Mr Butler's column runs in the Nassau Guardian on Mondays and he also hosts a weekly political talk show on Island FM. He is the grandson of Sir Milo Butler, the first governor-general of an independent Bahamas. Email: cfmilobutler@hotmail.com.
Nicolette Bethel is the author of Essays on Life in the Nassau Guardian Weekender, and currently serves as director of culture for the Bahamas government. She is a social anthropologist, a writer and researcher in the fields of Bahamian national identity and Junkanoo. Her plays have been produced locally, and her fiction and poetry have appeared in various collections. In her spare time she teaches English, Cultural Studies and Anthropology at the College of The Bahamas. Email: nico@nicobethel.com.


Prof Alan Mackay-Sim, who heads up the HSP research team at Griffith University, has received worldwide recognition and praise for his key role in the world's first successful restoration of walking ability in someone with a severed spinal cord.
Nasal stem cells, as used in HSP research, were employed in repairing the severed spinal cord in an operation performed in Poland. Prof Mackay-Sim is a recognised global authority on nasal stem cells and has brought his expertise to bear on the problem of repairing severed spinal cords over many years.
A quadriplegic Bulgarian man with no ability to walk since severing his spinal cord in an accident is now learning to walk again following the successful surgical procedure.
The story received global media coverage in late October. Here is an editorial focused on Prof Mackay-Sim from The Australian newspaper in late October.
Editors Note: There is no relevance of this successful treatment for severed spinal cords to treating HSP. This article is to highlight the standing and prominence of Prof Mackay-Sim and provide warranted confidence that the HSP research program supported by this Foundation is in the best hands available anywhere in the world.
Fantastic story which gives confidence the research for HSP is in very good hands!
I like a lot of other people will be watching to see how Prof Mackay and his team go with the next stage with HSP research program.


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ENERGY FROM FORESTS
Calophyllum inophyllum (Tamanu tree)
Illustration by Karyono, CIFOR
Home » Features » Energy from forests » Calophyllum inophyllum (Tamanu tree)
Also known as nyamplung or the tamanu tree, Calophyllum inophyllum is a pantropical species tree found in 38 countries stretching from East Africa to Southeast Asia, Oceania and the South Pacific. Originating in East Africa in Kenya, Tanzania, and Mozambique, the tree's native range is unclear, making it hard to fully define native and introduced localities for the species. Nyamplung is well-adapted to coastal areas and lowland forests, and can grow in sandy, clay and degraded soils.
Calophyllum inophyllum is often planted as a roadside tree to provide shade and as wind breaks with its large canopy. Informally known as mastwood, the tree has long been used in shipbuilding, such as for the construction of canoes and small boats, but also for masts, keels, knees and pulley blocks. Similar to other species with bioenergy potential, nyamplung is also used in medicine. In Mauritius, for example, liquifying roots by extracting the essence of a substance is used to treat ulcers, boils, and ophthalmia. The bark can also be used to treat orchitis, while the latex has been traditionally used on the skin to treat rheumatism and psoriasis, and a leaf extract is used to treat eye infections as an anti-inflammatory. The stones of the fruit are used as marbles.
Since 2017, experimental nyamplung plantations cultivated on degraded lands in the Indonesian provinces of Central Java, Central and East Kalimantan are showing promising potential. In these areas, the tamanu tree demonstrates an ability to restore and improve former forests and peatlands that have been burned or damaged from commercial actives or large-scale deforestation.
Nyamplung oil offers much potential for bioenergy conversion, with similar properties to diesel fuel. The species is a ready source of clean, carbon-neutral biodiesel, with a yield of 95 percent under optimal conditions. There are several technological options for the conversion of nyamplung oil into biodiesel. Further, the properties of Calophyllum inophyllum biodiesel meet American Society for Testing and Materials (ASTM) standards, an international standard, generally and yields more oil per hectare compared to other plants, with an average oil yield of 5,000 L/ha. These properties make the species an attractive alternate biodiesel feedstock.
Rahman, S. A., & Baral, H. (2018). Integrating bioenergy and food production on degraded landscapes in Indonesia.
Barstow, M. (2019). Calophyllum inophyllum. The IUCN Red List of Threatened Species 2019: e.T33196A67775081.
Arumugam, A., & Ponnusami, V. (2019). Biodiesel production from Calophyllum inophyllum oil a potential non-edible feedstock: An overview. Renewable Energy, 131, 459-471.
Lemmens, R.H.M.J. (2005). Calophyllum inophyllum L. [Internet] Record from PROTA4U. Louppe, D., Oteng-Amoako, A.A. & Brink, M. (Editors). PROTA (Plant Resources of Tropical Africa / Resources végétales de l'Afrique tropicale), Wageningen, Netherlands.
Iinuma, M., Tosa, H., Tanaka, T., & Yonemori, S. (1994). Two xanthones from root bark of Calophyllum inophyllum. Phytochemistry, 35(2), 527-532.
Vitex pubescens
Croton megalocarpus Hutchinson
Azadirachta indica
This work is supported by:
The information provided in this article is not peer-reviewed and is not intended as advice. It is based on current knowledge and the available literature. CIFOR and partner organizations do not provide any guarantees and recommend interested stakeholders undertake further independent assessments and verifications before making business and investment decisions.

