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Development of potency assays for a plasmid containing vascular endothelial growth factor 2 | Huang | Electronic Journal of Biotechnology
Full Text - Development of potency assays for a plasmid containing vascular endothelial growth factor 2
Biotechnology Industry
Electronic Journal of Biotechnology ISSN: 0717-3458 Vol. 13 No. 1, Issue of January 15, 2010
2010 by Pontificia Universidad Católica de Valparaíso -- Chile Received October 17, 2008 / Accepted August 4, 2009
DOI: 10.2225/vol13-issue1-fulltext-8
Development of potency assays for a plasmid containing vascular endothelial growth factor 2
Li-chun Huang*
Corautus Genetics Inc.
500 Forbes Blvd.
South San Francisco, CA94555
San José. California, USA
E-mail: Lhuang2006@gmail.com
Emily Chin
Yawen L. Chiang
Keywords: cell proliferation, plasmid DNA, potency assays, receptor binding, VEGF-2.
BrdU: 5-bromo-2'deoxyuridine
BSA: bovine serum albumin
CHO: Chinese hamster ovary
HLMVEC: human lymph microvascular endothelial cells
PBS: phosphate buffered saline
RSD: relative standard deviation
VEGF: vascular endothelial growth factor
VEGFR: vascular endothelial growth factor receptor
We have developed analytical methods to measure the biological functions of pVGI.1(VEGF2), a naked plasmid DNA product containing vascular endothelial growth factor 2 used in clinical trials for coronary artery diseases (CAD) and peripheral artery diseases (PAD). After being injected into muscles, vascular endothelial growth factor 2 (VEGF-2), presumably expressed in muscle tissues, binds to the endothelial cell receptors VEGFR2 or VEGFR3, triggering the downstream responses including cell proliferation and vascularization. As it is important to make sure clinical material is biological active, we developed a quantitative assay first to measure the receptor binding activity of the pVGI.1(VEGF2) gene product expressed by the transfected host cells, and then a qualitative assay to confirm the cell proliferation promoting activity of the expressed protein. In both assays the signals were plotted directly against input DNA concentrations used to transfect the host cells. We confirmed specificity for both assays. In addition, we demonstrated acceptable levels of spike recovery (86.7-116%), precision (largest relative standard deviation (RSD)=19.3%), linearity and range (60-140% relative potency, 15 - 35 µg/mL) for the quantitative assay. We intend to use the potency assays for routine lot release and stability studies.
The vector pVGI.1(VEGF2) is a 5283 base pair plasmid encoding the full length cDNA of the VEGF-2 (VEGF-C) gene. Preclinical studies and early phase clinical trials have demonstrated that injecting the pVGI.1(VEGF2) directly into muscles provided therapeutic angiogenic effects both in the heart and in the limb (Witzenbichler et al. 1998; Vale et al. 2001; Losordo et al. 2002; Fortuin et al. 2003; Kawamoto et al. 2004; Losordo and Dimmeler, 2004; Reilly et al. 2005; Shah and Losordo, 2005; Shintani et al. 2006). It was postulated that the pVGI.1(VEGF2) gene product, the secreted VEGF-2 protein, mediated proliferative and chemotactic responses in endothelial cells (Cao et al. 1998; Cao et al. 2004; Bauer et al. 2005), which resulted in formations of new blood vessels in the ischemic tissue.
The VEGF-2 protein is synthesized as a pre-propeptide that undergoes post-translational modifications including proteolysis and dimerization (Joukov et al. 1997). The mature VEGF-2 protein is a ligand for the vascular endothelial growth factor receptor (VEGFR)-2 and VEGFR-3, however, all intermediate forms of VEGF-2 binds to VEGFR-3 (Joukov et al. 1996; Jussila and Alitalo, 2002). The bound receptors then undergo auto-phosphorylation reactions that trigger downstream signal transduction cascades (Lee et al. 1996; Olofsson et al. 1999; Veikkola and Alitalo, 1999; Hamada et al. 2000; Veikkola et al. 2000; Makinen et al. 2001; Dixelius et al. 2003; Yong et al. 2005; Shibuya and Claesson-Welsh, 2006). The receptor binding is considered a first step for the pVGI.1(VEGF2) gene product to elicit its subsequent biological functions (Tomanek et al. 2002), which includes endothelial cell proliferation, a key process of angiogenesis (Tille et al. 2001; Persaud et al. 2004).
The objective of the work is to develop suitable potency assays for manufacturing lot release of pVGI.1(VEGF2). Ideally the potency assay must be able to measure clinically relevant biological function(s) of the gene product in a manner that fulfils the regulatory requirements in precision, accuracy, linearity and range (U.S. Food and Drug Administration, 2008). To achieve the goals we undertook the matrix approach by developing a combination of assays where the combined results constitute an acceptable potency assay. The potency assays consist of two methods: the quantitative relative potency assay, which utilizes receptor binding activity of the pVGI.1(VEGF2) gene product as the readout; and the qualitative cell proliferation assay, which confirms the downstream biological function of the expressed VEGF-2 protein by detecting the endothelial cell proliferation promoted by the protein secreted by the pVGI.1(VEGF2) transfected cells.
Here we report the development of both quantitative and qualitative potency assays to support pVGI.1(VEGF2) product release. The characterizations of the assays, including specificity of the qualitative assay, and accuracy, precision, linearity and specificity of the quantitative assay are also described.
Plasmid DNA (pVGI.1(VEGF2)) and pVGI.0 preparations
The plasmid pVGI.1(VEGF2) contained the cDNA of human vascular endothelial growth factor 2 (VEGF-2, also known as VEGF-C) under the direction of a mammalian viral promoter and enhancer. The DNA used in assay development was produced under GMP conditions in various preparations. As negative control plasmid, pVGI.0 had the same plasmid backbone as pVGI.1(VEGF2) but without the VEGF-2 cDNA insert. Both plasmids were purified to high homogeneity (> 90% supercoiled form).
Cell lines and cell culture reagents
The Chinese hamster ovary (CHO)-K1 cell line was purchased from american type culture collection (ATCC) (Catalog number CCL-61) and maintained in RPMI1640 with 10% Fetal Bovine Serum. The 293H cell line was purchased from Invitrogen (Catalog number 11668-019 San Carlsbad, CA), and cultured in DMEM with 10% Fetal Bovine Serum. The primary human lymph microvascular endothelial cells (HLMVEC) were purchased from Cambrex (Catalog number CC-2812 Walkersville, MD), and cultured in EGM-2MV (Cambrex). CHO-5, the reduced serum medium for CHO-K1 cells, was purchased from Kemp Biotechonologies (Catalog number S0185 Frederick, MD) under a license agreement.
Transfection method for receptor binding assay
The CHOK-1 cells were first plated out in 96 well plates at a density of 5 X 104 cells/well. Both plasmid DNA (pVGI.1(VEGF2)) reference standard and test articles were first diluted to 25 µg/ml and then serially diluted with equal amounts of formulation buffer in 96 well dilution plates. The dilutions were made in triplicates. The transfection mix was prepared following the same plate format on a second 96 well plate. For each well, 50 µL of the diluted DNA was mixed with an equal volume of 25 µL/ml lipofectamine-2000™ in Opti-MEM-I®. The mix was incubated at room temperature for 30 min before the addition of 0.1 ml per well of CHO-5 medium. Immediately before transfection, the growth medium in the CHO-K1 cell plate was completely removed, and then 0.175 ml of the DNA/lipofectamine-2000™/CHO-5 medium was added to each well to start the transfection process. The transfection plates were incubated at 37ºC for 4 hrs. For gene expression of the transfected cells, the transfection medium was replaced with 0.15 ml/well of CHO-5 with 2 mM Glutamax (Invitrogen, Carlsbad, CA), 20 µg/ml recombinant human insulin (Sigma, St. Louis, MO), 0.2% FBS and 1% penicillin-streptomycin (Invitrogen, Carlsbad, CA). The plates were then returned to 37ºC incubator with 5% CO2 for 24-48 hrs before the receptor binding assay.
Transfection method for cell proliferation detection
The 293H cells were seeded in 24 well plates at a density of 5 X 105 cells/well a day prior to transfection. Plasmid DNA was serially diluted in Opti-MEM I (Invitrogen, Carlsbad, CA) and then mixed with an equal volume of 1:10 diluted lipofectamine-2000. The DNA:lipofectamine complex was incubated at room temperature for 20 min. Just before the end of the incubation, the medium from every well containing 293H cells was replaced with 0.35 ml of OptiMEM-I. The DNA:lipofectamine-2000 complex was then added to the 293H wells containing the Opti-MEM-I medium. The plates were incubated at 37ºC with 5% CO2 for 4 hrs. At the end of the 4 hrs incubation period, 0.35 ml of DMEM with 20% FBS (Invitrogen) was added into each well. The plates were then further incubated at 37ºC with 5% CO2 for 24 hrs.
The proliferation of the endothelial cells was detected using 5-bromo-2'deoxyuridine (BrdU) cell proliferation kit (CalBiochem Catalog number QIA48-1000) according to the instruction of the manufacturer.
Receptor binding ELISA
High binding ELISA plates (NUNC) were coated with 0.625 µg/ml recombinant human VEGFR-2(Flt-4)/Fc (R and D systems, Minneapolis, MN) at 4ºC overnight. The plates were then blocked with 2% bovine serum albumin (BSA) (Sigma, St. Louis, MO) in phosphate buffered saline (PBS) for 2 hrs at room temperature. Test samples were loaded into plates at 0.1 ml/well and incubated at room temperature for 2 hrs. The plates were washed three times with 0.3 ml/well of washing buffer, which was PBS with 0.05% Tween-20. The binding of VEGF-2 (VEGF-C) to the coated receptor was detected with biotinylated anti VEGF-C antibody (R and D systems, Minneapolis, MN) at room temperature for 2 hrs. After being washed three times with the washing buffer, the plates were stained with horse radish peroxidase (HRP) conjugated streptavidine (R and D systems, Minneapolis, MN) at room temperature for 20 mins. After another three washes with the washing buffer, 0.1 ml of the coloring reagent (R and D systems, Minneapolis, MN) was added into each well. The reaction was stopped with 0.1 ml/well of 1N HCl. The plates were read at A450 and A650 by Spectra Max 190 plate reader (Molecular Devices, Sunnyvale, CA).
Quantitative assay characterizations: accuracy and precision
Accuracy of the quantitative assay was evaluated by pre-diluting the reference standard at 60%, 80%, 100%, 120% and 140% of the test concentration for the assay and determining the recovery (%), which was defined as the following:
% Recovery=observed value/theoretical value x 100%
Precision was evaluated by the relative standard deviation (RSD) of at least three replicates.
Curve fitting and statistics
The dose response curves were modeled using a four-parameter logistic fit Y=((A-D)/(1 + (x/C)B)) + D by the SoftMax®Pro software (v.3.1 Standard, Molecular Devices). For reference standard, the graph was plotted using the A450 - A650 in the Y axis and DNA concentration in the X axis; for the test article (as an unknown dilution sample), the Y axis was also A450 - A650 but the X axis was the dilution factor. Other statistical analyses were performed using the GraphPad Instat Software (v.3.06, GraphPad Software, San Diego, CA) or Microsoft Excel spread sheet.
Quantitative potency assay
The quantitative assay is based on the receptor binding activity of the pVGI.1(VEGF2) gene product. To simplify handling and minimize analyst errors, all operations, including serial dilutions, transfection and receptor binding detection were performed using a 96 well plate format. The transfection mix was prepared by mixing serially diluted pVGI.1(VEGF2) with a fixed amount of transfection reagent lipofectamine-2000™. The transfection mix was then transferred from the 96-well preparation plate to the corresponding location on the 96-well tissue culture plate containing the host cells to initiate transfection. The transfected cells were incubated for 24-48 hrs to allow sufficient time for VEGF-2 expression. The conditioned medium from each well was then collected and transferred to the corresponding location on the 96-well ELISA plate coated with a soluble VEGFR-3 receptor. The binding of the secreted VEGF-2 protein to the ELISA plate was detected by biotinylated anti-VEGF-2 antibody, and subsequent wash and incubation steps with horseradish peroxidase and coloring substrates. The raw data were analyzed by Softmax Pro® Version 3.0.
