text:
The lowest scoring teams in Premier League history – ranked
Frank Lampard could be sacked if Chelsea lose to Leicester
Barcelona set to appeal Lionel Messi red card
Vogts resigns from Azerbaijan post
Berti Vogts has resigned from his role as the head coach of the Azerbaijan national team after six years at the helm.
The German took over in 2008, but has failed to guide the side to qualification for any of the three major tournaments since then, falling well short on each occasion.
Azerbaijan's Euro 2016 qualifying campaign has got off to a dismal start, with 2-1 defeats to Bulgaria and Italy followed up by a 6-0 hammering at the hands of Croatia last time out.
It was announced on Friday that Vogts would not longer continue in his position, with the Association of Football Federations of Azerbaijan (AFFA) releasing a statement.
"AFFA President Rovnag Abdullayev met Azerbaijan national team coach Berti Vogts yesterday in Zurich, Switzerland," it read.
"During the meeting they discussed the matches played in the 2016 UEFA Euro qualifying round. Berti Vogts noted with regret that he couldn't see his future in the Azerbaijan national team. He said that the Azeri A-team did not show persistence during the last game.
"Furthermore, the problem of physical fitness, which has been repeatedly emphasised for several years, has not been resolved. Vogts said that he has already worked six years in Azerbaijan, and during some games he brought much joy to the fans.
"Berti Vogts thanked AFFA for their work and noted that he considers Azerbaijan his second home. AFFA President Rovnag Abdullayev said that Berti Vogts was requested to continue his work and his activities in the Azerbaijan national team is highly valued by AFFA.
"AFFA President noted that the work done by Berti Vogts, in general had a positive influence on the development of football in Azerbaijan.
"Despite the fact that this is not a great achievement, but it was under Berti Vogts that the Azerbaijani team was able to earn the most points in the qualifying rounds – nine points [during World Cup 2014 qualifying], and the Azerbaijan national team was able to leapfrog to the 73rd place in the FIFA Rankings.
"Mr. Abdullayev said that Berti Vogts will be remembered with appreciation for the work he done and he will always remain as a friend of Azerbaijani football."
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text:
POZ Index
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MAIN MENU POZ July 1998
Next Indelicate Balance
July 1, 1998 • By Dominic Hamilton-Little
Cumulative AIDS cases in the United States: 612,078
Cumulative AIDS cases in sub-Saharan Africa: 14,000,000
Documented cases of female-to-female transmission of HIV: 5
Number of Spice Girls: 5
Retail cost of one year's worth of syringes for average IV drug injection: $95
Wholesale cost of one year's worth of Combivir (AZT/3TC) before retail mark-up: $5,320
Cost of a new toilet on the space shuttle: $23,000,000
Cost of James Cameron's film Titanic: $200,000,000
Number of Starbucks coffee shops in San Francisco: 30
Number of needle exchanges in San Francisco: 10
Number of syringes dispensed each week by the San Francisco AIDS Foundation: 40,000
Total AIDS deaths in the United States through June 1997: 379,258
Total casualties due to the atom bombs dropped on Hiroshima and Nagasaki: 170,000
Total U.S. casualties in the Persian Gulf War: 265
Total U.S. casualties due to Legionnaire's Disease: 29
Total number of films made by Audrey Hepburn: 25
Number of weeks And the Band Played On was on The New York Times bestseller list: 5
Number of weeks The Bridges of Madison County was on The New York Times bestseller list: 164
Title of the George Orwell novel in which the totalitarian government outlawed orgasms: 1984
The year HIV was identified and condoms promoted to prevent transmission of HIV in semen: 1984
Percentage of the U.S. population that is African American: 12
Percentage of new U.S. AIDS cases that are African American: 42
Chance that an American will die in an automobile accident: 1 in 125
Chance that an American will test positive for HIV: 1 in 333
Chance of winning the jackpot in the average state lottery: 1 in 4,000,000
Amount raised by Carol Burnett in 1998's Jeopardy Celebrity Invitational for the Hereditary Disease Foundation: $15,000
Amount raised by Gay Men's Health Crisis' 1997 AIDS Walk: $4,800,000
Number of walkers at 1997's AIDS Walk: 36,000
Number of pills popped in the movie Valley of the Dolls (excluding two overdoses): 12
Number of pills taken by this writer each day: 38.5
Total U.S. box-office gross of Philadelphia: $77,324,000
Total U.S. box-office gross of Dumb and Dumber: $127,175,000
Number of ACT UP chapters in the United States as of March 1997: 12
Number of panels on the AIDS Memorial Quilt as of September 1997: 43,077
Number of panels dedicated to Mike, Mikey or Michael as of October 1996: 291
Number of panels dedicated to Deb, Debbie, Deborah or Debra as of October 1996: 37
Total National Institutes of Health budget allocated to AIDS in 1996: $1,400,000,000
Total amount that pet owners spent on health care for dogs, cats and birds in 1996: $11,100,000,000
Number of women incarcerated in New York state prisons as of January 1998: 3,700
Percentage of women in New York state prisons who are HIV positive as of January 1998: 18
Number of CD4 cells that POZ founder Sean Strub had in spring 1995: 1
Number of CD4s Sean had in spring 1998: 357

redpajama_set_name: RedPajamaCommonCrawl


text:
Q: Firefox Touchscreen in Ubuntu 20.04 with Wayland I am using Ubuntu 20.04 with Wayland (for fractional scaling) on a laptop with touchscreen. I would like to use firefox with touch-scrolling. In Ubuntu 19.10, I needed to modify the environment variable GDK_BACKEND=wayland.  In 20.04, this setting does not work and firefox does not start. I just get the terminal output:
(firefox:10489): Gtk-WARNING **: 16:44:22.416: cannot open display: :0

I also tried MOZ_USE_XINPUT2=1 and MOZ_ENABLE_WAYLAND=1 but firefox keeps using x11 as window protocol (shown in about:support). How can I make firefox use wayland?

A: I am unable to replicate this on Ubuntu 20.04, Gnome Session (Wayland), Firefox 77.0.1. I am however able to replicate it perfectly - the non-start, the terminal output - by logging in to the default Ubuntu session, which is still X-based.
The obvious question then is how sure you are that you really are on Wayland? Run the command echo $XDG_SESSION_TYPE from the command line. The response is wayland on the GNOME session (don't know if it works the Ubuntu on Wayland session).
You can install the "Gnome" session as the package gnome-session and select it from the settings pop menu in the lower right corner (assuming GDM), once you start typing in your password on the login screen. Or just try "Ubuntu on wayland" from the same menu.

redpajama_set_name: RedPajamaStackExchange


Beppe Severgnini has 1 media appearance.
Beppe Severgnini discusses Ciao, America!.
Beppe Severgnini is currently considered a "single author." If one or more works are by a distinct, homonymous authors, go ahead and split the author.
Beppe Severgnini is composed of 6 names. You can examine and separate out names.


More than 200 players flooded Tucson R48 V Stop #2 Memorial/Junior WPH Benefit at the Tucson Racquet Club on Friday, with the R48 qualifiers kicking off the action on Friday morning. The largest qualifier draw in Race 4 Eight history featured top big ball players from California "crossing over" into small ball play, top junior and collegiate players from Minnesota and Canada, as well as R48 tour veterans seeking the coveted eight spots in the Tucson R48 V Stop #2 Memorial/Junior WPH Benefit R48 main draw.
Top qualifier seeds Jon Iglesias, Vic Perez, Shorty Ruiz, Adam Bernhard, and Abe Montijo advanced to the main draw with straightforward victories. Killian Carroll qualified officially for the first time, while the LAAC's Carlos Chavez qualified for the first time with a victory against Sal Duenas. WR48 #1 Catriona Casey came within three points of becoming the first woman to earn a R48 ranking, losing in the first R48 17th place final to Ryan Bowler, 15-12. Both Bowler and Casey won three matches back-to-back to advance to the 17th place final.
The R48 #2 and Tucson Memorial #1 seed cruised in two games against Shorty Ruiz, Sean Lenning dominated Anthony Selestow, Emmett Peixoto overcame a slow start to defeat Carlos Chavez in straight sets and Tucson Memorial #4 seed and R48 #6 Mando Ortiz defeated his sparring partner Vic Perez in two games. Fifth seeded Daniel Cordova defeated Texas' Adam Bernhard in two, Luis Cordova defeated Tucson transplant Jonathan Iglesias in two, and Killian Carroll dominated SR48 #1 and Tucson Memorial #8 Marcos Chavez. Local favorite Abraham Montijo was defeated by Ireland's Stephen Cooney in two games in the final main draw match on Friday evening.
The R48 main draw and playoffs will highlight Saturday's action, while Big Ball singles and doubles, amateur singles and doubles, and more than 50 junior players competing in small ball and big ball and Junior WPH clinics will fill the courts at the Tucson Racquet Club on Saturday.
The World Players of Handball is a non-profit foundation aiming to grow the game of handball through innovation and inspiring the next generation of players through junior clinics with certified WPH coaches, junior tournaments and the Race 4 Eight tour. Donations are tax-deductible donation and support the growth of the game. 100% of the donations received by the WPH go towards junior handball development. Donate on-line at here or send your generous gifts through PayPal (extremehandball@cs.com) or by mailing checks to: WPH, 3561 E. Sunrise Dr. Suite 125, Tucson, AZ, 85718. You can also contact WPH Development Director David Fink at fink@race4eight.com.


