text:
Q: Node: Bootstrap's JavaScript requires jQuery I am going through the Manning Getting MEAN book.  I want to use bootstrap sass and have loaded bootstrap-loader.  When I run nodemon I get the following error:
/Users/mglaz/Projects/mean/theCMF/node_modules/bootstrap/dist/js/bootstrap.js:8
throw new Error('Bootstrap\'s JavaScript requires jQuery')

here's my app.js
var express = require('express');
var path = require('path');
var favicon = require('serve-favicon');
var logger = require('morgan');
var cookieParser = require('cookie-parser');
var bodyParser = require('body-parser');

require('jquery');
require('bootstrap');
require('bootstrap-loader');

var routes = require('./routes/index');
var users = require('./routes/users');

var app = express();

// view engine setup
app.set('views', path.join(__dirname, 'views'));
app.set('view engine', 'pug');

// uncomment after placing your favicon in /public
//app.use(favicon(__dirname + '/public/favicon.ico'));
app.use(logger('dev'));
app.use(bodyParser.json());
app.use(bodyParser.urlencoded({ extended: false }));
app.use(cookieParser());
app.use(express.static(path.join(__dirname, 'public')));

app.use('/', routes);
app.use('/users', users);

// catch 404 and forward to error handler
app.use(function(req, res, next) {
    var err = new Error('Not Found');
    err.status = 404;
    next(err);
});

// error handlers

// development error handler
// will print stacktrace
if (app.get('env') === 'development') {
    app.use(function(err, req, res, next) {
        res.status(err.status || 500);
        res.render('error', {
            message: err.message,
            error: err
        });
    });
}

// production error handler
// no stacktraces leaked to user
app.use(function(err, req, res, next) {
    res.status(err.status || 500);
    res.render('error', {
        message: err.message,
        error: {}
    });
});

module.exports = app;

and my package.json
{
  "name": "loc8r",
  "version": "0.0.0",
  "private": true,
  "scripts": {
    "start": "node ./bin/www"
  },
  "dependencies": {
    "body-parser": "~1.8.1",
    "cookie-parser": "~1.3.3",
    "debug": "~2.0.0",
    "express": "~4.9.0",
    "jquery": "^3.1.1",
    "morgan": "~1.3.0",
    "pug": "^2.0.0-beta6",
    "serve-favicon": "~2.1.3"
  },
  "devDependencies": {
    "bootstrap": "^4.0.0-alpha.4",
    "css-loader": "^0.26.1",
    "exports-loader": "^0.6.3",
    "extract-text-webpack-plugin": "^1.0.1",
    "file-loader": "^0.9.0",
    "imports-loader": "^0.7.0",
    "node-sass": "^3.13.1",
    "postcss-loader": "^1.2.0",
    "resolve-url-loader": "^1.6.1",
    "sass-loader": "^4.0.2",
    "style-loader": "^0.13.1",
    "url-loader": "^0.5.7",
    "webpack": "^1.14.0"
  }
}

A: Have a look at jQuery not define in bootstrap module and http://blog.npmjs.org/post/112064849860/using-jquery-plugins-with-npm
You need to set the global jQuery variable
global.jQuery = require('jquery');
require('bootstrap');


