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Synthesis, identification and in vivo studies of tumor-targeting agent peptide doxorubicin (PDOX) to treat peritoneal carcinomatosis of gastric cancer with similar efficacy but reduced toxicity
Molecular Cancer volume 13, Article number: 44 (2014)
This work aimed to synthesize a cathepsin B (CTSB)-cleavable tumor-targeting prodrug peptide doxorubicin (PDOX) and study the in vivo efficacy and toxicities on an animal model of gastric peritoneal carcinomatosis (PC).
PDOX was synthesized using doxorubicin (DOX) attaching to a CTSB-cleavable dipeptide Ac-Phe-Lys and a para-amino-benzyloxycarbonyl (PABC) spacer. PC model was established by injecting VX2 tumor cells into the gastric sub-mucosa of 40 rabbits, which then were randomized into 4 groups: the Control (n = 10) without treatment, the HIPEC (n = 10) receiving cytoreductive surgery (CRS) plus hyperthermic intraperitoneal chemotherapy (HIPEC), the PDOX (n = 10) and the DOX (n = 10) receiving systemic chemotherapy with PDOX 50.0 mg/kg or DOX 5.0 mg/kg, respectively, after CRS + HIPEC.
The median overall survivals (OS) were 23.0 d (95% CI: 19.9 d - 26.1 d) in the Control, 41.0 d (36.9 d - 45.1 d) in the HIPEC, 65.0 d (44.1 d - 71.9 d) in the PDOX, and 58.0 d (39.6 d - 54.4 d) in the DOX. Compared with the Control, the OS was extended by 70% in the HIPEC (p < 0.001) and further extended by 40% in the DOX (p = 0.029) and by 58% in the PDOX (p = 0.021), and the PC severity was decreased in the HIPEC and further decreased in the PDOX and DOX. Animals receiving DOX treatment showed hematological toxicities with marked reduction of white blood cells and platelets, as well as cardiac toxicities with significant increases in creatine kinase mb isoenzyme, evident myocardium coagulation necrosis, significant nuclear degeneration, peri-nucleus mitochondria deletion, mitochondria-pyknosis, and abnormal intercalated discs. But these toxicities were not evident in the PDOX.
PDOX is a newly synthesized tumor-targeting prodrug of DOX. Compared with DOX, PDOX has similar efficacy but reduced hematological and cardiac toxicities in treating rabbit model of gastric PC.
GC is one of the most common malignancies in developing countries, where it ranks second in terms of incidence rate and third in terms of mortality rate among the male population, and ranks fourth in terms of both incidence rate and mortality rate among the female population, according to the most recent global statistics [1, 2]. GC is also the third leading cause of cancer mortality in China , where over 70% of GC has already become clinically advanced by the time of surgical exploration, thus surgical resection alone is no longer curative .
PC in GC has long been considered as a fatal clinical entity. It is defined as the implantation of tumor cells throughout the peritoneal cavity and characterized by the presence of tumor nodules of various size, number, and distribution on the peritoneal surface as well as malignant ascites, frequently resulting in locoregional morbidity without broader systemic metastases. Patients with gastric PC face a dismal outcome, with a median survival of about 6 months .
There is no standard treatment for gastric PC. Current treatments for such PC are systemic chemotherapy, best support care and palliative therapy. In order to tackle this problem, a new treatment modality called CRS plus HIPEC has been developed over the past 3 decades, taking advantages of surgery to reduce visible tumor burden, and regional hyperthermic chemotherapy to eradicate micrometastases . Increasing evidence has suggested that the combination of CRS and HIPEC could bring survival benefit for selected patients with gastric PC. In our previous experimental study , we have also proved that CRS + HIPEC could indeed bring survival benefit with acceptable safety, providing evidence to support this combined strategy to treat selected patients with gastric PC.
During the development of PC, GC cells secrete enzymes to facilitate cancer cells seeding and colonization on the peritoneum. CTSB is one of the key enzymes in this critical process, over-expressed in GC as well as other cancers [7–9] and actively involved in cancer invasion [10–12]. On the other hand, it is extremely low expressed in normal cells and inactive or loses activity as soon as it is dispersed in aqueous media away from cells . Thus CTSB has long been considered as a candidate target in cancer therapy .
It has been established in the MAGIC trial that the anthracycline-contained regimen is a useful chemotherapy for GC . DOX is a typical representative of anthracyclines. Although DOX is an important drug in chemotherapy, its toxicities are evident, such as cardiac toxicities and bone marrow suppression. To retain the therapeutic effect while reducing the side effects, Dubowchik et al [16–18] designed a smart prodrug of DOX, PDOX (Figure 1). In this modified DOX, Ac-Phe-Lys is a dipeptide specific for CTSB, and PABC (para-aminobenzyloxycarbonyl) is a self-immolative spacer . The prodrug is inactive when there is little CTSB activity, such as normal tissues and peripheral blood, thus avoiding the side effects on normal tissue. During cancer invasion, activated CTSB is over expressed on the exterior membrane of the invading cancer cells [19, 20], which cleaves the Ac-Phe-Lys dipeptide at the Lys-PABC bond . Then the exposed PABC spacer can self-hydrolyze upon deacylation  and free DOX molecules are released, resulting in direct killing of the invading cancer cells .
In this study, we synthesized the PDOX and evaluated the efficacy and safety of CRS + HIPEC with molecular targeted therapeutic regimen PDOX for targeted treatment of rabbit model of gastric PC.
Synthesis and identification of PDOX
PDOX (Ac-Phe-Lys-PABC-Dox · HCl) was a red solid powder with molecular weight of 1046.51 (MS: m/z calculated for C52H60ClN5O16: 1046.52, found: 1046.50), chemical purity of 99.1% (by HPLC), structure of C52H60ClN5O16 (by both of H1-NMR and C13-NMR) (Figure 2) and melting point of 180°C (decomposition), which is stable at -5°C to 0°C for 36-54 months, ambient temperature for 24 months, solvable in water, partially solvable in methanol and ethanol. PDOX used in this study was stored in the dark, dry area at 4°C.
PC model construction and histopathological characteristics
Rabbit model of gastric PC was established in all animals (100%, 40/40). On d 8 after tumor cells inoculation, small, hard and transparent tumor nodules developed on the greater omentum, and typical ulcerative cancer about 0.5-1.0 cm in diameter formed on the antrum of the stomach. No ascites was observed. No obvious PC was found in other regions. There were no differences in the PC severity among all the rabbits. This could be equivalent to clinical stage I PC by Gilly criteria  (Figure 3A).
All investigated tumor specimens showed extensive invasive growth and tissue destruction. The tumors, on the greater curvature of the gastric antrum, penetrated the mucosal layer to form ulcers. Histopathological study showed tumor nests penetrating the entire stomach wall, with typical invasion into the muscle layer and the gastric glands (Figure 3B). The tumor cells are round, oval or atypical morphology with many pathological mitotic figures. There were also conspicuous infiltration of lymphocytes, plasma cells and other inflammatory cells (Figure 3C). Apoptotic and necrotic tumor cells were observed in the central region of the tumor nodules (Figure 3D). Typical PC presented as tumor nodules on the surface of the omentum (Figure 3E) and intraperitoneal lymph node metastases were also observed (Figure 3F).
Typical ulcerative cancer with PC was observed in post mortem pathological examinations of rabbits in the Control. The stomach wall was totally invaded by the tumor to create cancer ulcer encased by confluent nodules on the greater omentum, forming a big tumor block. The abdominal wall and diaphragm were totally invaded by the tumor. Many tumor nodules formed on the intestinal wall, the mesentery and the retroperitoneum. Bloody ascites could be more than 100 mL. All the features are similar to the clinicopathologic characteristics of gastric PC in patients (Figure 3G).
Animal was observed to record OS. Two long surviving rabbits (one in the PDOX and another in the DOX, living more than 100 d) were euthanized on the 100th day by an overdose injection of 2% pentobarbital sodium. The median (95% confidence interval, CI) OS was 23.0 d (19.9 - 26.1 d) in the Control, 41.0 d (36.9 - 45.1 d) in the HIPEC, 65.0 d (44.1 - 71.9 d) in the PDOX and 58.0 d (39.6-54.4 d) in the DOX. Compared with the Control, the OS was extended by at least 70% in the HIPEC (p < 0.001, log rank test). Compared with the HIPEC, the OS was further extended by 40% in the DOX (p = 0.029, log rank test) and by 58% in the PDOX (p = 0.021, log rank test) (Figure 4A).
Tumor burden at the survival endpoint
At the study endpoint, the tumor burden in the Control was heaviest, with tumor weight of (137.51 ± 16.09) g, tumor-bearing ratio (tumor weight to body weight) of (7.88 ± 0.85)%, bloody ascites of (65.50 ± 33.45) mL, and ePCI score of (9.50 ± 2.17). Compared with the Control, the HIPEC had significantly reduced tumor weight (76.50 ± 11.41) g (p = 0.007) and tumor-bearing ratio (3.94 ± 0.54)% (p = 0.001), but not ePCI score (9.50 ± 2.17)% (p = 0.420) and bloody ascites [(29.16 ± 15.30) mL, p = 0.085]. Compared with the HIPEC, PC severity was further decreased in both the PDOX and the DOX, with ePCI score (6.40 ± 2.07, p = 0.020 in both PDOX and DOX vs. HIPEC) and bloody ascites (0.00 mL, p < 0.001 in PDOX and DOX vs. HIPEC); while there were no significant differences in tumor weight [(65.30 ± 14.55) g, p = 0.552, PDOX vs. HIPEC; (48.74 ± 16.31) g, p = 0.180, DOX vs. HIPEC)] and tumor-bearing ratio[(2.91 ± 0.69)%, p = 0.552, PDOX vs. HIPEC; (2.13 ± 0.78)%, p = 0.180, DOX vs. HIPEC]. There were no significant differences in tumor burden between the PDOX and the DOX (Table 1).
Body weight changes
The body weight of each animal was recorded every 4 d. No significant differences were found in initial body weight of 4 groups before the treatment. Perioperative body weight decreased in all groups because of the overnight fasting. In the Control, the body weight recovered once food intake was resumed but again decreased progressively till the study endpoint. In the 3 treatment groups, postoperative body weight decreased considerably after model construction and to the lowest 4 d after CRS + HIPEC, and then stayed at a low level during the post-operative phase until chemotherapy had been completed. The body weight began to increase at d 44 in the HIPEC and d 56 in both the PDOX and the DOX, and gradually increased to a higher level, which was related to both tumor growth of the living tumor-bearing rabbits and death of the failure rabbits with lesser body weight. Thereafter, body weight decreased progressively again until the study endpoint in both the PDOX and the DOX (Figure 4B).
Postmortem pathological examinations
By the time of animal death, detailed information on postmortem pathological examinations was listed in Table 2.
In addition to systematic examinations of all anatomic sites for possible cancer metastases, particular attention was paid to lung and liver metastases. As shown in Figure 5, in the Control, extensive liver metastases were observed in 8 animals, but no pulmonary metastases were evident in all animals, because of the very aggressive growth behaviors of the tumor led to short OS in this group of animals (Figure 5 A2, B2). In the HIPEC, as animals lived much longer because of CRS + HIPEC, fewer liver metastases were observed in 4 animals, but considerably much more pulmonary metastases were observed in 7 animals (Figure 5 A3, B3). In the PDOX and DOX, animals developed much fewer metastases in both the livers and the lungs, although the OS was further improved remarkably, because the animals received not only CRS + HIPEC to control local regional metastases but also systemic chemotherapy to control hematogenous metastases and lymphatic metastases (Figure 5 A4, B4, A5, B5).
Peripheral blood profile and major biochemistry changes
There were no differences in peripheral blood cell counts on d 2, d 6 and d 14 among the 4 groups, but the white blood cells and platelets were significantly decreased on d 36 after systemic chemotherapy in the DOX (p < 0.05). There were no differences in the hepatic and renal functions throughout the experiment among the 4 groups. In terms of cardiac functions parameters, there were no differences in creatine kinase and creatine kinase mb isoenzyme on d 2, d 6 and d 14 among the 4 groups, but creatine kinase mb isoenzyme was significantly increased on d 36 after systemic chemotherapy in the DOX (p < 0.05). Of particular attention, 2 animals in the DOX with short OS of 47 d and 53 d had exceptionally higher serum creatine kinase mb isoenzyme levels, 467.5 U/L and 656.4 U/L, respectively (p < 0.05) (Table 3).
Detailed studies on myocardium toxicities
Histopathological studies under both light microscope and transmission electron microscope found no myocardium toxicities in the Control, the HIPEC and the PDOX, but significant histopathological changes of myocardium were found in all animals (100%, 10/10) in the DOX, including coagulation necrosis of the myocardium with deep red stained cytoplasm, and the karyopyknosis (Figure 6 C1, H1, P1, D1 black arrows). Electron transmission micrographs showed that normal myocardium with abundant mitochondria packed around normal nucleus in the Control and HIPEC, rich normal but with slightly dilated cristae mitochondria scattered around normal nucleus in the PDOX, nearly absent mitochondria around the morphologically senile and degenerative nucleus in the DOX. In addition, partial myocardium-lytic necrosis (yellow arrow) and mitochondria-pyknosis away from the nucleus (red arrow) were also evident in the DOX. Complete, clear and continuous lines of normal intercalated discs in the Control, the HIPEC and the PDOX, but obscure, loosing and discontinuous intercalated discs were found in the DOX (white arrows).
There is no standard treatment for gastric PC. CRS plus HIPEC represent a multidisciplinary approach to this problem. It was first reported in 1988 by Fujimoto et al  on 15 patients with PC secondary to advanced GC, with a mean survival of 7.2 ± 4.6 months with acceptable morbidity. This new treatment modality gradually gains acceptance in many countries. The reported studies use different PCI scoring system to evaluate the extent of PC and different HIPEC approaches, but they produce similar results that CRS + HIPEC provides treatment benefit. In order to more objectively evaluate such treatment, it is necessary to study this treatment modality under experimental conditions, in which most of the confounding factors could be well controlled.
In our previous study [24, 25], we have established a stable rabbit model of gastric PC by injecting VX2 cancer cells into the sub-mucosal layer of the stomach. The non-immuocompromised larger animal model is characterized by typical ulcerative GC with progressive PC, making it more suitable for surgical interventional studies to evaluate CRS and HIPEC against gastric PC. This rabbit model of gastric PC has provided us with suitable platform to evaluate different therapeutic approaches against PC. On this first large animal model of gastric PC, we proved that CRS + HIPEC with docetaxel and carboplatin could significantly prolong the OS by at least 60% (40 d vs. 23 d) with acceptable safety. The addition of molecular targeted therapy with PDOX could provide much better survival benefits with satisfactory drug safety.
This study provided new approach of CRS + HIPEC combined with molecular targeted therapeutic regimen PDOX to gastric PC. Compared with the Control, the OS was extended by at least 70% (18 d) in the HIPEC (p < 0.001). Compared with the HIPEC, the OS was extended by at least 40% (17 d) with DOX therapy (p = 0.029) and by 58% (24 d) with PDOX treatment (p = 0.021).
In currently available chemotherapeutic regimens, anthracyclines are important drugs, as is well demonstrated in the MAGIC trial [26–28]. Anthracyclines cause cell damage by intercalating into DNA, leading to chromatin unfolding and aggregation, which ultimately results in apoptosis . However, like many cytotoxic agents, anthracyclines can cause serious organ damages. With DOX, toxicity to the heart and bone marrow are usually dose-limiting, with the maximum tolerated dose (MTD) far below the minimum curative dose (MCD). Therefore, strategies to shield the heart and bone marrow by excluding DOX from them have long been a top priority. Another strategy has been to target the drug to the tumor by attaching it to some tumor-binding moiety, e.g. a tumor-specific monoclonal antibody (MAb) such as trastuzumab used in chemotherapy for HER2-positive GC . Drawbacks to the use of MAbs have been that (1) the limited diffusion of MAbs due to their large macromolecules into solid tumors could compromise their potency; (2) heterogeneous antigen expression in large tumors will evidently lead a portion of tumor cells in the solid tumor unresponsive to MAbs; (3) the tumor Ag that binds the MAb is never completely tumor-specific, so that some of the drug goes where it does harm; (4) foreign MAbs are often immunogenic; and (5) MAbs therapy is very expensive [31–33].
In the present study, normal organs are protected by masking the cytotoxic drug DOX with a simple dipeptide that renders it nontoxic. At the tumor the mask is removed by CTSB, a ubiquitous proteolytic enzyme that is so destructive to tissue that normally it occurs only within cells, encased in lysosomes . Only tumor cells secrete CTSB externally, confined to their plasma membranes, for the purpose of penetrating basement membrane and extracellular barriers as they spread . The prodrug PDOX is rapidly cleaved by CTSB at the Lys-PABC bond . The resulting PABC-DOX decomposes at once to para-aminobenzyl alcohol, CO2 and free DOX [16, 21]. The PABC self-immolating linker is necessary because the CTSB’s active site cannot accommodate the bulky DOX molecule, but the smaller PABC fits into the active site . Free DOX released right on the tumor cells penetrates them readily, killing them. To be sure, a certain portion of the free drug may drift away from the tumor, but the concentration ratio tumor/heart-bone marrow should be much higher than if the DOX is given as the free drug, when the expected ratio is about 1. In this way, even without a positive targeting agent like a MAb it is possible to raise the MTD without significantly raising the MCD. The goal is to raise the MTD above the MCD, where cures become possible, but even short of that, a rise in the MTD/MCD ratio will enhance the effect.
In the previous study , human plasma stabilities of PDOX and releasing rates of free DOX were measured. The results indicated that PDOX was not hydrolyzed (no observable changes) over 6 - 7 h in human plasma without CTSB, but hydrolyzed in 16 min in CTSB rich conditions, and the appearance of DOX correlated precisely with the disappearance of PDOX at 495 nm, suggesting that PDOX could be stable in blood circulation and effectively released at the CTSB-rich tumor site. The evidence implies that PDOX could stay relatively stable in human blood circulation for a reasonable time, but the rabbit plasma stability of PDOX was not measured in this study.
DOX is one of the most efficacious anticancer drugs with limited usage due to its dose-dependent toxicities to the heart, kidney, liver and bone marrow. Over the past years, a number of drug delivery strategies have been developed in an attempt to improve efficacy and reduce the toxicity profile of DOX. The most successful drug delivery strategy reported to date has been liposomal DOX, which favors accumulation of the drug at tumor sites because liposomes easily exit the bloodstream at sites of leaky vasculature but do not readily exit the circulation in healthy tissues, such as the heart, with its “tight” endothelial capillary junctions . Such theoretical advantages have been confirmed by phase III clinical studies on metastatic breast cancer, which demonstrated that liposomal DOX could achieve similar or slightly better clinical efficacy but with significantly reduced cardiac and hematological toxicities, in comparison with conventional DOX [37, 38]. Compared with liposomal DOX, PDOX is a structurally different molecule designed for different antitumor mechanisms. Since we did not perform direct comparisons of efficacies and side effects between these two agents, it is likely that PDOX would not have an advantage in cardiac toxicity if it were compared to liposomal DOX.
In this study, animals receiving DOX treatment showed significant hematological toxicities with marked reduction of white blood cells and platelets, as well as cardiac toxicities with significant increases in creatine kinase mb isoenzyme, which is sensitive marker of cardiac toxicities, evident myocardium coagulation necrosis under histopathological studies. No hepatic and renal toxicities were observed in the DOX. In contrast, these toxic effects were not evident in animals receiving PDOX treatment.