8.8 axle with 3:55 gears, new rebuilt with all new bearings and clutches 4 lug.
93 cobra rear disc brakes hawk pads, frpp prop valve and 1" master.
16 pony wheels powder coated white with new bfg comp 2 rubber.
front bumper and needs to have black mounding sanded and painted.
The wiper pump has no power and needs a wire repair.
I've added a few additional pics of the car, I will note in the door jambs are spotless. The corners have seam sealer they are not rusted.
The car is still available.
Very nice car, I wish I had the room to take it off your hands.


text:
package com.amazonaws.services.dynamodbv2.model.transform;

import javax.annotation.Generated;

import com.amazonaws.SdkClientException;
import com.amazonaws.services.dynamodbv2.model.*;

import com.amazonaws.protocol.*;
import com.amazonaws.annotation.SdkInternalApi;

/**
 * AttributeDefinitionMarshaller
 */
@Generated("com.amazonaws:aws-java-sdk-code-generator")
@SdkInternalApi
public class AttributeDefinitionMarshaller {

    private static final MarshallingInfo<String> ATTRIBUTENAME_BINDING = MarshallingInfo.builder(MarshallingType.STRING)
            .marshallLocation(MarshallLocation.PAYLOAD).marshallLocationName("AttributeName").build();
    private static final MarshallingInfo<String> ATTRIBUTETYPE_BINDING = MarshallingInfo.builder(MarshallingType.STRING)
            .marshallLocation(MarshallLocation.PAYLOAD).marshallLocationName("AttributeType").build();

    private static final AttributeDefinitionMarshaller instance = new AttributeDefinitionMarshaller();

    public static AttributeDefinitionMarshaller getInstance() {
        return instance;
    }

    /**
     * Marshall the given parameter object.
     */
    public void marshall(AttributeDefinition attributeDefinition, ProtocolMarshaller protocolMarshaller) {

        if (attributeDefinition == null) {
            throw new SdkClientException("Invalid argument passed to marshall(...)");
        }

        try {
            protocolMarshaller.marshall(attributeDefinition.getAttributeName(), ATTRIBUTENAME_BINDING);
            protocolMarshaller.marshall(attributeDefinition.getAttributeType(), ATTRIBUTETYPE_BINDING);
        } catch (Exception e) {
            throw new SdkClientException("Unable to marshall request to JSON: " + e.getMessage(), e);
        }
    }

}

redpajama_set_name: RedPajamaGithub


The East Hampton Library's Long Island Collection doesn't have any village maps showing individual homeowners or property lines before 1858 or even 1873 for most of the East End, which can be tricky when we are asked to identify where someone used to live.
There are, however, many deeds and wills available in the collection, and they frequently describe both the property being transferred and the name of the adjoining property owners.
The deed shown here, from Jan. 23, 1836, transfers land in East Hampton from two Southampton residents, Peleg and Ruth Rogers, to Aaron Osborn Jr., an East Hampton resident. While we don't know exactly which lot this was, the deed provides a general description of the property's location, along with a list of property owners.
Within 40 years of the time this deed was written, published atlas pages would begin to name property owners and mark where houses stood for the residents of East Hampton's Northwest Woods. The deed itself hints at the advances in printing technology and the gradual transitions to modern records. Large portions of the text of the deed are pre-printed, such as the itemized list of property rights a seller relinquishes. The pre-printed text is paired with blank lines for unique details, such as the property description and the owners' names. Even the date was printed, leaving space for the day, month, and the last digit of the year.