The A450 dose response curve (subtracted with the background noise by A650) was a function of DNA concentration used in the transfection (Figure 1a). The coefficient of determination (R2) was generally above 0.99 for the reference standard. The fitness of the curve was further examined by residual analysis, where the differences between the observed value and the theoretical value were plotted against the test concentrations. Visual inspection indicated that no obvious bias for any of the concentrations within the range (data not shown). The potency of unknown test article was then determined by comparing the curve of the unknown samples to that of the reference standard on the same plate without further data transformation.
The assay was characterized by accuracy and precision in the range of 60%-140% of the test concentration, which corresponded to 15-35 µg/mL of pVGI.1(VEGF2). The recovery ranged from 86.7% to 116% for 60%, 80%, 100%, 120% and 140% of target pVGI.1(VEGF2) concentration. The largest inter-day RSD was 19.3%. There was a linear correlation between the measured and expected values (Figure 1b) within this range with the coefficient of determination (R2) of the regression line of 0.976. The linearity was further improved with a narrower range of 60-120% of the test concentration, for which the coefficient of determination (R2) was 0.997. The result falls within the commonly-accepted range of about 25% RSD for cell based potency assays (Tuomela et al. 2005). Similar results have been obtained during assay validation after the assay was transferred to a QC laboratory (Waerner et al. 2007).
The assay was specific to the VEGF-2 expression of the pVGI.1(VEGF2) plasmid. When pVGI.0 was used for transfection, the receptor binding signals were not above the background (data not shown). The receptor binding detection system used for the potency assay was specific to the VEGF-2 protein. Other proteins, such as VEGF-A, VEGF-D and PIGF from the VEGF family (Yancopoulos et al. 2000; Jussila and Alitalo, 2002; Tammela et al. 2005) had no detectable binding in the system (Figure 1c).
The assay was capable of distinguishing DNA isolated from a strain different from that of the reference standard (Table 1). The reference standard was isolated from E. coli host DH5α. The same plasmid DNA produced from a non-DH5α host generated a curve with the parameter B of 17.521, which was drastically different from the normal distribution of parameter B of reference standards (3.383 ± 1.066). Therefore, the response curve of the DNA from a new strain was considered non-parallel to the response curve of the reference standard. Furthermore, the reduced potency values were produced when heat damaged or fine needle sheered pVGI.1(VEGF2) was used for transfection (Table 1), suggesting the assay was stability indicative.
Qualitative potency assay
The qualitative biological activity assay was designed to confirm the biological function of the gene product of pVGI.1(VEGF2) as a complementary method of the quantitative potency assay. The principle of the assay was based on the cell proliferation promotion activity of the expressed VEGF-2 protein. Although many proliferation assays have been developed for purified proteins, it would be impractical to incorporate protein purification steps in the analytical method designed for qualifying plasmid DNA. An alternative strategy was to screen commercially available endothelial lines for their response to VEGF-2 protein secreted by the host cells transfected with pVGI.1(VEGF2) in conditioned media. The human lymph microvascular endothelial cells were selected based on the specific response to the VEGF-2 expression of the host cells.
As in the quantitative assay, serially diluted plasmid DNA in the transfection mix produced DNA concentration dependent proliferation responses. The procedure was similar to that of the quantitative assay. After 24 hrs of incubation, the supernatants of the transfected cells were transferred to 96 well plates containing HLMVEC (Kriehuber et al. 2001) after the removal of the host cells. The proliferation of HLMVEC was determined by BrdU incorporation followed by staining with anti-BrdU antibody provided by a commercially available BrdU cell proliferation kit (Calbiochem). The transfection step of the assay was performed in triplicate wells, and supernatants from each well were used to treat triplicate wells plated with HLMVEC. Therefore, at each dilution level, the proliferation results by the plasmid DNA were collected from optical density readings of nine wells. A triplicate mock transfection was included in each plate.
Figure 2a shows typical results of the qualitative potency assay. Proliferation was dependent on the presence of pVGI.1(VEGF2) in the transfection mix; and furthermore, the extent of proliferation was dependent on the concentration of pVGI.1(VEGF2), with an optimum at 1.25 µg/ml of pVGI.1(VEGF2). On the contrary, transfecting host cells with same amounts of pVGI.0 DNA did not result in supernatants that promoted proliferation. We have confirmed that the level of cell proliferation correlated with the level of VEGF-2 expression (Figure 2b) except for the highest DNA concentration tested, where the level of proliferation appeared to drop although the host continued to make large quantity of VEGF-2 protein.
Many factors affect the outcome of the qualitative potency assay. In addition to the age of the HLMVEC, the age of the host cell line, the lot to lot variation of fetal bovine serum used in the medium and the types of the media used to grow the host cells after transfection were all critical for the level of cell proliferation detected by the assay. These factors not only influence the quantity of the VEGF-2 protein expressed in the medium, but also the profile of all protein species present in the conditioned media. For example, during the initial screening exercise, the HLMVEC grown in DMEM based conditioned medium produced larger proliferation signal than those grown in MCDB131 based medium, even though MCDB131 was optimized to support clonal growth of human microvascular endothelial cells (Knedler and Ham, 1987). However, the VEGF-2 expression level, measured by VEGFR3 binding activity, was much lower in DMEM based medium than in MCDB131 based medium (data not shown). Further investigation is required to understand the roles of interactions between various components in conditioned media in promoting HLMVEC proliferation.
In conclusion, the potency assay methods, both qualitative and quantitative assays, fulfil the basic requirements for potency measurement for biological products. The characteristics of the combined assays, considering specificity, sensitivity, accuracy and precision, meet the requirements of product release and stability monitoring.
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text:
Washington Promises Punitive Measures on Houthi Authorities
According to Washington, the Houthis are accountable for recent assaults on Saudi Arabia and the UAE.
They're also accountable for their involvement in prolonging the fight in Yemen, which is considered the world's greatest humanitarian disaster.
US officials at the State Department and the White House have used strong rhetoric in their speeches and diplomatic maneuvers.
A solution for Yemen must be found by international standards, notably UN Resolution 2216.
The Houthis just fired a missile on the city of #Taiz, I'm hearing of several casualties.
These daily attacks are rarely reported, that doesn't make them less true.
So before coming with ideas on how the Saudis need to give concessions to the Houthis, think of the Yemenis first.
— Baraa Shiban براء شيبان (@BShtwtr) January 27, 2022
A US State Department spokesman told Asharq Al-Awsat the review of such a decision continues, due to the bad conditions in Yemen and the inability to negotiate a truce.
Truce and Political Dialogue
Even though he didn't expand on the review process, he stated work is being done to improve Yemen's humanitarian situation.
An unnamed official said the US identified no good role for Iran in Yemen.
"For this aspect, we have advocated communication between it and the nations of the region for the sake of stability and security," the spokesperson told Asharq Al-Awsat.
The US government will continue to stand with regional allies against "upsetting Houthi attacks" and hold the Houthis accountable for their heinous deeds.
The official cited US Special Envoy to Yemen, Tim Lenderking's efforts to generate international agreement for an early and comprehensive ceasefire and political settlement.
Both Saudi Arabia and the Yemeni authorities backed a truce and political dialogue.
The source also complimented Oman for its "decisive and proactive regional role" in the Yemeni situation.
The US' Stance
US State Department spokesperson Ned Price said the government is "closely scrutinizing" the Houthi strikes.
We condemn the overnight attack by the Houthis on the UAE and Saudi Arabia. These attacks, as well as recent airstrikes in Yemen, represent a troubling escalation that only exacerbates the suffering of the Yemeni people. We call on all parties to commit to a cease fire. pic.twitter.com/OZj9S5vVsz
— Ned Price (@StateDeptSpox) January 24, 2022
Price stated "we will always stand with the UAE and Saudi Arabia."
Price said at a news conference that Washington would keep working with friends and partners to hold Houthis responsible.
"We will keep holding the Houthis accountable. We'll accomplish it in various ways. We've already utilized a few tools, and I expect we'll use more in the next days and weeks," Price said.
"To that end, we will not desist in our efforts to identify Houthi commanders and entities participating in military offensives that endanger people and regional security," he stressed.
Norman Roule, a former CIA officer and current senior counselor to the "United Against Nuclear Iran" group, said Washington intended to compel the Houthis to return to genuine dialogue for a peaceful settlement.
Houthi attacks and comments show a "terrorist group."
Roule told Asharq Al-Awsat Iran is aiding the Houthis. As a result, the gang has repeatedly denied diplomatic efforts and assaulted scores of innocent sites.
Relisting the Houthis as a terrorist organization will not be easy.
Several US government members feel the renaming will delay humanitarian efforts and prevent a political settlement.
"I don't think the government will be able to do anything in that direction," Roule added, citing the Houthis' willingness to explore a political settlement after military defeats.
He said the US would keep sending Lenderking to investigate political solutions.
Roule said the Iranians have always isolated nuclear negotiations from regional and missile problems.
"Maybe Iran's assaults on the UAE and Saudi Arabia are unrelated. However, the scale of the strike and the complexity of the weapons deployed by the Houthis necessitate Iranian involvement," Roule said.
He said the UAE and Saudi Arabia are still relatively safe, despite recent incidents. The air defense forces and security agencies are skilled, but the Iranian-backed Houthi strikes threaten the millions of expatriates residing in these nations.
"The Houthi strikes are an attack on the worldwide community," Roule noted.
Abdul-Rahman
SHOCKING: FBI Violating Free Speech
Biden Putting Air Travelers in Danger After This Controversial Decision
WATCH: Small City in Vermont Has Soaring Crime After Defunding Police
War About To Break Out Between India and China
Senator Joe Manchin Weighs in On Switching Parties
SICK MURDER CASE Shows How Far US Justice Has Fallen
Biden Made Unacceptable Compromises to Release Griner From Russia
IRAN NUKES: How Close Are We To A Nuclear Iran?
Kari Lake in Arizona CHALLENGES Midterm Election Results

