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Receptor-interacting protein kinase 3 as a predictive adjuvant chemotherapy marker after lung adenocarcinoma resection
Lung cancer is the leading cause of death worldwide (1). Surgical treatment is the best option for treating patients with early-stage (stage I or II) non-small cell lung cancer (NSCLC). However, the role of surgery in treating stage IIIA NSCLC remains controversial. The clinical approach for treating patients with stage IIIA NSCLC is often determined by a multidisciplinary team comprising thoracic surgeons who analyze the number of lymph nodes, tumor size, preoperative neoadjuvant therapy, and other challenging clinical problems (2). Even after complete tumor resection, patients are at a substantial risk of the recurrence of and death due to NSCLC. The use of adjuvant chemotherapy has shown survival benefits in patients who undergo complete resection of stage II–IIIA NSCLC. Moreover, advancements in cisplatin-based chemotherapy has widely increased its use as an adjuvant chemotherapy. A study by an NSCLC collaborative group compared the 5-year OS of patients undergoing surgery alone with that of patients undergoing surgery plus adjuvant chemotherapy and showed that the OS of the patients who underwent surgery plus adjuvant chemotherapy arm was longer than that of the patients who underwent surgery alone [hazard ratio (HR), 0.86; 95% confidence interval (CI), 0.81–0.92; P<0.0001] (3). Previous guidelines have attempted to prevent adverse effects such as toxicity in patients undergoing chemotherapy. However, toxicity remains a concern in patients undergoing chemotherapy, and not all patients benefit from chemotherapy. Therefore, it is important to determine predictive biomarkers of adjuvant chemotherapy to identify patients who will benefit the most from this therapy.
Receptor-interacting protein kinase 3 (RIPK3), a member of RIP serine/threonine family, is a key regulator of necroptosis. The human RIPK3 gene is located on chromosome 14q11.2 and is implicated in the tumorigenesis of several cancers (4,5). However, its clinical significance and potential role in the pathogenesis of lung cancer have been poorly studied. Recent studies have shown that treatment with hypomethylating agents restores silenced RIPK3 expression and increases sensitivity to chemotherapeutic agents in an RIPK3-dependent manner (6). Upregulation of silenced RIPK3 expression also restores cisplatin sensitivity, suggesting that RIPK3 contributes to cisplatin chemosensitivity (7). Based on these findings, we examined the role of RIPK3 as a potential predictive biomarker of cisplatin-based adjuvant chemotherapy in patients who previously underwent surgical resection of lung adenocarcinoma (LUAD).
Patients and clinical specimens
In all, 143 frozen tumor tissue specimens that were surgically resected from patients who underwent lobectomies for primary LUAD from 2009 to 2015 were provided by the Biobank of Pusan National University Yangsan Hospital. All the tumor specimens were histologically examined and were classified using the 2015 World Health Organization International Classification of Lung Tumors (8). Tumor tissue microarrays (TMAs) were constructed using two tissue cores (diameter, 1 mm) per tumor, specifically tissues obtained from central and peripheral tumor areas, after performing the histological examination of the 143 LUAD specimens at the Pathology Department of the Pusan National University Yangsan Hospital. As a part of staging work-up, all the patients underwent contrast-enhanced computed tomography (CT), molecular resonance brain metastasis scans, and positron emission-CT within 1 month before the surgery. Patients with invasive adenocarcinoma, including lepidic, acinar, papillary, mucinous, and solid adenocarcinomas, were included in a conventional adenocarcinoma group. Lymphatic and vascular invasion of tumor cells was observed in lymphatic and vascular vessel lumens, respectively. The following clinical characteristics of the patients were analyzed to determine their effect on patient survival: age, sex, smoking history, and tumor differentiation degree. The clinical and pathological data of all the patients were integrated. Of the 143 patients who underwent surgical resection for LUAD, 50 patients received a cisplatin-based regimen for at least three cycles (Table 1). CT scans were performed to evaluate the efficacy of the chemotherapy regimen after two to four cycles. This study was approved by the Institutional Review Board (05-2017-162) of the Pusan National University Yangsan Hospital.
Full table
Western blotting analysis
The lung tissue samples of the patients were homogenized and lysed using PRO-PREPTM Protein Extraction Solution (iNtRON Biotechnology, Korea). The lung homogenates then were centrifuged, and supernatant obtained was collected. Protein concentration of the supernatant was determined using a protein assay kit (Bio-Rad, Hercules, CA). Next, equal amounts of protein present in the supernatant were resolved by performing SDS-PAGE and were transferred onto PVDF membranes for performing immunoblotting analysis. The membranes were blocked with 5% skimmed milk in tris-buffered saline containing Tween-20 and were incubated with primary antibodies against RIPK3 (Cell Signaling Technology, Danvers, MA) and β-actin (Santa Cruz Biotechnology, Dallas, TX). The bound antibodies were detected using a horseradish peroxidase-conjugated secondary antibody (Santa Cruz Biotechnology, Dallas, TX), and antigen-antibody complexes were visualized using WesternBright ECL HRP substrate (Advansta Inc., Menlo Park, CA). Densitometric analysis was performed using FluorChem SP AlphaEase® FC (version 6.0.0) software (Alpha Innotech, San Leandro, CA), and data obtained were normalized using those obtained for β-actin.
Immunohistochemical analysis
Immunohistochemical analysis of the lung tumor specimens was performed using an anti-RIPK3 antibody (Thermo Fisher Scientific, Waltham, MA), according to the manufacturer's instructions. The tissue sections were counterstained with Mayer's hematoxylin, and immunostaining was visualized using cellSens standard (DP21; Olympus Life Sciences, Tokyo, Japan). Staining intensity in each slide was scored on a scale of 0 to 3+. RIPK3 expression was evaluated based on the staining intensity score, with a score of 0 to 1+ indicating low RIPK3 expression and a score of +2 to +3 indicating high RIPK3 expression. In case of discrepancy in the staining intensity scores, the tissue sections were re-examined by another pathologist.