redpajama_set_name: RedPajamaCommonCrawl


text:
Caspase-3-Generated Fragment of Gelsolin: Effector of Morphological Change in Apoptosis
Srinivas Kothakota*,
Toshifumi Azuma*,
Christoph Reinhard,
Anke Klippel,
Jay Tang,
Keting Chu,
Thomas J. McGarry,
Marc W. Kirschner,
Kirston Koths,
David J. Kwiatkowski†,
Lewis T. Williams
S. Kothakota, C. Reinhard, A. Klippel, K. Chu, K. Koths, L. T. Williams, Chiron Corporation, Emeryville, CA 94608, USA.
T. Azuma, J. Tang, D. J. Kwiatkowski, Division of Experimental Medicine, Brigham and Women's Hospital, Boston, MA 02115, USA.
T. J. McGarry and M. W. Kirschner, Department of Cell Biology, Harvard Medical School, Boston, MA 02115, USA.
DOI: 10.1126/science.278.5336.294
Srinivas Kothakota
Toshifumi Azuma
Christoph Reinhard
Anke Klippel
Jay Tang
Keting Chu
Thomas J. McGarry
Marc W. Kirschner
Kirston Koths
David J. Kwiatkowski
The caspase-3 (CPP32, apopain, YAMA) family of cysteinyl proteases has been implicated as key mediators of apoptosis in mammalian cells. Gelsolin was identified as a substrate for caspase-3 by screening the translation products of small complementary DNA pools for sensitivity to cleavage by caspase-3. Gelsolin was cleaved in vivo in a caspase-dependent manner in cells stimulated by Fas. Caspase-cleaved gelsolin severed actin filaments in vitro in a Ca2+-independent manner. Expression of the gelsolin cleavage product in multiple cell types caused the cells to round up, detach from the plate, and undergo nuclear fragmentation. Neutrophils isolated from mice lacking gelsolin had delayed onset of both blebbing and DNA fragmentation, following apoptosis induction, compared with wild-type neutrophils. Thus, cleaved gelsolin may be one physiological effector of morphologic change during apoptosis.
A conserved family of aspartate-specific cysteinyl proteases (caspases) has been identified as critical mediators of apoptosis in Caenorhabditis elegansand mammals (1, 2). Although multiple protein substrates of caspases have been found, the functional significance of the substrates is poorly understood (3). We reasoned that an unbiased approach to determine proteins that were the best substrates of caspase-3 in vitro would yield a physiologically relevant substrate. Therefore, we constructed a protein library by translating a murine embryo cDNA library in vitro (4) and tested the translated proteins for their sensitivity to caspase-3 cleavage. To facilitate screening, we separated the cDNAs into small pools before in vitro translation and incorporated [35S]methionine into the translation mix to label the proteins. The translated pools were separated into two parts; one portion was incubated with active caspase-3, and the other portion was incubated with caspase-3 inactivated by the addition ofN-acetyl-Asp-Glu-Val-Asp–fluoromethyl ketone (DEVD-fmk) (2). The reaction products were resolved by SDS–polyacrylamide gel electrophoresis (PAGE), and protein targets of caspase-3 were identified by comparing the pattern of35S-labeled proteins in the samples treated with active and inactive caspase-3. To avoid false positives, we found it necessary to carefully titrate the amount of enzyme. We used an amount sufficient to cleave 80% of a known substrate of caspase-3, baculovirus protein p35 (Fig. 1A) (5).
Download high-res image
Identification of gelsolin as a substrate of caspase-3. (A) Autoradiogram of [35S]methionine-labeled proteins treated with purified active (–) or inactivated [by addition of a caspase-3 inhibitor, DEVD-fmk (+)] recombinant caspase-3. Lanes 1 through 6 contain the translated proteins from six different representative pools of cDNAs (23). The arrows indicate a caspase-3–sensitive protein that is cleaved from an apparent size of 65 kD to 48 kD. This protein was identified as gelsolin. (B) Cleavage of bacterially expressed murine gelsolin (16) by caspase-3. Gelsolin (13.5 μg) and caspase-3 (6 ng/ml) were incubated at 37°C for the indicated times in 6 mM tris-HCl (pH 7.5), 1.2 mM CaCl2, 5 mM dithiothreitol (DTT), 1.5 mM MgCl2, and 1 mM KCl in a 25 μl volume. A volume of 2 μl was resolved on a gel and stained with Coomassie blue. Bands of apparent size of 48 and 40 kD, corresponding to actual molecular sizes of 41 and 39 kD, respectively, are seen.
One thousand cDNA pools, each containing 100 cDNA clones (100,000 cDNA clones total), were screened to identify substrates of caspase-3. In three different pools, incubation of active caspase-3 with labeled proteins reduced the intensity of a 65-kD band and generated a new band at 48 kD (Fig. 1A, pool 4). To identify the cDNA clone encoding the 65-kD protein, the DNA from the positive pools was used to transformEscherichia coli, and the DNA prepared from single colonies was screened again. DNA sequencing and polymerase chain reaction analyses of the single positive clone from all three pools identified the 65-kD protein as the partial sequence of gelsolin from residues 142 to 731.
Caspase-3 rapidly cleaved full-length recombinant murine gelsolin, confirming that gelsolin is a substrate for caspase-3 (Fig. 1B). Gelsolin was also cleaved upon incubation with cell extracts prepared from cells that were induced to undergo apoptosis by Fas activation (6). However, no gelsolin cleavage activity was observed in cell extracts prepared from untreated cells. Protein microsequencing by Edman degradation was used to determine the NH2-terminal sequences of the cleavage products and indicated a cleavage site between residues Asp352 and Gly353 of murine gelsolin, so that products had actual molecular masses of 39 (NH2-terminal) and 41 (COOH-terminal) kD. The sequence in this region, Asp349-Gln-Thr-Asp-Gly353, is consistent with the known requirements for efficient cleavage by caspase-3 (2) and is conserved between mouse, human, and porcine gelsolin (7). Cleavage of gelsolin also resulted in dissociation of the 39- and 41-kD cleavage products when assayed by size-exclusion chromatography (8).
To examine cleavage of gelsolin during apoptosis in vivo, initially we used a model cell system in which apoptosis is highly inducible (9). The assay used fibroblasts that expressed a chimeric receptor composed of the extracellular and transmembrane domains of murine CD4 and the cytoplasmic domain of Fas. Apoptosis was induced by antibody cross-linking of the extracellular CD4 domains. Gelsolin was cleaved into the predicted 39- and 41-kD products 30 min after apoptosis induction (Fig. 2A). Cleavage of gelsolin occurred early and was comparable to the time course of cleavage of poly–adenosine diphosphate ribose polymerase (PARP), a substrate of caspase-3 (Fig. 2A). Both gelsolin and PARP cleavage was blocked by the cell-permeable inhibitor of caspase-3,N-benzyloxycarbonyl-Val-Ala-Asp–fmk (zVAD-fmk) (10). Gelsolin cleavage was specific, because another cytoskeletal protein, filamin, was not cleaved under identical conditions (Fig. 2A). Thus, gelsolin was specifically cleaved by a caspase-3–like enzyme in vivo, and this cleavage is an early step in Fas-mediated apoptosis.
Gelsolin was cleaved in apoptosis induced by Fas and TNFα. (A) Apoptosis was induced by receptor cross-linking with and without 200 μM zVAD-fmk (caspase inhibitor) in murine L929 cells as described (9). The cell lysates from an equal number of cells were analyzed by SDS-PAGE and immunoblotted with anti-gelsolin [top panel; polyclonal antibody (16)], anti-PARP (middle panel; Enzyme Systems Products), and anti-filamin (bottom; Sigma). The immunoblots were visualized by ECL (Amersham). (B) Human neutrophils were isolated with the neutrophil isolation medium (Cardinal Associates) and resuspended in RPMI with 10% fetal bovine serum (FBS) (24). Equal numbers of cells were used to prepare cell lysates after incubation with FAS antibody (1 μg/ml, top) or 10 μg/ml TNF and 10 ng/ml cycloheximide (bottom). The lysates were separated by SDS-PAGE and analyzed by immunoblotting with a monoclonal antibody to gelsolin (Sigma), which detects only the COOH-terminal 41-kD fragment. (C) Human neutrophils were stained at different time points after isolation from blood, using annexin V apoptosis kit (Clontech), and the number of live cells was estimated by counting the number of annexin- and propidium iodide–negative cells (24).
Neutrophils purified from human blood express large amounts of gelsolin and undergo spontaneous apoptosis. The rate of neutrophil apoptosis can be further enhanced by cross-linking of Fas or treatment with tumor necrosis factor α (TNFα) and cycloheximide (11). To determine if gelsolin is also cleaved when neutrophils undergo apoptosis, we analyzed neutrophil lysates with a monoclonal antibody to gelsolin that recognizes an epitope in the COOH-terminal half of gelsolin. During spontaneous apoptosis in neutrophils, gelsolin amounts decreased and a 41-kD fragment appeared with time (Fig. 2B)—a size identical to fragments generated by cleavage of gelsolin by caspase-3. Increasing the rate of neutrophil apoptosis by cross-linking Fas with antibodies or treatment with TNFα and cycloheximide resulted in an increase in the rate of gelsolin cleavage and appearance of the 41-kD fragment (Fig. 2, B and C). Thus, the cleavage of gelsolin observed in vitro also occurred during apoptosis of neutrophils.
To determine the functional significance of gelsolin cleavage, we examined the activities of cleaved and native gelsolin, using pyrene-actin fluorimetry. In this assay, the conversion of filamentous (F) actin to monomeric (G) actin is monitored by the 25-fold difference in fluorescence between the two states. Native gelsolin severs actin polymers in a Ca2+-dependent manner (12, 13), whereas caspase-3–cleaved gelsolin severed actin polymers independent of Ca2+ (Fig. 3A, top). Gelsolin has both actin monomer–binding and F-actin–severing activities (12, 13). However, the cleaved gelsolin preferentially severed actin filaments even when briefly incubated with excess monomeric actin (Fig. 3A, bottom), suggesting that the cleaved gelsolin generated in cells during apoptosis may preferentially sever actin filaments rather than bind monomeric actin. Next, we used permeabilized fibroblasts to determine the ability of cleaved gelsolin to depolymerize the cytoskeletal actin filaments. Cleaved gelsolin depolymerized the actin cytoskeleton in a Ca2+-independent manner, whereas uncut gelsolin was inactive in the presence of EGTA (Fig. 3B).