text:
Optimizing the Preparation Procedure of Recombinant PSCA, as a Practical Biomarker in Prostate Cancer
Neda Saraygord-Afshari
Mahboube Shahrabi Farahani
Mohammad M Farajollahi
Department of Medical Biotechnology, Faculty of Allied Medical Sciences, Iran University of Medical Sciences, Tehran, Iran
Background: The unique expression pattern of prostate stem cell antigen (PSCA) in a number of prevalent neoplasms has made the antigen a great target for cancer researches, and many clinical methods have been developed based on the application of this tumor marker. Hence, optimal PSCA laboratory production can be considered a hallmark for many researchers. Objective: An analytical study was designed to improve the quality and quantity of PSCA production. Materials and Methods: The effects of different compositions of lysis buffers and some ultrasound durations were assessed by calculation of the protein recovery followed by PSCA specific blotting experiments. Then, based on the results of the web based characterization, interference removal, followed by re-solubilization of the protein in various buffers, was designed, applied, and assessed. Results: Since the selection of an appropriate methodology depends merely on the research purposes, we tried to discuss the pros and cons of the investigated methods according to the hydrophobic nature of PSCA as well as its dramatic tendency to aggregate in the form of inclusion bodies in the expression hosts. Conclusions: We introduced a newly designed method to fit the delicate immunological surveys and overcome some limiting factors in PSCA production.
Protein aggregation
Prostate stem cell antigen
Specific measurable indicators of cancers, known as biomarkers, can play critical roles in identifying the nature of diseases. Hence, the scope of molecular, cellular, and imaging biomarkers is rapidly increasing through the efforts made by researchers worldwide to provide more sophisticated surrogate markers. Effective cancer biomarkers are those which can efficiently help a clinician to diagnose and monitor the disease. They also must have the ability to predict metastasis risk and suggest an appropriate therapy ( 1 ). None of the present biomarkers can ideally meet all the aforementioned characteristics, but there are a number of biomarkers used efficiently for prostate cancer (PCa) such as prostate-specific antigen (PSA), prostatic acid phosphatase (PAP), and prostate stem cell antigen (PSCA). PSA has been extensively used for the diagnosis and monitoring of prostate cancer. Nevertheless, its low specificity may lead to a high number of false positives. PSCA is a cell surface molecular marker that has been proven to be overexpressed in several prevalent malignancies, including renal cell carcinoma, bladder, gastric and pancreatic neoplasms ( 2 , 3 ).
This valuable biomarker can also be monitored in liquid biopsies via detection of circulating tumor cells (CTCs) ( 4 ). Medical application of PSCA has gained more attention in the case of prostate malignancies as not only its up-regulation has been reported in the metastatic castration-resistant prostate cancer (mCRPC) but also its over-expression has been positively correlated with Gleason score. PSCA has been proven to act as both oncogenic and tumor-suppressing agents and is also widely used for prognosis and theranostics of prostate cancer (e.g. immunotherapy) ( 2 ). Nevertheless, its precise biochemical and physiological functions remain unclear ( 5 , 6 ). Hence, PSCA has been the subject of many types of research to decipher its biological and physicochemical properties to improve the efficacy of the present clinical methods.
To investigate the dark side of biochemical, physiological, and clinical characteristics of PSCA as well as the development of medical techniques, a sufficient amount of this antigen must be provided for research purposes. Generally, recombinant-DNA technology is used in many laboratories to clone and express PSCA corresponding genes. In this way, E. coli is an appropriate host which has been extensively used for the production of recombinant PSCA. After expression in the host, the protein must be recovered and purified; however, this procedure does not always yield a sufficient amount of the antigen, and sometimes the quality of the product does not satisfy the researcher ( 7 ). Formation of inclusion bodies (IBs) due to overexpression of the recombinant product and its hydrophobicity, in addition to lack of post-translational modifications in the prokaryotic host, can be considered the main problem associated with the method ( 5 , 8 ). Therefore, optimizing the procedure to increase the quality and quantity of the product can be of much interest, which leads to much saving in time and costs.
In the present study, we investigated the effects of ten different laboratory methods to extract and purify the recombinant PSCA produced in the E. coli host. The investigated methods are either the commonly used ones or in some cases are newly introduced. We aimed to present a more efficient method for PSCA production at a laboratory scale and examine its physicochemical behavior accordingly.
3.1. Biological and Chemical Materials
The BL21 (DE3) strain of E. coli was purchased from Pasteur Institute of Iran and used as an expression host. The expression constructs, pET-28a(+) containing the PSCA coding region with six-histidine (6His) tag at its two termini, was provided by GenScript (USA). Miniprep kit was purchased from Vivantis Technologies Sdn Bhd (Malaysia). All the other chemicals and reagents were of analytical grade from Merck (Darmstadt, Germany), Sigma-Aldrich (Taufkirchen, Germany), Biomedicals (Meckenheim, Germany) and InvitrogenTM|Thermo Fisher Scientific (Fremont, CA, USA).
3.2. Preparation of the Construct and Expression of the PSCA Recombinant Gene
The expression construct was prepared and tested according to the standard molecular biology protocols. The positive vectors were then transformed into the competent cells of E. coli, BL21 (DE3) strain, by thermal shocking to overexpress the recombinant PSCA protein in the Luria-Bertani (LB) medium supplemented with 50 μg.mL-1 kanamycin ( 9 , 10 ). The transformation was performed according to the BioLabs protocol with some modifications. The bacterial cells, carrying our recombinant vectors, were harvested by centrifugation at 1600 ×g for 20 min at 4 °C. They were then stored at -20 °C for further analysis.
3.3. Extraction and Purification of the Recombinant PSCA; Trying Different Conditions
3.3.1. Protein Extraction in Different Lysis Buffer Solutions
Five different lysis buffer solutions were prepared and applied (Table 1). The common RIPA buffer composed of 150 mM sodium chloride, 50 mM Tris pH 8.0, an appropriate amount of protease inhibitor cocktail, 0.5% (W/V) sodium deoxycholate, 0.1% (W/V) SDS, and 1.0% (V/V) Triton X-100, and the four others were the same in the 2-mercaptoethanol (0.05% (V/V)), Tris-HCl pH 8.0 (50 mM) and protease inhibitor cocktail concentrations but varied in detergent type and/or quantity. Accordingly, sodium deoxycholate (0.5% (W/V)), Triton X-100 (2.0% (V/V)) and SDS at 1.0%, 2.0% concentrations and 5.0 % (W/V) were selected and applied. Sufficient and equal amounts of the harvested bacterial cells were then resuspended and incubated in each buffer for 20 min on ice. This extraction procedure was also assisted by an ice-cold sonication protocol (5 min with 10 s pulses during 10 s intervals) in JENCONS (130 W, UK) instrument. Finally, the extracted lysates were recovered by 15000 ×g centrifugation of the mixture at 4 °C for 15 min, and the cell debris accumulated in the pellet was discarded. The lysates were then stored at -80 °C for further analysis.
Lysis buffers
Western blot analysis result
I RIPA (150 mM sodium chloride, 50 mM Tris pH 8.0, cocktail protease inhibitor, 0.5% sodium deoxycholate (W/V), 0.1% SDS (W/V) and 1.0% Triton X-100 (V/V)) No signal
II 0.05% 2-mercaptoethanol (V/V), 50 mM Tris.HCL pH 8.0, cocktail protease inhibitor and 2.0% Triton X-100 (V/V) No signal
III 0.05% 2-mercaptoethanol (V/V), 50 mM Tris.HCL pH 8.0, cocktail protease inhibitor and 1.0% SDS (W/V) Faint band
IV 0.05% 2-mercaptoethanol (V/V), 50 mM Tris.HCL pH 8.0, cocktail protease inhibitor and 2.0% SDS (W/V) Sharp band
V 0.05% 2-mercaptoethanol (V/V), 50 mM Tris.HCL pH 8.0, cocktail protease inhibitor and 5.0% SDS (W/V) Sharp band
Table1.The list of different lysis buffers used for recombinant PSCA extraction and their western blotting results.
3.3.2. Protein Extraction at Different Ultrasound Exposure Durations
Ultrasound-assisted extractions were assessed in three different durations, routinely used in the laboratories (1, 5, and 10 min) ( 11 ). Hence, sufficient and equal amounts of the harvested bacterial cells were re-suspended and incubated in the most efficient lysis buffer again, from section 2.3.1, for 20 min on ice. This lysis procedure was then assisted by sonication at the mentioned durations for each sample. The ultrasound energy was applied in 10 s pulses during 10 s intervals for all the defined durations, and the JENCONS instrument used here was 130 W power, 50% amplitude, and 20 kHz frequency. The remaining steps were the same as in section 2.3.1.
3.3.3. Elimination of Non-Protein Components
After removing the cellular debris from the lysates, non-protein components of the supernatants were eliminated via an ice-cold acetone precipitation procedure. For this purpose, four volumes of pre-chilled (-20 °C) acetone was added to the samples and vortexed. They were then incubated at -20 °C for an hour. Proteins were then recovered by centrifugation at 13000 ×g for 15 min at 4 °C. Purified protein pellets were washed with a volume of pre-chilled acetone once more, air-dried, and resolubilized in an appropriate buffer ( 12 ).
3.3.4. Resolubilization of the Purified Protein Pellets in Different Buffers
The effect of re-solubilizing buffers was assessed on the most efficient procedure, from the previous sessions (2.3.1 and 2.3.2). For this purpose, ultra-pure deionized water, phosphate-buffered saline (known as PBS), and Tris-HCl buffer containing 2% (W/V) SDS and 0.05% (V/V) 2-mercaptoethanol were used for re-solubilizing the protein pellets, recovered after the precipitation step. Complete resolubilization was checked out by centrifuging the solutions at 15000 ×g for 20 min at 4 °C, and if the resolubilization procedure was not fully obtained, the supernatant was picked up as the pure soluble protein. These purified protein solutions were stored at -80 °C for further analysis.
3.3.5. Purification of the Recombinant PSCA
Poly(His)-tagged PSCA recombinant protein was purified from the crude bacterial lysates or the total protein extracts through the His-tag affinity column (Ni-NTA agarose) according to the Invitrogen's instruction. The purified recombinant PSCA was collected and stored at -80 °C for further analysis.
3.4. Protein Assay
Protein concentrations were assessed in different steps of the procedure. Considering the diversity of the reagents and materials used and their interference with different methods of protein assay, ( 13 ) protein concentrations were measured by either UV-absorbance at 280 nm (in a NanoDrop 2000c spectrophotometer, Thermo Scientific) or BCA assay (using a Pierce BCA Protein Assay Kit, Thermo Scientific).
3.5. Examination of the Purified Recombinant PSCA: Assessment of Different Procedures
The quality and quantity of poly(His)-tagged recombinant PSCA was assessed by a wet western blot system. For this purpose, equal amounts of protein samples (obtained from each procedure) were separated on 12% SDS-PAGE mini-gels (Bio-Rad), according to Laemmli's protocol ( 5 , 8 ). Finally the poly(His)-tagged recombinant PSCA visualized via a luminescent reaction which was recorded on a Kodak radiology film.
3.6. Computer-Assisted Sequence Analysis
The biophysiochemical properties of recombinant PSCA sequence and its native form as well as some other well-known proteins were calculated with the aid of ExPASy online tools. Hence, the protein sequences were used to generate the GRAVY (grand average of hydropathy) score values via Expasy ProtParam server (http://web.expasy.org/cgi-bin/protparam/protparam). The hydrophobicity plots of the poly(His)-tagged recombinant PSCA and its native sequence were also traced by Kyte and Doolittle algorithm using Expasy ProtScale server (http://web.expasy.org/cgi-bin/protscale/protscale/) to broaden the information.
Finally, all the obtained data were recorded to further discuss the results of the experimental sections.
In an analytical in vitro study, to improve the quality and quantity of the recombinant PSCA protein production, ten different methods were applied and evaluated. These methods summarized in Figure 1 were either the well-established, commonly used methods or the newly introduced ones for PSCA production, designed based on the behavior of biophysiochemical protein in the experiments.
Figure 1. Summarized ten different methods used for the PSCA production and purification.
4.1. Confirmation of Successful Expression of PSCA Fusion Protein
Before starting the optimization procedure, the successful expression of PSCA fusion protein after IPTG induction was confirmed by simple SDS-PAGE analysis. The results of this confirmation test (Fig. 2A) showed overexpression of a protein band near the molecular weight of about 15.0 kDa, which corresponded to the predicted size of the poly(His)-tagged recombinant PSCA.
Figure 2. A: Results of SDS-PAGE analysis for recombinant PSCA expression before (lane 1)and after (lane 2) IPTG induction. B: Results of the western blot analysis, indicating the effect of the application of different lysis buffer solutions to obtain the crude cell extract from the E.coli cells. Lanes 1, 2, 3, 4, and 5 correspond to the I, II, III, IV, and V lysis buffer solutions (Table 1), respectively. C: Results of the western blot analysis showing the efficacy of the three different sonication durations. Lanes 1, 2, and 3 correspond to 1, 5, and 10 minutes sonication time, respectively. D: Results of the western blot analysis indicated the efficiency of the protein pellet resolubilization in three different solubilization buffers. Lanes 1, 2, and 3 are protein pellets resolubilized in distilled water, PBS, and Tris-HCL buffer containing 2% SDS (W/V) and 0.05% 2-ME (V/V), respectively.
4.2. Analysis of Five Different Lysis Buffer Solutions Effect on the Yield of the Recombinant PSCA