The Cancer Genome Atlas data analysis
In the present study, The Cancer Genome Atlas (TCGA) mRNA sequencing datasets of 506 patients with LUAD, which are available on cBioPortal online platform, were analyzed to determine whether RIPK3 expression level affected the prognosis of patients with LUAD.
Statistical analysis
OS and disease-free survival (DFS) were analyzed by obtaining Kaplan-Meier plots showing survival probability curves with different strata. Cox univariate proportional hazard regression models were fitted to identify the effect and significance of each factor of interest on survival patterns. In the univariate analysis, HRs with 95% CIs were determined for each clinical characteristic of the patients.
RIPK3 expression status in patients with LUAD
The predictive significance of RIPK3 was determined by performing western blotting analysis. No significant differences were observed between RIPK3 protein levels between the tumor tissues and paired normal tissues (n=11, P=0.877; Figure 1A). However, a variation in RIPK3 expression levels was observed in the TMAs constructed from the tumor specimens obtained from the 143 patients with LUAD (high RIPK3 expression, 53 patients; low RIPK3 expression, 90 patients; Table 2). The correlations between the clinical characteristics of the patients with LUAD and RIPK3 expression are summarized in Table 2.
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NSCLC, non-small cell lung cancer.
High RIPK3 expression levels are associated with good clinical outcomes of patients undergoing cisplatin-based adjuvant chemotherapy after complete tumor resection
To confirm the differential RIPK3 expression and its correlation with the clinical characteristics of the patients with LUAD, we performed immunohistochemical analysis of the TMAs constructed from the tumor specimens obtained from the 50 patients who underwent cisplatin-based adjuvant chemotherapy after LUAD (stage IB–IIIA) resection. Each adenocarcinoma tissue specimen was classified into high (RIPK3 staining intensity score of 2+ or 3+) or low (RIPK3 staining intensity score of <2+) RIPK3 expression groups (Figure 1B). High RIPK3 expression was detected in 48.0% (24/50) TMAs and low RIPK3 expression was detected in 52.0% (26/50) TMAs (Table 3).
Full table
The Kaplan-Meier survival analysis with a log-rank test showed a correlation between high RIPK3 expression and the prolonged OS; however, the log-rank test of the curves showed no significant correlation (Figure 2A). Moreover, the Kaplan-Meier analysis showed that RIPK3 expression was significantly associated with DFS rate (log-rank test, P=0.015; Figure 2B). However, analysis of the TCGA mRNA sequencing data showed that the OS rate was higher in the patients showing high RIPK3 expression than in those showing low RIPK3 expression (P=0.0061; Figure 2C). These results suggest that increased RIPK3 expression levels are a good predictor of survival in patients who undergo cisplatin-based adjuvant chemotherapy after surgical LUAD resection.
High RIPK3 expression is associated with the positive outcomes of cisplatin-based adjuvant chemotherapy
To examine whether RIPK3 was associated with prognosis based on chemotherapeutic responses, we examined adenocarcinoma samples obtained from the 50 patients who underwent the cisplatin-based adjuvant chemotherapy after complete LUAD resection (Table 4). RIPK3 immunostaining was significantly more intense in the TMAs of patients showing complete remission, partial remission, and stable disease than in the TMAs of patients showing a progressive disease (PD). However, the frequency of developing a PD was higher in the patients showing low RIPK3 expression (14/26, 53.8%) than in those showing high RIPK3 expression (6/24, 25.0%). Results of the univariate analysis showed that the age (OS: HR, 1.09; 95% CI, 1.03–1.16; P=0.004, DFS: HR, 1.09; 95% CI, 1.03–1.16; P=0.003) and N stage (OS: HR, 1.71; 95% CI, 1.02–2.85; P=0.04, DFS: HR, 1.80; 95% CI, 1.09–2.99; P=0.02) of the patients with LUAD were associated with the prolonged OS and DFS, whereas the RIPK3 expression status of the patients with LUAD was only associated with the prolonged DFS (HR, 0.32; 95% CI, 0.11–0.98; P=0.04; Table 5). These results indicate that RIPK3 expression is significantly associated with the prolonged DFS of patient undergoing cisplatin-based adjuvant chemotherapy after LUAD resection.
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Cisplatin-based adjuvant chemotherapy alleviates the risk of relapse after complete resection in patients with high-risk stage IB, II, and IIIA adeno-NSCLCs. International Adjuvant Lung Cancer Trial examined the effect of cisplatin-based adjuvant chemotherapy on the survival of patients undergoing complete NSCLC resection and reported a benefit of 4.1% in the 5-year OS of 1,867 randomized patients (9). Another clinical study (JBR.10 trial) assessed the benefit of cisplatin-based adjuvant chemotherapy on the 5-year OS of patients with early-stage IB or II NSCLC and reported that the 5-year OS of the patients receiving the cisplatin-based adjuvant chemotherapy improved by 15% compared with that of control patients (10). However, a long-term follow-up showed that the 5-year OS was not significantly different between patients who underwent surgical resection plus cisplatin-based adjuvant chemotherapy and those who underwent surgical resection alone (11). At present, no objective biomarkers besides clinical risk factors such as tumor size and lymphovascular invasion are available for selecting patients with stage IB adeno-NSCLC for administering adjuvant chemotherapy. Several biomarkers that predict the benefit of cisplatin-based adjuvant chemotherapy have been identified to date (12,13); however, none of these biomarkers are appropriate for clinical use. Therefore, there is a need for determining biomarkers that can identify patients who are most likely to benefit from adjuvant chemotherapy. In the present study, we elucidated the predictive significance of RIPK3 expression levels in the patients who underwent cisplatin-based adjuvant chemotherapy after the surgical resection of stage IB–IIIA NSCLCs.
