I mean really, check this out from Kevin Muir.
MMT sounds to me like just another cult. In fact, I see – and insulate myself from – cults everywhere. The gold "community" cult, the Keynesian cult, the cult of Evil Fed/Jekyll Island, the cult of Socialist Nirvana and the other of Conservative dogma held so strict it loses its moral compass.
But I digress. Kevin fleshes out this new cult of economics, talks about inflation and freeing governments from the limitations of whatever our existing monetary system is.
If I had told you four years ago that the following picture wasn't a photoshop, you would have probably told me I was nuts.
But it's serious business, having so many millions of people so pissed off. Ironically, the system under the supposedly socialist Obama was completely bent on making the rich richer and the poor poorer. Hence, the pic above.
Interesting stuff, anyway. Check it out.
We are definitely not in Kansas anymore.


text:
PALLADIUM:
USD $/oz | /oz
TSX: GENM
Board of Directors / Consultants
Generation PGM
Davidson Molybdenum Deposit
Darnley Bay Gravity Anomaly
Alberta Zinc Property
Palladium 101
Molybdenum 101
Zinc * USD/lb
Lead * USD/lb
Moly * USD/lb
Generation Mining Begins Trading On OTCQB Market Under Ticker GENMF
Toronto, Ontario – June 8, 2020 – Generation Mining Limited (CSE:GENM) ("Gen Mining", "Generation", or the "Company") is pleased to announce that its common shares are now trading on the OTCQB Venture Marketplace (the "OTCQB Venture"), a US trading platform that is operated by the OTC Markets Group in New York. The Company will trade on the OTCQB Venture under symbol "GENMF"; the Company's common shares will continue to trade on the CSE under the symbol "GENM". Nauth LPC acted as the Company's OTC Markets Sponsor.
"Listing on the OTCQB Venture is part of our strategy to introduce the Company to a broader audience," said Jamie Levy, President of Gen Mining. "We believe trading on the OTCQB will help us to introduce the company and enhance liquidity by providing our current and future American investors with a trading platform."
Currently, there are 130,505,937 common shares of the Company outstanding. Generation is well financed for the next phases of work, including the upcoming feasibility study and the restart of the permitting process at the Marathon Palladium Project, with approximately $14.3-million in cash.
About the Marathon Palladium Project
The Marathon Deposit is the largest undeveloped platinum group metal Mineral Resource in North America. The Marathon Property covers a land package of approximately 22,000 hectares, or 220 square kilometres. Generation Mining acquired a 51% interest in the Marathon Property from Sibanye Stillwater on July 10, 2019 and can increase its interest to 80 per cent by spending $10 million over a period of four years. As of Q1, 2020, approximately $4 million of the $10 million has already been spent. Sibanye Stillwater has certain back-in rights that can bring its interest in the Property back to 51 per cent after such time as Generation Mining has earned its 80-per-cent interest (see the company's press release of July 11, 2019, for more details).
Jamie Levy
President and Chief Executive
This news release includes certain information that may be deemed "forward-looking information" under applicable securities laws. All statements in this release, other than statements of historical facts, that address acquisition of the Property and future work thereon, Mineral Resource and Reserve potential, exploration activities and events or developments that the Company expects is forward-looking information. Although the Company believes the expectations expressed in such statements are based on reasonable assumptions, such statements are not guarantees of future performance and actual results or developments may differ materially from those in the statements. There are certain factors that could cause actual results to differ materially from those in the forward-looking information. These include the results of the Company's due diligence investigations, market prices, exploration successes, continued availability of capital and financing, and general economic, market or business conditions.
Investors are cautioned that any such statements are not guarantees of future performance and actual results or developments may differ materially from those projected in the forward-looking information. For more information on the Company, investors are encouraged to review the Company's public filings at www.sedar.com. The Company disclaims any intention or obligation to update or revise any forward- looking information, whether as a result of new information, future events or otherwise, other than as required by law.
© 2021 Generation Mining Limited All rights reserved. | Website by Adnet | Privacy Policy
© 2021 Generation Mining Limited

redpajama_set_name: RedPajamaCommonCrawl


Institutional investors are increasingly relying on specialized external investment managers for implementation of investment strategies. Important drivers of this development are: increasing efficiency of (parts of) financial markets, operational complexity, regulatory pressure, and cost awareness. Asset managers are meeting the business opportunity by offering a wide range of investment products, offering something for everyone. However, do these products really meet the investment objectives of asset owners, and do asset owners really know what they are looking for in the first place?
In this PE module, we place the process of selection & monitoring external managers in the context of the overall investment policy and governance structure. We also address important considerations such as: mandate structuring, the value (and costs) of active investment mandates, skill versus luck in generating outperformance, the pitfalls of quantitative performance analysis, legal and operational due diligence. The course provides a good balance between theoretical content and practical applications.
Wouter ten Brinke is Managing Director at Anthos Fund & Asset Management in Amsterdam. Responsibilities include: global macro and asset allocation, and client portfolio management. Wouter has over 20 years of experience in financial markets and asset management, of which 8 years in manager selection, due diligence and monitoring at hedge fund firm Theta Capital Management.
Richard Jacobs and Sven Smeets, are both Managing Directors at Kempen Capital Management in Amsterdam. Responsbilities include: private markets and external investment managers. Before joining Kempen Capital Management in 2017, Richard and Sven worked for Altis Investment Management, mananging client portfolios on the basis of manager and mandate selection.
The module is suitable for everyone who is working with external asset managers: as asset owner, investment consultant, portfolio manager, pension fund manager or as a board/committee member.
The first day we will start by placing the manager selection process in the context of the overall investment policy and investment program. We learn how to avoid performance pitfalls and make manager selection a forward-looking process. How much of the manager performance can be attributed to skills and implementation, and how much to the environment (capital market conditions or luck)? Participants will work through a stylized example of a typical manager selection workflow, with a prominent role for setting expectations regarding risk, return and role of the external manager in the overall investment portfolio.
On the second day, participants will be requested to present one of their own case studies. An important decision when designing an investment program is the choice between active and passive management: we look at the empirical evidence and factors that may improve the chances of success in active investment management. Blending quantitative risk-return analysis with a structured qualitative due diligence framework, asset owners will need a broad set of skills, capabilities and industry experience to support their process. Finally, during operational and legal due diligence, we assess the operational infrastructure and governance in the context of the investment strategy.
Everyone is welcome. To ensure a balanced interaction with the teacher and fellow students the number of course participants is limited to a maximum of 25.
The course material will be sent to you upon registration.
During the course preparation instructors are supervised by an academic coordinator to ensure the quality of the courses is in accordance with the objectives of CFA Society VBA Netherlands.
You will receive a certificate of participation.
Stichting VBA Opleidingsinstituut Morreau (Training Institute) is the education and training institute of CFA Society VBA Netherlands and as such organizes and executes the society's PE Modules.
Upon registration you will receive an invoice for the course fee, after which there is a 14 day period to change your mind. You will be informed immediately if the course is fully booked. You can cancel your registration without cost up to four weeks before the start. After four weeks the course will be charged in full.
The Training Institute reserves the right to make changes (including the content, format, instructors, course material, date, time and location) in the announced courses and / or to cancel the announced courses without providing a reason. The Training Institute mandates a minimum number of 10 participants. In case of cancellation full refund will take place. Refunds will be processed within 14 days. The Training Institute is never liable for lost student income.
The Training Institute holds the property rights for course materials. The Training Institute declares that all information provided by students is treated as confidential by the Training Institute, its employees and the lecturers.


text:
Valeriana catharinensis är en kaprifolväxtart som beskrevs av Karl Otto Robert Peter Paul Graebner. Valeriana catharinensis ingår i släktet vänderötter, och familjen kaprifolväxter. Inga underarter finns listade i Catalogue of Life.