In previous studies, TEM revealed DOX-induced following myocardial damages in rabbits: sarcoplasmic vacuolization, mitochondrial disruption, and myofibrillar lysis, which became more serious along with the increase of DOX dose , while demonstrated marked mitochondrial damage and vacuolization in DOX-treated mouse hearts . Disruption or loss of myofibrils and vacuolization of the cytoplasm, mitochondria changes, patchy necrosis, and inflammatory cells were observed in DOX-treated rats . Heavily contracted fibrils and condensed mitochondria were seen in TEM of heart cells in DOX-treated dogs . In this study, DOX induced mitochondria-centered cardiac injuries involving significant nuclear degenerative changes, mitochondria deletion in the peri-nucleus region, mitochondria-pyknosis, and discontinuation of intercalated discs under electron microscopy. These results were similar to the cardiac toxicity profiles in the above literatures. However, such toxic effects were not evident in the PDOX, even though the dosage of PDOX was 5.55-fold that of DOX. In addition, the hematologic, hepatic and renal toxicities were also less evident in PDOX than in DOX. Therefore, PDOX demonstrated reduced overall toxicity in comparison with DOX.
In conclusion, this study successfully synthesized PDOX with high chemical purity and good water solvability. PDOX is a newly synthesized molecular targeting prodrug of DOX, with defined chemical-physical properties. Compared with DOX, PDOX has similar efficacy but reduced hematological and cardiac toxicities in treating rabbit model of gastric PC.
Synthesis and identification of PDOX
All reactions were carried out in dried flasks under an atmosphere of dry nitrogen unless otherwise specified. The reactions monitored either on HPLC or TLC for completion. All reagents and solvents used were used as received without further purification unless otherwise stated. Chemical purity of the intermediates and final compound was determined on Agilent 1100 series instrument. LC-MS was performed on Agilent 1100 instrument with Zorbax column (4.6 × 150 mm) eluted with 0.1% trifluoroacetic acid water/0.1% trifluoroacetic acid acetonitrile at 1 mL/min flow rate. Column chromatography was performed on silica gel 60 (40-63 μm). Nuclear magnetic resonance (NMR) spectra were recorded on a Varian 300 instrument operating at 300 MHz (1H) or Bruker AC 500 instrument operating at 500 MHz (1H), 125 MHz (13C). Chemical shifts δ were reported in parts per million (ppm) from tetramethylsilane (TMS) as internal standards (0.00 ppm). Abbreviations for signal coupling are as follows: s = singlet, d = doublet, t = triplet, q = quartet, dd = double doublet, m = multiplet. High resolution mass spectrum (ES, positive) was determined on a Thermo LTQ FT Ultra Mass Spectrometer.
Step-1: Fmoc-Lys(MMT) 1
In a 2 L r.b. flask, Fmoc-L-Lysine hydrochloride (47.6 g, 112.8 mmol) was suspended in anhydrous dichloromethane (500 mL). Under nitrogen, at room temperature the trimethylchlorosilane (30 mL, 2.1 eq) was added, followed by diisopropylethylamine (20.6 mL, 1.05 eq). The reaction mixture was then heated to reflux for 1 h, and the clear solution formed during this time. The reaction mixture was then cooled to 0°C, and the diisopropylethylamine (62 mL, 3.1 eq) was added in, followed by p-anisyl-diphenylmethyl chloride (36.6 g, 1.05 eq). The reaction mixture was stirred at room temperature for 16 h. The solvent was evaporated and the residue was re-dissolved in ethyl acetate (600 mL). The solution was washed twice by pH 5 buffer solution (biphosphate, 300 mL × 2), and dried over sodium sulfate. After filtration and evaporation, a foam solid obtained (69.4 g 96%). H1-NMR (CDCl3, ppm) 1.26 (m, 2H), 1.68 (m, 4H), 2.45 (m, 2H), 3.71 (s, 3H), 4.05-4.40 (m, 4H), 6.81 (d, 2H), 7.15-7.77 (m, 20H).
Step-2: Lys(MMT) 2
In a 1 L flask, under nitrogen, Fmoc-Lys(MMT) 1 (26.3 g, 41 mmol) was dissolved in a mixture of 1:1 dichloromethane acetonitrile (400 mL). The diethylamine (400 mL) was added at room temperature and the result solution was stirred for 1.5 h. The solvent was evaporated to dryness, and the residue was treated with acetonitrile (200 mL) and the ether 400 mL. the solid was collected by filtration. The solid crude product was stirred with a mixture solvent of dichloromethane methanol (100 mL: 20 mL). The solid side product was filtrated off. After evaporation the solid product was dried by high vacuum for 10 h and obtained 16 g (94%). H1-NMR (DMSO-d6, ppm) 1.34-1.72 (m, 6H), 2.05 (m, 2H), 3.38 (m, 1H), 3.68 (s, 3H), 3.71 (d, 2H), 7.03-7.40 (m, 12H).
Step-3: Ac-Phe-Lys(MMT) 3
In a 1 L flask under nitrogen, the Lys(MMT) 2 (20.5 g, 48.9 mmol) was dissolved in dimethoxyethane (350 mL), and the solution of lithium hydroxide (2.05 g, 1 eq) in water (125 mL) was added in. A solution of Ac-Phe-OSu (13.9 g, 1 eq) in dimetoxyethane (350 mL) was added during 15 min, and the reaction mixture was stirred at room temperature for 16 h. The solvent was evaporated as much as possible, and the residue was taken by ethyl acetate (350 mL). This solution was washed by pH 4 buffer solutions (250 mL × 2) and brine (100 mL), and dried over sodium sulfate. After evaporation, the pale-yellow solid was dried by high vacuum for at least 5 h, and obtained 26.5 g (90%). H1-NMR (CDCl3/CD3OD), ppm) 1.22 (m, 2H), 1.58 (m, 3H), 1.71 (m, 1H), 1.82 (s, 3H), 2.49 (m, 2H), 3.00 (m, 2H), 3.75 (s, 3H), 4.26 (t, 1H), 4.63 (t, 1H), 6.82 (d, 2H), 7.10-7.43 (m, 17H).
Step-4: Ac-Phe-Lys(MMT)-PABOH 4
In a 1 L flask under nitrogen, Ac-Phe-Lys(MMT) 3 (26.5 g, 43.65 mmol) and di-t-butyl-pyrocarbonate (14.3 g, 1.5 eq) were dissolved in dichloromethane (600 mL), and at room temperature, the pyridine (3.7 mL, 1.05 eq) was added in. The resulting mixture was stirred for 15 min, and then, p-aminobenzyl alcohol (8 g, 1.5 eq) was added in. The reaction was stirred for 16 h, and the solvent was evaporated to dryness. The residue was treated with ether (200 mL), and the solid product was collected by filtration, and washed by fresh ether (50 mL × 2). After drying by high vacuum for 10 h, the off white solid obtained 26.2 g (84%). H1-NMR (CDCl3/CD3OD), ppm) 1.31 (m, 1H), 1.50 (m, 1H), 1.71-2.01 (m, 7H), 2.18 (m, 2H), 3.00 (m, 2H), 3.74 (s, 3H), 4.40 (t, 1H), 4.61 (s, 2H), 4.68 (m, 1H), 6.67 (d, 1H), 6.77 (d, 1H), 7.00-7.55 (m, 21H), 8.92 (br, 1H).
Step-5: Ac-Phe-Lys(MMT)-PABC-PNP 5
In a 1 L flask under nitrogen, Ac-Phe-Lys(MMT)-PABOH 4 (25.5 g, 35.7 mmol) and bis-p-nitrophenylcarbnate (32.5 g, 3 eq) was dissolved in anhydrous dichloromethane (600 mL). The fresh powdered molecular sieve 4 (50 g) was added followed by addition of diisopropylethylamine (18.5 mL, 3 eq). The reaction mixture was stirred at room temperature for 20 h, and filtrated through a pad of celite. After evaporation of the solvent, the crude compound was loaded to a silica gel column, and the desired product was eluted with hexane-ethyl acetate (50% to 100%). The pure product was obtained after evaporation of solvents, gives 10.5 g (34%). H1-NMR (DMF-d7, ppm) 1.43 (m, 2H), 1.58 (m, 2H), 1.72 (m, 1H), 1.87 (m, 4H), 2.09 (m, 2H), 2.4 (br, 1H), 3.05 (m, 1H), 3,78 (s, 3H), 4.52 (m, 1H), 4.72 (m, 1H), 5.36 (s, 2H), 6.90 (d, 2H), 6.77 (d, 1H), 7.29 (m, 16H), 7.41 (d, 2H), 7.50 (d, 4H), 7.68 (d, 2H), 7.81 (d, 2H), 8.13 (s, 1H), 8.19 (d, 1H), 8.41 (d, 2H), 10.11 (s, 1H).
Step-6: Ac-Phe-Lys(MMT)-PABC-DOX 6
In a 1 L flask under nitrogen, Ac-Phe-Lys(MMT)-PABC-PNP 5 (11 g, 12.5 mmol) and DOX hydrochloride (7.6 g, 1.05 eq) were dissolved in dimethylformamide (600 mL). The diisopropylethylamine (2.3 mL, 1.05 eq) was added, and the reaction mixture was stirred at room temperature for 48 h. The reaction mixture was diluted with ethyl acetate (4 L), and washed with water (1 L × 4), and dried over sodium sulfate. After evaporation, the crude product was purified by silica gel column and eluted dichloromethane-methanol (5% to 15%). After evaporation and drying, the pure product obtained 9.8 g (61%). H1-NMR (DMF-d7, ppm) 1.25 (d, 3H), 1.41 (m, 2H), 1.72 (m, 2H), 1.87 (m, 4H), 2.09 (m, 4H), 2.34 (m, 4H), 3.12 (m, 4H), 3.63 (br, 1H), 3,78 (s, 3H), 3.92 (m, 1H), 4,11 (s, 3H), 4.33 (m, 1H), 4.51 (m, 1H), 4.68 (m, 1H), 4.81 (s, 2H), 4.90 (m, 1H), 5.00 (br, 2H), 5.13 (br, 1H), 5.40 (br, 1H), 5.61 (s, 1H), 6.78 (d, 1H), 6.89 (d, 2H), 7.29 (m, 17H), 7.41 (d, 2H), 7.50 (d, 4H), 7.70 (m, 3H), 8.05 (m, 3H), 9.98 (s, 1H).
Step-7: Ac-Phe-Lys-PABC-DOX-HCl 7
In 1 L flask under nitrogen, Ac-Phe-Lys(MMT)-PABC-DOX 6 (9.8 g, 7.6 mmol) was suspended in a mixture of anisole (83 mL, 100 eq) and dichloromethane (260 mL), then dichloroacetic acid (6.2 mL, 10 eq) was added. The reaction mixture was stirred at room temperature for 1.5 h, and the ethyl acetate (2 L) was added, and keep stirring for 2 h. The solid product was collected by filtration, and the solid was re-dissolved in methanol (400 mL). The solution was loaded to a AG2-X8 ion resin (Cl form, 250 g) exchange column, and eluting with methanol. The colorful fraction, which content product, was collected and evaporated to dry. The solid was treated with dichloromethane (10 mL) and after filtration, the product was dried by high vacuum for constant weight produced 7.1 g (95%).
H1-NMR (DMSO-d6, ppm
1.12 (d, 3H), 1.47 (m, 4H), 1.75(m, 4H), 1.83 (m, 1H), 2.11 (d, 1H), 2.2 (d, 1H), 2.67-3.02 (m, 4H), 3.74 (m, 1H), 3.86 (s, 3H), 4.13 (m, 2H), 4.38, (m, 1H), 4.51 (m, 1H), 4.50 (br, 2H), 4.75 (d, 1H), 4.89 (br, 2H), 5.21 (br, 1H), 5.48 (s, 1H), 6.89 (d, 1H), 7.11-7.26 (m, 4H), 7.59 (d, 1H), 7.64 (dd, 1H), 7.85-7.90 (m, 3H), 8.16, (d, 1H), 8.32 (d, 1H), 10.11 (s, 1H), 13.16 (s, 1H).
C13-NMR (DMSO-d6, ppm)
δ 186.6, 186.5, 171.8, 170.8, 170.1, 169.8, 161.1, 156.4, 155.7, 154.9, 138.9, 138.7, 138.3, 137.8, 136.5, 135.7, 134.8, 134.4, 132.3, 129.6, 128.8, 126.7, 126.6, 120.2, 119.5, 119.6, 110.9, 110.8, 100.7, 75.3, 70.1, 68.4, 76.1, 65.3, 64.1, 56.8, 55.1, 54.5, 53.7, 48.9, 47.5, 39.2, 38.8, 36.8, 32.4, 31.6, 30.2, 26.9, 22.8, 22.4, 17.4.
Agents and cells
Apart from PDOX described above, other agents were obtained from commercial sources, including DOX hydrochloride for Injection (Pharmacia, Milan, Italy), Docetaxel (Wanle Pharmaceuticals Co., Ltd. Shenzhen, China) and Carboplatin (Qilu Pharmaceuticals Co., Ltd. Shandong, China).
The animal study protocol was approved by the Animal Welfare Committee of Wuhan University, complied with the Helsinki Declaration. The animal study was completed in the Animal Biosafety Level 3 Laboratory at the Animal Experimental Center of Wuhan University (Animal Study Certificate ID: No.00022310).
Forty male New Zealand white rabbits, body weight between 2.5-3.0 kg, were obtained from Animal Biosafety Level 3 Laboratory at the Animal Experimental Center of Wuhan University (Approval ID: SCXK 2008-0004). The animals were individually housed and allowed free access to standard laboratory food and water as well as 12 h of light and dark cycle per day.
Tumor strain and tumor cell preparation
Rabbit VX2 carcinoma was used to establish GC with PC in this study. The VX2 tumor is a transplantable rabbit squamous cell carcinoma, characterized by rapid tumor growth and early metastasis, established from a virus-induced papilloma by Rous and coworkers . The tumor was maintained by successive in vivo transplantation into the hind leg of a carrier rabbit used for every passage.
When the VX2 tumor grew to about 2 cm in diameter on the carrier rabbit, the animal was anesthetized by ear vein injection of 2% pentobarbital sodium (30 mg/kg). After skin preparation and disinfection, the tumor was excised from the carrier rabbit and placed in icy cold 0.9% sodium chloride solution. Tumor tissue was minced into approximately 1.0-2.0 mm3 fragments and suspended in 2 mL of normal saline, then drawn into a 2 mL injector. Other tumor tissues about 3.0-5.0 mm3 were placed into the homogenizer embedded in ice bath, to which 3 mL of icy cold normal saline was added, and the tumor cells suspension was made, with the tumor cells concentration adjusted to 5 × 1010 vial cells/L.
Construction of rabbit model of gastric PC
Rabbit VX2 carcinoma was used to establish gastric PC in this study. All rabbits had overnight fasting before experiment, but water was given ad libitum. The animals were anesthetized by ear vein injection of 2% pentobarbital sodium (30 mg/kg). The abdominal skin was cleaned and disinfected. Tumor cells were injected into the stomach submucosa layer to construct rabbit model of gastric PC as described previously [24, 25]. Briefly, a midline incision of 3 cm long was made beginning 2 cm below the xyphoid and the upper abdomen was open. The stomach was exposed, 0.1 mL of tumor cells (5 × 1010 vial cells/L) was injected into the submucosal layer of the stomach, through the serosal layer and the muscle layer, the injection site was pressed for 1 min to keep the injected tumor cells in place, and the abdomen was closed with a double layer 3-0 vicryl interrupted suture. After tumor inoculation, Penicillin G at the dose of 100,000 IU/d was intramuscularly injected to each animal for 3 d.
Randomization and treatment
When animal model construction has been confirmed successful on d 8 after operation, these rabbits were randomly divided into 4 groups according to a computer generated randomize number, 10 animals in each group: the Control (n = 10) was observed for natural course of disease progression without any intervention; the HIPEC (n = 10) receiving CRS plus HIPEC (docetaxel 10.0 mg and carboplatin 50.0 mg in 250 mL normal saline, at 42.5 ± 0.5°C for 30 min); the PDOX (n = 10) and the DOX (n = 10) receiving systemic chemotherapy with PDOX 50.0 mg/kg (10.0 mg/kg every 4 d for 5 cycles) or DOX 5.0 mg/kg (1.0 mg/kg every 4 d for 5 cycles) after CRS + HIPEC, respectively; The CRS + HIPEC was performed on d 8 while the systemic chemotherapy was initiated on d 16 after model construction.
CRS plus HIPEC
For the three treatment groups, CRS + HIPEC was performed on 8 d after tumor cells inoculation. Rabbits were given 2% pentobarbital sodium (30 mg/kg) intravenously (i.v) for anesthesia. The abdominal skin was cleaned and disinfected. The abdominal exploration was performed through a midline incision of 8 cm long beginning 1 cm below the xyphoid. Once the abdomen was open, detailed evaluation of the PC was conducted in different regions including the parietal peritoneum, visceral peritoneum, the omentum, stomach, liver, spleen, intestinal wall, bladder and other pelvic organs. An ePCI score system  was developed to record the experimental peritoneal carcinomatosis index of the animals in each group, which took into consideration of tumor nodule sizes, distributions and the characteristics of ascites. In this system, the abdominal cavity of rabbit was divided into 4 regions: region I, sub-diaphragm; region II, the liver, spleen, stomach and affiliated ligaments; region III, small intestine, colon, mesenterium and abdominal wall; and region IV, pelvic cavity urogenital system and rectum. The detailed scoring criteria were modified from similar reporting systems on rat and mice PC models and set as the following: score 0, no tumor nodules throughtout the region; score 1, nodule size less than 5 mm in greastest diameter; score 2, nodule size greater than 5 mm and up to 20 mm; score 3, nodule size greater than 20 mm. If bloody ascites occurred, it was set as score 1. The sum of all the scores was the ePCI of the animal (ranging from 0 to 13). Thereafter, maximal CRS was performed including a routine omentectomy, and optimal removal of tumor nodules. Unresectable tumors were cauterized. The gastric tumor itself, however, was not removed but treated by injection of absolute alcohol. HIPEC was performed just before the closure of abdominal cavity after completion of CRS, as this open technique was believed to provide optimal thermal homogeneity and spatial diffusion , with 250 mL of heated saline containing 10 mg of docetaxel and 40 mg of carboplatin for each animal. The abdominal cavity was rinsed twice with 250 mL of normal saline preheated to 42.5°C and perfusion tube was placed in pelvic cavity just before HIPEC. The perfusion equipment consisted of a miniature heat exchanger and a roller pump, allowing perfusion with a variable dynamic flow of 6-12 mL/min. An inflow catheter was inserted into the upper abdomen between the hepatic and diaphragmatic surface and an outflow catheter was placed at the pelvic floor. The perfusion solution was heated to 42.5 ± 0.5°C and infused into the peritoneal cavity at a rate of 10 mL/min through the inflow tube introduced from the automatic perfusion pump. The perfusion in the peritoneal cavity was stirred manually to make equal spatial distribution.
The temperature of the perfusion solution in peritoneal space was kept at 42.5 ± 0.5°C and monitored using a thermometer on real time. The total HIPEC time was 30 min, after which the perfusion solution in the abdominal cavity was removed. Twenty min before surgery, 100 mL of 0.9% sodium chloride solution with 1 g of ceftriaxone powder, 2 mL of 10% potassium chloride solution and 20 mL of 50% glucose solution was infused intravenously for rehydration, nutrition support and infection control in all the four groups of animals. Such treatment was continued for 3 d.
Chemotherapy with PDOX and DOX after CRS + HIPEC
After CRS + HIPEC, the PDOX (n = 10) received systemic chemotherapy with PDOX 50.0 mg/kg (10.0 mg/kg and 3.0 mL/kg every 4 d for 5 cycles) on d 16, d 20, d 24, d 28, and d 32 after tumor cells inoculation respectively; the DOX (n = 10) received systemic chemotherapy with DOX 5.0 mg/kg (1.0 mg/kg and 3.0 mL/kg every 4 d for 5 cycles) at the same time; the Control and the HIPEC received 3.0 mL/kg 5% dextrose instead of chemotherapy drugs at the same time. All drugs were slowly injected i.v through the ear vein of rabbits. For each animal, 2.5 mg of dexamethasone was injected i.v 20 min before chemotherapy, and an additional 2 mL of 5% dextrose was injected i.v after PDOX or DOX injection to reduce local vascular damage.
Animal observation and disease course monitoring
The general status of the animals was daily recorded in a standard form. For pathological studies, euthanasia was performed on the rabbits by overdose injection of 2% pentobarbital sodium through the ear vein. Post mortem pathological examinations included gross pathology such as tumor size and distributions; local tumor features of GC such as ulcer formation, intestinal obstruction and perforation; special features of PC such as bloody ascites, discrete or confluent tumor nodules on the peritoneum and omentum cake; metastases to major organs such as the livers and the lungs.
Blood routine tests and biochemical test results
For laboratory studies, 3 mL of blood was harvested from ear vein on 2 d before tumor cells inoculation as the baseline (D-2), 2 d before CRS + HIPEC (D6), 2 d before chemotherapy (D14), and 4 d after completion of chemotherapy (D36). The samples were used for routine peripheral blood test, liver and kidney functions tests and detection of creatine kinase and creatine kinase mb isoenzyme.
Histopathological studies under both optical microscope and electronic microscope
During the procedure of CRS, the resected omentum, tumor nodules, enlarged mesenteric lymph nodes were harvested for HE staining and observation under optical microscope. After post mortem pathological examinations, myocardial tissues of all the rabbits were harvested at the first time for the electronic microscope, and all the suspected organ metastasis tissues were sampled for routine histopathology study with sections stained by hematoxylin and eosin (HE stain).
All data were integrated into a central database. The numerical data were directly recorded, and the category data were recorded into different categories. The primary endpoint was OS, and secondary endpoint was toxicity profile. Survival was analyzed on Kaplan-Meier survival plots and statistical significance was analyzed with SPSS software version 15.0 (SPSS Inc., IL, USA), using the log-rank test. The differences in blood routine and biochemistry among different groups were tested using nonparametric test at each time point. Categorized variables were compared by chi square test (χ2) or Fisher’s exact test. Two-sided p < 0.05 was considered to be statistically significant.
Hyperthermic intraperitoneal chemotherapy
Experimental peritoneal carcinomatosis index
Nuclear magnetic resonance
Maximum tolerated dose
Minimum curative dose.
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We would like to express our special thanks to Collin HONG at Princeton High School, 151 Moore Street, Princeton, NJ 08540, U.S.A, for his active participation in part of the chemical analysis work.
This work was supported by grants for major scientific projects “Key New Drugs Preparation” (2009ZX09301), Innovative Research Teams of the National Natural Science Foundation of China (20621502), the National Program of Innovative Experimental Plan for Students of Higher Education (081048646), and the Independent Research and Development Projects for Postgraduates of Wuhan University.
PDOX has been filed for patent protection to the STATE INTELLECTUAL PROPERTY OFFICE OF P.R.C, with the application no: 201110300925.4 (inventors: Yan LI, Raymond A FIRESTONE, Ya-ping HONG and Li-hua SHAO).
Conceived and designed the experiments: YLI, LTANG. Performed the animal experiments: LT, Y-jZ, RD, H-lW. Chemical synthesis: Y-pH, J-gL, Y-cX, Analyzed the data: LT, RAF, Y-pH. Contributed reagents/materials/analysis tools: YLI, Y-pH, J-gL, Y-cX. Wrote the first draft of the manuscript: YLI, LT. Contributed to the writing of the manuscript: YL, LT. Y-pH. ICMJE criteria for authorship read and met: YL, LT. Agree with manuscript results and conclusions: YL, LT, Y-pH. Performed record linkage and selected data for analysis: LT. All authors read and approved the final manuscript.
Li Tang, Rui Duan contributed equally to this work.
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Tang, L., Duan, R., Zhong, Yj. et al. Synthesis, identification and in vivo studies of tumor-targeting agent peptide doxorubicin (PDOX) to treat peritoneal carcinomatosis of gastric cancer with similar efficacy but reduced toxicity. Mol Cancer 13, 44 (2014). https://doi.org/10.1186/1476-4598-13-44