Previous studies have shown that RIPK3 expression is markedly reduced in cancer tissues compared with that in normal tissues, which decreases necroptosis as an alternative to cell death in the presence of apoptosis resistance (14). Furthermore, the regulation of RIPK3 expression plays an important role in cancer survival by modulating the sensitivity of cancer cells to anticancer chemotherapy (15-18). The predictive role of RIPK3 by using clinical tissue analyses has not been published to date. The present study is the first to examine the RIPK3 expression status in LUAD tissues and adjacent normal tissues by using the TCGA mRNA sequencing data and to show no significant difference in RIPK3 expression levels between the LUAD tissues and adjacent normal tissues (data not shown). The results of immunohistochemical analysis performed in the present study are consistent with those obtained by analyzing the TCGA mRNA sequencing data (n=11, P=0.877; Figure 1A). A recent study also reported no statistical difference in RIPK3 expression between LUAD tissues and normal tissues (7). However, we observed a variation in RIPK3 expression level in the TMAs constructed using the surgical specimens obtained from the 143 patients with LUAD (Table 2).
We assessed the association of high and low RIPK3 expression with the clinical characteristics, DFS, and OS of the 50 patients with operative stage IB–IIIA adeno-NSCLC who underwent the cisplatin-based adjuvant chemotherapy. A recent study reported that mutant RIPK3 overexpression in esophageal cancer cells restores cisplatin sensitivity and that RIPK3 silencing is associated with a short OS rate (7). Another study suggested that RIPK3 is an important independent prognostic indicator of the OS and DFS of patients with colorectal cancer (19). The Kaplan-Meier curves obtained in the present study showed that high RIPK3 expression levels were associated with increased OS; however, the log-rank test of the curves showed no significant association (P=0.109; Figure 2A). Because the present study included a small number of patients with LUAD, the TCGA mRNA sequencing data were used to investigate the OS of patients with LUAD and its association with RIPK3 expression levels in these patients. Our results showed that the OS rate of the patients showing high RIPK3 expression was higher than that of the patients showing low RIPK3 expression (P=0.0061; Figure 2C). Moreover, our results showed that the DFS rate was significantly higher in the patients showing high RIPK3 expression than in those showing low RIPK3 expression (P=0.015; Figure 2B). We next analyzed whether RIPK3 expression was associated with the prolonged DFS rate by analyzing the chemotherapeutic responses of the patients and found that the patients with high RIPK3 expression showed lower incidence of PD than those with low RIPK3 expression (P=0.03; Table 4). In addition, the Cox univariate proportional hazards regression model indicated that age, N stage, and RIPK3 protein expression status were associated with the prolonged DFS rate of the patients with LUAD who underwent the cisplatin-based adjuvant chemotherapy after complete tumor resection (Table 5). These results suggest that RIPK3, at least partly, is a useful biomarker for predicting the response of patients with adeno-NSCLC to cisplatin-based adjuvant chemotherapy. However, additional large-scale studies should be performed to validate the results of our preliminary study and to assesses whether RIPK3 is a predictive biomarker associated with the long-term effectiveness of cisplatin-based adjuvant chemotherapy in patients with early-stage LUAD.
The authors would like to thank Dr. YS Kim (Ajou University, Korea) for kindly gifting experimental supplies. The biospecimens and data used in this study were provided by the Biobank of the Pusan National University Yangsan Hospital. This study was supported by a grant from the Research Institute for Convergence of Biomedical Science and Technology, Pusan National University Yangsan Hospital, to DH Kim.
Conflicts of Interest: The authors have no conflicts of interest to declare.
Ethical Statement: This study was approved by the Institutional Review Board (05-2017-162) of the Pusan National University Yangsan Hospital.
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- Arriagada R, Auperin A, Burdett S, et al. Adjuvant chemotherapy, with or without postoperative radiotherapy, in operable non-small-cell lung cancer: two meta-analyses of individual patient data. Lancet 2010;375:1267-77. [Crossref] [PubMed]
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- Koo GB, Morgan MJ, Lee DG, et al. Methylation-dependent loss of RIPK3 expression in cancer represses programmed necrosis in response to chemotherapeutics. Cell Res 2015;25:707-25. [Crossref] [PubMed]
- Sun Y, Zhai L, Ma S, et al. Down-regulation of RIPK3 potentiates cisplatin chemoresistance by triggering HSP90-ERK pathway mediated DNA repair in esophageal squamous cell carcinoma. Cancer Lett 2018;418:97-108. [Crossref] [PubMed]
- Travis WD, Brambilla E, Burke AP, et al. WHO Classification of Tumours of the Lung, Pleura, Thymus and Heart. Lyon: International Agency for Research on Cancer, 2015.