Källor

Vänderötter
catharinensis

redpajama_set_name: RedPajamaWikipedia


text:
Pentagon Chief Austin and Israel's Gallant Discuss Defense
Two Arrested After Kidnapping, Body Found in Wyong
GPs: Medicare On Life Support
Cities come together to face future challenges
Information sought after service stations targeted at Hamilton
Samuel De Champlain Bridge Illuminated for Quebec Mosque Attack Day
AFP records spike in financial sextortion reports over school holidays
U.S.-Egypt Relationship
NZ police are responding to fatal crash at Maramarua, Waikato
UN Chief Calls to Remember and Act: Guterres
3 Dec 2022 4:17 am AEDTDate Time
Scientists reveal encouraging results in first-in-human clinical trials evaluating HIV vaccine
Scripps Research Institute
NEW YORK and LA JOLLA, CA—While scientists have struggled in the past to create an effective vaccine against HIV, a novel vaccine design strategy being pursued by researchers at Scripps Research, IAVI, Fred Hutchinson Cancer Center (Fred Hutch) and the National Institutes of Health, National Institute of Allergy and Infectious Diseases (NIAID) Vaccine Research Center (VRC) shows new promise, according to data from a first-in-human clinical trial.
In a paper published in Science on December 2, 2022, the scientists reveal critical new insights into their novel vaccine strategy, which involves a stepwise approach to producing antibodies capable of targeting a wide range of HIV variants.
"The data we are publishing in Science demonstrates for the first time that one can design a vaccine that elicits made-to-order antibodies in humans. We specified in advance certain molecular properties of the antibodies that we wanted to elicit, and the results of this trial show that our vaccine antigen consistently induced precisely those types of antibodies," says co-senior author William Schief, PhD, a professor and immunologist at Scripps Research and executive director of vaccine design at IAVI's Neutralizing Antibody Center, whose laboratory developed the vaccine antigen. "We believe this vaccine design strategy will be essential to make an HIV vaccine and may help the field create vaccines for other difficult pathogens."
The Phase 1 trial, known as IAVI G001, tested the first stage in a multi-stage HIV vaccine regimen the researchers are developing. The trial results show that the vaccine had a favorable safety profile and induced the targeted response in 97% of people who were vaccinated. Importantly, the Science study also provides a detailed immunological analysis of the vaccine responses.
"HIV represents an area of dire unmet need across the world, which is what makes the findings from our Phase 1 clinical trial so encouraging," says Mark Feinberg, MD, PhD, president and CEO of IAVI. "Through the close-knit collaboration of many different scientists, disciplines and institutions, we are that much closer to designing an effective vaccine that could help end the HIV pandemic."
Priming the Immune System
Broadly neutralizing antibodies (bnAbs) are a rare type of antibody that can fight and protect against many different variants of a virus—including HIV. This is why scientists have tried to develop an HIV vaccine that induces bnAbs, but thus far without success.
The researchers in the study are using a strategy known as 'germline targeting' to eventually produce bnAbs that can protect against HIV. The first step of germline targeting involves stimulating the rare immune cells—known as bnAb-precursor B cells—that can eventually evolve into the cells that produce the bnAbs needed to block the virus. To accomplish this first step, the researchers designed a customized molecule—known as an immunogen—that would "prime" the immune system and elicit responses from these rare bnAb-precursor cells.
The overarching goal of the IAVI G001 trial was to determine if the vaccine had an acceptable safety profile and could induce responses from these bnAb-precursor B cells.
"Through extensive safety and tolerability monitoring during the trial, we showed the vaccine had a favorable safety profile, while still inducing the necessary target cells," says study author Dagna Laufer, MD, vice president and head of clinical development at IAVI. "This represents a large step forward in developing an HIV vaccine that is both safe and effective."
To determine if the targeted bnAb-precursor B cells were induced, the researchers carried out a sophisticated analytical process.
"The workflow of multidimensional immunological analyses has taken clinical trial evaluation to the next level," says co-senior author Adrian B. McDermott, PhD, former chief of the Vaccine Immunology Program at the NIAID VRC. "In evaluating these important immunological factors, we helped show why the vaccine antigen was able to induce the targeted response in 97% of vaccine recipients."
IAVI G001 was sponsored by IAVI and took place at two sites: George Washington University (GWU) in Washington, D.C., and Fred Hutch in Seattle, enrolling 48 healthy adult volunteers. Participants received either a placebo or two doses of the vaccine antigen, eOD-GT8 60mer, along with an adjuvant developed by the pharmaceutical company GSK. Julie McElrath, MD, PhD, co-senior author, senior vice president and director of Fred Hutch's Vaccine and Infectious Disease Division, and David Diemert, MD, professor of medicine at GWU School of Medicine and Health Sciences, were lead investigators at the trial sites.
A Deeper Immunological Dive
The study also carefully examined the properties of the antibodies and B cells induced by the vaccine antigen, in what Schief likens to "looking under the car hood" to understand how the immune system operated in response to the vaccine. One analysis showed that the vaccine antigen first stimulated an average of 30 to 65 different bnAb precursors per person vaccinated, and then caused those cells to multiply. This helped explain why the vaccine induced the desired response in almost all participants.
Other analyses delved into the specific mutations the bnAb-precursor B cells acquired over time and how tightly they bound to the vaccine antigen. These investigations showed that that after each dose of the vaccine, the bnAb-precursor B cells gained affinity and continued along favorable maturation pathways.
One concern for this type of vaccine approach is the notion of "competitors"—in other words, the B cells induced by the vaccine antigen that are not bnAb precursors. The researchers extensively studied the "competitor" responses, and the results were very encouraging. Although the majority of the B cells triggered by vaccination were, in fact, "competitors", these undesired B cells could not match the binding strength of the desired bnAb precursors and did not seem to impede maturation of the bnAb-precursor responses.
"These findings were very encouraging, as they indicated that immunogen design principles we used could be applied to many different epitopes, whether for HIV or even other pathogens," adds Schief.
With these promising data in hand spanning both safety and immune responses, the researchers will continue to iterate and design boosting immunogens that could eventually induce the desired bnAbs and provide protection against the virus. These findings also come shortly after two additional studies in Immunity published in September 2022, which helped validate the germline-targeting approach for vaccinating against HIV.
"Working together with IAVI, Scripps Research, the VRC, GWU, additional investigators at Fred Hutch and many others, this trial and additional analyses will help inform design of the remaining stages of a candidate HIV vaccine regimen—while also enabling others in the field to develop vaccine strategies for additional viruses," says McElrath of Fred Hutch.
IAVI, Scripps Research, NIAID, the Bill & Melinda Gates Foundation and the U.S. President's Emergency Plan for AIDS Relief (PEPFAR) through the United States Agency for International Development (USAID) are partnering with the biotechnology company Moderna to develop and test mRNA delivery of these HIV vaccine antigens. Two Phase I clinical trials are underway that build on IAVI G001, one (IAVI G002) at four sites in the U.S. and another (IAVI G003) at the Center for Family Health Research in Kigali, Rwanda, and The Aurum Institute in Tembisa, South Africa. Both are testing mRNA delivery of the eOD-GT8 60mer that was evaluated as recombinant protein in IAVI G001, and the U.S. trial includes a boost antigen designed by the Schief lab and delivered with Moderna mRNA technology. A third trial (HVTN302), at ten sites in the U.S., is testing mRNA delivery of three different stabilized HIV trimers designed in the Schief laboratory that are candidates for late-stage boosters in multi-stage vaccines aiming to induce bnAbs. Using mRNA technology could significantly accelerate the pace of HIV vaccine development as it allows for faster production of clinical trial material.
This work was supported by the Bill & Melinda Gates Foundation Collaboration for AIDS Vaccine Discovery; the IAVI Neutralizing Antibody Center; NIAID; Scripps Center for HIV/AIDS Vaccine Immunology and Immunogen Discovery and Scripps Consortium for HIV/AIDS Vaccine Development; and the Ragon Institute of MGH, MIT, and Harvard. Other collaborating organizations include Duke Human Vaccine Institute, Karolinska Institutet, and La Jolla Institute.
Research at the IAVI Neutralizing Antibody Center that contributed to the development of the vaccine antigen eOD-GT8 60mer was also made possible by the government of the Netherlands through the Minister of Foreign Trade & Development Cooperation and through the generous support of the American people through PEPFAR through USAID. The contents are the responsibility of IAVI and Scripps Research and do not necessarily reflect the views of PEPFAR, USAID, or the United States government.
About IAVI
Tags:Africa, aids, america, american, HIV, Karolinska Institutet, Kigali, MIT, Moderna, Netherlands, Rwanda, South Africa, U.S., United States, Washington, Washington University