The Marin County coroner’s office has identified 46-year-old Avril Patch Clary as the Sausalito woman who was found dead in her bedroom Saturday afternoon.An autopsy is scheduled for today. Sausalito police went to Clary’s home at 501 Bridgeway around 4:30 p.m. Saturday and knocked on the door. When no one answered, they entered and found Clary’s body in the bedroom.An investigation into the death began after Richmond police received information that Clary’s boyfriend Chet Turner, 48, may have killed his girlfriend, Sausalito Police Chief Scott Paulin said.Sausalito police then asked Richmond police to see if Turner was at 2017 San Mateo St. in Richmond. Turner was found there, detained by Richmond police and arrested by Sausalito police on suspicion of homicide.Turner is expected to be arraigned in Marin County Superior Court Tuesday, the Marin County District Attorney’s office said.
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The third session of the trial of Egypt's ousted dictator Hosni Mubarak has begun minutes ago.
This is the first session of the historic trial that would not be televised as presiding Judge Ahmed Refaat had decided at the end of last round to ban TV broadcasting.
Clashes errupted inside courtroom between the defendants' and victims' lawyers, resulting in six injuries.
A confirmation about the clashes came from a lawyer representing the families of the victims, Gamal Eid of the Arabic Network for Human Rights Information. Eid, who is Tweeting from inside the courtroom, added that the clashes disrupted the session, forcing the presiding judge, Ahmed Refaat , to recess the session after lawyers yelled at him.
Newsreporters confirmed that all defendants - Mubarak, his two sons, his former minister of interior and six of his top aides - have all appeared in Court.
Mubrak was wheeled in on a stretcher as he did in the previous two sessions. However, his two sons, Alaa and Gamal, entered the courtroom in handcuffs for the very first time during the trial.
Earlier this morning, scuffles have broken out between protesters and police officers outside courtroom, as the 3-judge panel prepares to hear from police witnesses who were in the headquarters control room during the crackdown on pro-democracy demonstrators during the January uprising against the former dictator.
Our Ahram Online reporter on the scene says that clashes began when some families of the martyrs were infuriated that police prevented them from entering the court room while allowing relatives of accused police officers to do so.
Police pushed back martyrs families and their supporters and things quieted down for a few moments.
However, our Ahram Online reporter adds, tens of Mubarak's supporters began throwing stones at families of martyrs and journalists.
At this point, all hell broke loose.
Families of the martyrs and their allies, who number around 100, picked up stones and started to throw back at Mubarak supporters.
Security forces and Mubarak supporters responded and scuffles escalated.
In the melee that ensued, at least 3 protesters and one police officer were injured.
Eye witnesses say more police were deployed this time compared to the last two sessions of the trial.
The third session of the trial of Mubarak, his two sons and his former minister of interior will feature for the first time testimonies of witnesses on charges that Mubarak used snipers to kill protesters during the January uprising against his 30 year rule.
Mubarak is standing trial with his two sons, Alaa and Gamal, as well as Habib al-Adli, a former interior minister, and six senior police officers on charges of corruption and killing peaceful protesters .
Lawyers have been looking forward to this session that will include the testimony of four significant witnesses. Among these is a very important general, Hussein Moussa, who is the former head of operations in the Central Security Forces who were on the frontline during the crackdown.
According to lawyer Gamal Eid, head of the Arabic Network for Human Rights Information,, Moussa is especially important as he was “formally accused of erasing all information and recorded phone calls from the official records”.
Ten Kuwaiti lawyers were expected to join the defence team for Mubarak on Monday, but no one has shown up in court yet.
Some of the lawyers said their role comes as a gesture of gratitude to Mubarak for his support for a US-led coalition that expelled Iraq from Kuwait in 1991.