- Arriagada R, Bergman B, Dunant A, et al. Cisplatin-based adjuvant chemotherapy in patients with completely resected non-small-cell lung cancer. N Engl J Med 2004;350:351-60. [Crossref] [PubMed]
- Butts CA, Ding K, Seymour L, et al. Randomized phase III trial of vinorelbine plus cisplatin compared with observation in completelyresected stage IB and II non-small- cell lung cancer: updated survival analysis of JBR-10. J Clin Oncol 2010;28:29-34. [Crossref] [PubMed]
- Kris MG, Gaspar LE, Chaft JE, et al. Adjuvant Systemic Therapy and Adjuvant Radiation Therapy for Stage I to IIIA Completely Resected Non-Small-Cell Lung Cancers: American Society of Clinical Oncology/Cancer Care Ontario Clinical Practice Guideline Update. J Clin Oncol 2017;35:2960-74. [Crossref] [PubMed]
- Bell EH, Chakraborty AR, Mo X, et al. SMARCA4/BRG1 Is a Novel Prognostic Biomarker Predictive of Cisplatin-Based Chemotherapy Outcomes in Resected Non–Small Cell Lung Cancer. Clin Cancer Res 2016;22:2396-404. [Crossref] [PubMed]
- Ma X, Le Teuff G, Lacas B, et al. Prognostic and Predictive Effect of TP53 Mutations in Patients with Non-Small Cell Lung Cancer from Adjuvant Cisplatin-Based Therapy Randomized Trials: A LACE-Bio Pooled Analysis. J Thorac Oncol 2016;11:850-61. [Crossref] [PubMed]
- Wang T, Jin Y, Yang W, et al. Necroptosis in cancer: An angel or a demon? Tumour Biol 2017;39:1010428317711539. [Crossref] [PubMed]
- Chen C, Xiao W, Huang L, et al. Shikonin induces apoptosis and necroptosis in pancreatic cancer via regulating the expression of RIP1/RIPK3 and synergizes the activity of gemcitabine. Am J Transl Res 2017;9:5507-17. [PubMed]
- He W, Wang Q, Srinivasan B, et al. A JNK-mediated autophagy pathway that triggers c-IAP degradation and necroptosis for anticancer chemotherapy. Oncogene 2014;33:3004-13. [Crossref] [PubMed]
- Yang H, Ma Y, Chen G, et al. Contribution of RIPK3 and MLKL to immunogenic cell death signaling in cancer chemotherapy. Oncoimmunology 2016;5:e1149673. [Crossref] [PubMed]
- Vandenabeele P, Declercq W, Van Herreweghe F, et al. The role of the kinases RIP1 and RIPK3 in TNF-induced necrosis. Sci Signal 2010;3:re4. [Crossref] [PubMed]
- Feng X, Song Q, Yu A, et al. Receptor-interacting protein kinase 3 is a predictor of survival and plays a tumor suppressive role in colorectal cancer. Neoplasma 2015;62:592-601. [Crossref] [PubMed]


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Ashen Green Tweed Cape with Lilac Horse lining
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A lovely pale green tweed cape with baby blue and pinky purple hue over-checks. Lined in a lovely lilac horse lining. Finished with our horse fasteners.
Collar can be worn up, down or simply draped.
All our capes are handmade and fully lined. Other material & colour combinations available on request. Contact Us to commission your own bespoke design for adult or child.
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subset: commoncrawl


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Schenectady Art Week!! Just $25! Double Noodle Points!
June 4 @ 7:00 pm - 9:00 pm$25.00
Lighthouse to creativity!!! Adorn with hues of yellow, orange and pink or choose your own!!
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It would not be wrong to speak of people as having a compulsion to photograph: to turn experience itself into a way of seeing. Ultimately, having an experience becomes identical with taking a photograph of it, and participating in a public event comes more and more to be equivalent to looking at it in photographed form. That most logical of nineteenth-century aesthetes, Mallarmè, said that everything in the world exists in order to end in a book. Today everything exists to end in a photograph. Susan Sontag, On Photography
And now we've got that out of the way, let me proceed to prove her right. Here's us at Backyard last night. Me, the Prabhakaran and the Ads.
And here is Mark playing (but of course, why else would I be at Backyard?)
So I was going to tell you about Penang. Well, we went to the museum but weren't allowed to take pictures so I can offer you no visual record of the place. Instead, here's a frontal view of the E&O.
Anyways, we woke up early the next day (despite our relatively late night out) because I wanted to have the roti kosong and kopi susu at this place in Teluk Bahang. I kept telling the other two how Jackie, Julie and I had stumbled upon it in our meanderings through Penang and just how wonderful it was. All I remembered was...near the roundabout.
The place was closed. We asked a guy hanging about...and he said...raya. It was Hari Raya Haji the next day, so these guys were celebrating early. When would it open? After Raya. By which time we would be outta there. So we took our sad selves away, trying to find another place for our roti kosong and kopi susu (the other two wanted kurang manis and as for me, hey, what is life without sugar, lots and lots of it...healthy, you say? slim, you say? yeah, well there's more in life than that...like sugar)
The mamak around the corner was not half or even quarter as good. There was no fler in his sarong tariking the coffee. And the roti lacked ooomph. Young Eve and Younger Mary were not satisfied. They needed their morning coffee. So we ended up at a restaurant in Ferringhi, having morning coffee. Just coffee. And then...
Fast forward to a couple of hours later (bout the only hours in the day when we weren't stuffing our faces) and as we wandered through the fascinating exhibits (I think Penang has the most interesting museum in all of Malaysia) the two of them were dragging apace and complaining of hunger headaches. There was only one thing to do. Whack banana leaf. Accordingly we made our way to Vellu Villas. Being naturally unable to follow directions or trust in the directions we were following, we made phone calls to ascertain the way and stopped a satisfied looking Indian man who said, yes, yes, you just go straight and turn right there.
Mary, being the astute chickadee she is, asked him: "Did you just come from there?"
Nodding vigorously, he replied: "Yes, yes."
This is us whacking banana leaf. When you're really hungry, there's nothing so satisfying. Geez, I wish I could have some now. But Dadda is making rice and we will be having it with pickles and prawn sambal shortly. Shiokalingam!
And here's Mary...I can't remember what she was asking...maybe for the fried bitter gourd (they didn't have that but their vegetables and curries were fantastic anyway) I had the mutton curry and it was too errie for me. So Mary had to take my mutton and I took her chicken. It would have been OK if I had slathered the whole thing in tairu. But I have been known to change places cos I can't take the smell. Can't stand tairu, so can't eat hot mutton curry.
We dragged our overfull carcasses outta there in search of Bru coffee. Apparently Vellu (or is it Velu) Villas doesn't serve coffee until the evening. But coffee was of the essence so we went to Woodlands nearby and had our fill.
And then Eve decided that it was time to see the Khoo Kongsi. If you think we were clueless before, you should have seen us then. I think we asked about 10 different people for directions (although we kept advancing, generally in the right direction). Because Mary looked like this...
...everyone took us for tourists. One particularly bright spark we stopped and asked directions from asked if we were from China. I mean to say, what?