text:
português (Portugese)
JoVE Journal Immunology and Infection
Heligmosomoides polygyrus is a murine nematode with powerful immunomodulatory capabilities that closely resemble those of highly-prevalent human helminth infection. Here we describe a protocol for the long-term maintenance of the H. polygyrus lifecycle.
Heligmosomoides polygyrus (formerly known as Nematospiroides dubius, and also referred to by some as H. bakeri) is a gastrointestinal helminth that employs multiple immunomodulatory mechanisms to establish chronic infection in mice and closely resembles prevalent human helminth infections. H. polygyrus has been studied extensively in the field of helminth-derived immune regulation and has been found to potently suppress experimental models of allergy and autoimmunity (both with active infection and isolated secreted products). The protocol described in this paper outlines management of the H. polygyrus life cycle for consistent production of L3 larvae, recovery of adult parasites, and collection of their excretory-secretory products (HES).
Heligmosomoides polygyrus is a natural murine gastrointestinal helminth that is closely related to highly-prevalent human nematode parasites 1. In contrast to other nematode models such as Nippostrongylus brasiliensis, H. polygyrus consistently establishes chronic infection in mice as a direct result of multiple powerful immunomodulatory mechanisms it employs to suppress the host immune response 2.
H. polygyrus has a direct life cycle: infective L3 larvae are ingested by feco-oral transmission (or administered by oral gavage in the laboratory setting), whereupon they migrate to the subserosal layer of the duodenum and encyst before returning to the intestinal lumen as adult worms approximately eight days after initial infection. Mating and egg production occurs by day 10 and it is possible to harvest adult worms for culture and collection of excretory-secretory products from day 14 onwards 3. H. polygyrus also interacts with the commensal microflora, with increased Lactobacilli present in infected susceptible mice 4-6, and increased levels of H. polygyrus infection following exposure of mice to Lactobacilli 6.
Active infection with H. polygyrus has been shown to protect against immunopathology in many animal models of autoimmunity 7-10, colitis 11,12 and allergy 13-16. There has consequently been great interest in the potential of Excretory-Secretory molecules from this parasite ("HES") to down-modulate pathology in vivo 17,18. Indeed, protective effects are seen following treatment of mice with HES products 19 through pathways which are now being identified 18,20. Here we describe a protocol for the reliable production of Heligmosomoides polygyrus and recovery of its secreted products that can be further utilized for a range of functional biochemical and immunological investigations.
NOTE: All procedures in this protocol are performed in accordance with guidelines set out by the United Kingdom Home Office and the University of Edinburgh Veterinary Services.
1. Infection of Mice by Gavage
Store Heligmosomoides polygyrus L3 larvae in distilled water for up to six months at 4 °C.
Before use, wash L3 larvae three times in distilled water: centrifuge at 300 x g for 10 min (with brake), remove all but 500 μl of water (to avoid disturbing pelleted L3 larvae) and resuspend the pellet each time.
For the third wash, add water to an exact volume (typically 40 ml) and aspirate 20 µl with a 200 µl tip cut to widen its aperture. Place two 20 µl samples on the surface of a 60 mm culture dish and count the L3 larvae (usually mobile, and best viewed under 50X magnification with a dissecting microscope). Resuspend the final pellet in distilled water to a concentration of 2,000 L3 larvae per ml.
For life cycle production, infect 8 week-old F1 (C57BL/6xCBA) mice with 400 H. polygyrus L3 larvae in 200 μl of distilled water by oral gavage (restrain mice in the upright position by the scruff of the neck and gently pass the blunt gavage needle through the mouth and esophagus into the stomach). Agitate thoroughly prior to each infection (larvae settle quickly in water) and aspirate 200 µl in a 1 ml syringe; use a dedicated gavage needle with a rounded end. For experimental infection of younger mice (6-8 weeks old), or other inbred strains (e.g. C57BL/6 or BALBc), infect mice with 200 L3 larvae.
2. Propagation and Maintenance of H. polygyrus
Place charcoal in the center of a large plastic tub and allow cold tap water to run over it for a minimum of 30 min (unwashed activated charcoal is toxic to L3 larvae). Drain the water from the tub and place the charcoal on two layers of absorbent paper, leaving it exposed to room air until completely dry.
If eggs are required, scrape feces first out of the colon with forceps (and scissors if necessary). If a large number of L3 larvae is needed, place mice on a wire grid and collect fecal pellets over several days.
Mix the feces with granulated charcoal at a ratio of at least a 1:1, to achieve a consistency just damp enough to adhere to filter paper. Smear a thin layer on the center of some dampened filter paper in a petri dish and place this in a humid box (add some damp paper towel and a dish of water) in the dark for 12-14 days.
Remove L3 larvae from day 7 onwards, and collect them on at least two occasions before the paper is discarded. The larvae form a ring around the edge of the filter paper; lift the filter paper out of the petri dish and rinse the larvae that are left of the plate (using a pipette and 5 ml of sterile water per plate) into a 50 ml tube.
Lift off the filter paper and harvest the remaining larvae left on the plate with distilled water into a 50 ml tube. Centrifuge the effluent solution at 300 x g for 10 min. Wash the larvae three times with distilled water and then store at 4 °C in up to 50 ml of distilled water until required.
NOTE: Larvae remain viable and infective for at least 6 months.
3. Collection of Adult H. polygyrus Worms
Prepare the modified Baermann Apparatus in advance as shown in Figure 1.
Cull mice fourteen days after infection.
Wash the abdomen with 70% ethanol. Cut the skin over the abdomen and pull back to reveal the anterior abdominal wall. Make midline incision to enter the peritoneal cavity.
Remove the entire small and large intestine (from proximal duodenum to distal rectum). Place into a dry Petri dish.
Straighten the gut along its entire length; excise the feces-containing colon; place this into a separate dish for egg preparations later.
Excise the proximal 20 cm of small intestine that contains the adult worms –identified by the relatively thick wall of the duodenum and often a red appearance due to the intra-luminal worms. Place into a (100 mm diameter) Petri dish with 5 ml of Hanks' Solution, warmed to 37 °C (two specimens per dish).
Open the worm-filled proximal gut portion longitudinally with scissors (round-ended scissors are best for this), and scrape down inside of gut lining with two glass slides to remove the worms. Then discard the clean gut wall.
Tip worms into small muslin bags, staple closed and secure with paperclips around the edge of the glass funnel (Figure 1).
Fill funnel with Hanks' Solution and add approximately 4 Petri dishes of worms into each funnel.
Place apparatus in 37 °C incubator for 1-2 hr, gently agitating half way through to dislodge debris from the gut preparation that may occlude the muslin filter. Take care to avoid spillage of debris outside the muslin bag – this will cause contamination of the final HES preparation. Adult worms should have slowly migrated through the muslin cloth and settled at the bottom of the glass test tube. Carefully detach the test tube from the connecting rubber hose over the sink (taking care to avoid losing worms at this point).
Using a plastic pipette, transfer the worms into a 50 ml tube and wash six times with Hanks' Solution (allow worms to settle with gravity, remove media with a stripette, add 40 ml of Hanks' Solution and repeat five times).
NOTE: worm culture must be kept sterile from this point onwards.
Move to a laminar flow hood and wash another six times in sterile Hanks' Solution supplemented with 100 U/ml penicillin and 100 µg/ml streptomycin, ready for in vitro culture.
Count the adult worms recovered by taking two samples of 20 µl taken up with a yellow tip cut to widen its aperture; expect approximately 50% of the quantity of inoculated larvae.
4. Setting Up Cultures for HES: Medium Preparation, Washing Adult Worms
Soak the worms from 3.11 in approximately 10 ml of RPMI supplemented with 10% Gentamycin for 20 min, leaving the tube resting at an angle to ensure worms are fully covered.
Perform this in a laminar flow hood: wash again six times with Hanks' Solution (supplemented with 5 U/ml penicillin and 5 μg/ml streptomycin).
Prepare H. polygrus media.
Maintaining sterility in a laminar flow hood, to 500 ml of RPMI1640, add 11.1 ml of 45% glucose (final concentration 1.2% as RPMI1640 contains 0.2% glucose), 5 ml of 100x Peniclllin-Streptomycin (final concentration 5 U/ml penicillin, 5 μg/ml streptomycin), 5 ml of L-glutamine (final concentration 2 mM), and 5 ml of Gentamycin (final concentration 1%). Do not add FCS.
Aliquot worms into vented T25 flasks, approx. 1,000 worms in 15 ml H. polygyrus media per flask, and place upright in 37 °C incubator (5% CO2) for 3 weeks.
5. Preparation of HES
Collect HES-containing culture media from cultures at intervals of no longer than twice per week, setting aside the first collection after 24 hr of culture (due to high levels of LPS contamination and host proteins – can be processed separately or discarded). Keep each collection separate and clearly labeled with the date and batch number. Replace with an equal volume of H. polygyrus media on each occasion.
Centrifuge HES-containing media at 400 x g for 5 min. Then filter sterilize through 0.2 µm low-protein-binding filters into 50 ml tubes. Store in the -20 °C freezer clearly labeled with date of worm harvest and date of HES collection.
After 21 days of HES collection from culture, discard worms.
Pool 500-1,000 ml of HES supernatant (usually from frozen stock, and not including the first 24 hr collection) and concentrate over a 3,000 MWCO filter in the ultrafiltration device under nitrogen pressure.
NOTE: Be very careful not to let the filter run dry.
To set up the filter device, first wash the 3 kDa membrane shiny side down in a 1 liter beaker with distilled water for 3 x 20 min whilst stirring. Assemble the ultrafiltration device as per manufacturer's instructions with filter membrane shiny side up. Place in cabinet at 4 °C and pass 50 ml of distilled water through before starting to concentrate pooled HES.
Add each tube of HES into the filtration device as required (100-140 ml per day), until the volume is concentrated down to 2-5 ml.
In order to remove contaminants from the HES-containing culture media, add 50 ml of pyrogen-free PBS to the filtration device and and then concentrate down to approximately 2 ml. Repeat this step twice (150 ml of PBS in total). Transfer HES into a 15 ml tube, filter sterilize (with a 0.2 μm filter) in a laminar flow hood and measure protein concentration using a spectrophotometer (E280 = 10) or by Bradford assay.
Aliquot, label with batch number and date, and freeze at -80 °C.