We started to optimize the yield of the recombinant PSCA extraction via a survey on the effect of five different lysis buffer solutions. As mentioned in section 2.3.1, the buffers were varied mainly in the detergent type and/or quantity. The results of these assays are depicted in Table 1, and Figure 2B shows that in the case of I and II buffers, no detectable PSCA bands could be observed; however, in the three remaining buffer solutions, PSCA specific bands appeared with different intensities. It can be deduced that in contrast to high concentrations of SDS, sodium deoxycholate, Triton X-100 and low concentration of SDS (0.1% (W/V)) failed to extract PSCA from bacterial lysates. Thus, strong detergent treatments are needed to extract PSCA from bacterial lysates. A positive correlation of the intensity of the PSCA specific band with the increased concentration of SDS, as a powerful ionic detergent, in the lysis buffer solutions confirms our idea. Moreover, the Western blot analysis, total protein yields as well as purified PSCA protein yields were also assessed for each of these five experiments (Table 2, part A). The data for these assessments indicated various results for the blotting analysis. Hence, strong detergent treatment not only increases the total protein yields (from 5.16 0.82 μg.mL-1 to 9.13 0.36 μg.mL-1) but also affects the yields of purified PSCA extraction (form 0.650.13 μg.mL-1 to 1.100.12 μg.mL-1).
Lysis buffer containing 2% V/V Triton X-100
Lysis buffer containing 1% W/V SDS
Lysis buffer containing 2 % W/V SDS
A Protein concentration before affinity purification (mg.mL-1) 5.16 0.82 3. 75 1.00 6.24 0.52 7.66 0.73 9.13 0.36
Protein concentration after affinity purification (mg.mL-1) 0.650.13 0.710.22 0.890.09 0.960.20 1.100.12
B Ultrasound exposure time 1 min 5 min 10 min --- ---
Protein concentration before affinity purification (mg/ml) 5.1 0.12 8.3 0.09 10.5 0.22 --- ---
Protein concentration after affinity purification (mg.mL-1) 0.57 0.21 0.98 0.25 1.25 0.31 --- ---
C Medium Distilled water PBS SDS buffer
Protein concentration after re-solubilization (mg.mL-1) 2.18 0.03 3.36 0.06 5.49 0.04 --- ---
Table2.Recovery of recombinant prostate stem cell antigen from E.coli after each assay, A; lysis buffer assay, B; sonication assay, C; solubilization buffer assay.
4.3 Analysis of Different Ultrasound Durations Effect on the Yield of the Recombinant PSCA
The most efficient lysis buffer solution was then applied to investigate the effect of different ultrasound durations on the yield of the recombinant PSCA extraction. The results of this assessment (Fig. 2C) indicated that increasing the duration of the ultrasound-assisted extractions from 1 to 10 min not only increased the intensity of PSCA specific bands in the blotting profiles but also significantly increased the total (from 5.1 0.12 μg.mL-1 to 10.5 0.22 μg.mL-1) and PSCA protein (0.57 0.21 μg.mL-1 to 1.25 0.31 μg.mL-1) concentrations (Table 2, part B).
4.4. Enhancing the Purity of the Recombinant PSCA
A noteworthy point in all of these seven experiments (five experiments for the assessment of lysis buffer solutions and two more for the ultrasound durations) is the presence of nonspecific bands (Fig. 2B and Fig. 2C), far from the expected PSCA molecular weight (about 15.0 kDa). To get rid of these nonspecific unexpected bands and enhance the purity of the product, three more experiments were designed and performed. The newly introduced methods were based on the elimination of non-protein interfering components via an ice-cold acetone precipitation procedure followed by re-solubilizing the protein pellets in water as well as the two other appropriate buffers (PBS and Tris-HCl containing 2% (W/V) SDS, as mentioned in section 2.3.4). As it is depicted in Figure 1, these alterations were applied on the most efficient procedure from the previous sessions (lysis buffer and ultrasound assays) just before purification of poly(His)-tagged PSCA via Ni-NTA affinity column. The results of checking the complete resolubilization of protein pellets confirmed the successful resolubilization of proteins in the Tris-HCl buffer, which was supplemented with a high concentration of SDS. In contrast, water and PBS buffer failed to completely resolubilize the pellets, and as expected, this failure was more obvious in the performance of pure water. These effects were also reflected in the results of the protein assay investigations (Table 2, part C). Therefore, as the total protein and PSCA concentrations (2.18 0.03, 0.35 0.06, 3.36 0.06 (mg.mg-1) and 0.53 0.03, 5.49 0.04, 0.86 0.05 (mg/ml) for water, PBS and Tris-HCl containing 2% (W/V) SDS respectively) increased, the polarity of the resolubilization buffer decreased. The most interesting feature of these three investigations was observed in the results of their western blot analysis (Fig. 2D), which occurred by complete elimination of the unexpected protein bands in water. These results were also accompanied by increased intensity of PSCA predicted and/or unexpected bands by decreasing the polarity of the resolubilization buffer.
PSCA is a glycosylphosphatidylinositol (GPI)-anchored cell membrane glycoprotein with 123-amino acid and is located on the outer surface of the cell membrane lipid rafts; special structures which are enriched in cholesterol, glycosphingolipids, and other lapidated proteins (Fig. 3). To discuss and interpret the results obtained here, we decided to assess the biophysiochemical properties of this Jekyll and Hyde molecule. Hence, computer-assisted sequence analysis was performed via ExPASy online tools according to section 2.6. The results of this analysis indicated that although the comparison of the GRAVY index for native PSCA protein (0.48) and the recombinant poly(His)-tagged PSCA (0.12) showed a 25% decrease in the hydrophobicity index, our fusion protein could still behave as a hydrophobic protein. This was more obvious when its GRAVY index was compared to the hypothetical protein EGK_19311 (Macaca mulatta) and Prostate stem cell antigen isoform X2 (Cavia porcellus) GRAVY indexes (0.31 and 0.20, respectively) as hydrophobic proteins, or with fibrin chain G with the GRAVY index of -1.3 as a hydrophilic protein according to GRAVY index using the Kyte & Doolittle hydrophobicity scale. By the addition of six hydrophilic histidine residues to the ends of the PSCA native structure and deletion of a signal peptide, the overall percentage of the hydrophobic amino acid residues in the native structure decreased from 47% to 41%. This alteration can be observed as negative shifts at the end of the hydrophobicity plot of the recombinant poly(His)-tagged PSCA in comparison with the native structure (Fig. 4). This phenomenon confirms the result of the hydrophobicity assay. Although the content of hydrophobic residues was decreased by 6%, our recombinant protein could still behave as a hydrophobic structure due to its significant percentage of hydrophobic residues.
Figure 3. The schematic view of PSCA as a GPI anchored protein, located in lipid rafts.
Figure 4. A; The hydrophobicity plot of PSCA in native form, B; The hydrophobicity plot of recombinant poly(His) tagged PSCA.
Overall, the results of computer-assisted sequence analysis make it clear that the recombinant poly(His)-tagged PSCA is indeed a hydrophobic protein. Furthermore, this protein contains ten cysteine residues in both native and recombinant structures ( 14 ); cysteine residues comprise 7.75% of the total amino acid content of the recombinant protein. All these properties will result in the increased tendency of the recombinant PSCA to form intermolecular interactions either via hydrophobic forces or the formation of disulfide bonds. Moreover, conditions accompanied by protein expression in a prokaryotic host will also increase the probability of the recombinant PSCA accumulation ( 15 , 16 ). For instance, the reduction of bacterial cytosol environment will promote the formation of protein aggregates by inhibiting the establishment of the intramolecular disulfide bonds (which prevents complete packing of the protein) ( 17 ). Further, overexpression of the recombinant protein will also result in improper protein folding, and a lack of post-translational modifications will increase the instability and hydrophobicity of the protein ( 18 ). Thus, there is a lot of evidence that suggests that the expression of recombinant poly(His)-tagged PSCA will be accompanied by the formation of inclusion bodies (IBs) in bacterial cells ( 5 , 19 , 20 ).
In the assessment of the effect of five different lysis buffer solutions, we showed that strong detergent treatments were needed to extract the recombinant PSCA from the expression host and solve the problem of negative results. This can be attributed to the tendency that originated from the hydrophobic nature of the fusion protein to precipitate with the hydrophobic cell derbies as well as the membrane fragments ( 5 , 21 , 22 ). Hence, replacement of the mild detergent conditions by high concentrations of SDS, as a powerful ionic detergent, would result in the protein extraction from the precipitated cellular debris into a soluble fraction ( 23 ). Results of the protein assays would also confirm the idea. Similarly, increased power of the ultrasound-assisted extraction via a prolonged application time can lead to an increased PSCA predicted band and concentration. This can be attributed to the hydrophobic nature of the recombinant protein, as well. In a polar ionic environment, ultrasound-assisted extraction facilitates momentary detachment of the hydrophobic protein compartment from the cellular debris, allowing the formation of the solvation layer and increasing the protein yield in the aqueous sections ( 24 , 25 ).
Another noteworthy point in the experiments associated with the lysis buffer and ultrasound assays was the presence of unpredicted bands. The blotting analysis indicated that the intensities of these unpredicted bands would also increase by increasing ultrasound power and the detergent (Fig. 2B and Fig. 2C). Since these nonspecific bands vanished when the pure protein contents were resolubilized in water in the last three experiments, we can deduce that the bands are strongly hydrophobic IBs, formed during the recombinant PSCA expression and extraction. It should be remembered that in the aforementioned experiment, complete resolubilization in water was negative and, the supernatant was picked up as the pure soluble protein in the blotting assay. Therefore, increasing the detergent and ultrasound power not only increased the PSCA specific and predicted band but also could help in more efficient isolation of IBS. Many other reports introduce sonication as a fast, simple, and inexpensive mechanical cell rupture method, which can also be used for isolation of IBs from bacterial cells, and confirms the results presented here ( 5 , 26 ). Nonetheless, results obtained here suggest that strong detergent power and prolonged ultrasound durations can help to increase the recombinant PSCA yields and unwanted IBs aggregates. Here, we designed some additional experiments to overcome the problem of IBs nonspecific bands. We specifically precipitated the total protein content obtained from the extraction procedure not only to reach the optimal purity of the product but also to replace the lysis buffer with other media which conversely push the more hydrophobic aggregates out of the solution and yield pure recombinant PSCA compartments. The present study introduced a more efficient procedure for the PSCA protein recovery and enhancing its purity based on the interpretation of the biophysiochemical behavior of the protein. The results obtained here showed that the hydrophobicity of the protein, in addition to its great tendency to accumulate in the form of insoluble IBs, strongly affected the outcomes of the experiments. We also introduced a method to eliminate the nonspecific aggregated protein bands by complete elimination of the unwanted aggregates in the highly polar and hydrophilic medium of water. Providing a sufficient amount of purified cancer biomarkers is very important for clinicians and researchers; hence, it facilitates the experimental and industrial process for the development of diagnostics tools (e.g. molecular imaging techniques) as well as biomarker-based drug development approaches (e.g. production of recombinant protein-based vaccines or other kinds of immunotherapy). Finally, it was shown that the newly introduced approach yielded a suitable product for the researchers working on the delicate investigations, including those with immunological or pharmaceutical nature.
The results presented in this work are based on the data set of an MSc. thesis, recorded in the Iran University of Medical Sciences.
Conflict of Interest: None.
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Avila CL, Chaves S, Socias SB, Vera-Pingitore E, González-Lizárraga F, Vera C, et al. Lessons learned from protein aggregation: toward technological and biomedical applications. Biophys. Rev. 2017; 9(5):501-515. DOI
Ehgartner D, Sagmeister P, Langemann T, Meitz A, Lubitz W, Herwig C. A novel method to recover inclusion body protein from recombinant E. coli fed-batch processes based on phage ΦX174-derived lysis protein E. Appl Microbiol Biotechnol. 2017; 101(14):5603-5614. DOI
Saraygord‐Afshari N, Naderi‐Manesh H, Naderi M. Enhanced reproducibility of the human gel‐based tear proteome maps in the presence of di‐(2‐hydroxyethyl) disulfide. Biotechnol Appl Bioc. 2014; 61(6):660-667. DOI
Martínez-Alonso M, González-Montalbán N, García-Fruitós E, Villaverde A. Learning about protein solubility from bacterial inclusion bodies. Microb Cell Fact. 2009; 8(1):4. DOI
Salem DP, Gong X, Lee H, Zeng A, Xue G, Schacherl J, et al. Characterization of Protein Aggregation Using Hydrogel-Encapsulated nIR Fluorescent Nanoparticle Sensors. ACS Sens. 2020; 5,2:327-337. DOI
He C, Ohnishi K. Efficient renaturation of inclusion body proteins denatured by SDS. Biochem Bioph Res Co. 2017; 490(4):1250-1253. DOI
Baker SL, Munasinghe A, Kaupbayeva B, Kang NR, Certiat M, Murata H, et al. Transforming protein-polymer conjugate purification by tuning protein solubility. Bull. Am. Phys. Soc. 2019; 10(4718):1-12. DOI
Xi G, Esfandiary R, Sacramento CB, Jouihan H, Sharma A, Roth R, et al. Refolding and purification of cGMP-grade recombinant human neurturin from Escherichia coli inclusion bodies. Protein Expres Purif. 2020; 168(105552):1-8. DOI
Zhao F, Song Q, Wang B, Han Y, Zhou Z. Purification and immobilization of α-amylase in one step by gram-positive enhancer matrix (GEM) particles from the soluble protein and the inclusion body. Appl Microbiol Biot. 2020; 104(2):643-652. DOI
Saraygord-Afshari, N., Shahrabi Farahani, M., & Farajollahi, M. M. (2021). Optimizing the Preparation Procedure of Recombinant PSCA, as a Practical Biomarker in Prostate Cancer. Iranian Journal of Biotechnology, 19(2), 79-88. doi: 10.30498/IJB.2021.2631
Neda Saraygord-Afshari; Mahboube Shahrabi Farahani; Mohammad M Farajollahi. "Optimizing the Preparation Procedure of Recombinant PSCA, as a Practical Biomarker in Prostate Cancer". Iranian Journal of Biotechnology, 19, 2, 2021, 79-88. doi: 10.30498/IJB.2021.2631
Saraygord-Afshari, N., Shahrabi Farahani, M., Farajollahi, M. M. (2021). 'Optimizing the Preparation Procedure of Recombinant PSCA, as a Practical Biomarker in Prostate Cancer', Iranian Journal of Biotechnology, 19(2), pp. 79-88. doi: 10.30498/IJB.2021.2631
Saraygord-Afshari, N., Shahrabi Farahani, M., Farajollahi, M. M. Optimizing the Preparation Procedure of Recombinant PSCA, as a Practical Biomarker in Prostate Cancer. Iranian Journal of Biotechnology, 2021; 19(2): 79-88. doi: 10.30498/IJB.2021.2631