text:
cell death & disease
Pseudogene RACGAP1P activates RACGAP1/Rho/ERK signalling axis as a competing endogenous RNA to promote hepatocellular carcinoma early recurrence
Meng-Yao Wang1,
Dong-Ping Chen1,
Bin Qi1,
Ming-Yi Li1,
Yan-Yi Zhu1,
Wen-Jing Yin1,
Lu He1,
Yi Yu1,
Zhou-Yu Li1,
Ling Lin1,
Fang Yang1,
Zhi-Rui Lin2 &
Jin-Quan Liu1
Cell Death & Diseasevolume 10, Article number: 426 (2019) | Download Citation
Tumour biomarkers
Accumulating evidence has indicated crucial roles for pseudogenes in human cancers. However, the roles played by pseudogenes in the pathogenesis of HCC, particularly HCC early recurrence, still incompletely elucidated. Herein, we identify a novel early recurrence related pseudogene RACGAP1P which was significantly upregulated in HCC and was associated with larger tumour size, advanced clinical stage, abnormal AFP level and shorter survival time. In vitro and in vivo experiments have shown that RACGAP1P is a prerequisite for the development of malignant characteristics of HCC cells, including cell growth and migration. Mechanistic investigations indicated that RACGAP1P elicits its oncogenic activity as a ceRNA to sequestrate miR-15-5p from its endogenous target RACGAP1, thereby leading to the upregulation of RACGAP1 and the activation of RhoA/ERK signalling. These results may provide new insights into the functional crosstalk of the pseudogene/miRNA/parent-gene genetic network during HCC early relapse and may contribute to improving the clinical intervention for this subset of HCC patients.
Hepatocellular carcinoma (HCC) is one of most frequent cause of cancer-related mortality worldwide, with more than half a million deaths per annum1,2. Early HCC recurrence is predominantly responsible for dismal outcomes and is the major obstacle to improving HCC patients' survival3,4. Therefore, molecular investigation of the early recurrence of HCC is the major challenge and there is a pressing need to develop new biomarkers to screen out HCC patients with the higher risk of recurrence, who would be extremely beneficial for the clinical intervention5.
In pursuit of the prediction of early recurrence for HCC patients, previous studies have highlighted valuable biomarkers such as Rac GTPase Activating Protein 1 (RACGAP1)6, Epithelial Cell Transforming 2 (ECT2)7, Protein Regulator Of Cytokinesis 1 (PRC1)8 et al., which are all protein-coding genes. Considering long non-coding RNAs (lncRNAs) make up the major component of human transcriptome, it is essential to determine whether lncRNAs may serve as diagnostic markers for HCC early relapse. Pseudogenes, defined as dysfunctional copies of protein-coding genes, represent an intriguing class of lncRNAs and were primitively considered as "genomic junk", but their well preservation during the evolutionary pressures imposed by natural selection argues that they may possess important functions and that their dyregulation could contribute to the development of diseases9,10. Indeed, mounting evidence has suggested that pseudogenes are playing key roles in oncogenesis or tumour suppression11,12. In HCC, several lines of evidence have indicated that pseudogenes are aberrantly expressed, providing new insights into the pathogenesis of HCC13,14,15. However, few HCC early recurrence associated pseudogenes have been characterised thus far.
In the current study, we first identified a novel HCC early recurrence associated pseudogene RACGAP1P, of which the biological function and expression pattern in cancer have not been illuminated. We found that RACGAP1P was significantly upregulated in HCC tissues and markedly upregulated in the HCC samples with early recurrence. In addition, high expression of RACGAP1P in HCC was associated with poor prognosis. Mechanistically, RACGAP1P acts as an endogenous sponge for miR-15-5p to prevent its association with RACGAP1, leading to activation of the RhoA/ERK signalling axis and enhancing liver cancer cell proliferation and migration. Collectively, the results of this study show that RACGAP1P is an oncogenic regulator of HCC early recurrence, and may promote the development of effective recurrence-targeted therapy and improve the overall prognosis of HCC patients.
Clinical specimens
Tumour and normal tissues from HCC patients that were histopathologically and clinically diagnosed, and did not receive neo-adjuvant treatment were obtained from the Affiliated Cancer Hospital and Institute of Guangzhou Medical University in Guangzhou. Written informed consent was obtained from each patient, and the study was approved by the Institute Research Ethics Committee at the Cancer Center.
Cell culture, proliferation assays and colony-formation assays
The human cancer cell lines were maintained in Dulbecco's modified Eagle's medium supplemented with 10% FBS at 37 °C and 5% CO2.
For the proliferation assays, 1000 cells/200 μl of medium were plated in a 96-well plate (Corning) in six copies and stained with MTS (Sigma, M2128) for 3.5 h determined at OD570 with a microplate reader every day for 6 days.
For the colony-formation assays, 500 cells/2 ml were seeded into a 6-well plate (Corning) in triplicates. The cells were washed with phosphate-buffered saline (PBS), fixed with methanol for 10 min at room temperature, and stained with 1% crystal violet for 20 min. The colonies were counted after 10 days.
Migration assay
For migration assays, 3 × 104 HCC cells in 200 μl of serum-free DMEM were seeded into 24-well plates upper chambers with an 8-mm pore size filter (Falcon), the lower chamber was filled with 600 μl of 10% FBS–DMEM. After 24 h of incubation, the cells on the lower surface of the filter were fixed and stained with 1% crystal violet. The number of migrated cells in five random optical fields (×100 magnification) from triplicate filters was averaged.
Animal experiments
All protocol was approved by the Institutional Animal Care and Use Committee of the Guangzhou medical University. For the tumourigenicity studies, Hep3B cells stably transfected with vector or RACGAP1P-3'UTR (1 × 106 cells/tumour in 150 mL DMEM with 25% matrigel) were subcutaneously injected into either side of flanks of the 5 female BALB/c athymic nude mice (3–4 weeks of age). Tumour diameters were measured in indicated day after tumours were established. Finally, the mice were euthanized, and the tumours were isolated and weighed, the tumour volume was calculated as volume = length × width2 × 0.52.
RNA extraction and real-time PCR
For real-time PCR, total RNA was extracted from cultured cell lines using Trizol reagent (Invitrogen) and subjected to reverse transcription using a cDNA Synthesis Kit (Thermo, K1622). Real-time PCR was performed using a SYBR FAST Universal qPCR Kit (KAPA, KK4602). The relative expression of the target mRNAs were calculated as two power values of ΔCt (the Ct of β-actin minus the Ct of the target cDNA). The sequences of the PCR primers used for amplification were as follows:
β-actin forward, 5′-AAGGTCATCC CTGAGCT GAA-3′;
β-actin reverse, 5′-TGACAAAGTG GTCGTTG AGG-3′;
RACGAP1 forward, 5′-GAAAGCAGAGACTGAGCGAAG-3′;
RACGAP1 reverse, 5′-GTTGAATGCTGCCAGATGTGT-3′.
RACGAP1P forward, 5′-GCACCTGTACTCTCTGCTCTAC-3′;
RACGAP1P reverse, 5′-TCCCAACAGTGACCAGAACA-3′.
DICER1 forward, 5′-TGGATAGTGGGATGTCACTGG-3′;
DICER1 reverse, 5′-CTCTGACCTTCCCGTCGTAAG-3′.
Small interfering RNA transfection
The negative control small interfering RNA (NC) and siRNA targeting human DICER1 are 5′-CAGCACCTTTACCCTTAGT-3′, were purchased from GenePharma, transient transfections of HCC cells were performed by using the Lipofectamine RNA iMAX Reagent (Invitrogen) protocol with 30 pmol siRNA in Opti-MEM Medium (Invitrogen) were mixed, and incubated at room temperature for 5 min, then the mixture was added to the cells.
MS2-RIP
The MS2bp-MS2bs-based RIP assay was performed according to previous reports16, with modifications for using the EZ-Magna RIP Kit (Millipore) in accordance with the manufacturer's instructions.
Public expression profiles
Human pseudogene list is obtained from NCBI gene. Gene expression profiling datasets between HCC tumour tissues and normal tissues, including GSE84005, GSE76297, GSE6404, GSE54236, and GSE5975 were downloaded from GEO and TCGA. The expression data of HCC cancer cell lines was downloaded from CCLE.
Dual luciferase reporter assay
A dual luciferase reporter assay was performed as previously described with slight modifications. Briefly, the treated HCC cells were lysed, and the activities of the firefly and Renilla luciferases were analysed using a dual luciferase assay kit (Promega, Madison, WI, USA) according to the manufacturer's instructions. All reporter gene assays were performed in triplicate and were repeated twice. The results are expressed as the means ± SEM.
Luciferase reporter gene constructs containing the RACGAP1P and RACGAP1 3' UTR region were first amplified by PCR with specific primers and then, the PCR products were cloned into pGL3-basic (Promega) digested by XbaI(NEB) using ClonExpress II One Step Cloning Kit (C112-02, Vazyme Biotech).
Full length or 3'UTR of RACGAP1P were amplified by PCR from DNA of Hep3B cells and were cloned into lentivirus vector pLVX-puro according to standard protocols. RACGAP1P and RACGAP1 3'UTR was also subcloned into the pMS2 vector for RIP analysis.
Immunoblotting
Immunoblotting was performed as described previously. The sources of the primary antibodies were as follows: anti-RACGAP1 (Proteintech, 1:1,000), anti-DICER1 (Proteintech, 1:1,000), and anti-β-actin (Proteintech, 1:1,000), p-ERK1/2 (CST, 1:1,000), total ERK (CST, 1:1,000), Active Rho Detection kit (#8820), anti-mouse and anti-rabbit peroxidase conjugated secondary antibodies were purchased from Cell Signalling Technology (Danvers, MA).
The two-tailed Student's t-test was used to compare two independent groups of data. One-way analysis of variance (ANOVA) was used to analyze the significance among multi-groups. Chi-squared tests were applied to analyze the relationship between RACGAP1P expression and clinicopathological status. Pearson correlation analysis, Long-rank test were performed as indicated. P value < 0.05 was considered statistically significant in all cases. Asterisk (*) means p < 0.05, asterisks (**) means p < 0.01, asterisks (***) means p < 0.001. All statistical analyses were performed using the GraphPad Prism Software 5.0 or SPSS 16.0.
Pseudogene RACGAP1P is consistently activated in early recurrent HCC and serves as a predictor of dismal outcome