Perform a chromogenic LAL assay according to the manufacturer's protocols on each batch of HES prior to use. If LPS levels are greater than 1 U LPS per 1 µg protein, consider not using this batch for in vivo experiments or in vitro cultures.
Process the HES collected at 24 hr separately in the same manner; it will contain LPS and some host proteins and, while not suitable for functional experiments, it is a useful source of individual molecules that may be isolated by monoclonal antibody affinity purification.
Representative Results
Susceptibility to infection with H. polygyrus is controlled to a large degree by the genetic background of the mouse strain (Table 1); C57BL/6 and CBA mice are highly susceptible 21,22. For maintenance of the parasite life cycle, the F1 hybrid between these two strains has been chosen for its ability to withstand much higher worm burdens without morbidity (excessive intestinal epithelial damage) compared to either parental strain. Oral gavage of 400 L3 larvae is used to maintain the life cycle in F1 mice (resulting in adult worm burdens shown in Figure 2), whilst a dose of 200 L3 larvae is generally used for experiments in homozygous inbred strains (e.g., C57BL/6 or BALBc). However, this dosage may need to be reduced depending on environmental co-factors that may differ between animal facilities, such as variations in gut flora.
Batches of HES have proven reproducible efficacy in functional assays and in protein composition; moreover, when supernatants from each successive week of culture were analysed, up to a total of 4 weeks, the protein profiles were found to be very similar (Figure 3). When HES concentration is measured by Bradford assay (see 5.5), the total protein is usually approximately 1 mg/ml (Figure 4).
An alternative method of concentrating HES is with a centrifugal concentrator (eg Vivaspin 3-kDa cut-off membrane), in which samples are spun at up to 4,000 g in a swing-out rotor centrifuge, removing buffer salts and low molecular weight components. Centrifugal concentration is best suited to small processing volumes (1-10 ml) and are limited to a maximum concentration factor of approximately 30x.
When collecting HES, avoidance of contamination is critical. To avoid contamination with host molecules, we discard HES-containing culture media from the first 24 hr after adult worm harvest from the mouse intestine. We also quantify the level of LPS contamination in each batch of HES with a Chromogenic LAL assay (see 5.7). 1 U of LPS equates to ~100 pg LPS and levels below this are considered negligible 23. In our hands, most batches of HES are significantly below this limit, the mean concentration of LPS in HES being 0.23 U/µg (Figure 5). The effects of LPS in in vivo models of pathology (for instance the suppression of asthmatic responses) requires at least 10 ng of LPS 23. Hence in vivo administration of 5 µg HES from a batch with 100 pg LPS/µg HES will include 500 pg LPS, well below the effective concentration where LPS becomes a problem.
Figure 1: Baermann Apparatus Baermann Apparatus setup for collection of adult H. polygyrus (as described in section 2).
Figure 2: Mean Worm Burden 14 Days After Infection with 400 L3 Larvae Mean worm burdens 14 days after infection with 400 L3 larvae. Data points shown are from 19 separate rounds of infection of C57BL/6xCBA mice. Mean ± SEM shown.
Figure 3: Protein Profiles of HES From Successive Weeks in Culture SDS-PAGE profile of HES proteins collected in successive weeks of culture.
Figure 4: Final Quantity of HES from 500 ml ES-Containing Media Yield of HES protein from 11 different batches derived from approximately 500 ml of culture supernatant. Mean ± SEM shown.
Figure 5: LPS Contamination of HES Levels of LPS contamination in 41 batches of HES measured by the Limulus ameobocyte assay. Median LPS concentration = 86 U per mg of HES.
Figure 6: Animated Schematic of H. polygyrus Life Cycle Summary of key life cycle stages from oral gavage of L3 larvae, through recovery of larvae and adult worms to isolation of HES.
Figure 7: Animated Schematic of HES and its Functions Key immunomodulatory effects of soluble mediators and exosomes contained within HES.
Genotype (and background strain) Primary Infection Phenotype Reference
Inbred strains
A/J, CBA, C3H Highly susceptible 22,29
C57BL/6 and C57BL/10 Susceptible 22,30
BALB/c, DBA/2, 129/J Intermediate 22,31
NIH, SJL, SWR Low susceptibility 22,32
Transgenic for cytokines or cytokine receptors
IL-1β-/- More susceptible 33
IL-1R-/- Less susceptible (a); no change in susceptibility but increased granulomas (b) (a) 33
(b) 34
IL-2Rβ Transgenic (C57BL/6) Resistant 35
IL-4-/- Higher fecundity 36
IL-4R-/- (C57BL/6 or BALB/c) Highly susceptible 22
IL-6–/– (BALB/c or C57BL/6) Highly Resistant 37
IL-9 Transgenic (FVB) Resistant 38
IL-21R –/– (C57BL/6) Deficient Th2, decreased granuloma gormation 39,40
IL-25-/- (BALB/c) More susceptible 33
IL-33R (T1/ST2) -/- (BALB/c) No change in susceptibility 33
TGFβRIIdn (C57BL/6) High Th1, Increased susceptibility 41,42
Transgenic for T cell markers
CD28–/– (BALB/c) Marginally higher fecundity 43
CD86 (B7-2)–/– (BALB/c) Higher fecundity 44
OX40L–/– (BALB/c) Higher fecundity 45
Transgenic for innate Immune loci
Type 1 interferon receptor (IFNAR)-/- (C57BL/6) Higher fecundity, more granulomas 34
MyD88–/– (C57BL/6) More resistant, more granulomas 34
C-KitW/Wv (WBB6) More susceptible 46,47
Table 1: Primary infections with H. polygyrus in genetically different and gene-targetted mouse strains.
The life cycle of H. polygyrus proceeds in a reliably consistent fashion. Following natural ingestion or oral gavage of L3 larvae on day 0, cysts begin to form under the duodenal serosa by day 5, progressing to larval moults and then emerging as adult worms into the gut lumen from day 10, eggs can be seen in feces from day 14 and granulomas are visible on the duodenal serosal surface from day 21. The protocol described above (and summarized in Figure 6) allows for high-throughput production of H. polygyrus excretory-secretory products (HES), in addition to reliable recovery of viable L3 larvae for future experimental and life cycle infections.
H. polygyrus infection has been shown to be protective in models of asthma dependent on OVA or Der p 1 (House dust mite allergen)14. Furthermore, suppression of airway inflammation could be transferred with CD4+CD25+ regulatory T cells 14 or CD19+CD23hi regulatory B cells 24 from non-sensitised, H. polygyrus-infected mice. In models of autoimmunity, H. polygyrus infection has been shown to be suppressive in the experimental autoimmune encephalomyelitis (EAE) model of multiple sclerosis 9, and suppression can be transferred with either CD4+ T cells or CD19+ B cells from infected mice 24.
Heligmosomoides polygyrus excretory-secretory products (HES) modulate the host immune response and suppress Th2-mediated inflammation by a number of mechanisms (outlined in Figure 7), including: a) Inhibition of dendritic cell responsiveness to stimulation 25, b) induction of CD4+Foxp3+ regulatory T cells 18 . and c) blockade of IL-33 production 20. HES has been shown to be protective when administered at sensitization or challenge in the OVA-alum model of asthma 19 and also when administered intranasally with Alternaria extract allergen 20, through suppression of early IL-33 release. Avoidance of LPS contamination of HES is often crucial for the success of future immunological experiments. In the protocol outlined here, the critical steps in achieving this are to ensure that debris contained within the muslin bags of the Baermann Apparatus does not enter the final HES preparation (see 2.10) and to set aside ES solution from the first 24 hr of culture (see 5.1).
Over recent years, HES has been thoroughly characterized at the proteomic level with over 370 distinct proteins identified 26,27. In addition, ES from the 4th stage larvae has been subjected to proteomic analysis 28. Ongoing work includes characterizing the major glycan components of HES, known to be strongly immunogenic 3, and secreted micro-RNAs that are encapsulated in 50-100 nm vesicles or exosomes (Buck et al., submitted for publication). With the establishment of reproducible protocols for the collection of ES from this highly immunomodulatory parasite, and with extensive proteomic data identifying the molecular components of HES, the stage is now set for the identification and therapeutic testing of individual molecules from H. polygyrus that can mediate key effects on the host immune system.
The authors have no conflicts of interest.
The authors are funded by the Wellcome Trust through the Edinburgh Clinical Academic Track (C.J.C.J.) and Programme Grant (Y.H., and R.M.M.) and Fellowship (J.R.G) funding, the BBSRC (G.C.), the American Asthma Foundation (E.R., H.J.McS. and R.M.M.), Asthma UK (H.J.McS.) and the Rainin Foundation (D.J.S. and R.M.M.).
Name Company Catalog Number Comments
Amicon concentrator Millipore 5112 Model 8050 50 ml
http://www.emdmillipore.com/INTL/en/product/Stirred-Cell-Model-8050,-50%C2%A0mL,MM
_NF-5122
Hanks' buffered salt solution Sigma H9394 500 ml
http://www.sigmaaldrich.com/catalog/product/sigma/h9394?lang=en&region=GB
Penicillin streptomycin Invitrogen 15070-063 100 ml
http://www.lifetechnologies.com/order/catalog/product/15070063
L-glutamine Invitrogen 25030-081 200 MM 100ml
https://www.lifetechnologies.com/order/catalog/product/25030081?ICID=search-25030081
Activated charcoal Merck 1.09624.0500 18-35 mesh
http://www.merckmillipore.com/GB/en/product/Charcoal-activated,MDA_CHEM-109624
Glucose solution Sigma G8769 100 ml 45% solution
http://www.sigmaaldrich.com/catalog/product/sigma/g8769?lang=en&region=GB
Falcon tubes Sarstedt 62.547.254 50 ml
http://www.scribd.com/doc/124320105/Sarstedt-Falcon
T25 Flasks Sigma CLS430639 25cm vented flask
http://www.sigmaaldrich.com/catalog/product/sigma/cls430639?lang=en&region=GB
Chromogenic LAL assay Lonza 50-647U QCL-1000 assay
http://www.lonza.com/products-services/pharma-biotech/endotoxin-detection/endotoxin-detection-assays/endpoint-chromogenic-lal-assay.aspx
Gentamicin Gibco 15710-049 100 ml
Roswell Park Memorial Institute (RPMI) 1640 Gibco 11875093 500 ml; without L-glutamine
DOWNLOAD MATERIALS LIST
Monroy, F. G., Enriquez, F. J. Heligmosomoides polygyrus : a model for chronic gastrointestinal helminthiasis. Parasitol. Today. 8, 49-54 (1992).
Maizels, R. M., et al. Immune modulation and modulators in Heligmosomoides polygyrus infection. Exp. Parasitol. 132, 76-89 (2012).
Hewitson, J. P., et al. Heligmosomoides polygyrus elicits a dominant nonprotective antibody response directed at restricted glycan and peptide epitopes. J Immunol. 187, 4764-4777 (2011).
Walk, S. T., Blum, A. M., Ewing, S. A., Weinstock, J. V., Young, V. B. Alteration of the murine gut microbiota during infection with the parasitic helminth Heligmosomoides polygyrus. Inflamm Bowel Dis. 16, 1841-1849 (2010).
Rausch, S., et al. Small intestinal nematode infection of mice Is associated with increased enterobacterial loads alongside the intestinal tract. PLoS ONE. 8, e74026 (2013).
Reynolds, L. A., et al. Commensal-pathogen interactions in the intestinal tract: Lactobacilli promote infection with, and are promoted by, helminth parasites. Gut Microbes. 5, 10-19 (2014).
Saunders, K. A., Raine, T., Cooke, A., Lawrence, C. E. Inhibition of autoimmune type 1 diabetes by gastrointestinal helminth infection. Infect Immun. 75, 397-407 (2007).