redpajama_set_name: RedPajamaCommonCrawl


Goner began in 1993, releasing essential records from Guitar Wolf and Oblivians.
20 years later, we're still at it.
as well as other groovy digital distributors.
Order available vinyl and cds from the Goner Records Store .
Also available via other groovy physical stores, too.
Cococoma - 6 1/4 - 125 7"
Head - Spend The Night Alone 7"
Magic Kids - Hey Boy 7"
Jay Reatard - Hammer I Miss You 7"
Ty Segall - Caesar 7"
Ty Segall - Cents 7"
Useless Eaters - Sucked In 7"


text:
Plectris similis är en skalbaggsart som beskrevs av Frey 1973. Plectris similis ingår i släktet Plectris och familjen Melolonthidae. Inga underarter finns listade i Catalogue of Life.

Källor

Skalbaggar
similis

redpajama_set_name: RedPajamaWikipedia


text:
Leeds - a city of culture
School of Performance and Cultural Industries event Friday 27 September 2019
Date: Friday 27 September 2019, 14:30 –
Location: Stage@Leeds
A showcase of the range and depth of culture and the arts in Leeds and the surrounding region.
This free symposium features a guest panel of leading figures and key practitioners from major cultural organisations, and is a chance to hear about the many exciting companies and projects in the Leeds city region.
This is a public event, booking is not required.
Sharon Watson (Artistic Director, Phoenix Dance Theatre; ex-chair, Leeds 2023)
Alan Lyddiard (Artistic Director, the Performance Ensemble; formerly Artistic Director, Northern Stage)
Joyce Nga Yu Lee (Resident Director, Mind the Gap)
Paul Wilshaw (Assistant Producer, Mind the Gap)
Sharon is the seventh and longest-standing Artistic Director of the award-winning Phoenix Dance Theatre. Trained at the London School of Contemporary Dance, she was one of the first female Principal Dancers invited to join what was originally an all-male company, touring with them from 1989 to 1997, during which she choreographed Never Still and Shaded Limits. Having left Phoenix to pursue a number of other ventures including setting up her own company ABCD, Sharon returned in 2009 as the new Artistic Director. Since then Sharon has choreographed Fast Lane, Melt, Never 2 Still, Repetition of Change, TearFall and Windrush: Movement of the People.
During her tenure at Phoenix Dance Theatre, Sharon has received a number of awards and accolades. In 2010, she was named as one of the Cultural Leadership Programme's 'Women to Watch', a list of 50 influential women working in arts and culture in the UK. She was awarded The Sue Ryder 'Yorkshire Women of Achievement in Business Award' and named 'Yorkshire Woman of the Year' in 2016. She recently received the Liverpool Institute of Performing Arts' Companionship Award from Sir Paul McCartney and was recognised as one of '100 faces of a vibrant economy 2017' by Grant Thornton. Acknowledged at the 2017 Northern Power Women Awards as part of the very first 'Top 50 Power List' and in 2018 Sharon won the First English Woman's Award for Arts and Culture and was presented with an Honorary Fellowship by Northern School of Contemporary Dance. In July, 2019 Sharon received an Honorary Doctorate from Leeds Beckett University for her contribution to the arts. Sharon's recognitions are testament to her continued passion and drive to contribute to the local and national creative industries through the medium of dance.
Sharon is a previous trustee of Matthew Bourne's New Adventures and current trustee of The Place, Leeds Playhouse, and an artistic adviser for Central School of Ballet. She is committed to improving diversity within the arts by supporting the creative voices of women and black and ethnic minority artists.
Alan is currently Artistic Director of The Performance Ensemble a company of performers over the age of 60 working in the space between community, amateur and professional arts practice. He developed a new performance piece 'Anniversary' in collaboration with West Yorkshire Playhouse (now Leeds Playhouse) in 2016. In 2017 he toured across Asia making work in Guangzhou, Hong Kong and Singapore. He is currently developing a large-scale piece for Leeds 2023 entitled The Bus Pass Project.
As Artistic Director of Northern Stage in Newcastle (1992-2005), Alan is best remembered for his very successful adaptations of George Orwell's works. His production of Animal Farm (1993) stayed in the repertoire of Northern Stage for 12 years and toured in Spain, France, Netherlands, Israel and across the UK. In 2001 he made a version of 1984, which has inspired subsequent productions in Paris, Freiburg, New York and New Zealand. In 2003 he co-produced with Calixto Bieito an adaptation of Orwell's memoir, Homage to Catalonia. A Clockwork Orange, made in the late 1990s, was another successful production with the Northern Stage Ensemble touring the UK for four years.
Alan has always been a strong advocate of internationalisation of the UK theatre scene. He worked closely and over many years with Lev Dodin, and repeatedly presented Robert Lepage's and Peter Brook's work in Newcastle. He has also collaborated with and presented the work of the Belgian director Alain Platel, the Hungarian director Gabor Tompa, and the Romani director from FRY Macedonia, Rahim Burhan.
In 2003, he initiated the Newcastle / Gateshead Gypsy Festival which presented the work of Eastern European and Spanish Romani artists as well as an original piece by the Northern Stage Ensemble, Black Eyed Roses, which he later reworked for a Romanian production.
Later that year, a Newcastle-based adaptation of Wim Wenders' film Wings of Desire was created by Alan with his ensemble and a community theatre group based at Northern Stage. He was subsequently invited to make a Copenhagen-based adaptation of the film using the same model at the Betty Nansen Theatre in 2005.
Mind the Gap - Joyce Nga Yu Lee and Paul Wilshaw
Mind the Gap is one of Europe's leading learning disability theatre companies, and creates work for UK and international audiences.
The company's vision is to work in an arts sector where there is equal opportunity for performers with learning disabilities: a world where performers are trained, respected and employed equally, and feature every day on our stages and screens.
The company works in partnership with learning-disabled artists to deliver a bold, cutting-edge and world-class artistic programme that impacts locally, nationally and internationally. Work that excites, surprises and challenges audiences.
"Our aim is to make great theatre that makes audiences think differently. By "theatre" we don't just mean shows that you can see in arts and theatre venues, but also performances and events in different spaces and places."
Mind the Gap was formed in 1988 by Tim Wheeler and Susan Brown, and is based in Bradford. The Mind the Gap team includes a Board of Trustees, Staff Members, Associate Artists, Resident Artists, Ambassadors, Students and Volunteers.
Garry Lyons (School of Performance and Cultural Industries)
See all School of Performance and Cultural Industries events
Emergence 2020
School of Performance and Cultural Industries - Tuesday 28 January 2020
Postgraduate open day
School of Performance and Cultural Industries - Friday 7 February 2020
Professor Paul Fryer: Viktor Simov – Stanislavsky's designer
School of Performance and Cultural Industries - Monday 10 February 2020


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Bodendenkmäler der Gemeinde Lautrach

Bodendenkmäler in der Gemarkung Lautrach

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Bodendenkmäler in der Gemarkung Legau

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Bodendenkmäler unbekannter Gemarkung

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On 29 January 2015, the EAF launched its first report, the EAF Report 29Jan2015-english. This report is based on the latest scientific knowledge about ADPKD and insights from leading experts and patient advocates. It also features results from recent surveys of patients. The report was launched in Brussels at a meeting attended by patients advocates, representatives of various health stakeholder organisations, EU officials and ADPKD experts.
• Explain ADPKD and raise awareness of the disease and its implications for patients, health services and economies in Europe - issues that are often under-recognised by many health stakeholders and policymakers.
• Recommend strategies to improve ADPKD care within the context of health policy development at the European and national levels.
• Encourage and facilitate collaboration between those individuals and groups involved in the management of people with ADPKD, including health policy-makers, healthcare providers, payers, patients, caregivers and industry.
The Report is endorsed by PKD International and EGAN, the Patient Network for Research and Health. It was developed in alignment with the Kidney Disease: Improving Global Outcomes (KDIGO) Controversies Conference Report on ADPKD (Kidney International, in press). It is accompanied by the "Brussels Declaration on ADPKD", which encaptulates the EAF recommendations and their rationale.