It cost us about RM10 each to get into the Khoo Kongsi and was totally not worth it. Going through renovations or something...I thought this was cute though:
And here's some more of it with my two superstars posing:
Well, by this time we were hot and sticky. Really hot and sticky. It was back to base for a shower, nap and out to very good dinner at Mario's...it was an interesting dinner. Especially interesting in that we didn't pay for it. In fact, most of our meals in Penang seemed to be sponsored by someone or other. (And except for breakfast, which we paid for ourselves) they were all VERY GOOD MEALS. When I tell anyone that we overate in Penang, they look surprised and say...but what else is Penang for? So yeah. And we didn't walk worth a darn because it was too hot and the humidity was another thing altogether.
But young Evelyn didn't get her char kueh teow at Gurney Drive (or wherever) because were shanghaied away for dinner...and we didn't get to walk along the beach or go to Flag Hill or to the nightmarket by the beach. All stuff to do next time around.
But there was one thing I had to do. Simply had to. When we visited the gift shop at E&O I had seen their drool-worthy coffee table book. I mean it was really something else. Ilsa Sharp drawing from multiple sources to tell us why E&O, and by extension Penang, was just the most interesting place, like ever! (Yes Nits, I know you agree...)
Here's me glued to the fancy vintage seat in the gift shop leafing through the book. Merlene and then her assistant (whose name I do not remember but who was a very nice sweet young girl) let me sit there for as long as I liked playing with the book. It was just too cool. But it was also RM188.50 and I would have to justify it to my conscience. Finally decided to give myself a birthday present. And guess what? Mary decided the same thing. So here was the book I really really wanted, compliments of my friend. Woo hoo!
Awwww, look at the the little pudge! Isn't she cute?
OK, young Evelyn took off for Langkawi the next morning. We did an early morning drive to airport (finding our way through a mixture of directions and road signs) and then came back via Balik Pulau which was a mistake as the road was windy and I felt like throwing up by the time we arrived back home and I crawled into bed for a nap.
Then it was off to Mary's where I was staying the night...and here's her with her cute little mommie...
So....that's the end of the Penang pictures....all good things must come to an end.
Btw Jack, lemmee introduce you to our new sofa. Dadda's Christmas present. The Shi Sho girl is presently breaking it in...she's curled up in her blankie on it, fast asleep.
Tuesday, November 23, 2010
2 comments:
That first picture is a real prize, wonderful smiles all around. When one whacks banana leaf (looks quite tasty, btw) is it taboo to eat with your left hand?
Gotta love that sign--it needs a smaller sign hanging beneath it, saying "Beware of the Leopard."
- Jenn said...
You know, since you're a mat salleh you could even request a fork and knife and it would be ok. I think it's perfectly ok to eat with your left hand. After all, banana leaf is about your ease rather than manners. But when you finish, remember to fold the leaf towards you rather than away from you. That way you signal that you enjoyed the meal.
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I sat in the corner of that L-shaped couch much longer than I originally intended. I hadn't planned on staying at all, not that night. I had too many thoughts racing through my head, and none of them were the good, uplifting, God inspired thoughts that I was supposed to be thinking. The thoughts I was thinking were more like, "See, you shouldn't have trusted them, you can't trust anyone", or "This is what always happens", or the ever so popular, "Never again. I'm never opening up to anyone ever again". I won't ask you to raise your hand or anything (mostly because I can't see it 😬) but I can bet that many of you have felt the same way. I felt stupid for thinking any of those things, but I couldn't help it. Why? Because at that time in my life, all that I knew was rejection. I'd been down the treacherous road of constantly giving love to people who were probably never going to love me back. Not because I knew that they were going to reject me. Each time, I sincerely hoped that this would be the time when someone finally loved me. And it wasn't because the love I was giving wasn't genuine, or that I wasn't giving enough of it. It wasn't even because there was something wrong with me. It all came down to one very simple thing, they didn't know how. Friends, family members, guys, didn't matter. I was on a vicious cycle of loving people, who didn't know how to receive it, and certainly didn't know how to love me back…(not that I would have known how to receive it either.)
How do you receive something, when you don't even know what it looks like? (Imagine an invisible football, flying through the air, that you are supposed to catch. Got me?) @LauraPLytle #truelove
Of course I didn't know what the problem was back then, so after years and years of that vicious cycle, I did what any self-preserving human being would do. I closed myself off to everyone. I had friends, and people I would hang out with, but I entered these relationships with the understanding that at some point, they were probably going to reject me, so I didn't let any of them get close enough to really do so. They probably thought that they were close enough, I may have even given them just enough personal information to reinforce that. But when it came to my heart, no one was getting anywhere near that.
Along with that, I adopted a very firm mindset; if anyone wanted to know me at all, they were going to have to find me in the emotional cave that I was hiding in, pursue me at every given opportunity, and shower me with love that I certainly wasn't going to return, until they proved to me that they were worthy of my loving them in return. (A view that many of us have long sense placed on God's love for us.)
I had only been saved for a few months at that point, so I had quite a bit to learn. But the thing that I learned that night on the L shaped couch, that was something that would change my life forever. It was a Bible study that I'd been going to for a couple months. And that night we'd been talking about family and relationships, (both things that I didn't always know how to talk about). It wasn't like my family wanted me around anyway, and besides, they were never going to forget who I used to be. I'd met Jesus, He had changed me, I KNEW that. But they didn't, and I had a feeling that they really didn't want to.
It was just a normal night as far as I was concerned. I wasn't expecting the leader to look at me. And I definitely wasn't expecting him to say what he said.
"Laura, what if they really don't love you?"
He paused, and I thought strongly about getting up and walking out. He was talking about my family, my friends, all of the people who had ever hurt me, everyone that I loved. He was reminding me of their rejection. What was his point? I knew that they didn't love me, only Jesus loved me. Only Jesus would or could ever love me. I was to "damaged" for everyone else. I was too much, or not enough. I knew it all too well. I tried to figure out why he would bring that up now. But then he read this….
" Love is patient and kind. Love is not jealous or boastful or proud or rude. It does not demand its own way. It is not irritable, and it keeps no record of being wronged. It does not rejoice about injustice but rejoices whenever the truth wins out. Love never gives up, never loses faith, is always hopeful, and endures through every circumstance. (1 Corinthians 13:4-7 NLT)"
"Are they any of those things, to you?"