text:
Hepion Pharmaceuticals (NASDAQ:HEPA) advanced to the third dose level of CRV431 in its ongoing Phase 1 multiple ascending dose trial.
The clinical trial management team determined the 150 mg dosing level to be safe and well tolerated, authorizing escalation to the next dosing level of 225 mg daily for 28 days.
The study is designed to assess safety, tolerability and pharmacokinetics of CRV431, administered orally to healthy volunteers, once daily for 28 days. The study is examining doses of 75 mg, 150 mg, 225 mg and 300 mg, or higher.
“We are pleased that CRV431 continues to demonstrate safety and tolerability at increasing doses, and look forward to additional data that will inform the design of our Phase 2 non-alcoholic steatohepatitis clinical trial, which we plan to initiate in the second half of 2020,” Dr. Stephen Harrison, Hepion’s consultant medical director, said in a statement.

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[WASHINGTON, D.C.] –U.S. Representative Robin Kelly (D-IL) today met with U.S. Senator Dick Durbin (D-IL) to discuss constituent services and economic development priorities in Illinois's 2nd congressional district, including job-creating infrastructure projects and efforts to train and educate workers in advanced manufacturing.
CONGRESSWOMAN KELLY STATEMENT ON ILLINOIS HOUSE PASSAGE OF CONCEALED CARRY BILL
WASHINGTON, DC – Congresswoman Robin Kelly (IL-02) voted today to keep college within reach for Illinois students by voting against the so-called Smarter Solutions for Students Act that would force students into loans with skyrocketing interest rates.
REPUBLICAN-BACKED REPEAL EFFORT WOULD HURT ILLINOIS’ WORKING FAMILIES
WASHINGTON, DC – Congresswoman Robin Kelly (IL-02) voted today to protect Illinois’ working families, women and seniors by voting against the Patients’ Rights Repeal Act.
Congresswoman Robin Kelly delivered remarks on the floor of the House of Representatives advocating for common sense gun legislation. Click here to watch the video.
Full text of the remarks:
WASHINGTON, D.C. – Congresswoman Robin Kelly (IL-02) issued the following statement today in reaction to the United States Senate’s failure to pass the gun background check amendment :