Caspase-3–cleaved gelsolin is activated and severs actin filaments in a Ca2+-independent manner. (A) (Top) Ca2+-independent depolymerization of actin filaments in vitro. We incubated 0.5 μM polymerized pyrene-labeled actin and 50 nM gelsolin (circles) or caspase-3–cleaved gelsolin (triangles) in 2 mM tris-HCl (pH 7.5), 0.5 mM adenosine triphosphate, 0.2 mM DTT, 2 mM MgCl2, and 150 mM KCl, with CaCl2 (0.2 mM; open circles and triangles) or with EGTA (1 mM; solid circles and triangles). The change in pyrene fluorescence with time is shown. (Bottom) Cleaved gelsolin severs F-actin faster than a complex with G-actin is formed. Cleaved gelsolin (20 nM final concentration) was directly added to 0.5 mM F-actin (triangles), or mixed with a twofold molar ratio of G-actin and immediately added (open diamonds) or incubated for 10 min (closed diamonds) and then added to 300 μl of 0.5 μM F-actin. (B) Cleaved gelsolin severs the actin cytoskeleton. Permeabilized embryonic fibroblasts (Gsn–/–) were incubated with gelsolin or cleaved gelsolin and the actin filaments were visualized by staining with TRITC-phalloidin, as described (25). (C) The NH2-terminal gelsolin cleavage fragment induces breakdown of the cytoskeleton. The cells were injected with DNA encoding the NH2-terminal (top) or COOH-terminal (bottom) fragments of gelsolin and with a plasmid encoding GFP, and were stained for actin filaments with TRITC-phalloidin 5 hours later. The arrows point to the injected cells. The right panel indicates GFP expression, and the left panel indicates actin staining (TRITC-phalloidin) of the same field (26). (D) Adenovirus expressing the NH2-terminal gelsolin fragment induces apoptosis. A7 cells (a human melanoma cell line) were infected with adenovirus, prepared as described (27), expressing the NH2-terminal caspase-3 cleavage fragment of gelsolin (residues 1 through 352, top) or full-length gelsolin (1 through 731, bottom). (E) The NH2-terminal gelsolin cleavage fragment induces changes in cell morphology and acts downstream of caspases. The cells were injected with DNA as in (C) and were incubated with dimethyl sulfoxide (left panels) or 100 μM zVAD-fmk (right panels). The injected cells appear green because of GFP expression.
The actin-severing activity of gelsolin resides in the NH2-terminal region, residues 1 to 160 (12). To test if the NH2-terminal fragment of gelsolin generated by caspase-3 cleavage could depolymerize actin in vivo, we microinjected DNA encoding the NH2-terminal fragment (1 to 352) or the COOH-terminal fragment (353 to 731) into fibroblasts. Expression of the NH2-terminal fragment caused a rapid depolymerization of the actin cytoskeleton, whereas the COOH-terminal fragment had no effect on actin filaments (Fig. 3C). The injection of full-length gelsolin also had no effect on the cell morphology. To determine if gelsolin acts directly to depolymerize actin filaments or indirectly by activating caspases, we incubated the microinjected cells with a caspase inhibitor, zVAD-fmk. Inhibition of caspases did not block the changes in cell morphology induced by the expression of the NH2-terminal fragment of gelsolin (Fig. 3E). We also constructed adenoviral vectors expressing the NH2-terminal fragment (1 to 352) or full-length gelsolin. The adenovirus vector expressing the NH2-terminal fragment caused rapid cell death in A7 melanoma cells, with both morphologic changes and nuclear fragmentation in the majority of cells within 48 hours of application [determined by terminal deoxytransferase (TdT)–mediated deoxyuridine triphosphate (dUTP) nick-end labeling (TUNEL) staining (14)], whereas the full-length construct had no effect (Fig. 3D). A similar result was observed in other cell types, including M2 melanoma cells and NIH 3T3 cells. Both p53+/+ and p53–/– murine embryo fibroblasts (15) also displayed morphologic changes of apoptosis within 48 hours of infection by the gelsolin NH2-terminal fragment-expressing adenovirus. Thus, cleavage of gelsolin generated an NH2-terminal fragment that depolymerizes the actin cytoskeleton in a Ca2+-independent manner, induces cell death, and may be a downstream effector of the morphological changes that are observed during apoptosis.
We used gelsolin null cells derived from Gsn–/–mice (16) to examine the importance of gelsolin in apoptosis occurring during physiological stimuli. The course of apoptosis in peritoneal neutrophils treated with TNF plus cycloheximide (TNF+CHX) was assessed by videomicroscopy. Wild-type neutrophils developed blebs as early as 48 min after TNF+CHX, and 52% of cells (32 of 61) had begun blebbing by 6 hours (Fig. 4A). In contrast (Fig. 4B), Gsn–/– neutrophils did not develop blebs until 2 hours 2 min after TNF+CHX, and at 6 hours, only 12% of cells (10 of 83, P < 0.001) had blebs. This delay in the progression of apoptosis appeared to contribute directly to a delay in DNA fragmentation in Gsn–/– cells compared with wild-type cells, as assessed in TUNEL assays (Fig. 4C) and DNA analysis by electrophoresis (17). No difference in caspase-3 activation was observed when Gsn–/– and wild-type neutrophils were compared, as assessed with the peptide substrate DEVD–7-amino-4-trifluoromethylcoumarin (DEVD-AFC) (18), suggesting that the differences seen were from a defect in apoptotic progression downstream of caspase-3 activation. Similar differences in apoptotic progression were seen in Gsn–/– neutrophils in response to treatment with a monoclonal antibody to Fas. Our results may explain the previously observed moderate neutrophilia of Gsn–/– mice, which have approximately twice the number of circulating neutrophils as wild-type mice (16).
Gelsolin null neutrophils have a reduced rate of cell death. Neutrophils were isolated from the peritoneal exudates of wild-type and Gsn–/– mice 5 hours after thioglycollate administration (16). Neutrophils were washed in PBS and then minimum essential medium, then were suspended in RPMI with 10% FBS at 106 cells/ml. mTNFα and cycloheximide (10 μg/ml) were used to induce apoptosis. (A and B) Comparison of blebbing onset in wild-type (A) versus Gsn–/– (B) neutrophils. Arrows indicate blebbing cells. Videomicroscopy was performed with differential interference contrast optics on a Zeiss Axiovert 405M inverted microscope. Images were collected with a Hamamatsu C2400 video camera (Photonic Microscopy, Bridgewater, New Jersey) and recorded on a Panasonic TQ-3038F video recorder. (C) Comparison of DNA fragmentation in Gsn–/– and wild-type neutrophils. The fraction of apoptotic cells as determined by FACS analysis in a TUNEL assay are shown (14). n = 3 with error bars.
To confirm the role of cleavage of gelsolin in the morphologic changes of apoptosis, we used the human malignant cell line HeLa, which does not express gelsolin and is relatively resistant to apoptosis (19). Gelsolin-expressing, stably transfected HeLa cell lines were more sensitive to human TNFα than were HeLa sublines transfected with control vector alone. Some 72%, 52%, and 38% of the cells from three different gelsolin-expressing HeLa sublines displayed cell rounding and blebbing after treatment with TNF+CHX for 12 hours, while only 8% of the cells from a control line showed these changes.
Caspase or caspase-like cleavage of multiple proteins has been described (3, 20), but few of these are known to have direct physiological significance in the morphologic changes and nuclear degradation that are hallmarks of apoptosis. Using an unbiased approach, we found that the actin-modulating protein gelsolin is the most prominent direct substrate of caspase-3 in murine embryos. Our data also indicate that gelsolin is a probable in vivo target of the apoptotic caspase-initiated cascade and that the gelsolin fragment mediates, in part, the morphologic changes of apoptosis. Blockade or enhancement of gelsolin cleavage might retard or enhance apoptosis in multiple cell types. Gelsolin is the founding member of an evolutionarily conserved family of proteins that extends toDictyostelium and Drosophila, and in humans consists of at least six proteins, whose expression is tissue-specific (21). Apoptotic cleavage of other gelsolin family members may also occur, and it is possible that these gelsolin homologs have similar roles in mediating apoptotic cytoskeletal changes in specific cell types and tissues. Gelsolin itself is widely expressed in adult mammalian tissues (13, 16), and its expression is specifically down-regulated in many human neoplastic lesions, including bladder, breast, and colon cancer (22). These observations on the role of gelsolin in apoptosis suggest that gelsolin down-regulation in tumors may be one mechanism by which tumors evade apoptotic signaling pathways.
↵* These authors contributed equally to this work.
↵† To whom correspondence should be addressed. E-mail: kwiatkowski{at}calvin.bwh.harvard.edu
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Neutrophils (105) were treated with TNF+CHX or antibody to Fas, rinsed with PBS, and incubated in 50 mM tris-HCl (pH 7.4), 1 mM EDTA, 10 mM EGTA, and 10 μM digitonin at 37°C for 10 min. After centrifugation at 15,000g for 10 min, the lysate was incubated in DEVD-AFC (Enzyme Systems Products) for 30 min. Released AFC was measured by spectrofluorimetry with excitation at a wavelength of 380 nm and emission at 460 nm.
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A cDNA library was prepared from mouse embryo (day 14 postcoitus) RNA. The cDNAs were cloned into the expression vector pCS2+, generating 200,000 independent clones. The library was fractionated into smaller groups of 100 different clones each. Each group of cDNAs was used to express [35S]methionine-labeled proteins, using a coupled transcription-translation system (TNT system, Promega). We incubated 2 ml of the translation mix with 0.5 ml of active or inactivated caspase-3 (0.15 ng/ml) at 30°C for 0.5 hour. Caspase-3 was inactivated by incubation with 50 mM DEVD-fmk for 30 min before addition to the translation products. Recombinant active caspase-3 was prepared by expressing the sequence of human caspase-3 (YAMA/CPP32, a gift of V. Dixit) in E. coli (2) . Purified caspase-3 (generously provided by A. L. Fear) was fully active when assayed on DEVD–7-amino-4-methylcoumarin as previously described (3). Caspase-3 was >80% pure, consisting of 17- and 11-kD subunits, as assayed by SDS-PAGE.
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] were seeded on acid-treated glass cover slips, and 70% density cells were microinjected with plasmid DNA pGG-NT (encoding residues 1 through 352 of gelsolin and tagged with EYMPME epitope at the COOH-terminus) or pGG-CT (encoding residues 353 through 731 of gelsolin and tagged with EYMPME epitope at the COOH-terminus) at 0.1 μg/ml in 100 mM Hepes (pH 7.4). A plasmid encoding green fluorescent protein (GFP) (pEGFPN1, Promega) at 1 ng/ml was coinjected as a marker. Cells were fixed in 4% paraformaldehyde/PBS, permeabilized with 0.1% Triton X-100, and stained for the expression of tagged proteins, using a mouse monoclonal antibody to EYMPME (50 μg/ml) and a secondary goat anti-mouse immunoglobulin G coupled to Texas Red (Amersham). Actin filaments were visualized with tetramethyl rhodamine isothiocyanate (TRITC)–Phalloidin (Sigma).
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We thank G. Pronk for p35 cDNA; V. Dixit for YAMA cDNA; A. L. Fear for active caspase-3; A. Creasey and T. Gesner for help with neutrophil isolation; T. Brown, M. Giedlin and T. Shi for fluorescence-activated cell sorter (FACS) analyses; M. Heinzel and G. Harrowe for microscopy; J. Tucker for DNA sequence analyses; and D. A. Fisher and T. Jacks for p53–/– cells. Supported by Chiron Corporation and NIH grants RO1 HL54188 and P01 HL48743 to D.J.K.
You are going to email the following Caspase-3-Generated Fragment of Gelsolin: Effector of Morphological Change in Apoptosis
By Srinivas Kothakota, Toshifumi Azuma, Christoph Reinhard, Anke Klippel, Jay Tang, Keting Chu, Thomas J. McGarry, Marc W. Kirschner, Kirston Koths, David J. Kwiatkowski, Lewis T. Williams