I wasn't exactly calming down at this point. I was certain that my olive skin was turning more of a bright shade of crimson. He was asking me to answer the same questions that I'd been unable to answer my entire life. What was wrong with me that my own family didn't want me? Why was I so unlovable that no one loved me? No one that I loved ever loved me back, why? I could feel the tears forming behind my eyes, and my heart felt like it was going to pound out of my chest. But then he kept going…
"They don't KNOW HOW to love you. There is nothing wrong with you. Did you hear me? Laura, there is NOTHING WRONG WITH YOU! They just don't know what love really is, so they don't know how to love you. GOD loves you."
I didn't say much in response, if anything. I wasn't exactly sure what had just happened, but I knew that something had. When I got home I re-read 1 Corinthians 13, over and over and over. Then God revealed something to me. God is love. So if someone doesn't really know God, if they don't know Love, then how can they truly love another person? And within that revelation came another revelation; God's love was never based on our love for Him. He has always loved us, and will always love us, whether we love Him back or not. The only way that we can love Him, is in response to His love for us. Period.
Over the next few months I began to notice something…I was no longer loving with the stipulation that someone HAD to love me in return. Love is a gift that is given freely. It can't be bought, or earned, and it does not demand to be loved in return. To truly love, is to love without condition. And to truly know God's love, is to be complete in Him, knowing that no matter what, He loves you. Once I really got this revelation, I was able to stop expecting love from people who didn't know how to love me. But it didn't mean that I stopped loving them. In fact I asked God to help me love them the way that He loves them. Sometimes it was up close and personal, and sometimes it was silently through prayer. But over time I learned that it was always given freely, and never demanded in return.
Now don't get me wrong, you need to GUARD YOUR HEART. Not everyone should have access to it. And I am all for being pursued, especially when it comes to my future husband. But unless you're planning on marrying a creature like Gollum from Lord of the Rings, or you want a bunch of bats to befriend you, I don't suggest the emotional cave route. I also don't recommend the stage 5 clinger method (aka: Don't stalk people. Stalking is creepy…and again, love does not demand it's own way.
When I love, I love very extravagantly…my friends, my family, and my future husband. If I miss them, I tell them, even if I miss them twice a day. If I want to encourage them, I do it, I don't stop to question whether they need said encouragement. Who the heck doesn't need encouragement? If I want to talk to them, I call. I don't sit for an hour thinking "They're probably too busy", or "Maybe they don't want to talk to me." If I want to see them, I make it happen. I don't use the excuse of "I'm too busy", even though my schedule leaves very little room for 'me time', let alone 'them time'. If something is truly important to you, you will find a way, and you will make time for it. Period. If I want to get them a gift, I do it, because who doesn't like getting gifts? See all of those excuses stem from rejection, (unless of course you don't actually love them, or care about them, which would lead me to ask why you are thinking about them while reading this blog post, then. Nevermind.) My point is, that if you truly love and care about someone, then what is stopping you from telling them, and showing them, and making sure that they know that you do? There's only one key to remember and and the rest is very simple. I never do so just to "get something in return ". Why? Because God set me free from rejection, and now I love with my arms wide open, not just my hands wide open waiting to receive something.
Some may think it's "too much" , but that isn't a problem with me, that is a problem with them. If someone goes running the other way, they obviously don't belong in my life. There is no such thing as too much love, if it is real love that we are talking about. True love always responds to true love, eventually. The problem is that most people don't know what true love really is, because they don't truly know who God really is. Obsession is not love. Control is not love. Lust is not love. Fear is not love. Perversion is not love. Selfishness is not love. And I could go on and on about what isn't love. But the Bible is very clear on what love IS. It is "patient and kind. It's not jealous or boastful or proud or rude. It does not demand its own way. It is not irritable, and it doesn't keep a record of wrongs. It does not rejoice about injustice but rejoices whenever the truth wins out. It never gives up, never loses faith, and is always hopeful. It endures through every circumstance." If it isn't those things, then it isn't love, because if it isn't those things, then it isn't God.
Understand that people rejecting the love that God has put in my heart to show them, it might hurt sometimes, but don't let it harden your heart. I don't. My heart is in God's hands and guarded by Him. There are places that few people have access to. I choose to love those people excessively, and make sure that they know how I feel about them. And I've allowed God to teach me how to love all people like my Jesus, tell them about Him, help them when I am able, and share His healing love with them, I will do that always.
Jesus didn't say to love with conditions, He just said to love. But first you have to know what love really is. What does it really look like? It looks like Jesus. Ask Him to show you, and He will! @LauraPLytle #truelove


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in which it drew the ire of the Right to Censor (a group of morally conservative wrestlers).
Initial police reports stated she probably died of an accidental-drug overdose or natural causes.
In order to better convince the audience that she was injured, color commentator Jerry Lawler left the commentators' booth and entered the ring to check on Laurer's condition, something he had not done since the in-ring accident that killed Owen Hart in 1999.He introduced her as Kurt Angle's business associate (she had been previously referred to as his "Mistress") and tag team partner at Sacrifice, where they would face Jeff Jarrett and Karen Jarrett.During the taping she also took part in a battle royal, from which she eliminated Jeff.As a result, then "head of authority" Stephanie Mc Mahon-Helmsley declared them co-champions. Not long after losing the Intercontinental title, Laurer became the on-screen girlfriend of Eddie Guerrero.Guerrero and Laurer, originally villains, later became fan favorites during the summer of 2000, with Guerrero dubbing her his "Mamacita".She last saw her mother at the age of 16, and she claimed that her father was never able to get over her decision not to pursue a career in law enforcement with the Federal Bureau of Investigation.


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Windstar Cruises
Windstar Cruises is a cruise ship line that operates a fleet of small luxury cruise ships. Its six yachts carry just 148 to 310 guests and cruise to 50 nations, calling at 150 ports throughout Europe, the South Pacific, the Caribbean, and Central America.