text:
The New Christmas Tradition
by Mark Steyn
Mark's Midweek Notebook
https://www.steynonline.com/9084/the-new-christmas-tradition
Yesterday, the usual "known wolf" - that's to say, known to the highest levels of the French security apparatus - killed three and wounded a dozen more in this year's first attack on a Christmas market - in Strasbourg. The jihadist yelled ...oh, go on, take a wild guess: Joyeux Noël? Bonnes Fêtes? No, he stuck to the traditional greeting.
Just over two years ago, in the summer of 2016, I met a delightful German lady who lives just across the border from Strasbourg and whose husband, in fact, was born and raised there. Along with her child, my friend, as I put it, "had found herself on the receiving end of some vibrant multicultural outreach from one of Mutti Merkel's boy charmers":
As a result, she no longer goes out after dark. She had also decided - with reluctance, because she enjoyed it - to cancel her participation in a local Christmas market, where she'd sung carols every year - in broad daylight.
'Why would you do that?' I asked.
'Because it's Christmas,' she said, 'and I'm worried Christmas will be a target.'
I have never forgotten those words - because they were at once both absurd and chilling. But four months later "a German citizen" attempted to blow up the Christmas market at Ludwigshafen, and a mere two years on it is now taken for granted that Christmas markets across Europe have to be held behind the ugly throttling barriers of "security" - which as we saw yesterday are, ultimately, never secure.
Because we won't put up security barriers where they count - at the nation's border - we have to have ever more intrusive and constricting security on everything within the border. I have written about it innumerable times; I even wrote a short story about it. But the men and women who govern us refuse to change the policies that enable Yuletide slaughter, and their citizens continue to vote for them. And so Strasbourg's mayor Roland Ries says that "life must go on" - which fatuous bromide ensures that the killing of his citizens will go on. Because he and the rest of a depraved political class lack the will to do anything about it.
Spare a thought for those killed yesterday. We hardly do. Not anymore, not unless the death toll's three figures. The life that's going on is diminished and degraded by such complacency and indifference.
~I made a rare Tuesday appearance with John Oakley on Toronto's Global News Radio 640 to talk about sundry topics in the news, including the CBC's restoration of "Baby, It's Cold Outside" to the seasonal playlist, Gitmo alumnus Omar Khadr's application for a Canadian passport, and a subject not unrelated to yesterday's killings: treason as just another vibrant swatch in the quilt of multiculturalism. Click below to listen:
In fairness to Omar Khadr and his "Canadian" family, they have mostly killed and wounded people on distant battlefields far away. Since their heyday, Canada and the rest of the west have continued to admit vast numbers of people - like yesterday's killer - who find it more convenient and satisfying to slaughter us in our own home towns.
~For that brave band who prefer me in vision, tomorrow evening, Thursday, I'll be with Tucker Carlson south of the border at 8pm Eastern/5pm Pacific.
For those Ontario listeners who enjoy my visits to the Oakley show, we hope to see a few of you at my forthcoming stage appearances with the great Dennis Miller in western New York and Pennsylvania. We know at least one Montrealer coming to the Rochester show, but, if you're anywhere along the QEW, it's a lot closer. And Kingston (or even Ottawa) isn't that inconvenient for the Miller/Steyn Syracuse gig.
And for those who prefer me in audio only I'll be back on Friday evening for the second of this year's festive Tales for Our Time.
Tales for Our Time is made possible through the support of members of The Mark Steyn Club. If you're interested in joining the Club, please see here. Likewise, don't forget our second Steyn Club cruise, setting sail on September 4th next year, or our special Christmas Gift Membership. The latter comes with a special personalized Christmas card from yours truly and a handsomely-engraved set of three of our most popular Tales for Our Time for your loved one to listen to in the car or perambulating through the wilderness or almost anywhere else. For more details on our special Christmas Gift Membership, see here. We are grateful for the support of all our Club members around the world - from London, Ontario to London, England, to London, Kiribati - and look forward to welcoming many more in the months ahead.
Mark's Most Wanted
Love in a Time of Plenty: Judy Holliday and The Solid Gold Cadillac
Tal Bachman: New Year's Day, part three


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All access and use of the information and resources on this server is at the risk of the user. Eva Burrows College does not accept any liability for any loss or damage suffered due to anyone directly or indirectly relying on any information stored on this server.
Eva Burrows College owns the copyright in the information provided on this site, unless stated otherwise. All course content contained within this site remains the intellectual property of Eva Burrows College and The Salvation Army and may not be reproduced or redistributed, whether in part or in their entirety, without the written permission of Eva Burrows College.
Please email us at enquiries@ebc.edu.au If you would like to bring to our attention any matter on the Eva Burrows College site.


It often gets very heated between drivers competing for world class status in the FIA GT1 Race Series, Yesterday (June 5) at Silverstone Raceway the driver of the GTR (Richard Westbrook) and DB9 (Stefan Mücke) got into it on the track, resulting in a sorry end for one of the drivers.
It begins with the DB9 being rear swiped and sent spinning off the track by the #22 Nissan GTR. From the in-car shots it really doesn't look like Westbrook did it on purpose, the DB9 recovers quickly and immediately catches up to the GTR again, when he tries to pass Stefan clips the GTR and sends both cars spinning off the track, this time into the wall, putting an end to his race. In racing, Revenge is not always the best strategy, and often leads to regretful outcomes.
The Series is debating disqualifying the driver for his act of revenge by suspending his License.


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redpajama_set_name: RedPajamaGithub


Discover our new 28 Day Divine Renewal Program, a tailor-made solution for overworked skin. It is designed to work in harmony with the skin's natural rhythms to enhance its natural ability to regenerate. This program combines, for the first time, superior-quality immortelle essential oil from Corsica with a blend of active ingredients from the Corsican maquis, selected for their renewing properties.


This weekend people will lose their minds, then behave poorly, over the opportunity to purchase inexpensively priced electronics, clothing, jewelry, and toys. As a result, you're probably expecting to hear a pastor or a church leader say something abrasive about a "consumer mindset" or human greed.
It's not that people aren't selfishly motivated to try to purchase their own satisfaction or that their ugly displays of entitlement don't rankle my middle class sensibilities.
I haven't participated in the Black Friday feeding frenzy for nearly 9 years, but it isn't because I'm taking a stand against something, or trying to make a point. I just don't buy people televisions sets, digital cameras, or video game systems for Christmas.
Which is interesting because if you look at the numbers in regards to consumer debt and bank card financing, neither do most of the other people who hit the stores early on Thanksgiving weekend in search of great deals.
While they may not have the money, they are willing to risk their future by buying on credit because Christmas is coming.
Christmas is the reason that they blow their budget. Christmas is the reason that they won't be able to send their kid to college. Christmas is the resason that they decide to show their love for someone by purchasing them a… wait for it… Blu-Ray player or cell phone.
Don't get me wrong, if you love me and you want to give me a Blu-Ray Player or a waffle maker or 150 $1 dvd's from Target, I am not going to stop you. Go ahead, wrap it up and send it my way.
Just don't pretend that it has anything to do with Jesus being born.
This year my yuletide season isn't going to be wasted on protesting the obvious and odious because I understand that it's possible to buy someone an HDTV and still be a "good Christian" who didn't trample someone in a parking lot or berate a customer service rep because their electronic hamster section didn't meet your electronic hamster desires.
We all know that it's possible to participate in the buying of merchandise and not be a Christmas sellout in the same way it's also possible to use Christmas as your excuse to "not let anybody down" or buy yourself that special treat that you so richly deserve.
But why use the day that Jesus came to save the world as an excuse to give someone a gift that you can't actually afford? Why use it as an excuse to hand someone something that actually keeps them from thinking about Jesus for the rest of HIS birthday? Why not save and spend your money on the people you love in an appropriate fashion on any other day of the year?
You know that it's possible to buy your kids a PS3 and give it to them on March 8th right?
It is perfectly acceptable, and much more effective gentlemen, for your wife to recieve diamond earrings on say, a Thursday.
You can buy your dad a bottle of Old Spice and a tie for Flag Day.
Buying your loved ones a tangible, technologically complex, piece of entertainment equipment to celebrate the most supernaturally mystic event in history is as big a stretch as believing that watching Ke$ha sing about drunken partying actually has something to with female empowerment.
And let Jesus be the reason for it all?
When you treat your family right over the course of the year, Christmas doesn't have to be an extravagant outlay of merchandise in an attempt to maintain fragile relationships. It gets to be a celebration of the Person who treated us right, even though we were greedy and consumer minded.
That has everything to do with Jesus being born.