text:
Young Jospeh: A sermon based on Genesis 37
This summer we will be journeying through the story of Joseph. I've been spending a lot of time getting to know Joseph over the last couple of months. I've been struck by what a profound and mysterious story this is. When reading the story of Joseph you almost can't help but see patterns at play over and over that recur throughout the rest of the Bible, the history of the church, and the life of faith. Throughout these chapters are stories of promise and sacrifice, death and resurrection, transformation and redemption. The stuff that comprises Joseph's story set the pattern for the way that God will continue to work in the lives and histories of his people. So, as we walk through the story of Joseph, I won't be surprised at all if each of us begin to see something of our own story at play in the midst of these pages. It's my hope that as we read this story about how God is at work in the midst of dying dreams, broken families, entrapment, and finding restoration, we might find hope for our own dreams to be restored through the same God who acted in the life of Joseph.
To begin with, let's orient ourselves as to where this story sits in Scripture. This story comes as the climax and conclusion of the first book of the Bible, Genesis. The book of Genesis basically sets the agenda for the rest of the Bible. (Slide 2) This agenda becomes clear when you read Genesis Chapter 12, where God says to Abraham, "Go, I will make you a great nation and I will bless you and make your name great, and you shall be a blessing. And I will bless those who bless you, and those who damn you I will curse, and all the clans of the earth through you shall be blessed." The rest of the book of Genesis, as well as the rest of Scripture, concerns how God will bless the world through this family.
The story of Joseph marks a new chapter, recording how this promise was not only carried forward, but grown and developed. For example, when they interpreted Joseph's dream, his brothers introduced the notion of Joseph ruling over them. This is new. Up to this point, the promise given to Abraham's children did not carry the concept of ruling over others. Joseph's dream prepares the way for the future promise of the Messianic King. So, the story of Joseph marks a new chapter, recording how God's promise to Abraham not only was carried forward, but gives new meaning to that promise.
(Slide 3) What will strike anyone anyone reading the story of Joseph is just how unprepared Joseph is for carrying on this vision. The first we hear about Joseph is how he rules over his brother by being a tattle tale. The second thing we learn about Joseph is that he is without a doubt the favourite child. Jacob dotes on Joseph, the child of his beloved but deceased wife, Rachel. Jacob doesn't see Joseph like the rest of his brothers do, spoiled, a tattletale, and arrogant. Jacob dotes on Joseph and gives him the robe, that splendid beautifully ornamented robe. The robe which once belonged to royalty. This robe which Joseph would shamelessly wear about, broadcasting the horrific truth to the brothers: Joseph was the chosen one. Joseph had taken the place of Reuben and Judah. And it appears Joseph was rubbing their noses in it. What a poor choice Joseph seems to carry on the blessing to Abraham. How hard it was for the brothers to live with this beautiful, intelligent, ornamented little brat. (Slide 4). The brothers saw it was he their father loved more than all his brothers, and they hated him and could not speak a kind word to him.
So, why did God choose Joseph for the dream? It seems almost careless of God to give the gift of that dream to Joseph. Joseph did not appear to be able to handle the extravagant gift of the ornamented robe, it alienated him from his brothers. Through flaunting the robe, he was at risk of their jealousy. If the robe went to his head, how could the dream not do the same? And we see that our fears were grounded. Joseph saw no reason to keep his dream a secret. (Slide 5) Listen, pray, to this dream that I dreamed. And, look, we were binding sheaves in the field, and, look, my sheaf arose and actually stood up, and look, your sheaves drew round and bowed down to my sheaf." "Look, look look" says Joseph. "What could this possibly mean?" "Isn't this so wonderful?" We shake our head wondering whys God would give such a powerful dream to such an immature boy.
God blesses Joseph with another dream, a dream more extravagant than the first. Again Joseph says, "Look, look, look!" (Slide 6). "Look, I dreamed a dream again, and, look, the sun and the moon and eleven stars were bowing down to me". The interpretation of the second dream was no different than the first, but this time even his father wonders along with the brothers, "Shall we really come, I and your mother and your brothers, to bow down before you to the ground?"
Surely it would have been better for God to withhold these dreams until Joseph was older, more mature, more deserving. Maybe then Joseph would have been prepared and equipped to receive such a lavish gift from God.
But who is ever deserving of God's gifts? Who is ever mature enough to receive a vision from God. Who is ever wise enough to discern the Word of God? Who of us is holy enough to receive the Holy Spirit. In spite of our immaturity, each of us received the extravagant gift of the Holy Spirit at the moment we believed. Who of us ever deserved such a gift? Thank God for his foolish ways of giving gifts to his undeserving children.
It can be tempting for older, more seasoned Christians to look in disdain on of those who are new in the faith. Its easy to write off people who are green in their faith. Sometimes new believers, believers newly charged with the Holy Spirit, can come across as sometimes naive, sometimes arrogant, sometimes simplistic. It can be tempting to write off their extravagant dreams. This story cautions us from being to hasty in writing off others in their immaturity. What we're seeing may be a dull vision that will take years for God to sharpen to a reality. But that's God's grace, in our youth, in our imperfection, God blesses us extravagantly. That's the encouragement we find in Joseph. No matter what our personal shortcomings, we are not beyond blessing.
For the brothers, this was too much. That the father would give Joseph the robe was unreasonable, but that God would give these dreams was unbearable. It was bad enough for the robe to be a visual reminder of their father's favouritism, but imagine the torment of the brothers upon hearing the dream and suspecting that God too was blind to this dreamer's shortcomings. Is it no wonder that in the next section we learn that the brothers moved their flocks 80 kilometres away to Shechem. It seems even that distance was not far enough away from the dreamer, so they journeyed a further 21 kilometres to Dothan.
Perhaps it was too soon for Jacob to send the tattler, Joseph, to check up on his brothers. Perhaps, the father was hoping for reconciliation, if so, why didn't he forbid Joseph from wearing the robe of blessing. Certainly he would have known that was a bad idea. We know Joseph didn't have the foresight to avoid that fatal error. He approached his brothers robed in splendour. The son that they were trying so desperately to avoid walked right into their midst. Their rage roared to the surface as they saw the Dreamer walking towards them. "Here comes that dreamer!" They said to each other. "Come now, let's kill him and throw him into one of these cisterns and say that a ferocious animal devoured him. Then we'll see what comes of his dreams." Reuben tried to divert his brothers from their intent to shed blood. Probably not an act of altruism, but a lame attempt at getting back in his dad's good graces after sleeping with his father's concubine. Joseph walks closer. Perhaps when he saw the look in their eyes, he realized that he should say nothing. Perhaps at that moment Joseph realized that all people didn't love him more than they loved themselves. Perhaps this was his moment of awakening from his dream. Perhaps that was the moment he was stripped of his own arrogance and immaturity as his 10 brothers stripped him of that precious ornamented robe.
And its here that we see the grim reality of what we spoke of last week. (Slide 7). Jesus said, "I am the true vine, and my Father is the gardener. He cuts off every branch in me that bears no fruit, while every branch that does bear fruit he prunes so that it will be even more fruitful." Last week Aria told us a story of vineyards in Hungary where they prune the plant beyond recognition for the sake of fruitfulness. In the story of Joseph we hear of a God who is not afraid to prune. God is certainly not ignorant of Joseph's shortcomings, God loves Joseph far too much to leave him be. God blesses us in our ignorance and arrogance, yet he is not content to let us stay there. He gives us a robe, and allows it be torn and dipped in blood. He lets us be cast into the well, only so that we might be born again as we are pulled back into the light. This is the pattern God often has for us. God blesses us extravagantly in our imperfections, and then puts us through the refiners fire on the way to perfection. This is called sanctification, or the process of being made holy.
Each of our stories will of course be different. God can use any circumstance for refining. The circumstances will be unique. For some, like Joseph, it might be family troubles. Joseph's family was not something that God didn't anticipate in his plan for him. Getting thrown into the bottom of the well was not something that God didn't know about. These very things that weren't good things in and of themselves, in fact his brothers meant to harm, but God used them to do his work of making Joseph holy, of pruning him of his arrogance, so that fruit might grown. God can use any circumstance that might look like it was intended for harm in this way. It might be the loss of a job. For another, God's refining might come through a serious illness, or coping with great loss. It could be just a feeling of emptiness. Like the character in Paul Simon's song, "America," who on a road trip suddenly is suddenly overcome with dissatisfaction with life; "Cathy, I'm lost, I said, though I knew she was sleeping. I'm empty and I'm aching and I don't know why."
I wonder how many of us here have endured this pruning, this refining. I think the challenging part of this is whenever we go through these difficult times, our experience is not in a story where we can easily see that God is using these painful circumstances for refining. We don't see the circumstances like Joseph, as a necessary step on the way to holiness. We often can't see our circumstances as step towards growing into God's purpose for our lives. Instead, hardship can just feel meaningless. Yet often hardship can grow and mature us into truly being able to use the gifts he has given us. Without the trials of his life, Joseph never would have been able to live out the dreams God had given him.
(Slide 9). That's one reason why the author of Hebrews reminds us to "endure hardship as discipline; God is treating you as his children… No discipline seems pleasant at the time, but painful. Later on, however, it produces a harvest of righteousness and peace for those who have been trained by it." We must constantly remind ourselves that God can use all things for the good of those who love him.
It must be said that there was nothing good in the brothers' violent actions. They were sinful through and through. Yet, years later, only after Joseph had been through this refining fire, was he able to say to his brothers, "You intended to harm me, but God meant it for good." The action of the brothers were evil, but through those actions God was able to prune Joseph so that he might bear much fruit. View hardship as discipline. For the one who has blessed you extravagantly, the one who has gifted you with dreams, will raise you up out of the well.
This was the pattern walked by Joseph. This was the pattern walked by Jesus. (Slide 10) "Son though as he was, he learned obedience from what he suffered and, once made perfect, he became the source of eternal salvation for all who obey him…. This is the pattern set for us, whether we are robed with blessing or cast naked into the pit. Through Jesus, we are never beyond His2 restoration.