Isaac, Eccott, Perridge, Pittman
Additional Information - Posters # 1995 Edinburgh
A detailed investigation has been carried out on the microstructure of various polyethylenes recently developed for applications in the pipe industry. Of particular interest have been the PElOO formulations, and samples of these have been studied both from commercial piping and from laboratory produced plaques. Attempts have been made to relate the crystallinity and morphology to the cooling rates. In order to achieve some control over the cooling rates, a special laboratory-based rig was constructed to allow the compression moulding of polyethylene plaques containing embedded thermocouples. This equipment was designed to enable plaques to be produced at the rapid cooling rates that occur industrially, as well as slower rates, and it was therefore possible to provide a broad range of samples for analysis and comparison with the through pipe microstructural variations. Numerous etching techniques were used to characterise the morphologies of the various polyethylenes, and some striking differences between the materials were observed. Some of the samples showed the typical spherulitic microstructure normally associated with polyethylenes crystallised from the melt. However, it was found that, despite having relatively high crystallinities, some materials, even at the slowest cooling rates, did not exhibit such a regular arrangement of the lamellae, but only displayed a wheatsheaf-like arrangement reminiscent of embryonic spherulites. Attempts have been made to understand this behaviour by collecting data from various other physical techniques such as DSC, molecular weight measurements etc.