Liu, Q., et al. Helminth infection can reduce insulitis and type 1 diabetes through CD25- and IL-10-independent mechanisms. Infect Immun. 77, 5347-5358 (2009).
Donskow-Łysoniewska, K., Krawczak, K., Doligalska, M. Heligmosomoides polygyrus: EAE remission is correlated with different systemic cytokine profiles provoked by L4 and adult nematodes. Exp Parasitol. 132, 243-248 (2012).
Mishra, P. K., Patel, N., Wu, W., Bleich, D., Gause, W. C. Prevention of type 1 diabetes through infection with an intestinal nematode parasite requires IL-10 in the absence of a Th2-type response. Mucosal Immunol. 6, 297-308 (2013).
Sutton, T. L., et al. Anti-Inflammatory mechanisms of enteric Heligmosomoides polygyrus infection against trinitrobenzene sulfonic acid-induced colitis in a murine model. Infect Immun. 76, 4772-4782 (2008).
Hang, L., et al. Heligmosomoides polygyrus bakeri infection activates colonic Foxp3+ T cells enhancing their capacity to prevent colitis. J Immunol. 191, 1927-1934 (2013).
Bashir, M. E., Andersen, P., Fuss, I. J., Shi, H. N., Nagler-Anderson, C. An enteric helminth infection protects against an allergic response to dietary antigen. J Immunol. 169, 3284-3292 (2002).
Wilson, M. S., et al. Suppression of allergic airway inflammation by helminth-induced regulatory T cells. J. Exp. Med. 202, 1199-1212 (2005).
Kitagaki, K., et al. Intestinal helminths protect in a murine model of asthma. J Immunol. 177, 1628-1635 (2006).
Hartmann, S., et al. Gastrointestinal nematode infection interferes with experimental allergic airway inflammation but not atopic dermatitis. Clin Exp Allergy. 39, 1585-1596 (2009).
Pritchard, D. I., Lawrence, C. E., Appleby, P., Gibb, I. A., Glover, K. Immunosuppressive proteins secreted by the gastrointestinal nematode parasite Heligmosomoides polygyrus. Int. J. Parasitol. 24, 495-500 (1994).
Grainger, J. R., et al. Helminth secretions induce de novo T cell Foxp3 expression and regulatory function through the TGF-β pathway. J Exp Med. 207, 2331-2341 (2010).
McSorley, H. J., et al. Suppression of type 2 immunity and allergic airway inflammation by secreted products of the helminth Heligmosomoides polygyrus. Eur. J. Immunol. 42, 2667-2682 (2012).
McSorley, H. J., Blair, N. F., Smith, K. A., McKenzie, A. N. J., Maizels, R. M. Blockade of IL-33 release and suppression of type 2 innate lymphoid cell responses by helminth secreted products in airway allergy. Mucosal Immunol. 7, 1068-1078 (2014).
Behnke, J. M., Menge, D. M., Noyes, H. Heligmosomoides bakeri: a model for exploring the biology and genetics of restance to chronic gastrointestinal nematode infections. Parasitology. 136, 1565-1580 (2009).
Filbey, K. J., et al. Innate and adaptive type 2 immune cell responses in genetically controlled resistance to intestinal helminth infection. Immunology and Cell Biology. 92, 436-448 (2014).
Bortolatto, J., et al. Toll-like receptor 4 agonists adsorbed to aluminium hydroxide adjuvant attenuate ovalbumin-specific allergic airway disease: role of MyD88 adaptor molecule and interleukin-12/interferon-gamma axis. Clin Exp Allergy. 38, 1668-1679 (2008).
Wilson, M. S., et al. Helminth-induced CD19+CD23hi B cells modulate experimental allergic and autoimmune inflammation. Eur J Immunol. 40, 1682-1696 (2010).
Segura, M., Su, Z., Piccirillo, C., Stevenson, M. M. Impairment of dendritic cell function by excretory-secretory products: A potential mechanism for nematode-induced immunosuppression. Eur J Immunol. 37, 1887-1904 (2007).
Hewitson, J. P., et al. Proteomic analysis of secretory products from the model gastrointestinal nematode Heligmosomoides polygyrus reveals dominance of Venom Allergen-Like (VAL) proteins. J Proteomics. 74, 1573-1594 (2011).
Moreno, Y., et al. Proteomic analysis of excretory-secretory products of Heligmosomoides polygyrus assessed with next-generation sequencing transcriptomic information. PLoS Negl Trop Dis. 5, e1370 (2011).
Hewitson, J. P., et al. Secretion of protective antigens by tissue-stage nematode larvae revealed by proteomic analysis and vaccination-induced sterile immunity. PLOS Pathogens. 9, e1003492 (2013).
Prowse, S. J., Mitchell, G. F. On the choice of mice for dissection of strain variations in the development of resistance to infection with Nematospiroides dubius. Austral J Exp Biol Med. 58, 603-605 (1980).
Behnke, J. M., Robinson, M. Genetic control of immunity to Nematospiroides dubius: a 9-day anthelmintic abbreviated immunizing regime which separates weak and strong responder strains of mice. Parasite Immunol. 7, 235-253 (1985).
Zhong, S., Dobson, C. Heligmosomoides polygyrus: resistance in inbred, outbred, and selected mice. Exp. Parasitol. 82, 122-131 (1996).
Behnke, J. M., et al. Genetic variation in resistance to repeated infections with Heligmosomoides polygyrus bakeri, in inbred mouse strains selected for the mouse genome project. Parasite Immunol. 28, 85-94 (2006).
Zaiss, M. M., et al. IL-1β suppresses innate IL-25 and IL-33 production and maintains helminth chronicity. PLoS Pathog. 9, e1003531 (2013).
Reynolds, L. A., et al. MyD88 signaling inhibits protective immunity to the gastrointestinal helminth parasite Heligmosomoides polygyrus. J Immunol. (2014).
Morimoto, M., Utsumiya, K. Enhanced protection against Heligmosomoides polygyrus in IL-2 receptor β-chain overexpressed transgenic mice with intestinal mastocytosis. J Vet Med Sci. 73, 849-851 (2011).
Urban, J. F., Katona, I. M., Paul, W. E., Finkelman, F. D. Interleukin 4 is important in protective immunity to a gastrointestinal nematode infection in mice. Proc. Natl. Acad. Sci. USA. 88, 5513-5517 (1991).
Smith, K. A., Maizels, R. M. IL-6 controls susceptibility to helminth infection by impeding Th2 responsiveness and altering the Treg phenotype in vivo. Eur J Immunol. 44, 150-161 (2014).
Hayes, K. S., Bancroft, A. J., Grencis, R. K. Immune-mediated regulation of chronic intestinal nematode infection. Immunol Rev. 201, 75-88 (2004).
Fröhlich, A., et al. IL-21 receptor signaling is integral to the development of Th2 effector responses in vivo. Blood. 109, 2023-2031 (2007).
King, I. L., Mohrs, K., Mohrs, M. A nonredundant role for IL-21 receptor signaling in plasma cell differentiation and protective type 2 immunity against gastrointestinal helminth infection. J Immunol. 185, 6138-6145 (2010).
Ince, M. N., et al. Role of T cell TGF-β signaling in intestinal cytokine responses and helminthic immune modulation. Eur J Immunol. 39, 1870-1878 (2009).
Reynolds, L. A., Maizels, R. M. Cutting Edge: In the absence of TGF-β signaling in T cells, fewer CD103+ regulatory T cells develop, but exuberant IFN-γ production renders mice more susceptible to helminth infection. J Immunol. 189, 1113-1117 (2012).
Ekkens, M. J., et al. Memory Th2 effector cells can develop in the absence of B7-1/B7-2, CD28 interactions, and effector Th cells after priming with an intestinal nematode parasite. J Immunol. 168, 6344-6351 (2002).
Greenwald, R. J., et al. B7-2 is required for the progression but not the initiation of the type 2 immune response to a gastrointestinal nematode parasite. J. Immunol. 162, 4133-4139 (1999).
Ekkens, M. J., et al. The role of OX40 ligand interactions in the development of the Th2 response to the gastrointestinal nematode parasite Heligmosomoides polygyrus. J. Immunol. 170, 384-393 (2003).
Hashimoto, K., et al. Immunity-mediated regulation of fecundity in the nematode Heligmosomoides polygyrus--the potential role of mast cells. Parasitology. 137, 881-887 (2010).
Hepworth, M. R., et al. Mast cells orchestrate type 2 immunity to helminths through regulation of tissue-derived cytokines. Proc Natl Acad Sci U S A. 109, 6644-6649 (2012).
https://www.jove.com/t/52412
PDF DOI DOWNLOAD MATERIALS LIST
Johnston, C. J. C., Robertson, E., Harcus, Y., Grainger, J. R., Coakley, G., Smyth, D. J., McSorley, H. J., Maizels, R. Cultivation of Heligmosomoides Polygyrus: An Immunomodulatory Nematode Parasite and its Secreted Products. J. Vis. Exp. (98), e52412, doi:10.3791/52412 (2015).… More
Johnston, C. J. C., Robertson, E., Harcus, Y., Grainger, J. R., Coakley, G., Smyth, D. J., McSorley, H. J., Maizels, R. Cultivation of Heligmosomoides Polygyrus: An Immunomodulatory Nematode Parasite and its Secreted Products. J. Vis. Exp. (98), e52412, doi:10.3791/52412 (2015).
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Great Lakes cruises don't have to be long affairs that cover hundreds or thousands of miles. They can be short cruises centered around scrumptious dinners or lunches, or simply designed to sample the amazing sunsets that the Great Lakes provide. One Great Lakes ship that provides such cruises is the Spirit of Chicago.
If you want a truly unique dinner cruise experience, the Spirit of Chicago offers a Thanksgiving Dinner cruise featuring Walnut crusted turkey breast, orange glazed ham and slow braised center cut sirloin. There are also two after Thanksgiving Champagne brunch and dinner cruises available on November 27th. There are few other small ship cruise lines that offer this level of dining, or trips as late into the fall as the Spirit of Chicago offers.
For those who like the night, the Spirit of Chicago offers a midnight moonlight cruise. This Lakes cruise includes a professional DJ, cash bar and light hors d'oeuvres. Sorry, this cruise is adults only (over 21) and you must be able to show a valid photo ID before boarding.
Another cruise offering is the Sweetest Day Dinner cruise which is offered October 17th. Offering live music, champagne and memorable views of the Sweetest Day fireworks, this cruise promises to be a winner. Fireworks cruises are always popular among dinner and evening cruisers and small cruise ships offer an outstanding platform for viewing fireworks unobstructed by either buildings or milling crowds. The Spirit of Chicago offers an upper level observation deck as well as several open patios on the other decks and should allow excellent viewing.
Fireworks cruises are available on the Spirit of Chicago on the fourth of July, Sweetest Day and News Years Eve. Since many Great Lakes cruise ships are closed for business during the winter season, this makes the Spirit of Chicago exceptional as it offers cruises most of the year.
The spirit of Chicago offers three dining decks and an observation deck and has a reported capacity of 572 passengers. They will take single passengers, small groups or even large group charters that occupy one or more decks. The decks are open and spacious and appear well lighted. The ship departs from Navy pier and the cruise provider reports that ample parking is available at the Navy Pier parking lot. There is also a continuous running (and free to ride) trolley that carries passengers from the parking a lot along Navy pier.
There is a dress code of sorts for the dinner cruises, so be certain to check with the cruise providers for exact recommendations. These cruises are generally not shirt and tie affairs, but the cruise line prefers passengers on the dinner cruises be a little more dressy. There are also enhancements you can add to your cruise to make to more memorable. These include private tables, balloons, roses, appetizer plates, open bar, champagne, wine, souvenir glasses, photographs, guaranteed window seats and other perks that add to the experience of a Great Lakes cruise on this small ship.