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    if (IvGame::mGame->mEventHandler->IsKeyDown('k'))
    {
        xlate.x -= 3.0f*dt;
    }
    if (IvGame::mGame->mEventHandler->IsKeyDown('i'))
    {
        xlate.x += 3.0f*dt;
    }
    if (IvGame::mGame->mEventHandler->IsKeyDown('l'))
    {
        xlate.y -= 3.0f*dt;
    }
    if (IvGame::mGame->mEventHandler->IsKeyDown('j'))
    {
        xlate.y += 3.0f*dt;
    }
    mTranslate += xlate;

    // clear transform
    if (IvGame::mGame->mEventHandler->IsKeyDown(' '))
    {
        mRotate.Identity();
        mScale = 1.0f;
        mTranslate.Zero();
    }
}   // End of Player::Update()

//-------------------------------------------------------------------------------
// @ Player::Render()
//-------------------------------------------------------------------------------
// Render stuff
//-------------------------------------------------------------------------------
void
Player::Render()
{
    // build 4x4 matrix
    IvMatrix44 transform(mRotate);

    transform(0,0) *= mScale;
    transform(1,0) *= mScale;
    transform(2,0) *= mScale;
    transform(0,1) *= mScale;
    transform(1,1) *= mScale;
    transform(2,1) *= mScale;
    transform(0,2) *= mScale;
    transform(1,2) *= mScale;
    transform(2,2) *= mScale;
    transform(0,3) = mTranslate.x;
    transform(1,3) = mTranslate.y;
    transform(2,3) = mTranslate.z;

    ::IvSetWorldMatrix(transform);

    // draw geometry
    DrawSphere();

}   // End of Player::Render()

//-------------------------------------------------------------------------------
// @ Player::DrawSphere()
//-------------------------------------------------------------------------------
// Draw vertex arrays for a sphere centered around the origin
//-------------------------------------------------------------------------------
void
Player::DrawSphere()
{
    IvRenderer::mRenderer->Draw(kTriangleStripPrim, mSphereVerts, mSphereIndices);
} // End of Player::DrawSphere()

//-------------------------------------------------------------------------------
// @ Player::CreateSphere()
//-------------------------------------------------------------------------------
// Create vertex arrays for a sphere centered around the origin
//-------------------------------------------------------------------------------
void
Player::CreateSphere()
{
    // Creates a grid of points, shaped into a sphere.  This is not an
    // efficient way to create a sphere (the verts are not evenly distributed),
    // but it shows how to set up arrays of vertices and normals
    const unsigned int steps = 32;
    const unsigned int verts = steps * steps;

    mSphereVerts = IvRenderer::mRenderer->GetResourceManager()->CreateVertexBuffer(
        kCPFormat, verts, nullptr, kDefaultUsage);