text:
Who we are
Australia reMADE is an independent alliance of engaged people and community leaders from all walks of life -- working to name, plan and act on the Australia of our dreams. We started as A24, a group of civil society leaders tired of working in isolation, tired of being so focused on putting out fires that we had little room left for long-term proactive agenda-setting and collaboration.
We set ourselves the task of actually mapping out what we’re for: a cohesive vision of the world we want; grounded in the voices, values and experiences of everyday people, not policy wonks. That research project took us into people’s hearts and homes, having rich conversations about the Australia of their dreams.
It was a brilliant exercise for restoring faith in humanity, and for recognising that what divides us is surface; what unites us is fundamental.
The inspiring vision of Australia reMADE is just the beginning of the world that needs to be done. Drawn from hundreds of conversations across the country, it is a statement that gives us all hope in the simplicity and possibility of a better tomorrow. Australia reMADE paints a picture of a future where people and planet come first, where values of love and community triumph and where justice and fairness for all become the new ‘normal’ – the new common sense.
And our work does not just stop at visioning, important though this is.
The next phase of our work is focusing on how we make our vision a reality. Naming the big ideas that will put people and planet first is our ultimate goal while honing our understanding of what needs to change to make this happen.
This is a big conversation and we are not alone in diving into it. There is growing momentum for a discussion that gets us off the back foot, out of our silos and joined up with others wanting focus on solutions and transformation.
We feel the work associated with Australia reMADE could be useful to these conversations and welcome the opportunity to share, collaborate, and connect diverse conversations focused on creating a socially and ecologically just society.
Unity with nature
We are part of nature.
We respect nature’s limits.
We consider looking after nature as a gift to ourselves, each other and future generations.
Equal worth of all people
We all have equal worth as human beings and an equal right to a good life.
Our value as people does not depend on our gender, the colour of our skin, where we live, who we love, our capabilities, our income or who we worship.
Interdependence and community
We depend on each other.
We are not just isolated individuals but social beings who can achieve more together than apart.
Love and care for each other and the planet is our primary motivator.
Cooperation not competition: We believe people can achieve more when we work together than when we compete.
Well-being not economic growth as a measure of progress: The core goal of our economy and society should be to maximise the well-being of people and planet.
Powers is shared not concentrated: We need to redistribute power to ensure no group dominates another and correct imbalances that systematically privilege some groups and identities.
Collective rules not freedom to harm: We need rules that help us all act fairly and protect the many from the selfish instincts of the few.
Sharing not self interest: We believe people instinctively want to share with and care for others. Our society needs to enable these sharing behaviours, rather than perpetuate the myth that greed and self-interest are inherent.
Preserving of nature not exploitation: Without a healthy natural home, we cannot flourish. Our economy, society and laws must enshrine all our rights to fresh air, clean water and a world of natural beauty.
Our way of working
Love and care is at the heart of how we work together
None of us can win alone
We seek to build common ground based on shared analysis, while acknowledging and respecting difference
We reject cynicism, the pursuit of the perfect, divisive behaviours and undemocratic solutions
We work in good faith and with positive intentions
We aspire to bring many diverse voices into our community – but within a clear framework of articulated values
We exist because this project and its approach is not occurring elsewhere in this moment
History and human nature are on our side. We expect to win.
Making Australia reMADE
The Short Version
In June 2017, the A24 Engagement Project began. The project sought to hear from ordinary Australians about the future they want for Australia. And wow! the 200+ people we listened to had a lot to say about how great they think this country could be, and a lot to say about what they think is currently wrong. People were asked to dream out loud about the Australia they want and to think with us about how such a transformation could happen. Their ideas and dreams have come to give life to a vision for Australia, Australia reMADE, adding depth, breadth, spirit and hope to previous draft vision statements developed in the 2016 by many others who attended a series of A24 Gatherings and considered the same questions.
Read more about how we wrote the vision and hear some of the incredible stories we heard along the way…

domain: sociology


text: This is just average at best death metal. Obviously I have 1000’s upon 1000’s of death metal throughout the years and this was just to me another faceless one. They play straight forward death metal the right with crushing riffs, deep dark vocals and mix up the speed with some wicked slower parts. This was just like Death “Leprosy” era, but obviously not as good. If your new to the scene this would be a worthy pick-up, but to a veteran like myself, it is a case of heard it all before and then some.

domain: artistic


Varner to Deliver Glass Science Lecture at Alfred University
James Varner, professor emeritus of ceramic engineering in the Inamori School of Engineering at Alfred University (AU), will deliver the annual Samuel R. Scholes Sr. Lecture on April 25. Varner’s talk, “Fractography–The Key to Reliability (It’s Elementary!),” focuses on the key role that fractography plays in ensuring mechanical reliability. Fractography, Varner explains, is the “means and methods for characterizing a fractured specimen or component.” Fractography involves examining and interpreting markings on surfaces created by fracture, as well as crack patterns.
He says his presentation focuses on the fractography of glasses, but the same general principles apply to the fractography of ceramics. Fracture markings and crack patterns will be shown and discussed. Quantitative fractography will be introduced. Examples and case studies will be provided that illustrate the essential role of fractography in understanding glass strength and in ensuring mechanical reliability.
For additional information, visit www.alfred.edu.


text:
ColouredFemmes is a collection of stories by black women. Through anecdotes, poetry and diary entries, this platform is a safe space to reveal thoughts and testimonies on love, race, feminism, relationships, health, family, career, etc.
Hey, I’m Kimani and I’m the woman behind ColouredFemmes. Some of the stories are mine, but most of them are from wonderful Black women who needed to share their stories and be heard. There is so much power in sharing your story and so we decided to do so in a safe space for and by Black women.

domain: library


text:
Carceri VII, 1760 (etching)
Piranesi, Giovanni Battista (1720-78)
View of the Colosseum in Rome by Piranesi, 1761 (engraving)
PIRANESI: PONTE FABRIZIO Sectional view of the Ponte Fabrizio, Rome, Italy. Line engravi...
PIRANESI: TIVOLI View of the Temple of the Sibyl in Tivoli, Italy. Etching by Giovanni B...
Interior of the Basilica of San Paulo Fuori le Mura, Rome, from 'Le Antichita Romane de ...
Piranesi, Giovanni Battista (1720-78) (after)
Trajan's Column (engraving)
Allegorical Frontispiece of Rome and its history, from 'Le Antichita Romane de G.B. Pira...
Gladiatorial Combat, Piranesi (engraving)
English School, (19th century)
View of the Temple of Isis, Pompeii, engraved by Francesco Piranesi, 1788 (etching)
Desprez, Louis Jean (1743-1804) (after)
Magnificent Bridge with Loggia , 1753-1837 (etching and engraving)
The Temple of Neptune at Paestum, etched by Francesco Piranesi, 1778 (etching)
Ruined Gallery of the Villa Adriana at Tivoli (engraving) (b/w photo)
Carcere V (the lion bas-reliefs), 1753...
Piranesi, Francesco (c.1756-1810)
The paving of the Via Appia Antica, fr...
Interior of St. Paul's basilica outside the walls, 1753-1837 (etching and engraving)
Imaginary prisons, by Giovanni Battist...
Illumination of the Cross in St. Peter...
View of the interior of the Coliseum, from 'Le Antichita Romane de G.B. Piranesi (1756)'...
Aerial view of the Colosseum in Rome from 'Views of Rome', first published in 1756, prin...
View of the Tavern of Pompeii with the Priapus Shop Sign, from 'Antiquites de Pompeia' b...
Veduta del Porto di Ripa Grande, c.1760 (engraving)
Piranesi, Prison, Engraving
View of Temple of Neptune at Paestum, 1778, engraving from Views of Paestum, by Giovanni...
Egyptian fireplace, engraving by Giova...
Bird's Eye Views of: the Balbo and Mar...
Architectural interior and staircase, ...
View of the Spanish Steps in Rome, Italy, 1750 engraving by Giovanni Battista Piranesi (...
Hommage à Piranesi (oil on canvas)
McKenna, Stephen (b.1939)
Italy, Rome, Temple of Vestal Virgins,...


text:
Darren Star to be honored at Austin Film Festival
BY Advocate.com Editors
September 21 2002 12:00 AM ET
The Dallas Morning News reports that openly gay Sex and the City and Melrose Place creator Darren Star will receive the Outstanding TV Writers Award at October's Austin Film Festival and Heart of Film Screenwriters Conference. Also honored will be Richard LaGravenese (Living Out Loud, Beloved, The Mirror Has Two Faces), with the Distinguished Screenwriters Award.
- Why Can't We Talk About Homophobia in the Black Community?
- Vatican Official Calls Irish Marriage Vote 'Defeat for Humanity'
- The Cities LGBTs Love And the Ones We Shun
- WATCH: Out Musician Mika Shares Video for 'Good Guys,' a Bittersweet Homage to Gay Men
- EXCLUSIVE: Watch the Official Trailer for 54: The Director's Cut
- Op-ed: The Amtrak Accident Isn't Part of the Gay Agenda

domain: entertainment


Adhesive glass tapes are supplied in three temperature classes; Class B 130°C, Class F 150°C and Class H 180°C and are available in a broad range of sizes from 6mm to 50mm. Adhesive glass tapes offers excellent thermal and mechanical properties and has of high tensile strength ensuring long-lasting durability. It’s also flexible, easy to apply and resistant to fraying and ageing.
The thermosetting adhesive can be further cured to offer resistance to solvents and oils. Adhesive glass tape is used in various applications in electrical insulation such as motor repair, harnessing and electro plating and offers excellent protection.