To evaluate the role of pseudogenes in HCC pathogenesis, we screened potential cancer-related pseudogenes, which were differently expressed between HCC samples and normal samples in 4 microarray datasets (GSE84005, GSE7629717, GSE6404118, GSE5423619) downloaded from Gene Expression Omnibus (GEO) and identified 7 candidates, of which 4 genes were upregulated and 3 genes were downregulated (Fig. 1a). We put our attention on RACGAP1P since its real gene, RACGAP1, was well-known to act as a oncogenic driver in HCC and importantly, played a key role in HCC early recurrence6. Similarly, assessment via data mining of RNA-seq data from The Cancer Genome Atlas (TCGA) database confirmed that RACGAP1P expression is significantly elevated in tumour tissues compared with non-tumour tissues. Of note, RACGAP1P expression was obviously upregulated in tumour from patients with early (<2 years) recurrent disease (HCC-R) compared with samples from patients with no obvious sign of recurrence within 4 years (HCC-NR) (Fig. 1b). The analogous pattern of RACGAP1P expression was further validated by QPCR (Fig. 1c). Moreover, in TCGA cohort, correlation analysis between RACGAP1P expression and clinicopathological characteristics shown that high level of RACGAP1P in primary tumours was significantly associated with the advanced T stage, clinical stage and abnormal AFP level (Table 1). Importantly, it was demonstrated that patients with high RACGAP1P expression had a significantly shorter overall (OS) and disease-free (DFS) duration compared with patients with low RACGAP1P expression (Fig. 1d, e). Taken together, these data sustain RACGAP1P as a novel prognostic factor in patients with HCC.
Fig. 1: Pseudogene RACGAP1P is elevated in hepatocellular carcinoma (HCC) and is related to early HCC recurrence.
a Venn diagram analysis of altered pseudogenes profiling in GSE84005, GSE76297, GSE6404, and GSE54236 datasets. b The RACGAP1P mRNA expression were analysed in normal liver tissues (n = 47), non-recurrent tumours (NR-HCC, n = 22) and early recurrent tumours (R-HCC, n = 141) from TCGA. The highest RACGAP1P mRNA expression were shown in human hepatocellular carcinoma early recurrent tissues. c The RACGAP1P mRNA expression were tested in human hepatocellular carcinoma tissues by QPCR, including normal liver tissues (n = 25), non-recurrent tumours (NR-HCC, n = 25), early recurrent tumours (R-HCC, n = 41). d, e Kaplan–Meier curve shows that the overall survival (OS) and disease-free survival (DFS) rate were significantly lower in the high RACGAP1P group in TCGA cohort. ***P < 0.001, **P < 0.01, *P < 0.05. B&C, the p values were calculated by one-way ANOVA. D&E, the p values were calculated by long-rank test
Table 1 Association of RACGAP1P expression with clinicopathological characteristics of HCC patients in the TCGA cohort
Pseudogene RACGAP1P modulates RACGAP1 level in a ceRNA manner
Recently, the competitive endogenous RNAs (ceRNA) hypothesis has been raised as a novel layer of gene expression modulation. The hypothesis proposed that RNA molecules share miRNA response elements (MREs) and sequester common microRNAs (miRNAs), thereby regulating expression between each other20,21,22. It is not difficult to speculate that pseudogenes may serve as ceRNAs for their parent genes, considering the high sequence similarity. To test the hypothesis that pseudogene RACGAP1P may regulate the expression of its parent gene as a ceRNA, we firstly assessed the correlation of RACGAP1P and RACGAP1 via mRNA expression data derived from the TCGA, GEO (GSE5975, GSE76297) and Cancer Cell Line Encyclopaedia (CCLE) and found the co-expression patterns between RACGAP1P and RACGAP1 in primary HCC tissues, as well as HCC cell lines (Fig. 2a, b). Then, we perform QPCR assay to validate the significantly positive correlation in HCC samples (R2 = 0.34, P = 0.0001; Fig. 2c). Next, to experimental examine whether RACGAP1P regulate the expression of RACGAP1, and whether the effect of the RACGAP1P on RACGAP1 expression was in a miRNA-dependent manner, we suppressed DICER1, a ribonuclease critical for miRNA biogenesis, in Hep3B and HCCLM3 cells using a specific siRNA (Fig. 2d, Supplementary Fig. 1a) and found that ectopic expression of RACGAP1P in wild type cells elevated the mRNA level of RACGAP1 but not in DICER1-KD cell lines (Fig. 2e). Similarly, RACGAP1 3'UTR luciferase reporter activity was significantly increased in DICER1-WT cells with RACGAP1P overexpression whereas these effects were abrogated in DICER1-KD cells (Fig. 2f). Thus, the RACGAP1 pseudogene-induced effects on the RACGAP1 level are 3'UTR and miRNA dependent in human HCC.
Fig. 2: RACGAP1P is co-expressed with RACGAP1 in HCC and regulates RACGAP1 expression dependent on mature miRNA.
a, b Graphs showing a positive correlation between RACGAP1P and RACGAP1 in HCC using public gene expression datasets, including TCGA (n = 423, R2 = 0.43, p < 0.0001), GSE5975 (n = 238, R2 = 26, p < 0.0001), GSE76279 (n = 153, R2 = 0.29, p < 0.0001) and CCLE (n = 25, R2 = 0.19, p = 0.029). A, HCC clinical specimens, B, HCC cell lines. c The co-expression between RACGAP1P and RACGAP1 mRNA level was measured in 38 primary HCC tissues via QPCR (n = 38, R2 = 0.34, p = 0.0001). d The efficiency of RACGAP1P overexpression and DICER1 knockdown in HCC cell lines were determined by QPCR. e In Hep3B and HCCLM3 cells, RACGAP1P upregulated RACGAP1 mRNA levels in a DICER1 dependent manner. f RACGAP1P increased RACGAP1 luciferase reporter activity in a DICER1 dependent manner. Error bars represent means ± SD. ***P < 0.001, **P < 0.01, *P < 0.05, the p values were calculated by the Student's t-test
Identification of the common miRNAs sharing between RACGAP1P and RACGAP1
The finding that the pseudogene RACGAP1 mediates its effect through 3'UTR and mature miRNAs suggests that it may function as a ceRNA. To further elucidate which of the miRNAs targeted both RACGAP1P and RACGAP1 to mediate the observed crosstalk, we overlaid the miRNAs predicted to target RACGAP1 and RACGAP1P in RegRNA2.023 and previously reported HCC tumour suppressive miRNAs24 and finally obtained 8 candidate miRNAs including miR-15a-5p, miR-23b-3p, miR-138-5p, miR-29c-3p, miR-125b-5p, miR-150-5p, miR-193b-3p, miR-320a (Fig. 3a). Then, Hep3B and HCCLM3 cells were delivered with these 8 potential miRNA mimics, respectively, and the expression of RACGAP1 was detected by western blot analysis. As shown in Fig. 3b, only miR-15a-5p was validated to significantly repress protein abundance of RACGAP1 in both cell lines. Therefore, miR-15a-5p was selected for further analysis and QPCR results revealed that miR-15a-5p overexpression strongly inhibited the mRNA level of RACGAP1 and RACGAP1P (Fig. 3c). Conversely, inhibition of miR-15a-5p using miRNA antagomirs could upregulated mRNA levels of both RACGAP1 and RACGAP1P (Fig. 3d).
Fig. 3: miR-15-5p targets both RACGAP1P and RACGAP1.
a Eight candidate miRNAs were identified to be associated with RACGAP1P and RACGAP1 in HCC from Venn diagram analysis of three datasets, including tumour suppressor miRNAs dataset in HCC, the miRNAs datasets predicted to bind RACGAP1P and RACGAP1. b The 8 candidate miRNAs differentially regulated the RACGAP1 protein expression in Hep3B and HCCLM3 cells shown by WB. c QPCR analysis of RACGAP1P and RACGAP1 with miR-15a-5p mimics. d QPCR analysis of RACGAP1P and RACGAP1 with miR-15a-5p antagomics. e miR-15-5p seed sequence is shown along with the binding sites in RACGAP1P and RACGAP1 (shown by red box), and the mutated miR-15-5p binding sites (shown by star). f luciferase reporter assays verified the miR-15a-5p binding sites in 3'UTR of RACGAP1P and RACGAP1. g MS2-RIP strategy used to confirm endogenous miR-15a-5p binding to RACGAP1P and RACGAP1. Error bars represent means ± SD. ***P < 0.001, **P < 0.01, Student's t-test
To further investigate whether miR-15a-5p could target both RACGAP1 and RACGAP1P, we performed the prediction of miR-15a-5p MREs for RACGAP1 and RACGAP1P transcripts and found two miR-15a-5p binding sites in RACGAP1 3'UTR and one in RACGAP1P 3'UTR (Fig. 3e). Then, we artificially introduced mutation of miR-15a-5p seed regions in luciferase reporters to disrupt the miRNA binding. The results of luciferase reporter assay demonstrated that overexpression of miR-15a-5p significantly decreased luciferase reporter activity of RACGAP1P-3'UTR and RACGAP1-3'UTR whereas the inhibition effect of miR-15a-5p was rescued on luciferase reporter with miR-15a-5p binding sites mutations (Fig. 3f). To validate the direct binding ability of the predicted miRNA response elements on these transcripts, we performed an MS2-RIP analysis using the 3'UTR regions of RACGAP1P and RACGAP1 as baits in Hep3B and HCCLM3 cells. Notably, miR-15a-5p was highly associated with the 3'UTRs of RACGAP1P and RACGAP1, and miR-21-5p as a control miRNA with no predicted MREs on RACGAP1P and RACGAP1 bound neither 3'UTR. However, the binding of miR-15a-5p on RACGAP1P and RACGAP1 3'UTR was negated in MREs mutation group (Fig. 3g). All together, these findings indicated that the ceRNA crosstalk between RACGAP1P and RACGAP1 may be mediated at least through miR-15a-5p in part.
RACGAP1P is an oncogenic driver in HCC dependent on miRNA
Considering increased RACGAP1 expression drives the tumourigenesis and progression of numerous types of cancer25,26,27,28, especially HCC, we determined whether aberrant control of RACGAP1 by RACGAP1P overexpression enhanced oncogenic transformation of HCC cells. Indeed, Stably exogenous expression of RACGAP1P 3'UTR in Hep3B and HCCLM3 resulted in a elevation of RACGAP1 protein, GTP-binding activity of RhoA, and ERK phosphorylation, indicating the activation of the RhoA/ERK pathway (Fig. 4a). These molecular observations were accompanied by growth promotion and a significant increase in the number of colonies, as well as high migration ability (Fig. 4b–d). Furthermore, to determine whether RACGAP1P affects tumour growth in vivo, we utilised a subcutaneous xenograft mouse model which were inoculated with Hep3B cells stably transfected with control vector or RACGAP1P 3'UTR. All of the mice developed tumours at the injection sites. However, the average size and weight of tumours generated by RACGAP1P 3'UTR-expressing cells were significantly larger than those generated from control cells (Fig. 4e–g). QPCR analysis confirmed that the average expression of RACGAP1P and RACGAP1 was much higher in RACGAP1P 3'UTR overexpressing tumour tissues compared with control tumours (Fig. 4h). Finally, immunoblotting showed that expression of RACGAP1 and phosphorylated ERK was higher in RACGAP1P 3'UTR-expressing tumours than control tumours (Fig. 4i). Importantly, the cancer cell proliferation and migration promotion effects were abrogated in DICER1-knock out HCC cells (Fig. 5a–d, Supplementary Fig. 1b), supporting the notion that the 3'UTR of RACGAP1P requires mature miRNAs for its function towards RACGAP1. Taken together, our results demonstrate that RACGAP1P, operating as a ceRNA, activates RACGAP1/RhoA/ERK pathway to favour malignant phenotypes of HCC (Fig. 5e).
Fig. 4: RACGAP1P regulates the growth and migration of HCC cells via RhoA/ERK signalling pathway.