redpajama_set_name: RedPajamaCommonCrawl


A seating cushion to improve posture and stop fidgeting.
Wobble cushions (sometimes called disc cushions or instability cushions) are small round inflatable cushions, strong enough to sit or stand on. Made from flexible PVC free & latex free material and measuring 35cm. The cushion can be sat on to create instability (a wobble!) encouraging the user to engage their back and core muscles. The cushion can be used on a chair or on the floor. They are strong enough to be stood on to build core strength and practise balance.
Sitting on a wobble cushion is similar to sitting on a therapy ball – only smaller, portable and more convenient. The instability engages the core muscles improving posture and core stability. The wobble effect makes sensory seeking children focus on how they sit, reducing fidgeting and helping to improve focus on the task in hand.
They have a small valve in the back so the cushion can easily be inflated by blowing directly into the valve (they don't take much to blow up) or using a bike pump or any other type of pump. The cushion can be inflated as much or as little as required. Don't over inflate the cushion – it requires enough air to make it wobble when you sit or stand on it.
Maximum weight 200kg / 32 stone.
Our wobble cushions are smooth on one side and have raised dimples on the other for additional sensory input. Wobble cushions are not meant to be too comfortable! They are meant to be unstable and slightly uncomfortable to make sitting down a physical exercise – they will never be as comfortable as slouching!
Sitting on a wobble cushion engages the core muscles helping to improve core stability – so it is effectively an exercise. It is recommended that the cushion is not sat on all day – about ½ hour at a time is long enough. Put it to one side for a while and then use it again at regular intervals throughout the day.
The cushion can also be great fun to stand on to build muscle tone and help balance and concentration. Stand on both legs, one leg, make a tree shape etc…..be creative and have fun!


Ticket is for day long car-pool tour of the LA River with your guide Jenny Price. Food and drink not included. If you'd like to be a designated driver during the tour, we will be eternally grateful and shower you with love.
Difficult as it is for Angelenos or anyone else to believe, the Los Angeles River was once a very real flowing river, along the banks of which the city was founded. In 1781, 44 people (Los Pobladores) traveled from Sonora, Mexico to settle along its West bank. For more than 100 years, this 51 mile long river was the sole source of water for the city that grew around it.
Have you ever *really* seen the LA River? Do you ACTUALLY know anything about it? Sure, maybe you've passed a concrete slab with an LA River sign... but have you seen where it flows? Do you know the *true* stories of the river's past and present beyond the fictional details you remember from seeing Chinatown? And what about its future? Sure, everyone in Los Angeles has seen the LA River, and many have heard that it's being revitalized. But who knows where and what exactly is happening on its banks? There's much more to the story than meets the eye as you pass from a speeding car while commuting.
On this tour, we'll walk and drive along the river, as we talk about its central role in the city's history and the necessity of the ambitious revitalization to LA's future.
The tour convenes at the Los Angeles River Center & Gardens (570 W Ave. 26, near the 5/110 - DIRECTIONS HERE) where we will form carpools, and stops at the gorgeous Sepulveda Basin (San Fernando Valley), new parks in the Glendale Narrows (across from Griffith Park), the historic Arroyo Seco confluence, the Los Angeles State Historic Park (aka the Cornfield) in Chinatown, and the industrial downtown location that everyone's seen in movies and on TV.
We'll eat lunch at an excellent taqueria en route (or you can bring your own) and end with an optional stop at the world's best cream puff shop, in Little Tokyo.
Led by Jenny Price. Group rate available.
Bring snacks and water and shoes that can get just a little wet (optional, but way more fun!). Dogs welcome. Spaces limited.


Wanneer wilt u in Sofia verblijven?
by Hostelworld customers has a unique setup in a totally renovated 19th century house.
Welcome to Sofia's friendliest, somewhat-artisticist, most warmly-welcoming little hostel!
Convenient and welcoming, Hello Sofia is a small boutique guesthouse in Sofia Downtown, surrounded by the most famous city attractions.
We offer variety of rooms to suit each situation: Singles, Doubles, Triples, Quadruple, Private rooms and dormitories.
We also have apartments for 2, 4, 6 and 8 people.
Peter Pan lodging is for solo travelers or couples, families and friends looking for budget and affordable lodging, but who will not sacrifice balanced quality and fair service.
H2O Hostel is conveniently located in the heart of the city. Especially suitable for young travellers.
Apartment of 18 sq.m. with personal bathroom, towels, TV, wi-fi, office desk, air conditioned, tea, coffee at disposal, welcome set of bottle of water, bottle of wine and fruits.
Canape Connection Guest House - Created to inspire travelers!
We are centrally located, about a kilometer away from the Sofia Central Railway and Central Bus Stations and about 10 km away from the Sofia Airport.
Hostelworld.com brengt je online boekingen bij de beste selectie van hostels in Sofia . Lees recensies van klanten van Sofia hostels, jeugdherbergen te bekijken op de kaart van Sofia en gebruik maken van speciale aanbiedingen, zoals gratis SMS bevestiging. En waarom bekijkt u onze keuze voor de beste dingen die te zien en te doen zijn in Sofia terwijl je er bent. Hostelworld.com - de enige gids voor Sofia backpackers hostels.