text:
Four Reasons Why Payday Loans Are Risky
We’ve all been there — living from paycheck to paycheck, money just being enough to pay off your bills, not being able to afford car insurance. So, if you feel like such a financial failure, don’t be. Most of us have been (or might even still be) exactly where you are now — struggling with finances and thinking about taking out a loan.
A loan is like the devil’s cake. You know it isn’t good for you, but you can’t help yourself because it’s sinfully good. It seems to be a quick fix and an easy solution. It’s available everywhere and comes in many forms, the most popular of which is payday loans. When it comes to this kind of loan, there are a number of risks involved that you have to know before you should take one. Knowing the risks that come with it will help you make a smart decision. So what are these risks and should you really be concerned with them?
High Interest Rates
This is one of its biggest risks. They are going to offer you a low interest rate to entice you to get one, but there is actually a catch (and a huge one at that!). An interest rate that low usually comes with the stipulation of you getting a certain amount. And the fact is, the interest rate will vary according to the amount of money that you will borrow.
So if you think about it, the interest rates are actually expensive. With a credit card, you get an interest rate of about 28% to 36%. How will an interest rate that is about ten times higher than that would make you feel? That is usually how expensive this type of loan is.
Cycle of Borrow-Pay-Borrow
Although there are some lenders who prevent you from becoming a habitual borrower by maintaining a strict database, there are some lenders that would leave you stuck in a payday loan cycle (more money for them after all). In fact, according to a research, a whopping 76% of payday loans are made to pay an existing payday loan and that even though most of this kind of loans are meant to be paid over a period of two weeks, a borrower can stay in debt for more than 6 months (that’s probably longer than the time most of us spent on our first relationship!).
Debt Grows Bigger and Faster
Because of the high interest rates and the option set that you can roll over a payment, you can end up paying about three, four, or even ten times the amount of money that was originally lent to you. A debt that is made through a payday loan often quadruples within just a year, and this kind of loan can bring you a lifelong debt (think imprisonment, only for your wallet).
Too Available and Easy
While other types of loans require serious paperwork and thorough deliberation for an application to be approved, a payday loan, on the other hand, can be easily available (you can probably get one on your lunch break). And if you think about it, they are actually designed in a clever way: giving borrowers little time to think the deal over. A payday loan also carries no right to rescission. This means if you change your mind even just right after you sign the papers, too bad because you can no longer back out. Know the saying, “Nothing worth having comes easy”? Whoever said that must have taken out a payday loan and regretted it.
We all find ourselves in a bind, and sometimes, we can’t help but take out a loan. And for the average-income employed individual, loans are a big part of our lives. And most of the time, they really do help us in times of need. However, keep in mind that there are good lenders and bad lenders, and it’s up to you to choose wisely. Do a thorough research first — whether that’s through online articles, a financial advisor, or friends and family members who have experience availing payday loans. It may seem time-consuming and tedious, but it’s worth the effort, especially since your finances are involved! Never take it for granted and assess everything carefully before making a final decision.
More in Loans & Mortgages
Five Property Loan Milestones You Need To Unlock To Get A Loan
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The Non-Conforming Loan May Be Just What You are Looking For!
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Private Loans Versus Bank Loans: Which One Should You Choose?
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domain: finance


Yocto(1) is a software framework for embedded Linux systems. We've looked at it in previous blog posts(2). It forms the basis of many embedded Linux distributions, including The Qt Company's boot2qt that is part of their commercial product Qt For Device Creation(3).
Knowledge of Yocto is a very desirable and marketable skill to have for anyone working with embedded systems. While you can use emulators that run on a desktop Linux system, there is no substitute for experience with a real embedded board.
Yocto supports a number of embedded hardware platforms, one of the most popular being the BeagleBone Black(4). At a retail price of around US$60 it is quite affordable.
Yocto can also be built for the Raspberry Pi, which is even lower in cost than the BeagleBone. You may already have one or two kicking around, and if not you can start with a board as inexpensive as the $5 Raspberry Pi Zero.
The Raspberry Pi is not an officially supported board but there are some references around that cover how to get Yocto up and running on it, including Qt. I'll cover a few of them here that I have had experience with.
This procedure(5) is for a Raspberry Pi 2 with the official touchscreen display. I ran through the procedure and it successfully worked for me with a Raspberry Pi 2 and touchscreen, including being able run the qmlscene program to execute QML-based Qt 5 programs. I tried updating it to use the more recent "krogoth" release of Yocto so I could build Qt 5 with support for the Raspberry Pi 3. I was able to get it to build with a few changes to some of the bitbake recipes. While the generated image built and booted on the Raspberry Pi 3, running the qmlscene program would hang without anything being shown on the display. More investigation is needed to see what the problem is.
I also tried this procedure(6), configuring it for a Raspberry Pi 3. The generated Qt 5 image built and ran on the Pi, and even included the driver for the onboard WiFi hardware. I could run qmake and compile a simple widgets example natively on the board and run it successfully. The image is missing support for the xcb and eglfs platform plugins for Qt, and only has linuxfb, so it is of limited use because it won’t run QML programs. Getting eglfs working would be a good area of future work.
The Qt Company has been contributing(7) to the Qt 5 bitbake recipes for Yocto(8) and recently made their bitbake recipes for boot2qt open source(9). I was able to use these recipes to create a boot2qt image with Qt 5 and run it on a Raspberry Pi 3. I did have to make a few changes to remove some of the software components that are only available in the commercial version of Qt. This version has support for eglfs so it can run QML programs. It also builds a toolchain, which can be used to cross-compile and debug from the Qt Creator IDE.
A simple QML example running on the Raspberry Pi 3 with touchscreen (image credit: Jeff Tranter)
My experiments with Yocto on the Raspberry Pi are still in progress. I hope to cover more details in a future blog post.
- Linux Foundation Yocto Project, website, last accessed 11 July 2016, https://www.yoctoproject.org
- The BeagleBone Black - A Low-Cost Embedded Platform Part 2, ICS blog post, last accessed 11 July 2016, http://www.ics.com/blog/beaglebone-black-low-cost-embedded-platform-part-2
- Qt for Device Creation, Qt website, last accessed 11 July 2016, https://www.qt.io/qt-for-device-creation
- BeagleBone Black, BeagleBoard website, last accessed 11 July 2016, https://beagleboard.org/black
- Setting up a Yocto Build with Qt 5 for Raspberry Pi 2 with Official Touchscreen, blog.unixshell.se blog post, last accessed 11 July 2016, http://blog.unixshell.se/?p=27
- Building Raspberry Pi Systems with Yocto, Jumpnow Technologies website, last accessed 11 July 2016, https://www.jumpnowtek.com/rpi/Raspberry-Pi-Systems-with-Yocto.html
- Aligning with the Yocto Project, Qt blog, last accessed 11 July 2016, https://blog.qt.io/blog/2016/07/01/aligning-with-the-yocto-project
- Meta-qt5 Qt 5 layer for Yocto, Yocto Project website, last accessed 11 July 2016
- Meta-boot2qt Boot2Qt Layer For Yocto, Qt git repository, last accessed 11 July 2016, https://code.qt.io/cgit/yocto/meta-boot2qt.git


text:
In The Street
His attractive face a bit pale,
his brown eyes looking tired, dazed,
twenty-five years old but could be taken for twenty,
with something of the artist in the way he dresses
-the colour of his tie, shape of his collar
he drifts aimlessly down the street,
as though still hypnotized by the illicit pleasure,
the very illicit pleasure he's just experienced.
Constantine P. Cavafy
The copyright of the poems published here are belong to their poets.
Internetpoem.com is a non-profit poetry portal. All information in here has been published only for educational and informational purposes.


text:
I find a lot of inspiration on Instagram. As most of us know from experience, it is so easy to go down a rabbit hole following one interesting video or photo after another. I love to watch the DIY and home improvement stuff, but, of course, the themes that really tick the boxes for me are related to gardening and floral art.
Here is a selection of accounts that I pop over to regularly. They inspire me, they inform me and they give me great tips that I can try out in my own garden and potager.
I highly suggest you give them a follow, and if you have any favourite accounts that you know of and would like to share, please leave a comment at the end of the post, merci!
Anya Lautenbach – aka the “Garden Fairy” has a wonderful and interesting account filled with really practical tips and information. She is especially good at reminding me which tasks I should tackle each season!
Claus Dalby – Does Claus even need an introduction?! His garden in Denmark is quite remarkable. He is a plantsman and has a rare gift for combining plants and creating borders. But maybe his most surprising thing is the way he juggles thousands (yes, literally) of pots planted up for seasonal colour and interest.
Irene – of Irenemylife lives in the UK and shares photos of her beautiful property with her fluffy white geese. Her potager is picture-perfect as well as highly productive. Yew topiary and beautiful architecture only add to the charm.
Lee – of Urban Veggie Patch lives in Australia and has created a wonderful potager in her urban yard. She shares her tips and ideas most generously.
Charlie McCormick – A great place to visit to get inspired. His dahlias are world-famous and his vegetable patch, a wonder to behold!
Edoardo Grassi – lives in France but is actually Italian. He cultivates dahlias and tulips and is very generous with tips on how to grow them. I did a live video with him last year and hope to get back there in the spring. Watch it here!
Erin Benzakein – of Floret Flower is simply amazing. As well as cultivating acres of perfect plants, she also writes books and runs excellent online courses for those wanting to follow in her footsteps!
Kate Coulson – Kate takes gardening to a totally different level. Her garden in England is spectacular, as is her property in the South of France. I am fascinated by her topiary, her continual research on design and her espalier trees growing up stone walls.
Linda Vater – of Potager Blog is great at sharing her tips and her garden design. Inspirational and practical perfection!
All photography credited to respective account owners.