a RACGAP1 expression, RhoA and ERK activation were measured by WB after RACGAP1P-3'UTR overexpression in Hep3B and HCCLM3 cells. b MTT assays shows that RACGAP1P-3'UTR overexpression promoted HCC cells growth. c Colony formation assays shows that RACGAP1P-3'UTR overexpression enhanced the proliferation of HCC cancer cells. Left: Representative images of colony formation. Right: Statistical results of colony number. d Transwell assays indicates that RACGAP1P-3'UTR overexpression augmented the migration ability of HCC cells. Left: Representative images of migrated cells. Right: statistical results of migrated cells. e–f Subcutaneous tumour growth in mice inoculated with RACGAP1P-3'UTR overexpression Hep3B cells or or control cells. e Photos of xenograft tumours, (f) growth curve of tumours, and (g) statistical analysis of tumour weight growth over 6 weeks. h QPCR reslults show that higher mRNA level of RACGAP1P and RACGAP1 in RACGAP1P-3'UTR overexpression xenograft tumours. i Western bloting indicates that the protein level of RACGAP1 and p-ERK were higher in RACGAP1P-3'UTR overexpression xenograft tumours. Error bars represent means ± SD. *P < 0.05, **P < 0.01, ***P < 0.001, the Student's t-test
Fig. 5: The oncogenctic activity of RACGAP1P depends on miRNA.
a RACGAP1 expression, RhoA and ERK activation were detected by immunoblotting after RACGAP1P-3'UTR overexpression treatment in HCC cells with DICER1 knockout. b–d The cell growth and migration was inhibited in RACGAP1P overexpression cells with DICER1 depletion. b MTT assays. c Colony formation assays. d Transwell assays. e Diagram depicting the underlying mechanism of RACGAP1P contributing to HCC recurrence. RACGAP1P activates RhoA/ERK signalling axis by modulating RACGAP1P/miR-15a/RACGAP1 ceRNA network
RACGAP1P crosstalks with RACGAP1 and associates with poor prognosis in various human cancers
To figure out whether the ceRNA regulation between RACGAP1P and RACGAP1 is HCC specific or common in pan-cancers, the analysis of data from TCGA was performed and found the highly positive correlation between RACGAP1P and RACGAP1 in various types of cancer (Supplementary Fig. 2). In addition, exogenous overexpression of RACGAP1P 3'UTR in different human cancer cell lines could induce RACGAP1 expression showed by QPCR results (Fig. 6a). Moreover, the mRNA level of RACGAP1P was significantly upregulated in a great many human cancers compared with the normal tissues (Fig. 6b). Further, a high level of RACGAP1P indicated the shorter overall survival in pan-cancers, such as BRCA, LUAD, LGG, LAML, HNSC, PAAD (Fig. 6c). Thus, high RACGAP1P level in human cancers significantly correlated with poor outcome.
Fig. 6: RACGAP1P is elevated in multiple human cancers and correlates with poor outcome.
a QPCR analysis indicates that overexpression of RACGAP1P 3'UTR induced the expression of RACGAP1 in various human cancer cell lines. b The mRNA Level of RACGAP1P is significantly upregulated in 8 types of human cancer. c High expression of RACGAP1P was related to worse overall survival (OS) in 6 types of cancer. BLCA bladder urothelial carcinoma, BRCA breast invasive carcinoma, CESC cervical squamous cell carcinoma and endocervical adenocarcinoma, COAD colon adenocarcinoma, HNSC head and neck squamous cell carcinoma, KIRC kidney renal clear cell carcinoma, LUAD lung adenocarcinoma, LUSC lung squamous cell carcinoma, PAAD pancreatic adenocarcinoma, PRAD prostate adenocarcinoma, LGG low grade glima, AML acute myeloid leukaemia. Error bars represent means ± SD. *P < 0.05, **P < 0.01, ***P < 0.001, the p values were calculated by Student's t-test or long-rank test
Early recurrence is the major obstacle in achieving long-term survival outcomes for the treatment of HCC via operative treatment. Our knowledge of the underlying cellular and molecular pathways driving HCC early relapse is rather finite. Previous studies have addressed the clinical implication of gene expression profiling in predicting the early recurrence of human HCC29. However, there is pretty limited information about ncRNA, especially pseudogenes involved in HCC early relapse. In the present study, we identified a novel liver cancer early recurrence associated lncRNA RACGAP1P, which is significantly upregulated in liver cancer tissues and mainly expressed much higher in recurrent HCC. Higher expression of RACGAP1P was correlated with late T stage, advanced clinical stage and abnormal AFP level. Moreover, increased RACGAP1P expression was associated with shorter OS and PFS of HCC patients. In vitro and in vivo assays demonstrated that ectogenic expression of RACGAP1P promoted cell proliferation and migration. These findings indicate that RACGAP1P has an oncogenic role in liver tumourigenesis and could be considered as a potential prognostic indicator for HCC.
RACGAP1, a member of GTPase activation protein family30, is the first gene reported to work as an independent informative biomarker for HCC early recurrent, as well as its silencing mainly affected genes in an interactome clinically relevant to early relapse. Given the significant effects of RACGAP1 on cancer recurrence, the regulatory network related to RACGAP1 urgently needed to be explored. Genetic or epigenetic alterations may directly lead to the aberrant expression of cancer-related genes. Notably, data analysing of the TCGA data found that the RACGAP1 locus undergoes copy number gains in 13.2% liver cancers and the methylation level in the promoter region is lower in cancer tissues than normal tissues and negatively associated with RACGAP1 mRNA level. However, the copy number and the methylation level were no obvious difference between recurrent and non-recurrent HCC (Supplementary Fig. 3). Therefore, the regulatory mechanisms of increasing expression of RACGAP1 in early recurrent HCC still remained to be elucidated. Herein, it was interesting to observe that the expression of pseudogene RACGAP1P significantly correlated with its cognate gene RACGAP1 expression and overexpression of RACGAP1P could increase the expression of RACGAP1 dependent on miRNA. Moreover, RACGAP1P is consistently upregulated in early recurrent HCC samples. All above suggest RACGAP1P may involve in the elevation of RACGAP1 during HCC early recurrence.
In recent years, ceRNA hypothesis provides a new crosstalk dimension between RNA moleculars that serve as miRNA sponges to regulate the level of the genes which harbour the common miRNA binding sites. Given the high sequence similarity between pseudogenes and their parent genes, pseudogenes are "perfect decoys" for their ancestral genes, critically impacting their parent protein-coding genes via the ceRNA network31,32. Based on the central of ceRNA hypothesis that miRNAs play as communication language, we identified miR-15a-5p, which is a well-known tumour suppressor in the pathogenesis of HCC, could target both RACGAP1P and RACGAP1. And the direct interaction between miR-15a-5p and these paired genes was confirmed by dual luciferase reporter assay and MS2-RIP assay, indicating that RACGAP1P regulates RACGAP1 expression at least partially through competitive binding with miR-15a-5p as a ceRNA. Similarly, the studies by the groups of Shen et al., Sharp et al. and Stoffel et al. focused on ceRNA regulation that is mediated by a single miRNA. Importantly, ceRNA pairs in general, and gene/pseudogene pairs in particular, share numerous miRNAs and it is reported that ceRNA crosstalk is enhanced when it is mediated by more miRNAs33,34. Therefore, more shared miRNAs that being present at crosstalk-favoring levels needed to be identified.
Pseudogenes could exert critical functions in biological processes and their dyregulation may conduce to the occurrence and progression of diseases12,35. Herein, our study establishes the RACGAP1P as a potent proto-oncogene that can activate RACGAP1/Rho/ERK axis to elicit malignant phenotypes, such as cell growth and migration. Mechanistically, RACGAP1P works as a ceRNA to compete with miR-15a-5p for binding to the 3'UTR region to relieve the inhibitory effects of miR-15a-5p on RACGAP1 expression, whereas MRE mutations impair the regulation of RACGAP1 by RCGAP1P. These findings provide strong evidence in support of a key role for RACGAP1P in the tumourigenesis of liver cancer. It is possible that RACGAP1P elicits its effects through more than one mechanism or pathway, so whether the oncogenic activity of RACGAP1P also requires additional ceRNA targets or non-ceRNA-related mechanisms will be the focus of future studies.
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This work was supported by grants from National Natural Science Foundation of China (No. 81702690), from Guangzhou Key Medical Discipline Construction Project fund, from Research fund for the Doctoral Program of Guangzhou Medical University (No. 2016C38). We apologise to all colleagues whose work could not be cited due to space constraints. We thank Prof. Yun-Fei Yuan for kindly providing HCC cancer cell lines.
Department of Radiation Oncology, Affiliated Cancer Hospital and Institute of Guangzhou Medical University, 510245, Guangzhou, China
Meng-Yao Wang
, Dong-Ping Chen
, Bin Qi
, Ming-Yi Li
, Yan-Yi Zhu
, Wen-Jing Yin
, Lu He
, Yi Yu
, Zhou-Yu Li
, Ling Lin
, Fang Yang
& Jin-Quan Liu
State Key Laboratory of Oncology in South China, Collaborative Innovation Center for Cancer Medicine, Sun Yat-sen University Cancer Center, 510245, Guangzhou, China
Zhi-Rui Lin
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Correspondence to Zhi-Rui Lin or Jin-Quan Liu.
Edited by G. Ciliberto
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'Fosters' Barreto makes directing debut at Hollyshorts
By Reel 360 Aug 8, 2019
Alexandra Barreto, who played Ana Gutierrez on Freeform's The Fosters, can now add director to her IMDB.
Her directorial debut, a comedic short film, Lady Hater, will screen at the 15th annual Oscar-Qualifyin HollyShorts Film Festival.
The short film premiered at 2019 Tribeca Film Festival and was called out by Forbes as "Five Not to be Missed Shorts from the Tribeca Film Festival."
Lady Hater, about a woman who thinks she's going to a yoga class, but instead gets stuck in an all-female goddess seminar, showcases a diverse group of women coming together to explore the idea of what it means to be a woman, and the ridiculous expectations that have been put upon them by society, but mostly by themselves. Watch the trailer below:
Lady Hater – Teaser from Alexandra Barreto on Vimeo.
ALSO READ: Colin Costello's 'Dreamwisher' among films at Hollyshorts
Alex wrote the feature version of the film partly in response to the 2016 Women's March when she started exploring the idea of what it really meant to call herself a "guy's girl." She had to admit that by using that term, she was distancing herself from women out of her own insecurities. And that's what she set out to explore in the film, using comedy as a unifying force.
This is not Alex's first time behind the camera. She produced the critically acclaimed feature film, Too Late, starring Academy Award Nominees, John Hawkes and Robert Forster, as well as six short films.
She is currently seeking funding for the feature film version of Lady Hater, which she has written and plans to direct.
Audiences can catch the film on Friday, August 16th at 10pm. Schedule times and tickets can be found here.
Source:M4PR