text:
package com.jdroid.android.sample.ui.http;

import android.support.v4.app.Fragment;

import com.jdroid.android.activity.FragmentContainerActivity;

public class HttpActivity extends FragmentContainerActivity {

	/**
	 * @see com.jdroid.android.activity.FragmentContainerActivity#getFragmentClass()
	 */
	@Override
	protected Class<? extends Fragment> getFragmentClass() {
		return HttpFragment.class;
	}
}

redpajama_set_name: RedPajamaGithub


text:
Guided Search: Hospitality and Promotion, North America, Philadelphia-Area Residents/Citizens
2022 Preservation Achievement Awards: TOGETHER AGAIN
6/8/2022 5:00pm - 8:00pm
600 N Broad Street
https://www.preservationalliance.com/202...
Join the Preservation Alliance in celebrating their annual Preservation Achievement Awards.
Historical Preservation / Archaeology
Philadelphia Taco Festival
7/31/2021 12:00pm - 8/1/2021 8:00pm
1100 Pattison Ave
Philadelphia, NJ 19148
https://www.upcomingevents.com/e/philade...
The 5th annual Philadelphia Taco Festival is taking over South Philly on Saturday & Sunday July 31 & August 1, 2021! Get ready for an action-packed day filled with a variety of tacos, margaritas, delicious indulgences, live music & entertainment and tons of taco themed activities! Come hungry and get ready for the Philadelphia Taco Festival!
Festivals/Parties
Walnut2Walnut River Challenge
9/12/2021 9:00am
https://phillyseaport.org/Walnut2Walnut
Are You Up for the Walnut2Walnut River Challenge? Join us on Sunday, September 12!
The excursion begins at Schuylkill Banks and ends at Independence Seaport Museum's boat basin.
The challenge course offers spectacular views of the city, while traveling almost 15 miles along both rivers. The Walnut2Walnut River Challenge is suitable for experienced paddlers looking for a new way to get in touch with Philadelphia's waterways.
11th Annual Virtual Shofuso Cherry Blossom 10k/5k: Spring Starts at Shofuso
https://runsignup.com/Race/PA/Philadelph...
Healthy Kids Running Series Spring 2021
4/18/2021 3:29pm - 5/23/2021 3:29pm
Subaru Park - Philadelphia Union 1 Stadium Drive. Chester, PA US 19013
Chester , PA 19013
https://runsignup.com/Race/PA/Chester/He...
T.H.E Summit
https://sthm.temple.edu/us-asia-center-f...
Due to the ongoing pandemic, Tourism, Hospitality, Events (T.H.E.) higher education institutions across the world are grappling with unprecedented disruption. This trying, but exciting time has caused the need for rapid, dramatic transformation in the T.H.E. higher education industry.
Solas 2021
https://welcomingcenter.org/solas/?mkt_t...
Join us on Wednesday, March 3rd at Solas 2021 as we celebrate the important work that YOU have helped make possible!
This year will be the first virtual Solas Awards and while it will be different from past events, we are excited to be able to welcome everyone to join us in experiencing The Welcoming Center. During Solas 2021, you will have the opportunity to virtually network with others in The Welcoming Center community, learn more about the impact your support has on immigrants in Philadelphia, and watch a live conversation with our 2021 Solas Award honoree, Dr. José Ramón Fernández-Peña hosted by our friend and former colleague, Amanda Bergson-Shilcock.
Central America/Carribean
Founder of Cossist
A technology enthusiast who is passionate about making life better for community-at-large. Many years of Big 5 consulting experience and currently working to provide a better experience for Tourists and Vacationers. Will realize this vision through Cossist, an unavoidable global mobile platform for Tourists and Businesses who serve to the Tourism industry. We serve Hotels Museums Vacation Rentals Airlines Tour Operators Local Experience Providers It is true that we see a slump due to Covid-19. But Travel and Tourism have to return.
People Type:
Business Person/Executive
Beer: The Fuel of Ancient Civilization
Science History Institute 315 Chestnut Street
https://www.sciencehistory.org/event/bee...
Long before beer became a pub favorite, it gave our nomadic ancestors fuel for survival. It was a safer source of hydration than water, provided ample nutrition, and even helped early humans make the transition from a nomadic lifestyle to village life. Evidence of brewing dates back to more than 14,000 years ago, and while we may not appreciate the taste of the ancient version, our ancestors brewed it and celebrated it much like we do today.
GlobalPhilly 2019 Events
CiderFest at the Historic Houses of Fairmount Park
9/21/2019 12:00pm - 5:00pm
Woodford Mansion 3400 West Dauphin Dr.
https://fairmountpark.ticketleap.com/cid...
Spend a sparkling autumn afternoon on the grounds of six historic houses in Fairmount Park sipping on cider samples from some of the best cideries Pennsylvania has to offer. Enjoy live music, pay as you go food trucks, food samples, and activities to complement your tastings.
: Hospitality and Promotion
: North America
: Philadelphia-Area Residents/Citizens
: all » Hospitality and Promotion
Food/Cuisine (52)
Indoor/Outdoor Recreation (30)
: all » North America
American (United States) (82)
Virgin Island (1)
: all » Philadelphia-Area Residents/Citizens


text:
Reimagining how we create relationships and communities.
EssaysSpeakingContact
You, Robot
A couple of years back, Jay Ruskey, developer of the "Stak Block" — a wonderful way to reduce a huge source of worldwide carbon emissions by turning waste rice straw into building blocks (BTW, someone needs to go and make a few million dollars selling this to the Indian government) — told me of a conversation he had when presenting his technology in China. From the US perspective, a major selling point was that the machinery they had developed, and the newer machinery that was under development, would increasingly reduce the manpower needed in the production of Stak Blocks. The Chinese representative protested, saying, "No, no, we want more people needed to do it, not less."
Where the US view was looking to more efficiency and profits, the Chinese view was looking to social stability.
Last week I attended a talk given by General Stanley McChrystal at the Westmont University President's breakfast. It was a thought-provoking talk.
The General touched on a fact that we are beginning to see play out more and more: that technology, in various ways, is taking over jobs traditionally done by people. It is a known fact that most jobs that have been lost in the US have not been lost to off-shoring, but to robots, efficiency and technology.
And it's worth noting that these changes are only just beginning to pick up steam.
For instance, think of any job that involves driving: truck drivers, bus drivers, train drivers, etc. Estimates are that those jobs are all going away, perhaps within ten years. (That is approximately 6 million truck and bus drivers.)
I think many people (yours truly included) have found refuge in the idea that their job could not easily be done by a robot (or more correctly, Artificial Intelligence).
But what happens when we are the robots? Because I believe that is where we are heading.
THE FOLLOWING IS NOT A POLITICAL STATEMENT. (How's that for the sign of the times!)
So our President made a speech to Congress recently. I think everyone agreed it was his most balanced, reasoned speech to date. Some of his previously most ardent fans worried that he was wavering on important issues; some of his bitterest opponents found hope in his words. (That may be the bell-weather for a good speech! :) What I found amazing is that the next day Van Jones completely reversed his opinion of Donald Trump based on that speech. I wanted to ask him, "So your recommendation is that our opinion of people should fluctuate based on how they acted for an hour yesterday, not on what they did for the previous year?"
What was glaring to me was that, in this "Kardashian" world of ours, we are more and more open to having our views and opinions moulded for us, and that the timeframe for our malleability is becoming very short.
The warm up act has been a general acceptance of product malfunctions in our current "technology age" that was never accepted in the engineering age that preceded it. Witness the almost blasé acceptance of fatal shortcomings in the Tesla self-driving technology vs. the backlash that was the exploding Ford Pinto or the crashing Chevrolet Corvair. And as AI gets more involved in our lives, with algorithms deciding which Google results pop up and what movies, songs or clothes are recommended for us, one might legitimately ask, "Who is in the driving seat, me or the AI?"
Is AI moving toward us, or are we moving toward it?
In the world of starlings (birds), they call it "murmuration" — when a flock of starlings change direction as one. They are not thinking the course changes; it's an automatic subconscious process. Similarly, our technology is encouraging us to hand over more and more of our cogent work to be computed external to our own brains. I know Netflix says their recommendations are based on our preferences, but are they? And is that all they are based on? — My son tells me that Dr. Dre is regularly in his "recommended" list on Apple Music. Yet he has never listened to Dre on Apple; but Dre is a business partner of Apple…
Remember, a small rudder can turn around a super-tanker, given enough time, and the change of direction is so gradual that in the open-sea you might not notice that it happened. Yet you end up chugging in the opposite direction to your previous course.
Do you really know what "being you" means? Because if you don't, then how could you possibly know whether "who you are" is being manipulated?
In 20 years, I probably won't need to influence you directly, I will just need to influence the person in charge of the AI that influences you — if it even is a "person" at that point…
Some say that in 50 years, most jobs will be gone and the "basic living wage" will be enacted. And most of our choices will be made for us. At which point the question will be:
What is the point?
And I must admit that there, I'm rather lost for an answer.
– Simon Dixon
Recent Essays
Evolve or revolve
Behavior change can come about from many motivators, including negative ones. However, as motivation wanes, the behavior change will go with it. If you're interested in true, long-lasting behavior change, the key is in identity.
My Way is the High Way
The way we imagine our personal and national identities makes us liable to act and think in certain ways and, just as importantly, makes us liable to interpret other people's actions through the prism of our own imagined identity – not through theirs.
See all essays →
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©2020–2022 Simon Dixon. All rights reserved.


text:
Томаш Соареш Дабо (; 20 октября 1993, Бисау) — гвинейский и португальский футболист, защитник сборной Гвинеи-Бисау.

Карьера

Клубная

Родился в Гвинее-Бисау, но в детстве переехал в Португалию. Воспитанник академии "Браги". Был одним из ведущих футболистов молодежной команды клуба и входил в состав молодежной сборной Португалии. Однако за основу "Браги" Дабо сыграл всего девять игр. В 2014 году он перебрался в "Ароку", но и в этой команде он не стал ведущим игроком. В 2017 году защитник оказался в коллективе третьей лиги "Фаренсе". В сентябре того же года защитник перебрался в Молдавию, где подписал контракт с Кишинёвской "Дачией". Однако за новый клуб он провел всего две игры. В декабре Дабо вместе с четырьмя другими легионерами "Дачии" покинули команду.