So today saw Ob-Gyn who is so so lovely. All is well. 25 and half weeks down and 14 to go!
I also had to make appointments for glucose testing tomorrow. I know hubbie is hoping I pass, just so he doesn't have to listen to my rant of how horrible and what a con the diabetes specialist folks are. Here's hoping!
Then made a dentist appt for me, a dentist appt for wee lad.
Seeing accupuncture lady again this week and Physical therapy Friday. Thankfully for now that's still just once a week.
Then made another appt to see surgeon for follow up appt next week to check on her fantastic work taking the softball lipoma out of my arm.
Then my primary care called me and she wants to see me re H1N1 aka swine flu shot. She doesn't have it but I have to go see her they said which baffles me but I will.


text:
FreeSpace Releases »
Mission & Campaign Releases »
Topic: RELEASE - The Procyon Insurgency
Pages: < Prev 1 2 3 [4] 5 6 7 8 9 10 11 ... 45 Next > Go Down
Author Topic: RELEASE - The Procyon Insurgency (Read 253030 times)
Snake on a Cain
Re: RELEASE - The Procyon Insurgency
Quote from: Sparky on April 30, 2007, 11:55:46 pm
Stealth mission pissed me off... took me about 50 trys (not kidding either...) I even put it on very easy. Bah, that blew.
That mission was easy, it was just like Playing Judas.
To consider the Earth as the only populated world in infinite space is as absurd as to assert that in an entire field of millet, only one grain will grow. - Metrodorus of Chios
I wept. Mysterious forces beyond my ken had reached into my beautiful mission and energized its pilots with inhuman bomb-firing abilities. I could only imagine the GTVA warriors giving a mighty KIAAIIIIIII shout as they worked their triggers, their biceps bulging with sinew after years of Ivan Drago-esque steroid therapy and weight training. - General Battuta
Dr. Evil
Reply #61 on: May 01, 2007, 12:00:35 pm
For those of you having trouble with 13, you want to spend a minute or two observing the fighters' paths before you head in. You will notice that heading straight in is not the optimal path to the target. Instead, you want to fly around the "sphere" of fighters and approach it from the top or bottom. This is described in the walkthrough.
In any case, the updated version of this mission will adapt to the difficulty setting. It should now become a walkover on very easy, well, unless you really suck.
When you say you've fixed the issues, will we have to redownload it?
It's a patch file containing only the modified data. You don't need to download the whole thing again, although I will update the main download as well for people who haven't downloaded the campaign yet.
I'm interested in Dasmar's plans. What was that Ancient Device for? What does he mean with return?
I'll post a detailed explanation in a day or two. I had written a bit of an essay in response to this last night, but didn't have time to finish it.
I plan on installing this new campaign tomorrow but I just want to make sure that all that's required is to download all of the links provided on the dl page?
Yeah, that's it. They don't replace any FS2 files. You might want to wait a bit though, as I'm planning to release the patch tonight, so you can just get the updated version.
Quote from: CP5670 on May 01, 2007, 12:00:35 pm
That's basically what I did, only I went left and then went though the left of the "sphere" (turning right), making adjustments when I got too close to the fighters. Took me about a minute to get in doing that, and about the same time to get out. Mission was too friggin' long though.
Alan Bolte
Deneb III
I found it much easier to get out than to get in. Right now I'm stuck in the nebula mission. Somehow I get through all that scripted stuff and then the shivan fighters blow me to bits. It's a bit irritating to have to fly around doing nothing for so long each time. Like WC Saga, heh.
Anything worth doing is worth analyzing to death -Iranon
Mission was too friggin' long though.
Yeah, I agree. The funny thing is that conversation originally used to be much longer, almost 20 minutes by itself. I removed a ton of stuff from it to cut down on the length as much as I could.
I was originally planning to have a big cutscene going on where all the stuff they talk about is actually happening. It would have made things much more interesting, but I had to drop that idea due to time constraints.
You said cutscene and it made me think something Independence War-ish. Maybe someone could change that...
I skipped mission 17 (well, I tried to...). the mission seems to be a 2 parter... it loads...then I'm stuck ,the screen flickers.... and i can't restart the the first part of the mission (except in the simulator) ,so i can't complete the campaign. Is there a skip-level cheat or a way to jump back a mission? ( or better: a patch )
Dude, holy crap! I recognize some of these music tracks you use!
Order of the New Dawn Theme - Ground Control
The Waiting (Or something like that) - Battlezone
There's another one you used too but I don't know where it comes from.
Also you make good use of the FS1 and Silent Threat tracks.
I'm generally not a fan of those either, but I figured that the rest of the campaign is hectic enough that a few "calm" missions wouldn't hurt too much at that point.
You want to close in on the Shivans (using the usual corkscrew maneuver) before attacking them. Their ships are pretty weak once you get past their weapons, and they don't fire much at close range.
Exactly right. The other one you're referring to is probably is the "neutral" debriefing track, which is from Earthsiege. They had to be modified a bit to make them suitable for PI. The rest are from FS1 and the Freespace masters archive that Nix sent me, and the intro track is a simple remix I made from some unfinished stuff in that archive. We'll release all that stuff once we have everything from Dan Wentz, as the archive is currently somewhat incomplete.
I skipped mission 17 (well, I tried to...). the mission seems to be a 2 parter... it loads...then I'm stuck ,the screen flickers.... and i can't restart the the first part of the mission (except in the simulator) ,so i can't complete the campaign. Is there a skip-level cheat or a way to jump back a mission? ( or better: a patch)
That doesn't sound good. It seems that this is a problem with FS2 itself rather than PI. I don't know exactly how the game behaves if you try to skip a mission preceding a red alert one after dying, but from what you're saying it sounds like it does not work. I am afraid your campaign progress may be hosed, but you can still play all of those missions in the mission simulator by hitting ctrl-shift-s in the selection screen. The only thing you will lose out on is your work on taking down the turrets in 15 (which stay dead in 17), but the mission still uses a default selection of destroyed turrets if played through the simulator.
Former JBX MegaRaptors
The problem for me on mission 13 wasn't flying TO the ship... but leaving. holy crap that was a pain. I knew how to get in no problem, just wait for Cancer 3 to fly in front of you and start to turn away. then just a straight shot. But leaving, why that was so hard I dunno. I think Cancer 2 kept spoiling it for me.
Yeah, kudos on making it work.
Actually, I was probably thinking about the FS(2) beta mainhall track. I didn't even notice the debriefing one.
[EDIT]Not to sound like a kiss-ass or anything, but also more kudos to making the speculation stuff seem plausible.
« Last Edit: May 01, 2007, 08:36:35 pm by Cobra »
Vretsu
Gah! Bloody mission 3.
The sentry guns fired beams at me.
I was all like, "oh, look, a pansy Vasudan Edjo gun. DIE!"
It was all, "WTFPWN!!"
*zapzapzapzapzapzapzapzapzapzapzapzapzap*
"AAAAAAAAAAUGH!!!"
*zapzapzapzapzapzapzapzapzapzapzapzapzapzap*
*cheesy explosion sounds*
Back where he started
Porto l'azzurro Dolce Stil Novo nella fantascienza
Why we don't voice act the campaign...?
The Lightblue Ribbon
Inferno: Nostos - Alliance
Series Resurrecta: {{FS Wiki Portal}} - Gehenna's Gate - The Spirit of Ptah - Serendipity (WIP) - <REDACTED> (WIP)
FreeSpace Campaign Restoration Project
A tribute to FreeSpace in my book: Riflessioni dall'Infinito
My interviews: [ 1 ] - [ 2 ] - [ 3 ]
Spoiler that, dammit. We need that to be a surprise.
And unless CP5670 says so, no.
This campaign would probably be better off without voiceacting.
1.) The constant Vasudan-English translating would get annoying.
2.) As a Vasudan pilot, you should be able to understand Vasudan without a translation, anyways!
I think the biggest challenge in getting voice acting in, apart from gathering community support, is to find a way to generate the Vasudan voices. The stuff in the default vasudantalk.wav isn't going to cut it in many places (the Dasmar speech in mission 4, for example, would sound strange if it started with that). There are several Vasudan voice files in FS1 that would be perfect, but they're mixed in with the translator.
There is also the fact that I'm currently using a bit of a hack to specify the debriefing music, which wouldn't work if voices were also present there. Although I could probably get the SCP team to add in a proper way of doing this.
Thanks, but...what speculation are you referring to?
The guy tells you about that in the briefing though. Those sentries are still fairly ineffective, but they can at least do something. The Edjo is pure cannon fodder.
Also, I'm uploading the 1.02 patch now. This should take care of all the bugs and stuff brought up earlier. If you find mission 13 too hard, you can now turn down the difficulty level to make it easier.
Quote from: CP5670 on May 02, 2007, 01:01:58 am
Well, not speculation per se, I mean speculation since it's not FS canon.
***SPOILER***
I meant that the talk of the ancients, Shivans, and GTI were all plausible. They could have happened. For a second I was thinking this was actually canon stuff too, that's how believable it was, in my opinion.
TrashMan
T-tower Avenger. srsly.
God-Emperor of your kind!
Nice campaign...Actually, great campaign!
But honestly, Im not much into all those conspiracy campaign, as they tend to suffer from the *eh???* syndrom as I liek to call it. Either the bad guy ends up looking like the total idiot, or the GTVA, for being so utterly blind.
Nobody dies as a virgin - the life ****s us all!
You're a wrongularity from which no right can escape!
Quote from: TrashMan on May 02, 2007, 07:27:20 am
I'm not sure what you're on about here. Care to explain?
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text:
Here's How the Events Industry Could Slash Greenhouse Gases
Author: Barbara Palmer
Serving portobello mushroom and pepper fajitas instead of beef fajitas reduces the amount of greenhouse gases generated by more than 75 percent, according to a new report by the Center for Biological Diversity.
When it comes to reducing the amount of energy it takes to produce the food served at events, even little tweaks can have a big impact.
Just replacing dairy-based cream cheese on the morning bagels with vegan cream cheese cuts the amount of greenhouse gases generated by nearly a half, according to a new report, Catering to the Climate, produced by the nonprofit Center for Biological Diversity. And serving fajitas made with portobello mushrooms and peppers instead of beef reduces the amount of greenhouse gases generated by more than 75 percent and uses 92 percent less land to produce.
The positive benefits of replacing meat and milk-based food products with plant-based menu items keeps growing along with event size and as more plant-based alternatives to meat and dairy are served. When feeding 500 people, a low-impact menu can save five acres of natural habitat, avoid the emissions equivalent to 50 people flying round-trip from New York City to Chicago, and conserve nearly 100,000 gallons of water from irrigation and food processing, according to the report's authors, Isaac Emery, founder of Informed Sustainability Consulting LLC, and Jennifer Molidor, a scientist at the Center for Biological Diversity
It's a critical issue, the report's authors add, because, according to the United Nations' Intergovernmental Panel on Climate Change, the world can't meet its climate goals without significantly reducing meat consumption and the emissions associated with animal agriculture. In the U.S., 80 percent of farmland is used for raising animals and animal-feed crops, and half of all consumed water goes toward meat production.
The impact of the $11 billion-a-year catering and events industry — including conferences, weddings, and corporate events, as well as school dining halls — is so large, that substituting plant-based menu items can have a "significant environmental impact," the study's authors write. Yet most venues don't offer them, they add.
What's Stopping Us?
In creating the report, the Center for Biological Diversity surveyed a small group of meeting planners and found that 83 percent would prefer a streamlined approach of offering an entirely plant-based menu to all attendees or offering meat and dairy as "opt-ins," compared to adding more plant-based options to conventional menus.
But 40 percent of respondents identified venue limitations or availability of food as obstacles to offering non-meat-based menus. If venues and caterers offered "greener" options, event planners would be more willing to switch to low-impact menus, wrote the report's authors.
RELATED: How the Virginia Beach Convention Center Created a 100% Plant-Based Menu
And the leading obstacle cited by planners was the perception that event attendees may not be satisfied with plant- based options — 37 percent of the planners pointed to attendee expectations and their response to alternatives as obstacles, along with donor and board objections (4 percent). Two percent cited higher costs as barriers.
The more positive experiences that attendees have with plant-based menus, and the better-trained that chefs become in preparing delicious plant-based options, the faster that will change, write the report's authors.
"Public demand for plant-based, low-carbon menus is growing quickly," Molidor said. "Even small changes in purchasing, like replacing dairy with plant-based milks and cheeses, can bring substantial benefits to suppliers and their clients. When the event and catering industry serves plant-based menus, it's an environmental and culinary success."
Barbara Palmer is deputy editor of Convene.
This page from the Catering to the Climate report compares the environmental costs of common catering items and those of plant-based alternatives. GHG stands for greenhouse gases. (Center for Biological Diversity)