    // temporary pointers that can be stepped along the arrays
    IvCPVertex* tempVerts = (IvCPVertex*)(mSphereVerts->BeginLoadData());

    // A double loop, walking around and down the sphere
    const float phiIncrement = kPI / (steps - 1);
    const float thetaIncrement = kTwoPI / steps;

    unsigned int j;
    for (j = 0; j < steps; j++)
    {
        float theta = thetaIncrement * j;

        float sinTheta, cosTheta;
        IvSinCos(theta, sinTheta, cosTheta);

        unsigned int i;
        for (i = 0; i < steps; i++)
        {
            float phi = phiIncrement * i - kHalfPI;

            float sinPhi, cosPhi;
            IvSinCos(phi, sinPhi, cosPhi);

            tempVerts->position.x = mRadius * cosTheta * cosPhi;
            tempVerts->position.y = mRadius * sinTheta * cosPhi;
            tempVerts->position.z = mRadius * sinPhi;

            tempVerts->color.mAlpha = 255;
            tempVerts->color.mRed = 255;
            tempVerts->color.mGreen = 255;
            tempVerts->color.mBlue = 255;

            tempVerts++;
        }
    }

    mSphereVerts->EndLoadData();

    // Create index arrays - just a 32x31-quad mesh of triangles
    // Each of the 32 strips has 31 * 2 triangles plus two dummy indices
    // This means that there are 31 * 2 + 2 + 2 (two extra to start the
    // strip, and two extra to end the previous strip) indices in each
    // strip, although we can avoid two indices in the first strip, as
    // there is no previous strip to be ended in that case.  Thus,
    // 64 + 66 * 31 indices for the entire sphere
    const unsigned int sphereIndexCount = steps * 2 + (steps - 1) * (steps * 2 + 2);

    mSphereIndices = IvRenderer::mRenderer->GetResourceManager()->
        CreateIndexBuffer(sphereIndexCount, nullptr, kDefaultUsage);

    UInt32* tempIndices = (UInt32*)(mSphereIndices->BeginLoadData());

    for (j = 0; j < steps; j++)
    {
        UInt32 baseIndex0 = steps * j;
        UInt32 baseIndex1 = steps * ((j + 1) % steps);

        // restart the strip by doubling the last and next indices
        if (j != 0)
        {
            *(tempIndices++) = tempIndices[-1];
            *(tempIndices++) = baseIndex0;
        }

        unsigned int i;
        for (i = 0; i < steps; i++)
        {
            *(tempIndices++) = baseIndex0;
            *(tempIndices++) = baseIndex1;

            baseIndex0++;
            baseIndex1++;
        }
    }

    mSphereIndices->EndLoadData();
}   // End of Player::CreateSphere()

redpajama_set_name: RedPajamaGithub


text:
Q: Any issues with increasing maximum mailbox size for Exchange 2010? The boss of a company for whome I am responsible for their exchange server has a full mailbox. The current mailbox limit is 2Gb (pretty standard).
He's asked if I can bump his mailbox up to something around 10Gb in size. The exchange server is located on the same physical network as he is on 95% of the time.
Is there any "gotchas" I should know about before agreeing to bump his mailbox to 10Gb? What issues may we encounter if he actually fills his account to 10Gb?

A: I'm going to add another consideration to Evan's answer. Users with large mailboxes lead to large databases. The larger the database, the longer the recovery time in the event of a disaster. That's something the boss should be made aware of. How many hours is it going to take to recover a database with 10GB mailboxes?
You could plan for high availability by keeping copies of the database on multiple servers. DAGs in Exchange 2010 totally rock! If the budget isn't available for that, consider moving into the cloud with hosted Exchange (make high availability somebody else's problem to worry about).

A: If he gets to 10GB, he may start to notice performance issues compared to what he has today, but I have had MANY users with 6-8GB mailboxes with no problems. That said, Exchange 2010 has improved the compression algorithms with Outlook 2010, so they are even more efficient than Exchange 2007, and way better than 2003. The only caveat: Don't do it if you are not running gigabit Ethernet.

A: To expand on my comment on Evan's answer, my experience is that mailboxes are never too large because of the messages, it's the attachments that cause them to grow.  Limiting message size helps, but like most places, people here are always sending around powerpoint files and documents with pictures and it doesn't take too many 20MB files to add up to a GB.  The problem is that email provides useful meta-data to many people: I sent this version of the file to these people on this date.
I haven't been terribly successful, but I'm always working on getting people to save their attachments, and then (or simply) delete them.  In a couple of cases, I was able to train someone with a big mailbox to use MessageDetach to extract attached files, but that's about a 1% success rate.

A: As long as you've got the disk space and backup capacity there's not really an issue. Exchange has been able to handle large mailboxes for a long time, and Exchange 2010 does a stellar job.
Outlook would be the only place where I'd have any cause for concern, and then only if the user is using an older version (pre-2007). Newer versions of Outlook aren't going to have problems with such a large mailbox (and older versions really won't, either, unless you're doing caching).
The only performance problems I've seen with Outlook and large mailboxes relate to large numbers of individual items in folders, but that's more of a count-of-items problem and not a size-of-items problem. The user shouldn't keep 20,000+ items in a single folder and expect it to perform well.
You might consider using the personal archives feature in Exchange 2010 if the user needs to keep a lot of old data around, but there's no reason you can't keep it in his mailbox, just as well.

A: According to Microsoft, it is not the size that matters, it's the number of items.


When Did Ocean Education Get Political?
I am incredibly lucky to get to live in Juneau and feel fortunate every day that my life, work, and passions overlap in Alaska. Life here is inextricably tied to the ocean. I get to see first-hand how it is changing, learn from people whose livelihoods, communities, and cultures depend on healthy oceans, and work toward sustainable management choices.


To buy or to sell business is a great deal and needs a right decision. You will find yourself in a big dilemma when you have to decide whether you are to buy a business or to sell your business. You can make a decision by yourself, but as the consequences, you sometimes end up by making a mistake. The wrong decision will make you lost your money. By using an acquisitions and professional merger, you will be able to make a proper decision because they will help you go through all the process smoothly.
The most important thing before you choose any companies that provide you with the service of selling and buying business is to make sure that the company you choose are honest, well credentialed, and very experienced. Several companies only concern in money making, they do not care whether you are going to success with the business or not. These companies only focus on the commission that they will earn after the sale.
The great and honest company will give you an honest advice. They will tell everything that you need, not tell everything you want to hear. If you find the company that can tell you everything that exactly what you need, it is the right time to buy or sell a business by using this company as your acquisitions and professional merger. Before you buy or sell a business, it is important to make business value estimation.
By making a business value estimation you can find the right price for the buildings, physical property, land, inventory on the building, furniture, including the potential income of the business. To measure the business value, you will need an acquisition and merger advisor or a professional business valuation. So it is important for you to get any merger advisor that can help you go through an important business deal.