В сборной
Зимой 2017 года Томаш Дабо принял предложение федерации футбола Гвинеи-Бисау и решил перейти в сборную своей исторической родины. За нее он отыграл на Кубке африканских наций в  Габоне. Защитник участвовал во всех трех матчах с участием национальной команды.

Примечания

Ссылки

Футболисты Гвинеи-Бисау
Футболисты Португалии
Игроки сборной Гвинеи-Бисау по футболу
Игроки ФК «Брага»
Игроки ФК «Брага» B
Игроки ФК «Арока»
Игроки ФК «Фаренсе»
Игроки ФК «Дачия» Кишинёв
Игроки ФК «Металоглобус»
Игроки ФК «Риети»
Игроки ФК «Середь»

redpajama_set_name: RedPajamaWikipedia


text:
Tag: Rose Byrne
The first minutes of Irresistible tell us the setting is "Rural America," followed by "Heartland, USA." One more intrusive line of text would establish the overkill as a joke. The opening credits might also have noted how the role of Deerlaken, Wisconsin is played by Rockmart, Georgia. Unfortunately, beloved writer/director Jon Stewart never becomes that acidic or… Continue reading Irresistible
Categorized as Movie Reviews Tagged Comedy, Filmed in Atlanta, Filmed in Georgia, Jon Stewart, Rose Byrne, Steve Carell
Mia (Tiffany Haddish) and Mel (Rose Byrne) are lifelong best friends that have formed their own cosmetics company together. Suddenly, Claire Luna (Salma Hayek), a millionaire cosmetics magnate, enters the picture, not only with an offer to expand their brand but to wipe out Mia and Mel's huge $500K debt. A generic, boilerplate plot we've… Continue reading Like a Boss
Categorized as Movie Reviews Tagged Comedy, Filmed in Atlanta, Filmed in Georgia, Rose Byrne, Salma Hayek, Tiffany Haddish
January Georgia-Filmed Release/Premiere Dates
Known as Limited Partners when it was filming in Atlanta, the comedy Like a Boss with Tiffany Haddish, Rose Byrne and Salma Hayek will run in theaters beginning Jan. 8… Bad Boys for Life with Will Smith and Martin Lawrence opens Jan. 16… Vietnam War movie The Last Full Measure, starring Sebastian Stan, Samuel L.… Continue reading January Georgia-Filmed Release/Premiere Dates
Categorized as Call Sheet Tagged Filmed in Atlanta, Filmed in Georgia, Martin Lawrence, Rose Byrne, Salma Hayek, Stephen King, Tiffany Haddish, Tyler Perry, Will Smith
Georgia Proves Irresistible for Stewart, Carell
Smug, unfunny dimwit Jon Stewart will be directing Steve Carell in the political satire film Irresistible, shooting in Georgia from April 22 through the end of May. Rose Byrne is also said to be starring in the story of a race for mayor between a slick Democrat and the incumbent Republican in a small, right-wing… Continue reading Georgia Proves Irresistible for Stewart, Carell
Categorized as Call Sheet Tagged Filmed in Georgia, Jon Stewart, Rose Byrne, Steve Carell
November Georgia-Filmed Release/Premiere Dates
Nobody's Fool, which filmed in Atlanta as The List, hits theaters on Nov. 2. Tyler Perry wrote, produced and directed, while Tiffany Haddish, Tika Sumpter and Whoopi Goldberg star… The Front Runner, with Hugh Jackman as Gary Hart, gets a limited release on Nov. 6, expanding on Nov. 16 and then a wide release on… Continue reading November Georgia-Filmed Release/Premiere Dates
Categorized as Call Sheet Tagged Filmed in Atlanta, Filmed in Georgia, Hugh Jackman, Mark Wahlberg, Rose Byrne, Tiffany Haddish, Tika Sumpter, Tyler Perry, Whoopi Goldberg
November Georgia Film/TV Quick Cuts
Shot with DXO ONE Camera Processed with VSCO with m5 preset
Taylor Black (Midnight, Texas, pictured) has joined the cast of The Banker, which is currently filming in the Atlanta area. Set in the 1950s, the fact-based story revolved around two black entrepreneurs who arrange for a working-class white guy to pose as the head of their thriving banking empire while they pretend to be a… Continue reading November Georgia Film/TV Quick Cuts
Categorized as Call Sheet Tagged Filmed in Atlanta, Filmed in Georgia, Lil Yachty, Rose Byrne, Salma Hayek, Tiffany Haddish, Tyler Perry
Tiffany Haddish Comedy to Produce in Atlanta
The suddenly unavoidable Tiffany Haddish (pictured) will star in Limited Partners, a comedy feature from Paramount for which Rose Byrne has been in talks to co-star. Miguel Arteta (Chuck & Buck) will direct the tale of two besties who build a successful company, then find their friendship tested when the venture is met with a lucrative… Continue reading Tiffany Haddish Comedy to Produce in Atlanta
Categorized as Call Sheet Tagged Filmed in Atlanta, Filmed in Georgia, Rose Byrne, Tiffany Haddish
Beatrix Potter is a punk rocker in this updated revenge epic that blends CGI animation and live action in its tale of the rebellious raids on McGregor's garden. Narrated by Margot Robbie, Peter is voiced by James Corden, whose manic antics cause the old man McGregor to die – after all, the guy did turn Peter's… Continue reading Peter Rabbit
Categorized as Movie Reviews Tagged Animation, Comedy, Margot Robbie, Rose Byrne, Sam Neill, Sia
Wahlberg, Byrne Making Instant Family
Mark Wahlberg and Rose Byrne will star in the Paramount Pictures comedy Instant Family, which looks to start filming in Atlanta by March. They'll play a couple at their wits end after adopting three outta control kids through the foster care system. Director Sean Anders co-wrote the script with John Morris, with whom he collaborated… Continue reading Wahlberg, Byrne Making Instant Family
Categorized as Call Sheet Tagged Filmed in Atlanta, Filmed in Georgia, Mark Wahlberg, Rose Byrne