The Windstar fleet has won the Condé Nast Traveler award for "World's Best Small Ship Cruise Line."
Jobs at Windstar Cruises
Windstar are looking for new talent to join their team and now is a great time to step on board. Windstar are the first choice for exciting career opportunities within the luxury small cruise ship sector. Working with a talented, international team on board – Windstar is a perfect place to move your career to the next level.
They are seeking qualified Deck Officers, Marine Engineers and Hotel Professionals to join their team. They offer an attractive employment package, including spouse travel (after a qualifying time). Previous cruise experience is not essential; however for Deck and Engine positions experience of single handed watching is desired. Previous experience from the cruise industry is desired for candidates applying for the Higher Hotel Department positions.
Viking Crew is the exclusive hiring partner and recruiter for Windstar Cruises.
For more information visit their careers website.
For current vacancies with Windstar Cruises on All Cruise Jobs:


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By Rustin Burr on Mon 09/24/2018 08:27am
Posted in
The Sheridan Fulmer Library will be one of the final stops on local author Craig Johnson's newest book tour Wednesday, as he will be in the Inner Circle for an event that evening. Sheridan County Library System Programming Coordinator Zola Shockley explains further.
Shockley said the free event is open to the community and it will start at 6:30 Wednesday evening. For more information visit www.sheridanwyolibrary.org.


text: yes this means that there is a infrastructural decision to make sure that UI can be controlled. >>> There are also some basic things that would need some adjustment as I >>> see it. I think core or basic classes and models shouldn't make any >>> assumptions about the UI or even if there is an UI at all. >> >> YESSSssssssss.... and a minicore will help /force us to do that (imagine an hudson script blocked because of a nice popup) > > Consider that this is not the UI framework that should decide to open > a dialog or not. If somebody tells morphic to open a window it should > open it otherwise you break all kinds of assumptions. > > If a user wants to support multiple UI frameworks (or the possibility > to disable certain UI actions), then he needs to take appropriate > action himself. OB for example uses notifications for all UI > interactions that are captured either by a morphic handler (by > default), a web handler (if you use the web interface), or by a test > handler (that builds a fake UI that doesn't display anything but gives > you a nice model to exercise the UI). > > Lukas > > -- > Lukas Renggli > www.lukas-renggli.ch > > _______________________________________________ > Pharo-project mailing list > Pharo-project at lists.gforge.inria.fr > http://lists.gforge.inria.fr/cgi-bin/mailman/listinfo/pharo-project


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1970 Plymouth Hemi Cuda Races For $3 Million Sale
https://www.yahoo.com/autos/1970-plymou ... 81718.html
Yeah, it's not a 1971, but that's why I like it better! I love that '70 front end.With the market for all types of collector cars running at near-record levels, many collectors have chosen to put their prized cars on the market. And while such sales are often dominated by foreign brands, a high-profile exception will be this 1970 Plymouth Hemi Cuda convertible in California, which Mecum Auctions expects to bring between $2.5 million and $3 million.
Built as the personal car of John Herlitz, who designed the exterior of the 1970-71 Barracudas at the ripe age of 27, the 425-hp Hemi Cuda convertible is just one of 19 such models built in 1970. While its not quite as rare as the 1971 Hemi Cuda convertible that Mecum sold last year for $3.78 million, it would still count among the most expensive Detroit vehicles ever sold at auction.


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The flat cut metal signs use waterjets and lasers to precisely cut metal letters and logos. Offering superior detail to the router-cut alternative, these cutting technologies allow any font or vector image to be replicated. With five major metals and over 50 standard finishes available, flat cut metal signs are durable enough for the outdoors yet stylish enough for any boardroom. The customization options for color, size and shape make flat cut metal an excellent choice for any signage project.
Flat Cut Metal
- Sizes range from 1/2″ to 60″
- Finishes include: painting, anodizing, oxidizing & polishing
- Detailed custom painting
- 5 metal types in multiple gauges
- Lifetime Guarantee
- Anodized Aluminum: Anodized finishes are not just coatings – they are part of the finished aluminum letter. Anodizing is an electrochemical process that hardens aluminum and allows it to be colorized. Benefits of anodized aluminum include abrasion and weather resistance; increased hardness and durability; will not crack or peel; unaffected by sunlight/UV exposure; environmentally friendly; cost effective
- Patina: By their very nature, patina finishes on bronze will vary from letter to letter and order to order. These finishes will change in appearance over time depending on contact with moisture and temperature. Patinas can "bleed" onto the installation surface when installed outside.
- Oxidized: Bronze, brass & copper are chemically treated, then hand rubbed, to produce an antique look on the letter face. Letter returns are painted black and the letter is sprayed with a low gloss clear coating.
- Brushed: All metals can receive a variety of brushed finishes for a range of visual effects.
- Polished: All metals can receive a polished finish.
- Painted: Choose from over 40 standard colors for painting aluminum letters.
If you are an active Navitor dealer, request a quote or place your order manually by submitting an email to Customer Service at firstname.lastname@example.com.
To become a Navitor dealer, follow this link to enter your information.
NSP is a trade printer and does not sell directly to consumers.


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Come out and celebrate Mardi Gras! The Zydeco Trail Riders, formed in 2004, have performed predominantly in upstate New York at regional festivals and at nightclubs, wineries and eateries. They play Zydeco, Creole, Cajun and Blues from Louisiana. The band is led by Greg Grodem, on vocals and accordions, with Jim Reidy on guitar, Ted Oberhaus on bass, Stuart Krasnoff on drums, and Peter Wolfanger on rubboard.
Check them out at The Brewery and at: http://zydecotrailriders.com/


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Brr.... It's getting cold. And you can help us freeze this!
Brr.... It's getting cold. And you can help us freeze this!
It's been a week since our last RSTop40 Special and you'd think we'd just relax and take some time to chill and cool off before our next, right? Right!
The next special edition of RadioSEGA's Top 40 Countdown, as announced during this week's show, will be the WinterFest Edition! WinterFest? Is that even a thing?