text:
HOLLYWOOD. BURNING MAN. EDINBURGH FRINGE.
I am moving out of Hollywood.
I went to my last Burning Man in 2005.
I have just decided to go back to Edinburgh Fringe this summer.
Sometimes people ask me if they should take a show to Edinburgh. I don't know. Below is something I wrote right after I got back last fall.
I just made a T-Chart of pro's/con's in order to actually decide that my Edinburgh Fringe experience was a good one. We all say "roller coaster" to each other during the month. Oh yeah, well, it's a roller coaster isn't it? which is to say that we understand that even if we had a really good show today, it's bound to be a shit show tomorrow, or something in between, or whatever man, the point is, there is no such thing as having a "good time" or a "bad time" at edinburgh fringe. It's just a fucking thing. It's no thing. It's every thing.
"You're addicted now," said Barry Church-Woods, who is one of the main administrators of the festival, and also—weirdly, considering it's the biggest festival in the universe—a super-approachable guy. Also clearly a maniac.
But maybe it is a kind of addiction. The Edinburgh Fringe is an incredible cocktail of Hollywood and Burning Man in a gorgeous old Gothic city. Maybe, sure, maybe I'm addicted now. Or maybe I'm just going again.
WHY IT'S LIKE HOLLYWOOD: Industry. Whatever That Means.
Edinburgh Fringe has Industry. People who make money off of performing artists! Mythical beasts! They come to Edinburgh to see shows and meet people! It could be YOU they meet, at one of the needy-grabby Fringe bars you need a secret card to get into. It could be YOUR show they wander into. And then BAMMO! LIFELONG TOUR!
That happens to some people, probably, maybe. Or it happens to differing degrees, but not the way you imagine. Or maybe it totally happens a lot.
The smartest advice I got was to be specific in goal-setting. Do some research, find some names, introduce yourself as much in advance as possible, follow up. Just try to move a teeny bit further along in your plans for world domination, in a specific, down-to-earth, case-by-case way. Keep your head down, except to notice the moody Scottish sky, hanging delicately above all that massive stone and chimney.
Being specific worked wonders, because on the "pro" side of my T-chart were actual people I invited to see my show, people who came. So they came! That's a goal met! Mazel tov! You don't know what/where/how may lead to gigs/gigs/gigs. The more specific you are in terms of whom you want to meet, the more clear your results or lack of results will be to you. And that is useful information for one's emotional brain, which during this festival is all over the goddamn map.
WHY IT'S LIKE HOLLYWOOD: Annoying Personalities Everywhere (Including Your Own).
Because this festival is aware of its own Star-Making-Potential Myth, stakes feel high for everyone, and everyone becomes potentially the worst version of him/herself imaginable. Or maybe it was just me. I was moody as hell, all month long. Some of those moods were great moods. I cried at BEAUTY, all month long. I was deeply moved by positivity, by hope, by art. I moved in a ethereal way, floating above a month-long Achilles tendon issue. How did I survive all those cobblestones with all that heel pain? Euphoria, my friends. There are euphoric moments.
But the bad moods were horrible. Feeling un-cool, worst of all. My wise flatmates talked me off many a metaphorical fire escape. (Seriously, living with friends was goddamn genius. I did make friends, but it's just not the kind of festival to go at alone.) It was a month in which I was somewhat embarrassed to be me, in terms of the levels of self-involvement that feels like it's in the water. And I drink a lot of water.
A month-dose of that kind of behavior is not horrible, though, when you consider that so many artists in Hollywood have to deal with that bullcrap every day of their life. You can take a month of Hollywood, right? Provided you can spend good portions of the rest of the year feeling more like a positively contributing member of society and not like a needy desperate devil-spawn of pipsqueak shitsackdom. Which Edinburgh did make me feel. Real highs and lows, is what I'm saying. Big ones.
It may be an addiction, Barry Church-Woods, but it's an addiction that can be managed.
WHY IT'S LIKE BURNING MAN: ARTISTIC ABUNDANCE!
Say what you will about Burning Man, it is full of creativity. There is creativity everywhere, not all good creativity, a lot of it inebriated creativity, but some of it genius creativity, plus plenty of damn fine creativity, and all of it inspirational. It does do something to you to be surrounded by that much creativity. And the hustle-bustle of Edinburgh when it's full of performers getting their shindig on certainly feels like something. You do feel part of something larger, that drop-in-the-bucket feeling, yes, but you know you love the bucket.
At Burning Man, there is no way you can see/experience everything. You will hear about some amazing fur-lined trailer where virgins drool cucumber water into your belly button to a chorus of singing bowls, and you can't find the cross streets or someone says it moved or left or whatever it's finished you lose.
So you learn, at Burning Man, that whatever journey you're on, it's the right one to have. And you have your adventures and paths, and you realize that there is an infinite amount of paths that were probably equally awesome, and the one you were on was uniquely yours.
There are far more shows at Edinburgh than you can possibly see. You want to see a lot of shows, and your capacity to see them is compromised. So you become okay with your unique journey.
One dear flatmate of mine experienced at least two dead raw chicken bodies in two different shows she saw, and she realized that dead raw chicken bodies were a theme in her Edinburgh experience.
Themes in my Edinburgh experience: catchy music snippets, spotlights, tiny shorts, drag, tea, crying, weight-shifting, cobblestones, cobblestones, cobblestones.
WHY IT'S LIKE BURNING MAN: IT TAKES ALL KINDS!
There are people who get up early at Burning Man, and sleep at night. When I did my time in Black Rock City, I did not meet these people. But we all heard about them. Apparently, they all made pancakes for breakfast.
Just like that, there are a lot of different ways to do Edinburgh Fringe. I know I mentioned the omnipresence of Industry, but I met plenty of performers who gave a lot less of a fuck about it. Why they were so sang-frois? Because they come to Edinburgh Fringe every year, the way some kids go to summer camp. They bring a new show, new material, every year. In this way, they get good at the Beast, they have perspective about the Beast, and they are far more fun to hang out with than, say, me. Guess what else: they're all fricking British. Or a lot of them are. There are some toughass not-Brits who go every year, and they are epically heroic and inexplicable. But of course it's mostly Brits who go every year, because it's a fricking train ride away. Why shouldn't they go?
And just like that, they get to be better artists. A month'll do that to you. It's a really good system, that way. Brits are lucky to have a Hollywood Burning Man so close at hand.
WHY IT'S NOT LIKE HOLLYWOOD OR BURNING MAN:
A lot of people, myself included, wound up caring a whole lot about how many stars were printed next to reviews of our shows. Did I get loads of 5-star reviews? No I didn't. And I officially think stars should go fuck themselves.
I don't think I'd feel differently if I had 5-star reviews plastered all over my face like so much foundation. Stars are an insult to reviewers, artists and audiences. Who wants this rating system based on no generalized criteria and no objectivity? Doesn't everyone want readers to actually read the reviews? GET RID OF THOSE LITTLE BULLSHITS.
Alright I'm off the podium.
I don't think my show is the son of Christ, but I think it's good enough.
Edinburgh reminded me I have a lot of faith in it, and love it.
That's a nice thing to know.
Edinburgh is toughening; toughening is healthy. Yeah, okay, do Edinburgh. At some point.
When you love the shit out of your show and don't give a fuck who likes it, because you love the fuck out it. When you love your show so goddamn much you want to give it a big crazy present. When you want to marry your goddamn show, you are that much soul mates. It is you; you are it. It is a once-in-a-lifetime show for you. Maybe twice.
Pack those bags for August. It's almost the honeymoon. Forget the crappy moments, and go love your show into the sunset. I LOVE THE FUCK OUTTA YOU, SHOW, you'll whisper, I'MA GIVE YOU SOMETHING YOU NEVER FORGET.
Edinburgh Fringe: pretty good, overall. It might be worth checking out. Three stars.
ASK THE CRITICS


Brand new 2018 Naish Hover 120 Foiling SUP board, 7'6 x 30" 120vol, tough light full sandwich construction, comes with two US box reinforced foiling boxes and Quad boxes with glass fins. NO FOIL with this board we do have plenty of options of foils on the shelf from Naish and other brands. The Naish Hover 120 is the perfect first time foiling SUP board, gives an amazing sensation once up and foiling, super fast and silent, but does require an experienced SUP'er to ride it. Suitable for riders to learn to SUP foil on, super stable for it's size. Last couple now on deal £500 off.