text:
Disarm and Conquer
By Miguel A. Faria, Jr., M.D. - November 26, 2012
Before a standing army can rule, the people must be disarmed; as they are in almost every kingdom of Europe. The supreme power in America cannot enforce unjust laws by the sword; because the whole body of the people are armed, and constitute a force superior to any band of regular troops that can be, on any pretence, raised in the United States. Noah Webster, An Examination of the Leading Principles of the Federal Constitution, 1787
DATELINE: Warsaw, Poland, 1943
In the Spring of 1943, the inhabitants of the Warsaw Ghetto, having become aware the Nazis were deporting the remaining Jews to the gas chambers of Treblinka, took up arms, whatever they could nd, and rebelled against the German occupiers. These determined insurgents had only homemade Molotov cocktails and a handful of small arms, revolvers, pistols, and a few military or hunting ri es. It took vastly superior Nazi forces to subdue the rebels, and the Germans suffered up to 300 casualties in pacifying the city.(1)
There were several other armed struggles and Ghetto uprisings, and the Poles and Warsaw remained a serious problem for the Germans during the remainder of the war. The Polish Jews continued to procure whatever arms they could find to defend themselves in their struggle, repeatedly disrupting the timetable of the Nazi high command and the German war effort.
The last major Polish uprising in August 1944 was to be launched in concert with the liberating Soviet Red Army. But the advancing Red juggernaut suddenly and inexplicably halted. Advised by General Georgi Zhukov, Stalin rejected the appeals of the Western allies to assist the insurgents. For two months the courageous Poles fought the Nazis in heroic, urban warfare without any assistance. Stalin had simply halted the advance of the giant Red Army to allow the Germans to destroy the non-communist freedom fighters: 200,000 Poles perished and 800,000 were deported to the death camps.(2) Warsaw was erased by orders of Adolf Hitler, and the Soviets at the outskirts of the city did nothing.(3)
DATELINE: Budapest, Hungary, 1944
During the summer of 1944 in the ongoing carnage of World War II, German troops were retreating on multiple fronts. Hungarian allied troops were defending the motherland from the onslaught of the Soviet Red Army, yet in the midst of the chaos, nearly 500,000 Hungarian Jews were rounded up and deported to the Nazi death camp Auschwitz–Birkenau, where 90% of them were summarily exterminated in the gas chambers. A reign of terror ensued for those who remained in Hungary; thousands of them were tortured, robbed, or murdered; Jewish women were raped; property was looted or con scated.(4) These atrocities were carried out with a minimum of Nazi troops. While seated in a restored synagogue in Budapest a few months ago, a friend of mine listened in utter disbelief as this story was recounted. How could this have happened? How could these atrocities have been carried out with only a minimum of Nazi troops? The Hungarian state had outlawed the possession of firearms for its citizens. Simply put, this happened without resistance because the Hungarian people had been disarmed.
DATELINE: Havana, Cuba, 1959
And here is a story with which I am intimately familiar. After the triumph of the revolution in 1959, Fidel Castro reneged on his promise to establish democracy in Cuba. In response, the Revolutionary Directorate (RD), the opposing, non-communist group, threatened to renew the insurrection against the Castro brothers' revolutionary government. To back their demands with substance or perhaps even expecting a showdown, the RD seized arms from the San Antonio de los Baños Air Force Base in the western part of the island.
Fidel Castro reacted characteristically in a speech at the Maestre Barracks of San Ambrosio, artfully commanding the situation by accusing the RD of seizing the arms and then, in a masterpiece of rhetoric, asking: Armas ¿para que? ¿Para luchar contra quién? ¿Contra el gobierno revolucionario, que tiene el apoyo del pueblo? ("Guns, for what? To fight against whom? Against the Revolutionary government that has the support of the people?")
Fidel Castro defused the situation and neutralized the defiance of the RD. Shortly thereafter, Castro commenced his long-term campaign to disarm not only his confreres in the Revolutionary Directorate who had not joined him, but also, in due time, all Cubans. A 100,000-member "militia" was organized to seek out the political opposition and actively disarm it.
Unfortunately, they had a well-drawn blueprint to follow  the local firearm registration lists that the former dictator, Fulgencio Batista, had established. All the milicianoshad to do was to seize the registration (licensing) lists and then go door to door searching for and confiscating firearms. The militia tried to disarm my father, a physician, and the episode is recounted in my book Cuba in Revolution  Escape from a Lost Paradise (2002).(5)
During the Batista dictatorship (1953-1958), Cubans were free to leave the country with family, personal possessions, and their wealth, anytime they wished, but not so in communist Cuba.
Since Fidel Castro took over the island in 1959, the best figures we can glean are that between 30,000 and 40,000 Cubans were either executed en los paredones de fusilamiento(on the firing squad wall) or died at the hands of their communist jailers. Between 1960-1965, hundreds of anti-communist rebels, many of them former RD members, went back to the hills of the Escambray Mountains in my native Las Villas province to fight the new communist dictatorship of the Castro brothers. The mostly peasant insurgents, who were defending their lands, lacked sufficient weapons to overcome the well-armed, communist Cuban military forces. Thus, most of these peasant insurgents were annihilated by 1966.(4-6)
Moreover, between 1960 and 1993, 36,000 Cubans perished at sea trying to escape the Castro brothers' communist inferno. If we include all of those who died escaping the regime, those who were shot or died in custody, the figure well exceeds 100,000. In fact, the late scholar Armando Lago arrived at a death toll of 105,000 victims directly attributed to the regime of Fidel Castro.(4,5)
Countless thousands of other Cubans have died indirectly as a result of Fidel Castro's collectivist policies, unspeakable privations, malnutrition, and the general desolation of a once prosperous island  the island Christopher Columbus called the "Pearl of the Antilles."
LESSONS to be LEARNED
One day, as it happened in the Soviet Union, we will learn the truth, and more accurate numbers will be available, recording the full extent of the brutality and destruction wrought by communism in Cuba. One reason this happened and Cuba today remains a totalitarian state despite the collapse of Soviet communism is because the Cuban people were disarmed.(4-6)
In Part III of this essay will conclude with the troubling historical relationship of civilian disarmament to the development of tyrannical governments and genocide. In a better light, Part III of this essay will also end with a discussion of how armed citizens can preserve their freedom and republican form of self-government, more commonly referred to as representative democracy.
1. Edelman M. The Ghetto Fights: Warsaw, 1941–43. London, England, Bookmarks Publications, 1990.
2. Blejwas S. A heroic uprising in Poland and Kojder A, Wegierski M. The Role of Poland and the Poles in World War II. The Polish American Journal, August 15, 2004. Available from: http://www.polamjournal.com/Library/APHistory/Warsaw_Uprising/warsaw_uprising.html.
3. Wdowi ski D. And We Are Not Saved. New York, NY, Philosophical Library, 1963.
4. Faria MA Jr. Cuba in Revolution  Escape From a Lost Paradise. Macon, GA, Hacienda Publishing, 2002. http://www.haciendapub.com/articles/interview-dr-miguel-faria-part-i-myles-b-kantor.
5. Clark J. Cuba, Mito y Realidad  Testimonios de un Pueblo. Saeta Ediciones, 2ndedition, 1992.
6. Encinosa E. Cuba en Guerra  Historia de la Oposicion Anti Castrista 1959 1993. Miami, FL, The Endowment for Cuban American Studies, 1994.
Miguel A. Faria, Jr., M.D. is Clinical Professor of Surgery (Neurosurgery, ret.) and Adjunct Professor of Medical History (ret.) Mercer University School of Medicine. He is an Associate Editor in Chief and World Affairs Editor of Surgical Neurology International (SNI), and an Ex-member of the Injury Research Grant Review Committee of the Centers for Disease Control and Prevention (CDC). 2002-05; Former Editor-in-Chief of the Medical Sentinel (1996-2002), Editor Emeritus, the Association of American Physicians and Surgeons (AAPS); Author, Vandals at the Gates of Medicine (1995); Medical Warrior: Fighting Corporate Socialized Medicine (1997); and Cuba in Revolution: Escape From a Lost Paradise (2002).
This series of articles is based on the two-part essay entitled "America, Guns and Freedom" originally published by Surgical Neurology International(SNI), an on-line, peer review journal for neurosurgeons and neuroscientists. The illustrated essays were published in October and November 2012, but have been edited for an American audience and published for readers of GOPUSA in a three part series.
http://www.surgicalneurologyint.com/text.asp?2012/3/1/133/102951
Guns and Freedom: Historical Vignettes on Civilian Disarmament
Posted by JWRoss at 10:20 AM
Concealed Carry -- Only for a few?


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HomeNewsLyme Regis NewsVolunteer lifeboat crew reviews successful year
Volunteer lifeboat crew reviews successful year
THREE rescues due to be screened by the BBC, plus a special Christmas film featuring the crew and their families, and an all-time record of £40,000 raised during Lifeboat Week – these were the highlights of 2019 for the volunteers of Lyme Regis lifeboat crew.
The man at the helm of the lifeboat station, operations manager Nick Marks, said: "Once again teamwork and training by the crew and tremendous fundraising efforts by the Lifeboat Supporters Group have contributed to a very successful year for our volunteers."
The rescue of two 14-year-old boys being swept out to sea from the River Axe in June is due to be seen in the BBC's 'Saving Lives at Sea' series early in the New Year.
Video using a helmet camera worn by a crew member shows the boys' dramatic rescue as they clung to each other at sea.
Senior helm Tim Edwards, who was aboard the lifeboat when the two schoolboys were rescued and who features in the BBC film of the incident, said: "It was a generally a safe and successful 2019 but the rescue of those boys will remain with me for many a year.
"It was a wonderful end to an incident that could have been so different. It's what makes all our volunteer work and training so worthwhile."
Footage shot during two other 'shouts' is also expected to be featured in the series later this year. They are the rescue of two ex-professional sailors whose dinghy capsized and the successful search for a woman trapped on cliffs with her two dogs.
The RNLI also released a special festive film featuring the Lyme Regis crew and their families just before Christmas.
The Lyme Regis lifeboat crew answered 33 emergency calls during 2019, a third of them to people cut off by the tide. The first two 'shouts' of the year were to such incidents in the Charmouth area.
Mr Marks said: "We always urge people to check tide times and general conditions when out walking.
"Although the year was largely successful there was also sadness when an extensive search by our crew, Exmouth lifeboat and the coastguard helicopter failed to find a man who, it transpired, died in the Freshwater Bay area."
Lyme Regis Regatta & Carnival raises more than £12,000
25th August 2021 Francesca Evans
THIS year's Regatta and Carnival Week has been hailed a huge success with record numbers attending events and more than £12,000 raised for local good causes. […]
Damage revealed after Rousdon Bakery blaze
21st January 2020 Francesca Evans
UP to 10 fire engines tackled a blaze at the Rousdon Village Bakery on Tuesday morning. […]
Cancer Research UK finding new ways to fundraise during pandemic
WITH events cancelled throughout 2020, the Lyme Regis branch of Cancer Research UK has been coming up with alternative ways to continue its fundraising efforts in recent months. […]
Why is Lyme Regis published in Freshwater Lifeboat Station photograph?
Francesca Evans says:
This is a photo of Lyme Regis lifeboat crew.