text:
Q: Python 3 tkinter destroy all child windows at exit of root SO i have been working on this for a while now and i just have no idea what i am doing wrong. I have a main window my 'root' but i also have a 'helpmenu'. right now, when you close the root helpmenu will remain opened if you opened it. i want helpmenu to close when the root closes.
i have tried to cutout all the lines of code from my program which i believed had no baring on the situation but forgive me if it is still long.
    import atexit
    from tkinter import *

    root = Tk()

    def quit():
        helpmenu.destroy()

    atexit.register(quit)

    def helpmenu():
        helpmenu = Tk()
        helpmenu.geometry('800x600')
        helpmenu.title('Help')
        help_Label = Label(helpmenu, text='Welcome to the Boring Program!')
        help_Label.pack()

    class GUI(object):
        def __init__(self, master):
            self.master = master

            def activateCheck1():
                if c1Var.get() == 1:          #whenever checked
                    c3.config(state=NORMAL)
                elif c1Var.get() == 0:        #whenever unchecked
                    c3.config(state=DISABLED)

            def activateCheck2():
                if c2Var.get() == 1:          #whenever checked
                    c4.config(state=NORMAL)
                elif c2Var.get() == 0:        #whenever unchecked
                    c4.config(state=DISABLED)

            c1Var = IntVar()
            c2Var = IntVar()
            c1_Label = Label(root, text="Select Which Edges to Program")
            c1_Label.grid(row=2, columnspan=2)
            c1 = Checkbutton(root, text="Left Edge", variable=c1Var, command=activateCheck1)
            if parser.get('CONFIGURATION', 'LeftEdgeProgram') == '1':
                c1.select()
            c1.grid(row=3, column=0)
            c2 = Checkbutton(root, text="Right Edge", variable=c2Var, command=activateCheck2)
            if parser.get('CONFIGURATION', 'RightEdgeProgram') == '1':
                c2.select()
            c2.grid(row=3, column=1)

            c3Var = IntVar()
            c4Var = IntVar()
            c3_Label = Label(root, text="Select Which Edges to Insert Glue and Dowels")
            c3_Label.grid(row=4, columnspan=2)
            c3 = Checkbutton(root, text="Left Edge", variable=c3Var)
            if parser.get('CONFIGURATION', 'LeftEdgeDowel') == '1':
                c3.select()
            c3.grid(row=5, column=0)
            c4 = Checkbutton(root, text="Right Edge", variable=c4Var)
            if parser.get('CONFIGURATION', 'RightEdgeDowel') == '1':
                c4.select()
            c4.grid(row=5, column=1)

            menubar = Menu(root)
            filemenu = Menu(menubar, tearoff=0)
            filemenu.add_command(label="Print", state='disabled')
            filemenu.add_command(label="Save Configuration", command=save_config)
            filemenu.add_command(label="Help", command=helpmenu)
            filemenu.add_command(label="Restore Defaults", state='disabled')
            filemenu.add_command(label="About", command=restore_config)
            menubar.add_cascade(label="Options", menu=filemenu)

            root.config(menu=menubar)

    myGUI = GUI(root)
    root.mainloop()

A: Firstly you are creating two instances of a tk() windows. Creating two tk() windows almost never should be done, this is because two tk windows cannot communicate with eachother. Instead you should use a TopLevel window.
You should also change your HelpMenu to a class object which inherits the TopLevel Class
This should look something like this:
class HelpMenu(Toplevel):
    def __init__(self, master, *args, **kwargs):
        super(HelpMenu, self).__init__(master, *args, **kwargs)
        self.geometry('800x600')
        self.title('Help')
        help_Label = Label(self, text='Welcome to the Boring Program!')
        help_Label.pack()

The only thing that now needs to be changed is the command for the help option in the toolbar
Something like this (notice I am using a lambda function to make this change):
filemenu.add_command(label="Help", command= lambda:HelpMenu(self.master) )

This will fix your problem, because when the master window of a TopLevel window is closed the TopLevel window is automatically closed.


text:
Turkey launches mass coronavirus vaccination
Saudi Arabia to appoint women as court judges 'very soon'
Yemen says ballistic missiles used to attack Aden airport
Yemen says ballistic missiles used to attack Aden airport /node/1792476/middle-east
Smoke billows at the Aden Airport on Dec. 30, 2020, after explosions rocked the Yemeni airport shortly after the arrival of a plane carrying members of a new unity government. (File/AFP)
https://arab.news/58byk
They were launched from a site 100 kilometers away from the airport
The Ministry said "Iranian and Lebanese experts" are behind the missiles
DUBAI: The Yemeni Interior Ministry said the missile which were used to attack Aden's airport last month were ballistic, Al-Arabiya TV reported.
They were launched from a site 100 kilometer away from the airport, in Houthi-controlled areas, the report added.
#WATCH: Dramatic footage captured the moment the rockets smashed into #Aden airport in #Yemen killing several and injuring dozens more https://t.co/ZMu7XJuwj2 pic.twitter.com/48xjeLADoW
— Arab News (@arabnews) December 30, 2020
The Ministry said "Iranian and Lebanese experts" are behind the missiles which were used in the attack.
On Dec. 30, a deadly rocket attack shook Aden airport moments after a plane carrying members of Yemen's cabinet arrived on a flight from the Saudi Arabian capital, Riyadh.
No one has claimed responsibility. But the Yemeni government promptly accused Houthi rebels and their backers, the Iranian government, of being behind the airport attack and a drone assault on the Mashiq Palace in Aden shortly after the prime minister and his Cabinet were transferred there.
Prime Minister Maeen Abdulmalik Saeed said the deadly blast was carried out using three precision-guided missiles.
Houthi officials have denied being behind the attack and have sought to blame unspecified groups in the Arab-led coalition. Houthi leaders have not offered any evidence or answered requests for comment.
Topics: Yemen Aden Airport
Saudi cabinet welcomes US plan to designate Yemen's Houthis a terrorist organization
Abbas announces long-awaited Palestinian elections /node/1793536/middle-east
The last Palestinian parliamentary elections in 2006 saw Hamas win an unexpected landslide
The 2005 Palestinian presidential vote saw Abbas elected with 62 percent support to replace the late Yasser Arafat
RAMALLAH: Palestinian president Mahmud Abbas on Friday announced dates for the first Palestinian elections in more than 15 years, setting legislative polls for May 22 and a July 31 presidential vote.
Abbas's Fatah party, which controls the Palestinian Authority based in the occupied West Bank, and the Hamas group, who hold power in Gaza, have for years expressed interest in taking Palestinians back to the polls.
A long-standing rivalry between the two main Palestinian factions was seen as a leading factor in stalling progress toward a new vote.
But Fatah and Hamas have lately been engaged in unity talks, reaching an agreement in principle in September to hold elections in 2021.
Hamas on Friday welcomed Abbas's announcement.
"In recent months, we have worked to overcome obstacles in order to reach this day," it said in a statement.
It added that it looked to "free elections in which voters can express themselves without pressure and without restrictions, in all fairness and transparency."
A statement on the official Palestinian Wafa news agency said Abbas has signed "a presidential decree concerning elections," specifying the May and July dates.
"This announcement was eagerly awaited," Palestinian analyst Arif Jaffal, head of the Arab World Democracy and Electoral Monitor, told AFP.
"It is a very important step," he said.
The 2005 Palestinian presidential vote saw Abbas elected with 62 percent support to replace the late Yasser Arafat.
There has been no indication from Fatah as to whether the 85-year-old Abbas intends to seek re-election.
A rare poll by the Palestinian Center for Policy and Research carried out last year said Hamas leader Ismail Haniyeh would beat Abbas in a presidential election.
The statement from Abbas said he expects polls will be held "in all governorates of Palestine, including east Jerusalem," which was annexed by Israel following the 1967 Six-Day War but is considered occupied territory.
Israel bans all Palestinian Authority activity in east Jerusalem, and there was no indication the Jewish state would allow a Palestinian vote within the city.
Israel's right-wing Prime Minister Benjamin Netanyahu, who faces his own re-election contest in March, describes Jerusalem as Israel's "undivided capital."
Palestinian Prime Minister Mohammed Shtayyeh said his government "was ready to get things going to facilitate the electoral process, in total transparency, while waiting for pluralism."
Some 2.8 million Palestinians live in the West Bank, while the densely populated Gaza Strip is home to two million.
The last Palestinian parliamentary elections in 2006 saw Hamas win an unexpected landslide.
The polls resulted in a brief unity government but it soon collapsed and in 2007, bloody clashes erupted in the Gaza Strip between the two principal Palestinian factions, with Hamas ultimately seizing control of Gaza.
Numerous attempts at reconciliation, including a prisoner exchange agreement in 2012 and a short-lived coalition government two years later, have failed to close the rift.
But experts have said intra-Palestinian reconciliation talks have taken on greater urgency following a series of US-brokered normalization agreements signed between Israel and four Arab states.
The deals to normalize ties with Israel signed by the United Arab Emirates, Bahrain, Morocco and Sudan were condemned across the Palestinian political spectrum.
They also broke with decades of Arab League consensus against recognition of Israel until it reached an agreement to end the Palestinian conflict that included the creation of Palestinian state, with a capital in east Jerusalem.
Palestinian leaders have also voiced hope that the incoming administration of US President-elect Joe Biden will lead to renewed diplomacy on the Palestinian cause.
The PA cut ties with President Donald Trump's administration, accusing it of egregious bias toward Israel.
Topics: Palestine elections
Palestinian President Abbas congratulates Joe Biden
US-Palestine nationals denounce Pompeo visit