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Barnsjukhuset Samariten är ett tidigare barnsjukhus i Stockholm som existerade mellan 1890 och 1974. Sjukhuset låg vid nuvarande Ringvägen 21 på Södermalm.

Barnsjukhuset öppnades 1890 då under namnet Nya barnsjukhuset. Behovet av sjukvård för barn var under slutet av 1800-talet stort på grund av den snabba inflyttningen till huvudstaden; de små och ofta dåliga lägenheterna och den snabba spridningen av vissa epidemiska barnsjukdomar. 1895 ändrades namnet till Samariten, och 1909 uppfördes en ny huvudbyggnad. Detta var möjligt tack vare en större summa pengar som sångerskan Jenny Lind testamenterat till sjukhuset. Barnsjukhuset byggdes sedan ut i omgångar, och kom att spela en viktig roll i Stockholms barnsjukvård.

År 1972 beslutades att sjukhuset skulle läggas ned, eftersom man ville integrera barnsjukvården i den allmänna medicinska verksamheten. 1974 stängdes Barnsjukhuset Samariten efter att ha existerat i 84 år. Merparten av verksamheten övertogs av Huddinge sjukhus. Alla sjukhusbyggnaderna revs sedan, med undantag från administrationsbyggnaden. I administrationsbyggnaden låg den kommunala förskolan Samariten fram till 2015, då den kommunala verksamheten flyttade till andra lokaler.

Sedan 2016 bedriver Jensen education  förskoleverksamhet i samma lokaler. Namnet har dock ändrats till Jensen Förskola Zinkensdamm.

Referenser

Litteratur
 Svartz-Malmberg, G., Westlin-Argenziano,G-B., Öberg, L. Barnsjukhuset Samariten - självstyrande sjukvårdsenhet i Stockholm 1890-1974. Läkartidningen 1992; 89 : 2690 - /94?/.

Sjukhus i Stockholms län
Södermalm


Hilton Madrid, Spain @ Barajas International : Fascinatingly Modern, Elegant Hospitality!
Hilton Madrid Airport will have one of the group's most representative products, the "Relaxation Room". A new concept of premium room, which reflects high-end travellers' change in aesthetic preferences and work habits, the "Relaxation Room" is divided into three distinct areas to help guests differentiate their workspace from the relaxation area and the bathroom.
Created by designers and ergonomics with luxurious furnishings and the latest technological innovations, their focal point is maximum comfort and relaxation and contemporary design. The bathroom, for example, has a hydro spa bath and a powerful body shower providing the benefits of a jacuzzi or massage.
Hilton Madrid Airport will also have Executive Floor rooms with private lounge and an indoor/outdoor swimming pool.
Complimentary Shuttle Bus Services to // from Barajas and Hotel plus services into the City of Madrid.
Relaxation room // Complimentary upgrade // incl. ultra modern design, jacuzzi in bathroom, shower, large sizes of Peter Thomas Roth amenities and gorgeous luxury bedding, Sony Bravia LCD TV plus ample and elegant space incl. wooden floors.
A great second stay for us, a fantastic hotel, great services and always very friendly staff encountered plus pure spanish design and presentation, this is the fine art of modern hospitality. Gorgeous lines from the atrium to the water features surrounding the Hilton hotel @ Madrid Barajas.
Gentle Moments Of Great Hospitality @ The Hilton Madrid // Barajas International Airport // Madrid, Spain : Enjoy!