Yesterday on my birthday, a colleague brought in a blooming periwinkle colored hyacinth as a gift for me. As it was the coldest day on record in years and springtime seems so very far away, it brought hope and a beautiful fragrance to my day. Thank you Rebecca! With the memory of that heavenly fragrance still permeating my being, I have chosen a white jasmine tea for my morning cuppa. It is called Jasmine Downy White Pekoe.
A white tea is scented with jasmine blossoms to create the ethereal beauty of this tea. Early in the springtime, tea buds are gently plucked from the tea bushes. These buds are so new that they are still covered with the white down of the just emerged growth. They are newly born. The tea buds are dried and then heated to halt the oxidation process that would turn them dark.
I steeped the delicate leaf in 180 degree F water for 3 minutes.
It is important to remember that this tea is not flavored, it is scented. Since the tea leaf is so sensitive to picking up the scents and aromas to anything that is placed by it, it is perfect for scenting with flower blossoms. Once the jasmine is blooming in the summertime, the leaves are laid out and the jasmine blossoms are laid over them. This process is repeated until the tea master is satisfied with the level of scenting. There are no dried blossoms mixed with this tea. Its fragrant jasmine aroma and flavor come directly from the scented leaf. The delicate herbaceous quality of the white tea blends perfectly with the jasmine. Truly an aromatherapy experience!
I treated myself to a big bouquet of orange carnations this week. In my healing process, I am trying to bring as much warmth into my life as possible to thaw out this frozen shoulder of mine. The beautiful salmon orange color of the flowers brings color and sunshine to my spirit.
As I focus on healing, I have been, unfortunately, away from my studio and my artwork. I yearn to feel my materials in my hands, weaving them together to express what is present in my soul. I am hoping to return to my studio soon and my Studio Wednesday posts as the pain heals and I regain more mobility. A big thank you to all of my readers for their patience as I continue to step back from my creative side just now.
This entry was posted in Life Musings, Photography, Poetry, Tea, White Tea and tagged jasmine tea, Jasmine White tea, tea review.
We absolutely love this site. It's so personal and warm. We hope you don't mind, but we've added your blog to our blogroll! Thanks!
Hope you're feeling better soon. Looking forward to the addition of your art posts once you're back in the studio again.
Happy belated birthday, Karen! Gosh, I learn more about tea from reading your wonderful posts than I have in a lifetime of drinking the stuff.
Hang in there — I just know you will be soon back to creating beautiful and wondrous things . . .
Thanks, Jamie! I'm looking forward to getting back in my studio soon.
Thank you Kathy! I'm so glad to hear that I'm helping you learn more about tea.


Where are you going? Where have you been? Have you left your ♥ there??
Any ring size - any! Just custom order the size you need at check out. Whole sizes, half sizes, quarter sizes, odd sizes! Let me know if you need help determining your size.
Can be stamped inside as well. See last photo.
Cuff bracelets also available with coordinates.
Please select slanted or upright numbers, and choose to have your numbers showing outside or secretly hidden inside.
Comes in velvet drawstring pouch ready for gift giving.


Interstate 40 Grand Canyon RV Park and Campground invites you to enjoy an unforgettable camping experience when you come to play in one of Northern Arizona's premier vacation destinations: The Grand Canyon!
Ash Fork, AZ is a quaint little town located on the eastern end of the longest unbroken stretch of Historic Route 66. Part of what was once Route 66 still runs directly through Ash Fork, though as a divided highway, with Park Avenue running east and Lewis Avenue running west, both serving as a main thoroughfare.
Route 66 provided a boost to the town's economy in the 1950s, but construction of the divided highway through the town resulted in the destruction of many of the storefronts, sidewalks, and residential streets, forever. This action altered the aesthetic qualities of the downtown area. Some of the historic buildings, and old railroad company houses can still be seen today along these streets.
The majority of the town's commercial establishments, include the Oasis Lounge and the Ranch House Cafe and Family dollar can be found along Park Avenue.
Certain areas of the town were selected to be used as sets in the filming of 1992's Universal Soldier, supposedly due to the extremely low purchase price and poor condition of several buildings, including an old motel, which were blown up for cinematic effect. The surrounding geographical area and settlements also served as inspiration for the 2006 Pixar film Cars.
This historic community features the Ash Fork Route 66 Museum, located at 901 W. Old Route 66, (across the street) which displays not only Route 66 memorabilia, but, more exhibits and information on area history.
Many of Northern Arizona's most visited attractions are located within driving distance of this friendly little town. We ( I-40 GCRV Park) are also a great base camp for hunting and fishing. This includes, but not limited to, Cataract Lake, Dog town and many more.
Spaces fill up fast! Call to reserve your spot today!


text:
Home » Report launch: "Controlling the Message: Challenges for independent reporting in Greece"
Athens, Greece: Police forces block journalists from taking picures. picture alliance / ZUMAPRESS.com | Nikolas Georgiou
Report launch: "Controlling the Message: Challenges for independent reporting in Greece"
The MFRR has conducted a media freedom mission in Greece and the report consisting of findings and recommendations will be launched on 28 March, 2022 with an online event.
On 28 March 2022, the Media Freedom Rapid Response will publish the report of its online fact-finding mission to Greece that took place in December.
Under the title 'Controlling the Message: Challenges for independent reporting in Greece', the report reflects the mission's findings and recommendations on:
The assassination of Giorgos Karaivaz;
Polarisation of a fragmented media landscape;
Reporting on migration;
Reporting on protests; and,
Legal threats.
The report will be launched with an online panel on 28 March at 2pm CEST (=3pm EEST) with:
Laurens Hueting, Senior Advocacy Officer of the European Centre for Press and Media Freedom (ECPMF)
Iliana Papangeli, Managing Director of Solomon
Renate Schroeder, Director of the European Federation of Journalists (EFJ)
Nikos Smyrnaios, Associate Professor at the University of Toulouse
Anne ter Rele, Advocacy Officer at the International Press Institute
Please register for the event.
The report will be made available on mfrr.eu and the websites of the MFRR partner organisations at the time of the launch event.
For interview requests and media inquiries, please contact laurens.hueting@ecpmf.eu.
The fact-finding mission to Greece was coordinated by the Media Freedom Rapid Response (MFRR), a Europe-wide mechanism which tracks, monitors and responds to violations of press and media freedom in EU Member States and Candidate Countries.
IPI report shines light on hidden alliances and vested interests behind media capture in Bulgaria
Controlling the Message: Challenges for independent reporting in Greece
Greece Reports


All reviews are the subjective opinions of third parties and not of the website or its owners. Reviews are placed by members of the public that have held a function or event at Camborne Conservative Club.
At needadisco.com we only accept reviews from confirmed clients that have booked a DJ to play at Camborne Conservative Club via our website, with the arrangements independently verified by both the DJ and the client before the event takes place - no "fake reviews" here!
Helpful information for DJs and entertainers visiting Camborne Conservative Club provided by trusted members of needadisco.com.
ACCESS: The best access is at the back, via a small ramp, which you can reverse a large van up. At the top of the ramp is a door leading directly into the venue. The setup area is a just 3 metres away.
STAFF: Staff quite friendly, although they appear to spend all night in another room where the bar area is.
ADDITIONAL COMMENTS: "Quite a large room, with the bar located outside of this room, Ideally suited for larger parties birthdays etc."
Information is based on enquiries and bookings handled via needadisco.com so whilst it's a great insight, if you have any questions or concerns about holding a function or event at Camborne Conservative Club please contact the venue.
Camborne Conservative Club has previously been hired as a venue for private parties such as birthday celebrations, anniversaries, engagements etc.
Camborne Conservative Club is in our Sports, Social & Members' Clubs category. The map below shows other local venues in the same category.
Get FREE no-obligation disco quotes from SoundONE Disco Sound and Lighting and other DJs covering your venue. Take 2 minutes to fill in some basic details and we'll do the rest!


The Alternative for Germany (AfD) has been sitting in Germany's federal parliament since September 2017, having won 12.6 percent of the popular vote. In considering this young party's recent development, researchers have focussed on its rhetorical strategies (i.e., populism) and its radicalization. Until now, much less attention has been paid to antisemitism within the AfD— also because the party would prefer to keep this out of public debate. By investigating its treatment of antisemitism, Nazism, and the politics of remembrance, it can be shown that the AfD has the features of a far-right party, to a much clearer extent than might be guessed from its media image, particularly inside Germany.
Herr or Hüter of the Constitution?