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Located just east of the town centre of Denmark, the WA College of Agriculture is a 560 hectare working farm and educational facility for Year 10, 11 and 12 students, offering a wide variety of studies with an agricultural focus. WA College of Agriculture – Denmark operates at full capacity with 130 students (91 residential and 39 day students) and 60 staff members which support three main areas: the classroom program, the farm program and the residential program. The College is considered one of the leading lights in the achievement of educational and training outcomes for young men and women in a diversity of agricultural related fields.
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From Israel: "Is There a Chance??"
Arlene Kushner 06/07/2021 Comments Off on From Israel: "Is There a Chance??" 737 Views
Before I look at various events concerning Israel, I note the date: the Fourth of July. From here in Jerusalem, I remember.
But, as the title of this posting suggests, I am at a loss as to what will ultimately be for America.
I know only that the country in which I was born, and grew up, and raised my children is no more. I have watched with horror as free speech has been restricted, and race relations have been set back more than half a century; as violence has increased shockingly and antisemitism has grown exponentially; as those who imagine themselves "woke" make everything about race; as pride in an exceptional America has morphed in progressive circles to some notion of a nation born in sin and American history is trashed.
Is there a chance? Not so long ago, I would have said no, that America was self-destructing irrevocably. But what I am seeing of late is that there are many good Americans who still have their heads screwed on properly; they are fed up and furious, and are now pushing back. Can they restore America to a place of sanity and decency, a place where citizens are proud? Can they defeat the progressive-socialist agenda to bring America down? Perhaps. The roots of America – the noble concepts on which it was founded 245 years ago – still stand.
And so, on this Fourth of July I pray for America: May the Almighty grant those who are fighting to restore her, the strength and wisdom to do so.
Before I leave the topic of July 4, I mention another event that happened on that date in 1976: The rescue at Entebbe. Doubly amazing because it was on America's 200th birthday.
An Air France plane flying from Tel Aviv to Paris was hijacked by terrorists with 248 passengers on board and diverted to Entebbe, Uganda. They released the non-Jews and held the Jews for a number of demands – a large sum of money and the release of numerous terrorists, most of whom were in Israeli prisons. Ultimately, the Israeli government decided not to negotiate with the hijackers and instead set up a covert rescue operation.
Yoni (Yonatan) Netanyahu, Binyamin Netanyahu's brother, was leader of the operation. "Follow me!!" was his famous cry to his team. He lost his life early during the execution of the operation.
The bold and innovative operation, however, was a resounding success.
For a closer look at what went on:
It is difficult for me to avoid lamenting the difference between the mindset of Israeli leaders 45 years ago and what is happening today.
It was during the war with Hamas in 2014 that two of our soldiers – Oron Shaul and Hadar Goldin – were killed in Gaza and their bodies taken. In the months following, two Israeli civilians – Ethiopian Israeli Avner Mengistu and Israeli Arab Hisham al-Sayed, both said to be mentally unstable – wandered into Gaza of their own volition and were taken hostage by Hamas. Israel has not secured the release of either the bodies or the live hostages, about whom very little seems to be known with regard to location or condition. (Obviously, I am not privy to intelligence Israel may possess about them.)
Clearly, the goal of Hamas in holding these hostages is to secure the release of a large number of terrorists who have been convicted and are serving time in Israeli prisons. In my opinion, and the opinion of many others, that sort of exchange, which has happened on several occasions, should never take place again.
With the recent mini-war with Hamas there was some hope that Hamas might be squeezed in a manner that would convince them to release the hostages. But nothing of the sort happened.
When ceasefire negotiations began after fighting ended, again there was hope that Israel might stand strong. Israel began by saying crossings into Gaza would not be opened for many products and the area in which Gaza fisherman would be allowed to fish would be restricted until the hostages were released. It didn't take long before more was permitted through the crossing and the area in which fisherman would be permitted to operate was extended.
I find this very troublesome, for when threats are made and then withdrawn, it simply makes Israel look weaker.
Then it was said that Israel would not allow reconstruction to proceed until the hostages were released. Other countries that will be involved in reconstruction are undoubtedly putting pressure on Israel with regard to this stipulation: There are reports that the Biden administration is opposed to conditioning reconstruction on return of the hostages, which does not come as a surprise. Hamas continues to insist that ceasefire negotiations and arrangements for the release of the hostages are two separate issues and should be negotiated separately.
So far we are holding strong, more or less. On July 1, Defense Minister Gantz declared:
"The Hamas terror group in Gaza needs to understand: We are determined. If Hamas wants reconstruction and economic development, the time has come that it takes concrete steps to maintain the calm, to halt the rearmament and to return the boys home." And reports are that Prime Minster Bennett is determined to link reconstruction with release of the hostages.
https://www.timesofisrael.com/israel-hamas-fail-to-make-progress-in-cairo-on-prisoner-swap-deal/
This I salute. However…at the very same time an Israeli delegation went to Cairo last week to participate in indirect negotiations on a "prisoner exchange." The delegation returned without having made any agreement because Hamas was demanding release of prisoners who had been convicted murdering Israelis.
According to Kan news, the demands made by Hamas in Cairo "were dramatic and unrealistic," but the Israeli delegation continued to hope that Egypt would be able to exert enough pressure on Hamas to make an agreement possible.
And, pray tell, what sort of agreement would be "possible"?
I know I am a hardliner here, but this is how I see it:
Israel is a sovereign nation that has initiated no hostilities, but fights in defensive actions.
Hamas is a terrorist organization that stockpiles weaponry for the sake of attack and every so often initiates offensive actions – launching of rockets and more. Its control of Gaza is not legitimate, as it wrested it from the Palestinian Authority by force.
Holding of hostages is not permitted according to International Customary Law.
"The UN Commission on Human Rights has stated that hostage-taking, wherever and by whoever committed, is an illegal act aimed at the destruction of human rights and is never justifiable."
"The International Convention against the Taking of Hostages defines the offence as the seizure or detention of a person (the hostage), combined with threatening to kill, to injure or to continue to detain the hostage, in order to compel a third party to do or to abstain from doing any act as an explicit or implicit condition for the release of the hostage."
https://ihl-databases.icrc.org/customary-ihl/eng/docs/v1_rul_rule96
And Israel is expected to give Hamas something in order to secure release of our hostages? Whatever we gave would allow Hamas to declare a win and further reduce what minuscule deterrence we now maintain. It would also motive further hostage taking in the future.
The truly tragic part of this story is the fact that the international community is willing to close its eyes to what is happening here. Not a word from the UN.
And so the dictum for Israel must be: "If I am not for myself…" We must stand up.
Before moving to other issues, I note that incendiary balloons are still being launched from Gaza. Twice now, we have attacked in response, but clearly the attack is insufficient.
Credit: MFA
And yet Prime Minister Bennett is talking tough, which is a step in the right direction. He must act in accordance with his words, spoken at today's Cabinet meeting (emphasis added):
"I would like to clarify again: Things have changed.
"Israel is interested in quiet and we have no desire to harm the people in Gaza. However, violence—balloons, marches, harassment of any kind—will be met with a severe response.
"We are also working on a solution that would provide humanitarian assistance to the people of Gaza, but it will not include suitcases full of dollars [previously brought into Gaza by Qatar for distribution]. These suitcases of cash are something we inherited and must end once and for all."
https://www.israelnationalnews.com/News/News.aspx/309218
The tone was anything but tough when Ruby Rivlin, soon to retire from his seven-year term as president, visited President Biden, with whom he has had a relationship for decades, on his last official trip abroad.
Rivlin carried multiple messages to the US, the most pertinent being an expression of reservations about resumption of the Iran agreement and a request for additional precision weapons to utilize as necessary in Gaza – the precision being necessary because Hamas embeds itself within the civilian population.
I was decidedly underwhelmed by Rivlin's boasting with pride about the Islamist Arab party that is part of our new coalition. Is this the same Ruby Rivlin I followed over the years? Whatever Rivlin truly thinks about the Ra'am party, I imagine he knew full well how positively it would play in progressive America. This does not impress me.
Credit: Haim Zach/GPO
Nor is Biden's emphasizing "the importance of Israel taking steps to ensure calm, stability, and to support greater economic opportunities for the Palestinian people" impressive.
What made the most press was Rivlin's expressed assurance that Biden was on Israel's side. Biden's words, intended to comfort, were:
"What I can say to you is that Iran will never get a nuclear weapon on my watch."
Rivlin declared himself "very much satisfied" with this promise, but a number of analysts quickly picked up on the fact that it was a totally meaningless statement. There are serious doubts as to whether the US can even manoeuvrer Iran into going back to the 2015 JCPOA agreement (which would not really be the same agreement, as Iran has progressed in technology and in stockpiling of uranium since then). But the twilight clause that would allow Iran to develop nuclear weaponry would not kick in until after Biden's term of office had expired. He didn't promise that Iran would never have nuclear weapons, only that it wouldn't have them in the next three-and-a-half years.
This is called kicking the can down the road: Delaying the problem until someone else is in charge.
There is an additional plan that the Americans are promoting: after the JCPOA is signed, and billions in sanctions have been lifted, there would be a "follow-up agreement" that would address problems such as Iran's support of terrorism. The notion that Iran could be sweet-talked into agreeing to additional restrictions after the US had already surrendered the leverage of sanctions, is left-wing "pie-in-the-sky" nonsense. See, the US will be nice, and Iran will reciprocate by cooperating.
Perhaps Rivlin had a need to hear good things on this, his last official visit, but I assure you that Israeli military and security personnel put absolutely no stock in this, nor should any of us. Israel reserves the right to act independently in self-defense, and Biden knows this.
See David Weinberg, Vice President of the Jerusalem Institute for Strategy and Security, on this:
https://www.jewishpress.com/indepth/opinions/kicking-the-can-down-the-road-again/2021/07/04/
I will discuss results regarding an agreement on Evyatar shortly – matters are not entirely clear but more promising than I had expected.
Arlene from Israel website. https://www.arlenefromisrael.info/
Tags Entebbe Fourth of July hostages President Ruby Rivlin Prime Minister Bennett Yoni Netanyahu
Previous Evyatar, the window into the new government.
Next From Israel: "The One Foot Theory!"


Lost and Found is a 278-page visual recording of the of the time we live in, and the issues we face, expressed through David LaChapelle's unique and distinctive vision.
Featuring a monumental curation of images which have never before been published in book form, Lost and Found chronicles LaChapelle's emancipation from editorial photography, and his passionate return to fine art.
This fourth volume also introduces a decade of unseen work from LaChapelle's creative renaissance, where he offers a bridge into paradise and the sublime, leading viewers into Good News.