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FS Arizona Plus
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Suns' style outpaces Indiana
PHOENIX — A crucial philosophical saw commandeered from the sweet science goes very much as such:
"Styles make fights."
More specifically, contrasting styles are known to make confrontations a lot more compelling.
It certainly helps if — after said styles are applied — the actual fight turns out competitive.
That didn't exactly happen late Wednesday night at U.S. Airways Center during a showdown co-starring the Suns and the mighty Indiana Pacers. This matchup of two teams with disparate approaches was a 124-100 knockout for the home team.
"Well, yeah … I'd like it if they'd play like that all the time," Suns coach Jeff Hornacek said when asked if he wouldn't mind more performances like this from his confounding young players.
In the most basic interpretation of this difference in methodology, it should be known the Suns like to play fast, and the Pacers — whose successful implementation of their style has been accompanied by the NBA's best record — prefer the basketball equivalent of a waltz.
"I don't know if you can get them in that style," Hornacek, speaking before the game, said in regard to baiting the Pacers into more of a fox trot. "They're very good at slowing things down.
Well, usually. Unable to control the Suns' fancy footwork this time, they were blistered at deep-tissue levels they hadn't felt all season.
In the league's tempo rankings, Indiana (33-8) checks in at a yawning 22nd. The Suns (now 24-17), with Hornacek attempting to restore the franchise's speed-kills reputation, are a frisky ninth.
With 7-foot-2 Roy Hibbert and tough-guy David West lurking near the rim, the Pacers are a conservative 24th in 3-pointers attempted. The Suns, with Hornacek providing his blessing on more green lights than a funeral procession, are a plucky fifth.
Oh, and it figures the Suns lead the league in fast-break points, while the Pacers are 22nd in scoring via transition.
It should be noted that both teams are really good at defending the 3-point line; the Pacers lead the league in that department, while the Suns are third.
But all bets, statistic and analytic, were off in this one.
The Suns shot 68 percent (11 of 16) from 3-point range against a team giving up a stingy 32. Their overall shooting percentage was a gaudy 54.2.
"Their shot-making was off the charts," Pacers Coach Frank Vogel said.
Gerald Green rises up for a slam on Indiana's Roy Hibbert.
Hornacek said it helps when the shot-taking is selective.
"It wasn't like we were jacking them up," he said. "We took good shots. Against a good team, you have to take good shots. You can't take crazy ones."
The Suns' rout from behind the arc included 3 of 5 from Gerald Green, who along with Miles Plumlee was acquired by Phoenix in an offseason trade with Indy that also brought a first-round pick to the desert.
Green, who admittedly did little to justify the first year of a three-year deal he signed with the Pacers before last season, finished with a team-high 23 points. Plumlee, whose offensive production included a pair of alley-oop passes from Green, had 11 and seven rebounds.
When asked about the motivation of playing against a former team, Green — no slouch when it comes to changing teams — said, "This is my seventh team already, I've been down this road before."
Point guard Goran Dragic, the Pacers' most severe matchup deficit against Phoenix, made 8 of 10 shots and scored 21 points in only 22:18. For the second consecutive game, the performance of even-quicker Suns guard Ish Smith enabled Dragic to sit the entire fourth quarter.
While the Pacers were failing to slow things down, they were hit with highs for points allowed any half (62) this season and for an entire game (the previous high was 118).
Indiana, which leads the league in defensive efficiency and points per game allowed, had permitted only one team, the Philadelphia 76ers, to score 30 or more points in two different quarters. The Suns did it three times Wednesday and with the bench cleared knocked in 28 in the fourth.
"That's a good team over there," Pacers guard George Hill said of the Suns. "No matter where they are in the West, it's a team you've got to respect."
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Lessons from the Woman Who Built Squarespace's Customer Care Team from 1 to 184
If building a company were like planning a wedding, customer service is frequently the second cousin who's invited when slots open up on the attendee list. Many believe that it's not as critical to success as sales and product development. The reality is that customer care is your most direct line to customers and has the power to define your brand. Users equate its voice with your company's voice. At peak times, it represents your organization up to 60 times an hour — literally every minute. Christa Collins, ShopKeep's VP of Customer Care understands this value; if her experience stands for anything, it's that customer care is a vital early priority.
Collins was the first customer service employee at website builder Squarespace. Over her tenure, she built the customer care team from one employee (her) to a division of 184 people operating across three time zones. She designed a workflow to triage 25,000 customer support requests per week — and that was only via its email channel. Now Collins is applying the lessons she learned to build the customer care division at ShopKeep, a cloud-based iPad point-of-sale system.
People who work in customer care need to be adept ambassadors; they must speak on behalf of their company to the customer and flip the model to represent the customer back to the company. The best in customer care are functional chameleons, becoming conversant as product managers, marketers and salespeople to bring resolution to customers. In this exclusive interview, Collins shares her sage advice on assembling a great customer care team, best practices for keeping customers happy and measuring performance in this essential but (too) often overlooked area.
Why Customer Care Matters Early
For Collins, the trigger for establishing customer care is simple. The moment you've got a product that people are going to use, you'll have people who will want talk to you about it. "So much startup advice is growth stage specific, but launching customer care in parallel with a product is universal," says Collins. "It doesn't matter if you're a really small startup. Someone must own the relationship and the information that the customer will relay to you."
This notion holds true for even the most intuitive, human-centric products. This year, Apple retail chief Angela Ahrendts is reportedly revamping Apple's Genius Bars to include walk-ins, SMS progress updates and other updates to its customer service, marking one of the more significant overhauls to Apple's retail operations in years. "The lifespan of customer care should be that of the company's first product — it begins when it does and doesn't stop," says Collins.
Establishing customer care early is a very public gesture. It broadcasts: we plan to invest as much when the product is in your hands, as we do when it's in ours.
Throughout her career, Collins has observed that most initial customer care hires land between employee #5 and #10. It might seem premature, but if you're pre-product, you should start identifying who'll be the first member of your customer care team, with a goal to have the hire in place when your product launches.
"This does several things," says Collins. "First, it creates a core team. You've got engineers, product and the bridge to the customer: customer service. It also creates a customer-centric culture from the beginning. Many companies launch a product and then ask how to be more customer service-oriented. Do it from the start, and you won't deal with that question down the line."
How to Catch-up If You Begin Customer Care Later
If you're like most early-stage companies, you might not have considered or acted on building your customer care function in parallel with your product. If you've just brought on your first hire or early team members, encourage them to use this cheat sheet from Collins:
Ask why like a three-year old.
Instinct comes from habit. Wisdom stems from context. Your customer care team will need both, and the only way they'll get them is by channeling Simon Sinek and asking why first and often. Here's a starter kit of "why" questions to get the background needed to serve customers:
To everyone: Why did this change? Why is this changing? Why didn't that change?
To product/engineering: Why is this better for the customer? Why is this important now? Why is this the direction we're going?
To sales/marketing: Why have we sold it in this way? Why do we target this customer and not that customer? Why do we now promote it the way we do?
To founders: Why is this our mission? Why is this our vision? Why is this the problem we solve? Why are we best equipped to deliver a solution?
Though she was a customer care veteran at Squarespace, Collins is still figuring out the history of and horizon for ShopKeep and its products. "Fearlessly and genuinely express your desire to understand," says Collins, "Otherwise, people will assume that you're up to speed and expect you to act knowledgeably. Asking why so often can be annoying if it's challenging in nature rather than curious in spirit. But if you ask with respect and intent, you'll get the history and hindsight you need without having had the experience firsthand."
Ask why internally until you can start each answer to customers with: "Here's why we did/do/will do this."
Set up your primary channel and nail it.
There are countless ways to connect with customers, from email to live chat, texts to tweets and phone to in-person support. "It's especially hard to achieve excellent customer care across several channels as a small startup," says Collins. "Pick a primary channel and provide great support experiences to your customers via that avenue alone. If you can consistently deliver to your standard for 3-6 months along that channel, then select and add another."
Though Collins was a pioneering member of customer care at both Squarespace and ShopKeep, she developed different primary support channels at each.
"At Squarespace, we started with email communications and then live chat was introduced," she says. "Here at ShopKeep it's primarily phone support and, now, email exists. We'll continue to introduce more ways to serve customers down the road, but we're methodical about attaining great service levels via a channel before adding another. Ultimately, whether it's one or ten channels available, customers need to know that they'll have a great experience regardless."
After you choose your primary channel, it's paramount to concurrently build in self-service. "Regardless of the primary channel you choose, always pair it with self-service. That should be your number one focus at all times," says Collins. "Having a self-service option can address at least half of all requests in the early days. What's more is that building it in conjunction with your primary channel is incredibly helpful, as self-service is fleshed out and fine-tuned through conversations with customers."
Members of the ShopKeep Customer Care Team
Hire from your community.
The first two hires after Collins joined Squarespace were active members in their online support forum and community. "There's no better demonstration of qualifications," she says. "In the forums, they showed great product knowledge, enthusiasm for our offering and a desire to interact with our team and other customers. To me, that was the interview. They were dialed in before we hired them. And I believe they're both still with the company today."
Those who share their time and expertise voluntarily typically come with a level of commitment that extends beyond the standard call of duty. "I knew the two of them would not only do their job well, but also be able to train new hires and be natural role models," says Collins. "Also, they are seismic support when the foundation's trembling. Once, I remember when our average inbound customer care requests instantly doubled after a product release. They were among the first to volunteer for a 14-hour shift, starting at the end of the work day. They just didn't clock out."
Start tiering when your team hits 20.
As the company, product suite and number of features grow, so will your customer care team. Collins led a very informal, flat structure for a long time, but once her team hit twenty people, she started to put in a hierarchy. "One day, I walked into our part of the office and didn't know everybody's name. Can you imagine how mortifying that is to a customer care professional who prides herself on being personable and community-oriented? That's when I formally created a level of managers."
As products become more numerous and sophisticated, customer service must grow to mirror that complexity. Also, as the customer base becomes more familiar with the product, the requests and questions start to change. All these shifts require a more specialized and flexible customer care structure. "It's the same with other departments. Customer care is a living, breathing creature that continues to evolve," says Collins.
To build a tiered team to respond to these changes, Collins recommends the following structure:
Tier 1: These team members are your entry level customer support agents. They are on the front line, interacting directly with customers.
Tier 2: This is a level for specialists that deal with escalated issues. They also interface directly with product, engineering and other teams in the company.
Tier 3: This layer is for customer service managers, who oversee and optimize the performance of the other two tiers. As products shift and specialities are needed, this group can re-organize teams to make sure all key areas are covered.
As each company's customer care resources and needs vary, Collins' rule of thumb is to add someone to the team when you find yourself at a breaking point. "The reality for most companies will be a reactive approach to hiring for customer service. So when you're operating at maximum pain and you can't continue, it's time to add more people," says Collins. "However, when your customer service headcount is between 20-40 staff, you've gone through enough product and hiring cycles that you can start to reliably project how many you'll need by quarter and year."
Superb customer care translates into great word of mouth. The trust of your customer is not only what's at stake; it's the potential conviction of each customer's network, too.
What It Means To Truly Care for Customers
There are standouts in customer service — Zappos, Amazon, Starbucks, and Eventbrite — but for Collins, her line of work starts and ends with how her team communicates with customers. Her school of thought prioritizes relationships built over resolutions tallied, becoming an expert decoder of customer cues and establishing a standard tone with an individual's texture.
Care or Don't Care. There is no Try.
Calling it customer care, customer support, customer happiness or customer success is semantics in the end. However, whether you are primarily focused on customers OR their challenges is a nuance with important ramifications. "Those who ask how to create the feeling of caring for customers are typically focused primarily on issue resolution, while customers are more of an afterthought," says Collins. "It's easy to get hung up on metrics and KPIs, but I try to remind myself to shape the interaction with a customer to be a conversation, versus a response. It's about listening to what they're telling you, not waiting until you can give them an answer."
For example, two years into her job at Squarespace, Collins felt in command of her work. She clocked fast response times because she was a quick typist and knew the answers to most every question she'd get. One day, a customer wrote in regarding a past due bill for his account, explaining that, as a freelancer, he was waiting on his client to complete his invoice to get paid. "He politely asked for an extension and it hit a chord. I know what it's like to be tight on money," says Collins. "And not only that, but he's taking the time to write in and tell me his site is important to him. He's asking for a couple of extra days versus just defaulting on his payment."
Take a moment to ask yourself: will I take the opportunity to address more than what I'm being asked? In this case, Collins empathized with the customer, which allowed her to think more broadly and flexibly with her solution. She was not constrained by the problem presented to her. She could have just extended the due date, but instead said, "I'm just going to take care of your invoice. Don't worry about it. It's a free month. It's fine." Collins didn't think much of it, but the customer wrote back, effusively expressing his gratitude. "It wasn't because I gave him a free month. It was because I said, 'Hey, fellow human being, I care and can relate to your situation in some small way. I'd like to do something that would make your day a little bit easier."
Not everyone has someone ready to help who's just an email or call away. I'm astonished by what people have shared with me. It's a privilege to be entrusted with their vulnerability.
For all the Squarespace customers who have been touched by handwritten thank you cards or been sent flowers in the hospital, there are those who don't want a relationship with a startup. How do you proceed then? "Sometimes they just want an answer. That happens," says Collins. "And that's totally, absolutely fine. I think that you can create a relationship by not creating a relationship. The job really is being there in a way that the customer needs you to be when they need you to be there. So if they don't, then it's your role to answer and then step back."
Pinpoint and act on customer cues.
Consistent and comprehensive customer care requires understanding subtle cues in both a written and verbal setting. Here are a few pointers from Collins on being mindful of what isn't being said in your conversations with customers.
Lots of background noise. "If there's a lot of ambient sound — a barking dog, footsteps, the rumble of a train or children's voices, for example — please don't take up their time telling jokes or asking about their grandkids. They may be being polite, but they're busy. Their time is as important as the answer. Speak clearly and move quickly."
Non sequiturs and personal follow-up. "Verbose customers may be trying to soften you up to ask for a favor, but they also could be aiming for a real connection. It could be out of loneliness or interest. I recall customers that would always contact me before and after their vacations because we had talked about travel. It was great!"
Raised voice or curt notes. "Anger or frustration is generally easy to identify, but much harder to diffuse. I always try to think about anger as being passionate about a problem that a person can't control but cares deeply about. When you see or hear it, you know. It's best to provide structure to your response: playback what you've heard said and fix it immediately. If you can't get it fixed quickly, lay out why and what you'll do about it."
That said, there are cases in which picking up on cues and caring about the customer will fail. Those instances — though rare — may require extracting yourself.
"Early in my career, when I was a manager at Nordstroms, a customer got very belligerent and started screaming in my face. It's very intense emotionally when that happens. You can really hear a person's anger and malice towards you," says Collins. "I froze. I wanted to make sure he was okay, talk him down and resolve the situation, but it was so abusive. I'll never forget when the store manager on call arrived and calmly asked me to go to the back room."
The takeaway: if a line's crossed and it's no longer a productive conversation, then there's no need to continue. The manager removed me from the situation and dealt with the angry customer. She knew I was not okay, but asked if I was, then said, "I want you to know, that's never okay. You do not deserve to be spoken to or treated that way. You always have the resource of calling a supervisor if the situation becomes uncomfortable. That's what we're here for." Collins has taken that lesson with her and ensures there's always a lifeline for her customer service team. Content customers start with a happy customer care team — sometimes that takes shielding and protecting people from toxic customers.
Members of the Squarespace Customer Care Team
The company sets message and tone — individuals bring texture.
Like many teams in most early-stage companies, customer service is closely influenced by the founders early on. "Looking back, especially at Squarespace, our customer service communications took on the voice of [founder and CEO] Anthony [Casalena] and our early engineers," says Collins. "Then, over time, it evolved and the customer care team standardized how to support our customers and with which content. We direct the message and tone."
For straightforward or strict actions like a DMCA takedown request or a terms of service violation, customer care teams shouldn't deviate from prescribed language. But in many cases, Collins encourages her team members to communicate content with their own flair and personality. "That authenticity is essential to establishing an early connection with customers — and gives ownership to customer care representatives. Some may use more exclamation points, add in a personal greeting or sign-off in a formal way. The goal is to handle the communication so it's in line with our script and tone, but also sounds like the individual."
Collins' background as a legal secretary has informed her personal communication style. "Back when I communicated with customers, my tone was very dry. My words were simple and to the point, and I was never excessive with punctuation," says Collins. "It's what you'd expect from someone who wrote emails to attorneys for years. It was comfortable for me, and I want the same for each person on my team. I'll tell them: "Hey, this is how we answer this question. This is how we address this issue. Have at it. Just reply and use your voice."
Various voices can be equally effective, but treat the customer's replies as a mirror. "You can use an exclamation point to show enthusiasm without looking silly, but stay alert to how your customer receives it," Collins says. "Can you connect with this person and speak in a conversationally casual way or are they expecting something else from you? Are they seeking a really quick response that's straight to the point and cut and dry? You need to make that determination and calibrate to your customer."
Measuring the Effectiveness of Your Customer Care
When it comes to tools to evaluate customer support, there's an alphabet soup of tools: NPS (Net Promoter Score), CSAT (Customer Satisfaction Score) and CES (Customer Effort Score), to name a few. Collins recommends ramping up slowly to more systematic methodologies and running your own tests to understand the makeup of your specific customer base. Here's how:
Make it lightweight to start.
There can be a tendency to get intricate with surveying customers about their experience, but Collins recommends keeping it simple at the beginning. "At the start, try to measure success in a very lightweight fashion — literally, click thumbs up or thumbs down to share if we helped you. This will help you get a sense of the value you're giving and the rhythm with which you're providing it," she says.
It can become a lot more sophisticated down the road. Allowing for more free-form feedback will get you a bit more color, and you can start to figure out how to focus your resources. "If they say that it was a great interaction, but ask why they needed to write in to get it, it's time to build out your FAQ. If they say you don't know what you're talking about it, perhaps you should look into ongoing training for the team," says Collins. "Maybe they'll comment that the interface is terrible and you've made it difficult. While that's not necessarily reflective of the customer service performance itself, that's valuable feedback to pass along to Product or Engineering. Remember, you receive the report card for the entire class, not just you."
Think beyond what you measure. Prioritize where and how, too.
Collins sees a place for NPS and customer satisfaction measurement tools, but for different parts of the organization. "I think NPS is owned by the company, while customer satisfaction belongs to each department, including customer care. It tells us how if and how we are handling your requests," she says. "However, NPS — if you'd recommend this product to a friend — receives a rating, in which customer service only plays a part."
It's equally important to run tests that measure your unique take on customer support. For example, while at Squarespace, Collins emphasized conversations over transactions. She had a hypothesis that, as in life, a conversation — and relationship — would become more valuable after many interactions. So, if customers wanted to keep asking questions, let them. Her team measured customer satisfaction by number of interactions. As this was over email support channel, each response on an email thread constituted one interaction. Here's what they found:
2-4 interactions: low customer satisfaction
5-9 interactions: high customer satisfaction
10-18 interactions: low customer satisfaction
19+ interactions: moderate customer satisfaction
"Taken with the qualitative data, we found that a quick correspondence — even if we solved the problem — felt transactional or didn't provide a long enough runway to show our value-add," says Collins. "Between 5-9 interactions, customers felt that they were engaged in an experience that was informative and conversational. We could demonstrate that weren't robots or outsourced support — we were real humans taking time to help."
The biggest lesson emerged from the cases that involved ten or more interactions. "With the longer conversations, we realized that, by and large, we had messed up," says Collins. "Something happened and we missed something. Maybe we didn't pay attention. Or we overlooked a key piece of information or weren't reading properly. Obviously, those cases were really bad."
Then, for the outlier segment of 19+ interactions, someone senior from customer service would typically step in to own the case. "They were really long emails, and the support lead would eventually turn the ship around," says Collins. "They'd get it resolved. It might take five more back and forth emails, but the customer would get an answer and say something like, 'You know, I was really, really mad at you, but you fixed it. Thank you and I'm happy. Next time, do better but at least you put in the effort. You took the time to own it and to make sure that it was resolved.'"
Squarespace's Customer Care Team
For a long time, customer care was deprioritized and often built in haste when the need arose. Then it was acknowledged, but seen as a necessary evil. Now, from Warby Parker to Zappos, companies have increasingly made customer care indispensable to — and sometimes synonymous with — their product and reputation. It's a wise path forward that many companies still neglect. Savvy startups will launch customer care in parallel with their first product. If not, they'll heed Collins' tips in order to catch up quickly. Their departmental mission and metrics will value relationships over resolutions and conversations over transactions.
"At the end of the day, a customer care professional is benevolent double agent, acting on behalf of both the company and customer," says Collins. "If forced to choose a side, they won't. They'll go rogue and represent the product because, like them, they were formed by the company for the benefit of the customer. They'll always be of and between those worlds."