You are kindly invited to visit us at the International Boat Show INTERNAUTICA 2018.
Internautica will take place at Marina Portorož, Slovenia, on 17.05.2018 – 20.05.2018. It is one of the most influential boat shows in the region, presenting novelties from the nautical industry.
Read more … You are kindly invited to visit us at the International Boat Show INTERNAUTICA 2018.
Cranchi E52F received the Adriatic boat of the year 2017 award.
Successful premiere of a new exciting Cranchi Zaffiro 35 and a nomination for Boat of the Year 2016 award.
© copyright 2019, www.bby.si - All rights reserved!


text:
Dolly Parton Net Worth
Dolly Parton is, without any doubt, one of the most influential personalities in country music history. Her tremendous contribution served as a source of inspiration for many singers who chose to make a name in this genre.
Dolly Parton Net Worth – $500 MILLION
Birth Name – Dolly Rebecca Parton
Date of birth – January 19, 1946
Zodiac Sign – Capricorn
Birth Place – Sevier County, Tennessee, United States
Profession – Record producer, Screenwriter, Multi-instrumentalist, Television producer, Singer-songwriter, Film producer, Voice Actor, Author, Philanthropist, Composer, Businessperson
Her career as a singer, songwriter and actress spans over more than 5 decades, and brought her public recognition through the impressive number of awards she won over time. The fortune she accumulated from selling her albums, acting in various cinematographic and TV productions and from her numerous tours places Dolly Parton among the richest country singers.
A Brief History of a Brilliant Career: Even when she was only a little girl, the future legend of country music showed that she was born for stardom. From the young age of ten, she already became famous in her native area, Sevier County, from the Eastern Tennessee. She participated in different radio shows, singing gospel songs at first, but gradually shifting to the genre that was going to bring her wide recognition, country music.
Although she released her own singles and album from the very young age of 13, the road to fame began in 1967. Then, she appeared in a TV show, singing Tom Paxton's The Last Thing of My Mind, in duet with Porter Wagoner.
Jolene, The Bargain Store, Please Don't Stop Loving Me or Love Is Like a Butterfly are just a few of her great hits. All these songs, and many more, stationed for numerous weeks on the charts' number one position, taking Dolly Parton net worth to astronomic figures.
Successful Actress and Songwriter: The success she reported in her musical career encouraged the pop-country singer to exploit other sides of her artistic personality. Her film debut was in the comedy movie From 9 to 5 (1980), where she portrayed a secretary.
The remarkable performance brought her a Golden Globe Award in the New Star of the Year category. She was also nominated for the Golden Globe Awards in the Best Actress – Motion Picture Musical or Comedy category.
Steel Magnolias, a drama based on the Robert Harling's eponymous play, Rhinestone, with Sylvester Stallone as co-star, and the musical The Best Little Whorehouse in Texas are also on the list of commercial and critical successful movies Dolly Parton played in.
As a songwriter, she is known for composing the soundtracks for two of the films she starred in, From 9 to 5 and The Best Little Whorehouse in Texas. The albums featuring the songs became rapidly famous, selling tens of millions of copies worldwide.
She also composed I Will Always Love You, one of the melodies with the most cover versions in the history of music. Among the great artists who contributed to the song's success, there were Kenny Rogers and Whitney Houston, whose version was included on The Bodyguard movie's soundtrack.
Categories Producers, Richest Businessmen


text:
Here's the 2019 Lakefront Festival Of Art lineup
Remembering the time (June) when MPD extolled the life-preserving virtues of tear gas and rubber bullets
Here's a list of upcoming Milwaukee ice bars. Learn it well, for it's the chilling sound of your doom
Stacy Pawlowski to be "even livelier" in second one-woman show
The making of 'Markie In Milwaukee'
Carmella D'Acquisto
Gaze upon the four new murals being painted underneath I-794 along North Broadway
Great Job, Wisconsin! Green Bay is the best city for football fans
On The Record Episode 45: Milwaukee makers, buying local
Per development law, old Hamburger Mary's in Bay View will likely become mixed-use apartment building
Back in 2016, the owners of the Bay View Hamburger Mary's, 2130 S. Kinnickinnic Ave., packed up and moved shop to the former La Perla building, 734 S. 5th St. Since then, the Walker's Point Hamburger Mary's has become a bright, bold, and welcome addition to the rapidly changing neighborhood. But what of the old Hamburger Mary's? Well, per Milwaukee development law, it appears the location will likely become—wait for it—a mixed-use apartment building.
The news comes courtesy of Bay View Alderman Tony Zielinski:
"Plans seem imminent for a mixed use development proposal at the old Hamburger Mary's location," Zielinski says in a Facebook post. "The first floor would be retail and the upper floors would be housing. We should be ready for a community meeting in about a month or so."
The owners of the old Hamburger Mary's building, Phoenix Burger LLC (an affiliate of Milwaukee-based Phoenix Investors), bought the La Perla building in 2016 for $1 million. Milwaukee Record has reached out to the company for comment.
Bay View is currently in the throes of ye olde Milwaukee development, with a five-story apartment building taking the place of the old Faust music store, 288 apartments popping up a few blocks west, and more. Meanwhile, over on the East Side, the newly shuttered BBC awaits its marching orders.
UPDATE: BizTimes reports that the $21 million project is being proposed by Milwaukee-based New Land Enterprises, and will include "144 apartments and up to 15,000 square feet of retail space, parking and other amenity space, according to a zoning change application submitted to the city."
Hamburger Mary'sNews


Happy Thursday! I am excited that the weekend is almost here! I have the next week off work and I am excited to celebrate the 4th of July. My poor dogs are not as excited.
I picked up this striped top a few weeks ago when I was in Chicago. St. Louis does not have a Zara…so of course I had to go there!
Spotlight: Our spotlight of the week is Audrey of Five Foot Nothing Shopping! We love this color dress that Audrey is wearing! The colors are perfect for summer. Every detail is perfect! Be sure to check out her blog!
Such a cute tee, I love Zara too.
Love the bell sleeves on this striped top, Zara is one of my faves!
You're ready for the 4th cutie pie! Such a sweet striped top!
The cutest outfits! I love the stripes with your white jeans.
The cutest outfit! I love the stripes with your white jeans.
This is such a cute outfit! I love your bag!
Love this whole outfit!! Perfect for the 4th! Happy week off!
These stripes + bell sleeves are so good together! Great summer look, lady!
This look is perfection! The striped top is so cute and I love that bag.
This is exactly how I like dressing for the holiday, Jacqueline!!
This top is SO cute! I actually just got a couple of tops with sleeves like this and I love them. Have a great 4th!
Such a cute top!!! Perfect for the 4th!!
I love the sleeves on your stripe top! Such a cute 4th of July look!
That round bag is adorable! Perfect for a 4th of July look!
Love everything about this!! Your top is so pretty, love the round bag and you look amazing with red lipstick!
Thank you so much for the feature! I am so excited! I also absolutely love your outfit, that bag especially is adorable!
I love your cute sleeve detail on the top and the round straw bag is so summer and perfect!
Love your navy style t-shirt. With white jeans is so perfect!
Love everything about this look! The striped t-shirt is super cute and that bag is perfect!
Simply Beautiful! Love the combo!
I am loving this super chic but simple outfit. That top is so adorable and the ruffled sleeves are a bonus!


Découvrez 13 vitamines indispensables pour tous, en particulier pour les sportifs, dont les besoins sont importants.
Commentez l'article "Rôles, sources et apports journaliers des vitamines pour le sportif"
1 commentaire pour "Rôles, sources et apports journaliers des vitamines pour le sportif"
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"Linear regression models for prediction of annual heating and cooling " by Navid Aghdaei, Georgios Kokogiannakis et al.
Aghdaei, N., Kokogiannakis, G., Daly, D. J. & McCarthy, T. (2017). Linear regression models for prediction of annual heating and cooling demand in representative Australian residential dwellings. Energy Procedia, 121 79-86.
This paper presents the development methodology of linear regression models that were developed for the prediction of annual thermal loads in representative residential buildings across three major climates in New South Wales, Australia, and the assessment of the impact of building envelope upgrades. A differential sensitivity analysis was undertaken for sixteen building envelope parameters, with six parameters being identified as significant. These six parameters were then explored using EnergyPlus simulation, and a number of linear regression models developed from the simulation outputs. Random values for design parameters were generated, and the results of EnergyPlus simulations using these parameters were used to verify the outputs of the regression models. The differences between regression-predicted and EnergyPlus-simulated annual thermal energy requirements were of order 10%-15%. The coefficient of determination (R2) was over 0.90, indicating a good agreement between simulation and the regression models, and suggesting that the annual heating and cooling energy requirements can be forecasted with an acceptable accuracy using the regression models. It is envisaged that the regression models developed can be used as a quick alternative to building simulation for residential buildings of the area and the climate covered by our study, and can serve to rapidly estimate the likely energy savings/penalty during the retrofitting design stage when different building schemes and design concepts are being considered.