January 2011 Issue
Bringing Ocean Science To the
Forefront of National Policy
By Robert B. Gagosian
President and Chief Executive Officer
The Consortium for Ocean Leadership
The past year was an exceptionally active one for ocean science. From the executive order establishing a National Ocean Policy to the devastating Deepwater Horizon oil spill, ocean science has been front and center in 2010. As an organization striving to shape the future of ocean science and technology through discovery, understanding and action, the Consortium for Ocean Leadership, representing 96 of the leading public and private ocean research, education and industry groups, has in turn had an extremely busy 12 months.
The above image shows Dr. Niel Bruce of the Museum of Tropical Queensland studying specimens in a lighted aquarium on Lizard Island Reef as part of the Census of Marine Life.
Deepwater Horizon Oil Spill
The BP plc (London, England) oil spill in the Gulf of Mexico proved to be a stark reminder of both the importance of the marine environment to our nation and its vulnerability. Furthermore, the fluctuating and conflicting reports on the size, scope and impact of the spill highlighted the complexity of understanding the dynamic ocean environment.
Serving as a trusted broker for ocean science and policy, the White House Office of Science and Technology Policy asked Ocean Leadership to be the liaison for a meeting in late May with the academic research community on the fates and effects of the oil spill. Sensing the need for further engagement of the academic community, Ocean Leadership then hosted a science symposium at Louisiana State University on June 3. More than 200 scientists from around the country attended, including key agency officials such as NOAA administrator Dr. Jane Lubchenco, U.S. Geological Survey director Dr. Marcia McNutt and National Science Foundation assistant director Dr. Tim Killeen, with many more participating via a live webcast stream, which appeared on CNN. The dialogue between the government and nonfederal scientists highlighted the unique capabilities of the academic research community and identified opportunities for partnerships to address the key scientific and monitoring gaps. Ocean Leadership published a meeting summary, which included research priorities for determining the magnitude of the spill, tracking the fate and transport of the oil and evaluating the long-term ecological and human effects. This summary is available on the Ocean Leadership website.
While we worked to promote a comprehensive national science program, our member institutions were on the front lines dealing with the catastrophe, from collecting water samples to predicting the oil slick's track to assessing the potential for future environmental damage. Academic researchers performed much of the most critical work immediately, often without waiting for federal or industry support, clearly demonstrating the dedication of the gulf science community and its duty to public service. As Washington policy makers probed to ascertain the federal response and future impacts of the spill, several representatives from our member institutions also shared their science expertise and knowledge through congressional testimony.
The enormous quantities of crude oil and dispersants that fouled the ocean will have long-lasting effects on the gulf's ecosystems, economies and society. In order to understand the long-term fate and effects of this oil and the dispersants used, it is clear that a comprehensive, long-term scientific research effort is needed. This effort will take several decades, and the credibility of its conclusions is dependent on thoroughly peer-reviewed scientific processes.
Ocean Leadership has advised BP about the best practices to promote a comprehensive, peer-reviewed science initiative, as we have many years of experience, including managing the National Oceanographic Partnership Program, the Integrated Ocean Drilling Program, the Census of Marine Life and the Ocean Observatories Initiative. We are also strongly advocating for the establishment of a comprehensive, peer-reviewed federal science program.
The Deepwater Horizon oil spill has been a tragedy of epic proportions, and we must now take this opportunity to learn as much from it as possible—not only to understand the fates and effects of this spill, but to prevent and mitigate the impact of future spills.
National Ocean Policy
Another landmark moment for ocean science occurred when President Barack Obama signed an executive order in July establishing a national policy for the stewardship of the oceans, coasts and Great Lakes. The order also established a cabinet-level National Ocean Council and a framework for coastal and marine spatial planning. Ocean Leadership's efforts advocating that science remain at the heart of the policy were successful, as science is incorporated throughout the national priority objectives, as well as being recognized in its own right.
Objectives in 2011
In the next year, our organization's attention will be focused on the implementation of the National Ocean Policy, with an emphasis on a comprehensive, science-based approach toward the ocean and coastal management.
Proper implementation will require significant new scientific investments to improve understanding of the ocean system, its interaction with climate and the sustainable use of marine resources.
As Obama said when he entered office, it is time to 'restore science back to its rightful place.' So 2011 looks to be an interesting year. We plan on continuing to advocate for the importance of keeping the highest quality ocean science front and center in the nation's ocean policy.


text:
Sci-fi anime Dr. Stone is getting a sequel!
The sequel was announced after the conclusion of the TV anime series' second season on March 25. The announcement was accompanied by a special video teasing Senku's next adventures in the Age of Exploration arc.
Details on the sequel anime's release date, length, and format are yet to be revealed.
Based on Inagaki Riichiro and Boichi's manga of the same name, Dr. Stone is set in a world in which humanity has been turned to stone. After waking up alone, a young genius named Senku tries to bring back the world around him through the power of science.
©Kome Studio, Boichi / Shueisha, Dr.STONE Production Committee


text:
SO, it seems I have a poor yeast Viability issue, the 1214 I am using for my Backup Plan, is not running, it started bubbling about 6 hours after pitching, then 8 hours late that was it, so i left it and left it, hoping it was still doing something, it stopped at 1.020, yet another stuck fermentation on a small 12 litre batch!
I figured out that the viability may not be quite so good on the yeast and I guess that my own fault, as there wasn't a whole lotta activity, during the starter time.
So, I re-pitched a Dry Yeast packet of T-58, and it seems to be doing something, which confirms the yeast were a bit pooped, it'll be interesting to see what it tastes like at the end, I'm fermenting on the upper scale of this T-58 yeast.
Basically, its not going to taste anything like the commercial version of Plan B, but here's hoping it has a slight taste or something close-ish.
Next up, anotherchange of plans: I'll be making a belgian Red Ale, I've been working on a recipe for it, sessionable at around 5.5-5.7 % abv. With a new packet of 1214, I'm hoping this will work and I'll finally get my yeast cake for the KWAK Clone recipe I've got on the cards, Hopefully before Surgery. I'm still waiting for ACC anyways, So, I kinda figure I can get another brew or two in. In particular If I can get the KWAK clone down and maybe I'll get a helping hand from a local brewer to do some heavy lifting. once its racked over to another vessel.

domain: hobby


text:
Wednesday, September 3, 2014
I probably post way too many pictures of food.
This was our lunch today and it was beyond delicious. I felt like I kept cramming veggies into it and really thought it was going to be a huge wrap but when I rolled it up it was so little. Still. Delicious. Ever since Trey told me about these spinach herb tortillas for wraps, it's the only thing I will buy. I always have them on my kitchen cart just waiting to be eaten. They're delicious and you can put anything on them. I love to put black olives, spinach, tomatoes and especially avocado with all of the usual stuff like turkey or ham and then maybe cream cheese. I also really like the colby-pepper jack cheese on this. Actually, I like the colby-pepper jack cheese on anything that requires sliced cheese. The actual death of me ;) my love of cheeses.
I hope you guys will try these tortillas out for your wraps if you haven't already. I'm probably a wrap rookie compared to others. Given that I'm not one for change, hopefully my love of wraps will help me branch out and try anything you guys and gals might suggest.
I hope your day is incredible. Wednesday, middle of the week or not - you always have the opportunity to make your time matter. Arrivederci!

domain: food


Parkinson, John B
Model No. 11-A was designed as an improvement over N.A.C.A. Model No. 11, a complete test of which is described in N.A.C.A. Technical Note No. 464. In contrast with the longitudinal upward curvature in the planing bottom forward of the main step on Model 11-A was made as flat as practicable. Otherwise, the two models have very nearly the same form. The results of towing tests made on Model 11-A in the N.A.C.A. tank over a wide range of speed, load on the water, and trim angle are presented, both as original test data and as non dimensional coefficients. A comparison is made with similar results from the test of Model No. 11. The practical significance of the improvement obtained is demonstrated by applying the data from the new form to the illustrative design problem use in the note on Model NO. 11.
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What were the basic shifts of the industrial revolution?
The technological changes included the following: (1) the use of new basic materials, chiefly iron and steel, (2) the use of new energy sources, including both fuels and motive power, such as coal, the steam engine, electricity, petroleum, and the internal-combustion engine, (3) the invention of new machines, such as …
What year did the 8 hour work day become common?
September 3, 1916: Congress passed the Adamson Act, a federal law that established an eight-hour workday for interstate railroad workers. The Supreme Court constitutionalized the act in 1917.
How did workers respond to the Industrial Revolution?
The Industrial Revolution led to rapid changes in people’s living and working conditions. In response to poor working conditions, labor movements organized alliances known as unions and pushed for reforms.
Who made the 8 hour work day?
Industrial revolution In the early 19th century, Robert Owen raised the demand for a ten-hour day in 1810, and instituted it in his “socialist” enterprise at New Lanark. By 1817 he had formulated the goal of the eight-hour day and coined the slogan: “Eight hours’ labour, Eight hours’ recreation, Eight hours’ rest”.
What was before the 5 day work week?
In 1926, Henry Ford, the man at the helm of the Ford Motor Company, shut down his seven-day automotive factories for two days a week — giving rise to the foundation of the five-day workweek in North America. In 1940, the 40-hour workweek was mandated nationally across the United States along with the two-day weekend.
How many hours did people work in the Industrial Revolution?
With a long line of people willing to work, employers could set wages as low as they wanted because people were willing to do work as long as they got paid. People worked fourteen to sixteen hours a day for six days a week.
How did working conditions change during the Industrial Revolution?
Simply, the working conditions were terrible during the Industrial Revolution. As factories were being built, businesses were in need of workers. With a long line of people willing to work, employers could set wages as low as they wanted because people were willing to do work as long as they got paid.
What was the typical work day in the 19th century?
With little representation, education, or options, factory workers also tended to work in horrible working conditions to go along with the bad hours. The typical work day at this time lasted anywhere from 10-18 hours per day, six days a week. This all began to change in the 19th century.
What did men do in the Industrial Revolution?
While the Industrial Revolution eventually put great pressure on men to engage in paid work outside the home continuously from adulthood to retirement, some men, particularly older men, refused to work in the factories and preferred to engage in spot labor and work around the home.

domain: history