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text:
Statements on corridors around communication routes unacceptable for us – Iranian Ambassador to Armenia
YEREVAN, APRIL 30, ARMENPRESS. Governor of Syunik province Robert Ghukasyan received Ambassador of Iran to Armenia Abbas Badakhshan Zohouri. The meeting was also attended by Mayor of Kapan Gevorg Parsyan and head of the Syunik department of Police Alexander Mamikonyan, the Governor's Office said.
The main topic discussed at the meeting was the Consulate of Iran that is going to open in Kapan and in this context the actions to be taken to deepen and strengthen the cooperation between Syunik and Iran in different areas.
Governor Robert Ghukasyan thanked the diplomat for the visit, stating that the relations with Iran are very important for Armenia, especially for Syunik, and the governorate is ready to do everything to make the bilateral ties more effective.
In his turn the Iranian Ambassador said that despite the close cultural ties, the economic relations between the two countries are not at a desirable level, stressing the need for efforts on this direction. He also thanked for helping Iranian citizens who appeared in a difficult situation in the inter-state highway because of heavy snows this year in March.
The Ambassador reminded the position of Iran on the inviolability of Armenia's internationally recognized borders and said that all countries should respect it. "Statements on corridors around communication routes are unacceptable for us, we believe that Armenia should preserve its territorial integrity", he said.
The Governor of Syunik and the Mayor of Kapan thanked the Ambassador for Iran's official position on "corridor". "Your opinion is important and is appreciated by each resident of Syunik", Robert Ghukasyan said.
9 months ago the Iranian side reached an agreement to open a consulate in Kapan, and the Ambassador said that the purpose of this is to develop the relations in all areas. He also introduced the consul at the meeting, adding that they will soon present the staff of the consulate to the Armenian foreign ministry.
The Iranian side also talked about its interest to the section of the North-South highway passing via Syunik and informed that there are Iranian companies who expressed readiness to participate in the construction of that section.
The sides also discussed economic, cultural, educational and other issues.
Gevorg Parsyan informed that the teaching of Persian will start in the community's educational institution from September.
At the end of the visit the Iranian guests observed sites for the future consulate building in Kapan.
Tags: ArmeniaIran
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Appointment of a Service Provider(s) to collect data for the 2022 annual round of the South African Social Attitudes Survey (SASAS)
Bid Description: Appointment of a Service Provider(s) to collect data for the 2022 annual round of the South African Social Attitudes Survey (SASAS) – with the possibility of an extension for 2 additional survey rounds (conditional on the performance of the service provider and HSRC funding status).
Bid Number: HSRC/09/2022/23
The Human Sciences Research Council (HSRC) is a research organisation that advances social sciences and humanities for public use. The HSRC subscribes to the values of good governance, accountable public administration and sound management of the resources as reflected in the Public Financial Management Act of 1999 (PFMA as amended by Act 29 of 1999) and other laws and regulations applicable to public administration.
Established in 1968 as South Africa's statutory research agency the HSRC has grown to become the largest dedicated research institute in the social sciences and humanities on the African continent, doing cutting-edge public research in areas that are crucial to development.
Our mandate is to inform the effective formulation and monitoring of government policy; to evaluate policy implementation; to stimulate public debate through the effective dissemination of research-based data and fact-based research results; to foster research collaboration; and to help build research capacity and infrastructure for human and social sciences.
Preference Points System: 80/20 (Price: 80 and B-BBEE: 20).
NB: The minimum threshold on functionality is 70 points to qualify for further evaluation stages.
Access to Bid Documents:
Bid documents will be free of charge and available on the HSRC website (www.hsrc.ac.za). Bid documents must be deposited into the HSRC Tender Box at HSRC building, 134 Pretorius Street, Pretoria.
NB: The HSRC shares the building with the Department of Social Development. Bidders are requested to ensure that their proposals are deposited into the correct tender box.
4. Bid closing date: 23 August 2022 at 11:00.
5. Contact Details:
Bid process and document enquiries:
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The HSRC is committed to cutting-edge research that supports development nationally, in the Southern African Development Community (SADC) and in Africa.
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I attended my very first festival… the 2013 Rubber Duck Regatta on Friday and Saturday, September 20th and 21st. Friday was a raining day so there were many people at the event with the exception of us vendors! This being my first event, the rain was not really a bad thing as I found was not totally prepared! Let's just say there were several trips made to the local grocery store as well as office supply store!
Saturday, turned out to be a gorgeous day and I was fully prepared for my customers !
As you can see from the pictures my helper's were all eager to sale mini cakes and cobblers! Overall the experience was great! We all had a lot of fun and met some interesting and wonderful people who appreciate great desserts! I look forward to see you out at my next event…..until next time…Keep smiling!


text:
Regional Woman Falls To Her Death Inside World Trade Center Oculus
Woman Falls To Her Death Inside World Trade Center Oculus
Saturday, February 11, 2017 at 7:48 pm | ט"ו שבט תשע"ז
NEW YORK (AP) -
A 29-year-old New Jersey woman plunged about 30 feet to her death off an escalator inside the famed World Trade Center transit hub known as the Oculus on Saturday morning, police said.
Jenny Santos, of Kearny, was trying to retrieve a hat dropped by her twin sister while the two were on an escalator at about 5:30 a.m. when she reached too far over the railing and tumbled over the edge, a law enforcement official said. The official wasn't authorized to speak publicly about the incident and spoke to The Associated Press on condition of anonymity.
Port Authority police spokesman Joseph Pentangelo said the woman was taken to a hospital where she was pronounced dead. Police are investigating.
The striking $3.9 billion transportation hub was designed by architect Santiago Calatrava and provides connections between New Jersey's PATH trains and New York City's subways. It opened about a year ago and is used daily by more than 300,000 commuters. Light beams in from the windows in the dizzying, soaring platform to the shopping mall below.
New York Briefs – August 28, 2014
Founding Chairman of 9/11 Museum Dies
One World Trade Center Lit Up To Honor Wiesel
Expedited Autopsy Rules Garner Death a Homicide
Pioneer in Lamaze Dies


Surprise, Women's History Museum Actually Devoted to Jack the Ripper!
Surprise, Women's History Museum Actually Devoted to Jack the Ripper!
The founder of the controversial, soon-to-open Jack the Ripper Museum — initially presented as a museum that celebrates women in London's East End — is defending his vision of it as a site to explore local women's history and stories. Reports, however, suggest that he had always intended to establish a museum devoted to London's most notorious serial killer, misleading officials and East End locals along the way.
Yesterday, the London Evening Standard first reported that former Google diversity chief Mark Palmer-Edgecumbe submitted a proposal last July promising his institution would open as "the first women's museum in the UK." Tower Hamlet council approved his plan later last year. The museum instead has revealed itself as one devoted to the unsolved, serial crimes of Jack the Ripper, who brutally murdered a number of women in the late 19th century. Uproar has followed the revelation of the museum's true subject, with residents saying they were "completely hoodwinked and deceived" and others accusing the institution of glorifying gendered violence.
However, Palmer-Edgecumbe had previously co-registered an office on March 23, 2012 under the name "Jack the Ripper Museum Limited," then located in Kent; its status is now listed as dissolved as of March 25, 2014, with net assets of negative £738 (~$1,150). Tower Hamlet council received Palmer-Edgecumbe's application for the museum of East End women's history just five months later. His plan at the time promised "a world class museum that celebrates the historic, current, and future contribution of the women of the East End." The 35-page proposal also included several examples of proposed content, from a survey of the development of women's working conditions to an examination of the Suffragette movement.
Apparently the history of London's women most in need of understanding is the one associated with the city's most notorious murderer, who butchered his female victims in grizzly settings and circumstances. The museum, whose logo is the silhouette of a man in a frock coat and top hat with a pool of blood by his feet, is now slated to feature four floors and a basement mortuary of original and representative of artifacts. Newspaper clippings of the crimes provide the narrative, while medical instruments and knives allude to the violent murders. On view in the basement, "for over 16s only," are the original, explicit autopsy photos of the victims. Somehow, Palmer-Edgecumbe claims that this deviates only slightly from his proposal of July 2014.
On the museum's website, a message from Palmer-Edgecumbe also cites his original intentions, claiming that the museum "in no way glorifies or glamorises Jack the Ripper, quite the opposite, it presents the women of the East End's story for the first time." The Ripper crimes, Palmer-Edgecumbe explains, will shed light on the social context of East End during that era, touching upon issues such as poor living conditions and economic hardship.
Palmer-Edgecumbe assured the Evening Standard that his museum "is absolutely not celebrating the crime of Jack the Ripper but looking at why and how the women got in that situation in the first place." Only the first visitors exploring the museum after it opens ("next Tuesday," according to the Evening Standard) will be able to tell if it achieves that goal — that is, if it opens. Locals are reportedly attempting to overturn the council's decision, although Tower Hamlets said it ultimately has no control over how the museum plans its exhibitions. It is, however, "investigating the extent to which unauthorised works may have been carried out." Hyperallergic reached out to Palmer-Edgecumbe but has not yet received a response.
Notably, the museum intends to donate a portion of its profits to "local charities that promote women's rights and challenge child trafficking, domestic violence, and sexual exploitation. The first charity we will donate to is www.evesforwomen.org.uk." The link (which has since been removed) was broken, and although I assume Palmer-Edgecumbe had good intentions — as he allegedly did for his museum — and meant to share the website for Eaves Housing for Women, somehow such efforts seem to be implemented more for the sake of public appeasement than out of a sense of genuine goodwill or philanthropy. Meanwhile, a group dedicated to the history of East London Suffragettes hopes to make good on Palmer-Edgecumbe's alleged project by establishing an actual East End women's museum, calling for support on social media.