File: 1a_1_u18chan.jpg - (245.99kb, 419x620, 1a.jpg) Thumbnail displayed, click image for full size.
There must a lot of people love this comic, otherwise we won't see so many version of this comic.
And I remember someone even made a video and dubbed. Can't remember where though.
and another idiot that flaps his mouth instead of trying to prove he could do better...as if.
May we have the link?


Start checking the balance of any gift cards for The Container Store and get organized today! Raise lets you check the balance right here. So whether you're looking to buy new bins, file folders and more or sell your gift card to The Container Store for cash, we can help!


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Tag Archives: #COVIDandschools
The rules of schooling have been rewritten. Let's not go back to normal.
By Jason Weeby, Guest Blogger on February 1, 2021
Former Bellwarian Jason Weeby, who helped to develop and lead our work around education innovation, offers a series for Ahead of the Heard that makes the case for maintaining some pandemic-era education innovations. Learn more about Bellwether's work here. Read more posts in this series here.
When the COVID-19 pandemic closed school doors in March, the rules of grammar that underpin our collective beliefs about what "real schools" look like were erased. In a matter of months, school system leaders and teachers were required to break down complex and notoriously unalterable systems into their component parts and rearrange them so kids could keep learning.
An invisible virus accomplished in months what throngs of reformers with billions of dollars in philanthropic support couldn't.
Suddenly, waivers from rigid regulations and renegotiated labor contracts freed schools to try new ways of using their time, space, staffing, student groupings, and resources. Devices, broadband internet access, and online learning platforms, which have long been accessory to in-person instruction, became critical means for teaching and learning. Parents who are typically treated as customers were enlisted as co-educators.
Millions of students are now learning in ways that look nothing like school at all.
Yet, nearly a year on, districts are still in crisis response mode. And after months of physical isolation, prolonged stress and anxiety, upended routines, financial uncertainty, and, for some, sickness, death, and grieving, people just want to go back to normal.
For kids who thrive in a traditional setting, getting them back into school buildings may be the best thing. However, for many low-income students returning to "normal" means returning to dilapidated school buildings in segregated neighborhoods staffed with the least experienced teachers and operating on a fraction of the funds other schools enjoy. Lakisha Young, founder of Oakland Reach summed it up this way, "Our Black children have long been failed by in-person learning, so we don't want a return to the status quo." The status quo might also not be the best solution for students who are targets of bullies, suffer from anxiety, or simply prefer remote or hybrid learning environments. For students that were undereducated by their schools during "normal" times it would be a mistake to return to schooling the way it was and ignore the lessons that the pandemic has taught us.
Instead of going from the status quo to crisis education and back to the status quo again, what if we went from the status quo to crisis education to a more equitable and effective public school system?
Some district leaders envision post-pandemic learning to be fundamentally different from what schools were doing in March. For instance, Karen Quanbeck, superintendent of Clear Creek School District RE-1 in Idaho Springs, Colorado, said at a recent event: "We are navigating this seismic shift in education and my gut is, we're not going back to pre-pandemic learning, and there's pros and cons to that." Quanbeck isn't alone in her thinking. A recent Rand survey of over 300 district and charter leaders found that "…remote learning, in at least some form, will outlast the COVID-19 pandemic. One in five districts were considering, planning to adopt, or had already adopted a virtual school or fully online option, while about one in ten have adopted or are planning to adopt a blended or hybrid form of instruction." Innovation researchers are beginning to wonder whether the lessons that we've learned will change schools forever. Teachers are thinking about what pandemic-era classroom practices they want to carry with them into the future. Even parents expect schools to look different in the future. After taking time away from work to support his two kids' distance learning pod, a colleague returned certain that school would never look the same saying, "parents can't unsee all of the things they've experienced during the pandemic."
Of course, not everyone sees this moment as a quantum leap in education innovation. Education historian Larry Cuban, for instance, points to the limited effect of past efforts to create "the classroom of the future" and sees students heading back to school as soon as we hit herd immunity. Others rightly point out to the millions of students who are struggling with online learning or not showing up at all as proof that online learning is a failing experiment. Surely, much of what's been tried hasn't worked. There's value there too. We have as much to learn from the negative effects of the pandemic on schooling as the ingenuity that's been marshaled to respond to them.
For those of us stubborn optimists who work at the intersection of education and innovation, this moment is an important one. The nation's 13,000 school districts, 130,000 schools, and 3.7 million teachers have participated in what Sujata Bhatt, a senior fellow at Transcend Education called the "largest educational innovation experiment in the history of mankind" as they shifted to distance and hybrid learning. More accurately, the pandemic hasn't prompted one experiment, it's spun up millions of small and large experiments in a wide range of settings, some of which are succeeding while others are failing. Regardless, the experiments have generated countless lessons that can strengthen, diversify, and augment traditional schooling if — and only if — innovators, system leaders, and policymakers prepare for the rapidly approaching future when students can go back to schools safely.
Over the next few days, I'll make the case in a series of blog posts here on Ahead of the Heard that we should be actively looking for pandemic-era education innovations that increase equity and effectiveness. Even though there will be challenging headwinds, there are ways we can tack against them to create conditions where promising new ideas can be tested and shared.
All students, but most urgently low-income and BIPOC students, and students with special needs, should benefit from the countless lessons generated by the nation's 130,000 schools over the last 11 months.
You can read more from this series here.
This entry was posted in COVID-19, Education Innovation, Uncategorized and tagged #COVIDandschools, #InnovationAfterCOVID, covid, COVID-19 on February 1, 2021 by Jason Weeby, Guest Blogger.
Considerations For Private Schools and Their Allies Amid Budget Cuts
By Juliet Squire on October 22, 2020
In August, I wrote for Education Next about how the pandemic was affecting private schools — especially those dedicated to serving high-need students — and the factors that can influence whether a school is able to pivot during uncertain and disruptive times.
But as state budgets tighten, private schools that rely on vouchers or tax-credit scholarship programs to serve high-need students are not wholly in control of their own destiny. Rather, just as school responses have varied widely, policymaker responses will vary as well.
After the 2008 recession, for instance, state school choice policies sometimes lost — and sometimes gained — significant funding. For example, funding for Pennsylvania's Educational Improvement Tax Credit Program dropped 20% after 2008 before rebounding a couple of years later. Meanwhile, funding for Wisconsin's Milwaukee Parental Choice Program increased by 12%, and policymakers in the Sunshine State increased the ceiling on the Florida Tax Credit Scholarship Program nearly 50%.
There are limitations on what school choice proponents can predict in uncertain times. Here are three questions they should continue to revisit as the pandemic evolves this fall and new state legislatures prepare to convene in January:
What are the projections for your state's economy?
The more constrained the resources, the harder it will be for policymakers to preserve and expand private school choice programs. First, it will be important to understand the state tax revenues that fund state voucher programs. State revenues often come from a combination of income and sales taxes, and, as Jennifer Schiess explained, these can be hard to predict. They could be less vulnerable to downturns, since the pandemic hit low-wage workers hardest and low-wage workers make up a smaller share of income tax revenue. Internet shopping could also offset declines in sales tax revenue from brick-and-mortar retail. Continue reading →
This entry was posted in COVID-19, School Choice and tagged #COVIDandschools, Private Schools, tax credit scholarship program on October 22, 2020 by Juliet Squire.
Preventing a "Lost Generation" of Community College Learners
By Jeff Schulz on October 15, 2020
Photo by David Maiolo via Wikimedia
Community colleges have long served as an accessible and affordable post-secondary pathway to better jobs for high school graduates and adults looking to upskill. For example, the median weekly earnings for someone with an associate's degree are 17% higher than for those with only a high school diploma or a GED. Community colleges are particularly important for traditionally underserved students: compared to students at four-year colleges, community college students are more likely to be the first in their families to attend college, to be from a low-income family, and to be members of racial or ethnic minority groups.
This is one reason why the steep declines in community college enrollment this fall are especially troubling. According to newly released data from the National Student Clearinghouse Research Center (NSCRC), undergraduate enrollments across the country are down 4.0 percent compared to the same time last year, with the biggest losses being at community colleges, where enrollments declined by 9.4 percent, on fall enrollments.
Before the pandemic, there were approximately 5.5 million students enrolled in community college nationally. A 9.4 percent enrollment decrease equals about 520,000 students that have "stopped out" of community college, at least temporarily. This group of students is at risk of being a "lost generation" of learners. Dr. Karen Stout, president and CEO of Achieving the Dream, a nonprofit organization dedicated to helping community colleges become leaders in their communities, told The Well News: "The pandemic, if we are unable to find students we lost and keep students we serve, and open up new access points for new students, will result in a lost generation of learners that will hurt the economic and civic fabrics of the communities our colleges serve."
Why are more community college students "stopping out" of college relative to four-year students? We know from our work that, in aggregate, community college students often face more barriers on their road to completion. This excerpt from an April NEA Today article sheds some light:
Students who are first-generation, who are on financial aid, who face challenges around hunger, housing, transportation, and who are balancing work, childcare, and more, "they are all being adversely affected in ways that make those inequities more stark and more exacerbated," says [Kurt Meyer, an English professor at Irvine Valley College in California and president of the South Orange County Community College District Faculty Association], who notes a five-fold increase in South Orange County college students recently applying for emergency funds to pay rent, fix cars, and more.
One major challenge is that many community colleges were struggling financially even before the pandemic and are likely to see further financial challenges as a result of the pandemic. Not only are many schools facing losses of income from reduced enrollment and unexpected refunds, but many are facing additional financial challenges caused by cuts in state funding. Lakeland Community College in Ohio, for example, will lose over $780,000 in state funding by the end of June 2020 and expects to lose over $4 million for the fiscal year ending in 2021. Similar cases can be found around the country, and many community colleges have been forced to furlough and lay off faculty and staff.
So, what can leaders do to ensure that we don't lose a generation of students to COVID disruptions and increasingly tight financial constraints? Continue reading →
This entry was posted in COVID-19, Post-Secondary and tagged #COVIDandschools, community college, stopouts on October 15, 2020 by Jeff Schulz.