I figured why not stray away from the series, games, or composer-focused countdowns and do something different this time? And since we're doing this next Special in December, a winter-themed countdown seemed appropriate. Yes, a whole show to play your favourite 40 Winter-themed SEGA tracks! The big question is... How in the world are we gonna choose which tracks are gonna be in the poll???
That's where you come in: In the next month or so, you're gonna have the chance to suggest your favourite Winter-themed SEGA tracks and I'll add them (or most of them) to the final poll going live 2 weeks before the show airs, as usual. I'll probably be asking other community websites for their picks as well, so we can have some more variety.
How can you take part? Simple. Use the comments section below this exact blog post to suggest your favourites or send me an e-mail to kc [at] radiosega [dot] net. Alternatively, you can also use the official RSTop40 forum thread here.
So, go nuts! You've got a month to send me your suggestions and don't limit yourself to just Sonic tracks. You know what I think about that! :) Also, keep on listening to RadioSEGA's Top 40 Countdown, every Monday night at 8pm GMT!
Twinny 21st October, 2014 - 04:16Alright I got a few wintry-favs: Super Monkey Ball: Banana Blitz - Ice "Smooth Sherbet," Christmas NiGHTS - Dream Bells, NiGHTS Into Dreams - The Mantle, Sonic Advance 2 - Ice Paradise Act 1, SONIC The Hedgehog (2006) - White Acropolis: The Base
Rexy 23rd October, 2014 - 19:12We can nominate all we want, but I'm calling it now - Icecap(piest days of my life) will take the top stop :P
Willster 25th October, 2014 - 15:39I have a Hatsune Miku Wintry track: Project DIVA Arcade - The Snow White Princess is
Requests between 9th Sep and 15th Sep


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Incredibly windy (which is fine as a links), however the fairways were not in a good condition rough was far too long. The greens, however, were in very good condition.
Wimereux's classic seaside layout is truly spectacular, with views across the Channel to the white cliffs of Dover, and is a golf course of international repute. Many prestigious professional events have been staged at the 6,726 yard layout, which is punctuated by devilish pot bunkers, and is an exceptional golfing experience as part of an escape to France's nearest coast.
Golf de Wimereux is ranked as one of the best courses in Northern France
Incredibly windy (which is fine as a links), however the fairways were not in a good condition rough was far too long. The greens, however, were in very good condition.
It was a good weather day so perfect. If very windy it would be difficult. Many people playing this day including a golf society of about 30. There was a little waiting but OK. Menu for lunch was a bit limited. Course was perfect and so easily accessible from Calais.
This was a great course. Everything was in great condition. Would highly recommend. Watch the water on the right of the 18th
Very good
Our round was early in the morning 9.00am and was clear and bright but really windy (so really cold). Played link a true links - really windy but have to say the greens were really good even though had been sanded. If it wasn't so windy it would have been the best overall
Very uninspiring and didn't know where fairway and rough started but greens quick and true
A very good course which I would think in the wind would be difficult but we had a sunny still day. Busy place but we started at ten just before the rush
Great links style course, fabulous greens, it was calm and sunny when we played so felt comparatively easy, I'm sure it could be a real beast if the wind got up as its right on the cliff tops over looking the channel. This is evidenced with all the west facing fairways being low stroke indexes! Will definatly return here
Nice links course. The fairways were fast but the greens were perfection - find them and your ball would hold. The wind made some of the holes a real challenge. It was a bit slow going but not too bad. Great views of the sea and some interesting WWII bunkers to investigate!
Nice round to begin with. Fairways weren't in perfect nick so needed to pick and place on occasion
Lovely views as you progress through the round and greens played fantastic most holes were a little one dimensional though and I couldn't decide if the course wants to a links style course or downland but scores can be made if avoiding the rough
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text: Trina Reynolds-Tyler is an organizer, community builder, restorative justice facilitator, and graduate student at UChicago's Harris School of Public Policy. She also works at the Invisible Institute, a journalism production studio founded by Jamie Kalven (who was on the show a couple weeks back). She talks about the University's relationship to her childhood and neighborhood, learning from experience, wrestling with defining success, and much more.


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Kindy Houinato
Monday, 01 January 0001
I am a fourth-year student completing an Honour in Economics and a Major in Environmental Science. This summer, I am interested in understanding the advantage and disadvantages associated with the different chains formed by diatoms. Literature suggests that the shape and length of chains appear to be determined by several environmental factors, such as effectiveness of grazer defense (Sournia 1982; Smetacek et al. 2004; Bergkvist et al. 2012), adjustment of buoyancy (Smayda & Boleyn 1966; Smayda 1970; Takabayashi et al. 2006) , optimization nutrient uptake through micro shear (Reynolds 1984, Karp-Boss et al. 1996), and in pennate planktonic diatoms, a possibility for entanglement of chains to retain proximity for sexual reproduction (Kooistra 2007). My goal will be to gather available data on diatom traits from the literature. This can then be used to formulate new hypothesis about the purpose of chains or provide better insight on the current ones.


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OverviewProjects in the first funding period (B1-B18)B19 Ageing in a Transcultural ContextB20 Rethinking ArtB21 Transcultural Dynamics of PentecostalismMC2 SufismMC2.1 Sufi ShrinesMC2.2 Listening CommunitiesMC4.1 ObjectsMC4.2 DidacticsMC6 Global FascismMC12 Floating SpacesMC15 MediaNet1 Arts and TransculturalHERA
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Dr. Heather D. Baker (University of Vienna): June/July 2010.
Dr. Deniz Kaptan (University of Nevada, Reno): June/July 2010; February 2011.


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BMW M3 F80 with M Performance Exhaust
video note
540HP BMW M3 F80 Sedan fitted with a M Performance exhaust system and a stylish wrap. Top speed that day with rolling 50 km/h start after 1000meters was 257,82 km/h.
303 total views, 1 views todayUnited States of America, United States, America, USA, US, Canada