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Middle school principals accept Schools to Watch awards
Principals Dan O'Rourke and Joseph DeMar accepted the Schools to Watch Awards on behalf of Boyce Middle School and Fort Couch Middle School, respectively. The schools were recognized at the Pennsylvania Association for Middle Level Education State Conference on Feb. 26, 2017, in State College. This marks the fourth time that both schools have earned this honor.
In addition, Mr. O'Rourke and Matt Henderson, STEAM teacher at Boyce, delivered a presentation on the STEAM Design course and the Innovation Hub @ Boyce. Mr. DeMar and a team from Fort Couch - including Erin Peterson, assistant principal; Lauren Madia, school counselor; and Mike Davia, special education teacher - gave a presentation on the school's program of customized learning.
Boyce and Fort Couch middle schools in the Upper St. Clair School District have been named Pennsylvania Don Eichhorn Schools to Watch. This marks the fourth time that the USC middle schools have earned this honor – a feat accomplished by only two other Pennsylvania schools.
Schools to Watch is an initiative of the National Forum to Accelerate Middle-Grades Reform. Boyce and Fort Couch are among 38 schools in Pennsylvania that have earned this distinction since the program's inception in 1999.
"We are honored and excited for our middle schools to receive their fourth Schools to Watch designation," Dr. Patrick T. O'Toole, superintendent of schools, said. "This is an outstanding award for our staff, students and the entire Upper St. Clair community."
Schools are recognized for a three-year period, and at the end of three years, they must demonstrate progress on specific goals in order to be re-designated. Unlike the Blue Ribbon recognition program, "Schools to Watch" requires schools to not just identify strengths, but to also focus on areas for continuous improvement; thus the three year re-designation.
"The Schools to Watch process allows us to look deeper into our school practices educationally, athletically and within the arts," Joseph DeMar, Fort Couch Middle School principal, said. "We have an amazing school community that is so deserving of this recognition for its dedication, desire and commitment to excellence."
The National Forum identifies Schools to Watch based on three overriding principles of high-performing middle-grades schools:
They are academically excellent — these schools challenge all students to use their minds well.
They are developmentally responsive — these schools are sensitive to the unique developmental challenges of early adolescence.
They are socially equitable — these schools are democratic and fair, providing every student with high-quality teachers, resources, and supports.
"We congratulate these schools for being places that do great things for all of their students. These schools demonstrate that high-performing middle grades schools are places that focus on academic growth and achievement. They are also places that recognize the importance of meeting the needs of all of their students and ensure that every child has access to a challenging, high-quality education," Bruce Vosburgh, president of the National Forum, said. "In addition each school has strong leadership, teachers who work together to improve curriculum and instruction, and a commitment to assessment and accountability to bring about continuous improvement."
Boyce and Fort Couch first earned Schools to Watch honors in 2008. The schools were re-designated as Schools To Watch in 2011, 2014 and 2017. Selection was based on a written application that required schools to show how they met criteria developed by the National Forum. Schools that appeared to meet the criteria were then visited by state teams, which observed classrooms, interviewed administrators, teachers, students, and parents, and looked at achievement data, suspension rates, quality of lessons, and student work.
"The application process involves all staff members, and is a great way to reflect on the things we are doing well and to identify areas for potential growth," Dan O'Rourke, Boyce Middle School principal, said. "It provides an opportunity for us to really focus on middle level education and systems to support middle school students."
In addition to being named Schools to Watch, Boyce and Fort Couch are National Blue Ribbon Schools. Boyce was recognized with the 1991-92 Blue Ribbon Award. Fort Couch is one of only eight schools in the nation to have earned four Blue Ribbon Awards: 1986-87, 1992-93, 2001-02 and 2012-13.
"Honors such as Schools to Watch are important because our families, staff members and students get an opportunity to have their efforts rewarded and recognized," Mr. O'Rourke said. "These types of honors solidify the outstanding work that is done in our schools, and provides an opportunity for us to celebrate our successes."
A total of 11 Pennsylvania middle schools met the Schools to Watch criteria for 2017 and will be recognized at the Pennsylvania Association for Middle Level Education State Conference on Feb. 26, 2017, in State College. In addition, they will be recognized nationally at the National Forum's National Schools to Watch Conference on June 22-24, 2017, in Washington, D.C.
Launched in 1999, Schools to Watch began as a national program to identify middle-grades schools across the country that were meeting or exceeding 37 researched based criteria developed by the National Forum. Seventeen states have trained Schools to Watch State Teams, with more than 400 schools recognized across the country.
"We are pleased that our Schools to Watch program has shown that schools can meet high academic expectations while preserving a commitment of healthy development and equity for all students," Ericka Uskali, National Forum executive director, said. "These schools have proven that it is possible to overcome barriers to achieving excellence, and any middle-level school in any state can truly learn from their examples."
The National Forum sponsors the Schools to Watch program along with the Association for Middle Level Education, the National Association for Elementary School Principals, the National Association of Secondary School Principals, Learning Forward, ASCD, and State Schools to Watch affiliates.


The 2016 race has turned the battle of the sexes into an all-out war.
Even before the vulgar debut of the phrase "locker-room talk," or the front-page accounts of sexual assault on airplanes, or the national debate over what constitutes an improper amount of pageant-winner weight gain, there has been much that's split American men and women apart this year when it comes to electing a president.
To some degree, this was inevitable. The 2012 election saw a historic 20-point gap between the sexes as it concerned support for President Obama and former Governor Mitt Romney. But what makes this year's election different—beyond the unimaginably lewd headlines and pervasive sense of doom—is the fact that Trump and Clinton have divided not just men and women, but men and women who are married to each other.
By way of some background: As far as John and Jane Doe are concerned, 1972 and 1976 were the last truly harmonious years for the prototypical American male and female voter. According to Pew, Richard Nixon and Jimmy Carter were overwhelmingly the presidential choices for both.
But by 1980, men and women were split over the election of Ronald Reagan by 10 points: Gallup's final pre-election poll showed that men preferred Reagan over his Democratic challenger Jimmy Carter by 15 points, while women chose Reagan by just five.
In 2000, with the election of George W. Bush, the sexes were split by a whopping 15 points: Men supported Dubya by seven percent, while women opted for Gore by eight.
This year, the split between men and women is going to be epic: "We will have a gender gap the size of the Grand Canyon," said Republican pollster Whit Ayers.
Digging deeper into this gap between men and women, what becomes evident is that the political bonds that come with marriage have been broken—insofar as they are something that have held men and women together at the voting booth.
Throughout modern American elections, married couples—unlike single men and single women—have tended to vote in unison. A 1989 study by Laura Stoker and M. Kent Jennings observed that "marriage gives rise to a new and shared set of social and economic circumstances," as well as opportunities to "learn from and influence" each other.
Married couples have also tended to vote Republican (even after controlling for things like income and race)—whether because the GOP has traditionally been seen as the party of "family" values, or because men tend to vote Republican, and their wives have historically voted (according to a 2006 study from the Institute for Social and Economic Research) on the basis of their husband's economic interests. It might also be that married people, by virtue of entering into the (until recently) heterosexual and traditional institution of marriage, tend to be a more conservative bunch.
Either way, it's been working for the Grand Ole Party: In 2012, Mitt Romney beat President Obama among married voters by 10 points. Romney, further, won married women by four points, as did his Republican predecessor John McCain, by six points. The last Democrat to win married women was Bill Clinton in 1996—by just one point.
But because of the incredibly pitched and personal nature of the 2016 race, and in particular the numerous marital controversies surrounding both nominees, large numbers of American spouses are not necessarily on the same page this year.
Counter to the last 20 years of American politics, married women now are looking toward the Democrat in the race. An NBC News poll from early October—four days before the Access Hollywood tape was released—showed married women breaking 48 percent for Clinton, 40 percent for Trump. Among married men, however, Trump beats Clinton 52 to 35. That puts a chasm of 25 points between married men and married women. (To be clear, this spousal split may be a largely white phenomenon––voters of color overwhelmingly support Clinton).
Luntz explained that his research has shown that "men still define success by their money and careers. Women—even CEOs and opinion movers—still define success by their kids' health and happiness. [Trump] hasn't connected to any of that." The first lady, however, aimed to do just that: frame the argument against Trump as a matter of parenting.
If Clinton does manage to carry married women by large margins come November, it's unclear whether the Democratic Party will finally have changed the voting norms of (heterosexual, white) married Americans to its benefit in the medium-to-long term. Luntz, in fact, argued that the split between husbands and wives has as much to do with Trump's problems with women as it does Clinton's problems with men.
But then there's this: Independent of electoral gains and losses, what of the cultural fallout from this fractious American moment?
The 2016 presidential race has raised questions about many American institutions—whether media, finance, or government. Marriage, in this decade, has already undergone a radical shift in terms of its basic parameters: Merely to refer to it as a union between a man and a woman is now outdated. And yet, after November, it's possible that the primary assumptions undergirding the nuptial agreement itself may be in question: The 2016 election may change our expectations about what Americans want out of their marriages—or not. Perhaps it only makes sense that in this season of rancor and discontent, the institution charged with securing partnership between disparate factions may itself be challenged by the vicissitudes of the moment.


text:
Palmarès

Club

Maiorca: 2002-2003

Nazionale

1995

Collegamenti esterni


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