I hope this plane will find clear skies. I've been in emotional turbulence. False expectations, I suppose. Summer is supposed to be my good season. My break from the constant pain, fatigue, and general malaise that haunts me. Not this summer. Could be the baby. Almost two years old baby. And when Anika was this age, I had my big fall-apart. Then again, it could be the medication-adjustments. Summer is the season for trying new things. And the fatigue medication works okay, but it spins me out, too. I didn't take any today and felt a lot better than the past week. And I dropped the pain-meds thinking I wouldn't need them. Wrong there. The arthritis in my feet has four years now of aggravation. The bones are not happy. And they let me know big time when I quit medication. So I'm taking a low dose of the pain meds and I quit the fatigue meds.
Not to mention that I am on my feet a lot. Even in summer. Chores and projects and keeping up with the toddler. These are not laid-back summer haze-days.
Degenerative symptoms galore. I worry about the mad-tumble of troubles, like when Anika was a toddler. Back then it was the arthritis diagnosis and the weakness/numbness in my hands. I worry about what may be next.
Then I breathe. Worry won't change the path around the corner. I slow down. Listen to my girl. Watch her dance. Read my boy stories. And try to stay here now instead of worrying about the future. We could all worry.
I've also been contending with a new frustration. Anger. It could be a side effect of the medications, but I've been bee-spitting mad. The bees whirl around in me with stirring whirring fury. Why is everything such a struggle?
I'm not angry at anyone. Not at God. Not at myself. Just a decentralized anger. Why can't I fix this? I'm clever and creative. Why don't I have a solution or two? Why is waking up in the morning like a zombie climbing from the mud? Why the 13 thousand days of negotiation and compromise and frustration?
I found articles online that had clinical answers about disability and anger. Management issues. A stage in greiving.
And maybe that's part of the degenerative piece, the constant losses. So the anger resurfaces. But I feel an intense and unfamiliar resentment this time. I want to be better. Not better in my health, necessarily. But a better mom. A better writer. A better teacher. A better house-cleaner :). I see where I could go, who I could be, if only I had the energy.


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