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Competitors shine with bright ideas at Won in One 2018
Feb 5, 2018 | Business, Engineering and Science, General News
A low-cost medical product used for blood salvaging during surgery was a two-time winner at the recent Won in One competition at Louisiana Tech's Rawle Enterprise Center.
HemoCycle's winning team, left to right: Steven Pirvu, Caitlin Snell, Joshua Jacobs, Ryan Botts.
The annual cash-prize competition is a chance for community members and students to share their innovative ideas. Each contestant or team has 60 seconds to present their concept or venture in an elevator-pitch format. After all 15 participants presented — there were 15 this year —evaluations were made by both the audience and judges.
HemoCycle, presented by Caitlin Snell, a senior biomedical engineering major, won the People's Choice category and was also the judges' first-place pick. Created by Snell and senior teammates Steven Pirvu, Joshua Jacobs, and Ryan Botts, the medical device would be designed to be more accessible than current options for rural hospitals and developing nations.
In second place was LifeDrop, a device presented by senior biomedical engineering major Kayla Farrar. The goal of the team behind LifeDrop was to provide low-cost, easy-to-operate water quality testing for those in developing nations and those affected by natural disasters.
Third place went to Trenchless Technology Center (TTC) Smart Concrete, a green construction material proposed by Mohammadamin Azimi.
Tech's chief research and innovation officer Davy Norris recognizes Ryan Botts, a senior industrial engineering major, during the University Innovation Fellows pinning ceremony.
For the best presentation by a student, the audience selected Garrett Allen for the Checkplate concept, which detailed a vacuum fixture that could reduce setup time for machining processes. Allen, a senior mechanical engineering major, said the event was a beneficial experience for the team.
"Up to now, we didn't have a real elevator pitch with this thing," Allen said. "It took five minutes to explain this and how it works and that kind of thing. Now, I have a concise way to tell people about it, so that is really valuable."
The evening also included a pinning ceremony for Botts, a senior industrial engineering major and member of the HemoCycle winning team. The University Innovation Fellows (UIF) pinning was performed by Dave Norris, the University's chief research and innovation officer and leader of Tech's Innovation Enterprise. Botts and 300 other college students from around the world attended a seminar at Stanford University to learn more about being an agent of change at their respective universities. The pinning acknowledged his efforts with that program and also at Tech, including his work with the Innovation Enterprise and student organizations such as OpTech.
"I think we are to a point where Tech is starting to grow, and I think that we are about to get into some really good days of design, innovation, and entrepreneurship on campus," Botts said.
New Frontiers to focus on health disparities in Feb. 1 lecture
Student research published in Journal of Neurotoxicology
College of Business to host virtual Diversity and Inclusion in the Workplace forum
Assistant professor of civil engineering to be awarded professionalism award by LEF
More than $171 million raised through Louisiana Tech's Forever Loyal Campaign
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How many have come away from Italy memorized by the unique allure of this land? From the might of Rome to the marvels of the Renaissance, it's no wonder the capital is called the Eternal City when one is continually confronted with the origins of history.
What's more, the labyrinthine streets of cities such as Siena, Genova, & San Gimignano have always elicited something of the subconscious in me, like venturing into a dream replete with the paradoxical passageways of an Escher etching.
Today, as the world leaps into the technological age a new panorama of experience is becoming available. With this in mind, we've worked together with neuroscientists to develop a revolutionary vacation holiday that explores the connections between mind, culture, beauty, and wellness. This new project utilizes non-invasive neurostimulation that's scientifically proven to enhance well-being.
First conceived in Italy, tDCS technology (transcranial direct current stimulation) is now used worldwide by the likes of Olympic & professional athletes, and even by the US Military because of its unique ability to augment physical and cognitive faculties. It is a painless, easy procedure that stimulates the specific areas of the brain in control of mental, emotional and physical motor skills. Various studies have shown its ability to enhance learning, attention, memorization, mood, creativity, reasoning, endurance and coordination, both in healthy individuals and even in those with clinical conditions.
Like the tale of Amor & Psyche, our journey begins with a fairytale in the homeland of Romance and the Renaissance; Tuscany. The tale of Tuscany is hidden in the maze of its precious vineyards, the serene shadows of its olive groves, the spirituality of its abbeys, the warmth and affability of its people and the beauty of its countryside.
While encountering an enchanted past, we extend a hand to an enchanted future, as travellers are greeted by neuro-psychologists from a specialist brain-stimulation clinic that officially represents Italy's Top Pro Cycling Team. Guests undergo personalized neuro-stimulation procedures, harnessing the power of neuroplasticity & neurofeedback to enhance mood, creativity, attention, memory, perception of beauty, learning, cognitive capacity and overall well-being.
Neurofeedback is a physiological self-monitoring technique that uses EEG (electroencephalography) and allows subjects to observe their own neurological processes through audio or visual display. This has shown to facilitate neural connectivity, allowing individuals to become more intune with the mechanisms of their own minds. In addition to tDCS, neurofeedback has been shown to be another technique capable of boosting concentration, well-being, and ph.
What's more, studies on ordinary healthy subjects, such as actors, musicians, and dancers, have shown that neurofeedback techniques enhance cognitive creativity, flow states, artistic performance, musicality, expressiveness, and communication technique. One particularly interesting application is the ability to induce "borderline waking states associated with creative insights", a state known as hypnagogia.
Cerebral stimulation used in conjunction with neurofeedback represents the next frontier of advancement in these technologies. It's this frontier that guests will benefit from, so as to elevate cognitive ability and overall well-being, all together wrapped within a culturally authentic vacation experience.


Unissued Vietnam era petrol stove as issued to Australian troops. These stoves were known as choofers due to the sound they made when starting. This stove comes in its original box of issue with the original stripping tool.


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Our delicious Kettle Corn is available in full cases of 36 per flavor.
Our Kettle Corn is available in full cases of 36 per flavor.
Price for each case of 36 is $44.95 delivered to continental USA.