text:
Q: Kubernetes service communication isse - Kubedns I have two pods mapped to two services up and running using virtual box vm's in my laptop. I have kube dns working. One pod is a webservice and the other is a mongodb.
The spec of webapp pod is below
spec:
  containers:
    - resources:
        limits:
          cpu: 0.5
          .
          .
      name: wsemp
      ports:
        - containerPort: 8080
  #     name: wsemp
  #command: ["java","-Dspring.data.mongodb.uri=mongodb://192.168.6.103:30061/microservices", "-Djava.security.egd=file:/dev/./urandom","-jar","/app.jar"]
  command: ["java","-Dspring.data.mongodb.uri=mongodb://mongoservice/microservices", "-Djava.security.egd=file:/dev/./urandom","-jar","/app.jar"]

The spec of corresponding service
apiVersion: v1
kind: Service
metadata:
  labels:
    name: webappservice
  name: webappservice
spec:
  ports:
   - port: 8080
     nodePort: 30062
     targetPort: 8080
     protocol: TCP
  type: NodePort
  selector:
    name: webapp

Mongodb pod spec
apiVersion: v1
kind: Pod
metadata:
  name: mongodb
  labels:
    name: mongodb
spec:
  containers:
    .
    .
  name: mongodb
  ports:
    - containerPort: 27017

Mongodb service spec
apiVersion: v1
kind: Service
metadata:
  labels:
    name: mongodb
  name: mongoservice
spec:
  ports:
   - port: 27017
     nodePort: 30061
     targetPort: 27017
     protocol: TCP
  type: NodePort
  selector:
    name: mongodb

UPDATED TARGET PORTS IN SERVICE AFTER COMMENT
Issue
The webapp when it starts is not able to connect with the mongoservice port and gives this error on start
Exception in monitor thread while connecting to server mongoservice:27017
com.mongodb.MongoSocketOpenException: Exception opening socket
at com.mongodb.connection.SocketStream.open(SocketStream.java:63) ~[mongodb-driver-core-3.2.2.jar!/:na]
at        com.mongodb.connection.InternalStreamConnection.open(InternalStreamConnection.java:114) ~[mongodb-driver-core-3.2.2.jar!/:na]
at com.mongodb.connection.DefaultServerMonitor$ServerMonitorRunnable.run(DefaultServerMonitor.java:128) ~[mongodb-driver-core-3.2.2.jar!/:na]
at java.lang.Thread.run(Thread.java:745) [na:1.8.0_111]
Caused by: java.net.ConnectException: Connection refused (Connection refused)
at java.net.PlainSocketImpl.socketConnect(Native Method) ~[na:1.8.0_111]

describe svc
kubectl describe svc mongoservice
Name:           mongoservice
Namespace:      default
Labels:         name=mongodb
Selector:       name=mongodb
Type:           NodePort
IP:         10.254.146.189
Port:           <unset> 27017/TCP
NodePort:       <unset> 30061/TCP
Endpoints:      172.17.99.2:27017
Session Affinity:   None
No events.

kubectl describe svc webappservice
Name:           webappservice
Namespace:      default
Labels:         name=webappservice
Selector:       name=webapp
Type:           NodePort
IP:         10.254.112.121
Port:           <unset> 8080/TCP
NodePort:       <unset> 30062/TCP
Endpoints:      172.17.99.3:8080
Session Affinity:   None
No events.

Debugging
root@webapp:/# nslookup mongoservice
Server:     10.254.0.2
Address:    10.254.0.2#53

Non-authoritative answer:
Name:   mongoservice.default.svc.cluster.local
Address: 10.254.146.189

root@webapp:/# curl 10.254.146.189:27017
curl: (7) Failed to connect to 10.254.146.189 port 27017: Connection refused
root@webapp:/# curl mongoservice:27017
curl: (7) Failed to connect to mongoservice port 27017: Connection refused

sudo iptables-save | grep webapp

-A KUBE-NODEPORTS -p tcp -m comment --comment "default/webappservice:" -m tcp --dport 30062 -j KUBE-MARK-MASQ
-A KUBE-NODEPORTS -p tcp -m comment --comment "default/webappservice:" -m tcp --dport 30062 -j KUBE-SVC-NQBDRRKQULANV7O3
-A KUBE-SEP-IE7EBTQCN7T6HXC4 -s 172.17.99.3/32 -m comment --comment "default/webappservice:" -j KUBE-MARK-MASQ
-A KUBE-SEP-IE7EBTQCN7T6HXC4 -p tcp -m comment --comment "default/webappservice:" -m tcp -j DNAT --to-destination 172.17.99.3:8080
-A KUBE-SERVICES -d 10.254.217.24/32 -p tcp -m comment --comment "default/webappservice: cluster IP" -m tcp --dport 8080 -j KUBE-SVC-NQBDRRKQULANV7O3
-A KUBE-SVC-NQBDRRKQULANV7O3 -m comment --comment "default/webappservice:" -j KUBE-SEP-IE7EBTQCN7T6HXC4
$ curl 10.254.217.24:8080
{"timestamp":1486678423757,"status":404,"error":"Not Found","message":"No message available","path":"/"}[osboxes@kube-node1 ~]$

sudo iptables-save | grep mongodb
[osboxes@osboxes ~]$ sudo iptables-save | grep mongo
-A KUBE-NODEPORTS -p tcp -m comment --comment "default/mongoservice:" -m tcp --dport 30061 -j KUBE-MARK-MASQ
-A KUBE-NODEPORTS -p tcp -m comment --comment "default/mongoservice:" -m tcp --dport 30061 -j KUBE-SVC-2HQWGC3WSIBZF7CN
-A KUBE-SEP-FVWOWAWXXVAVIQ5O -s 172.17.99.2/32 -m comment --comment "default/mongoservice:" -j KUBE-MARK-MASQ
-A KUBE-SEP-FVWOWAWXXVAVIQ5O -p tcp -m comment --comment "default/mongoservice:" -m tcp -j DNAT --to-destination 172.17.99.2:27017
-A KUBE-SERVICES -d 10.254.146.189/32 -p tcp -m comment --comment "default/mongoservice: cluster IP" -m tcp --dport 27017 -j KUBE-SVC-2HQWGC3WSIBZF7CN
-A KUBE-SVC-2HQWGC3WSIBZF7CN -m comment --comment "default/mongoservice:" -j KUBE-SEP-FVWOWAWXXVAVIQ5O
[osboxes@osboxes ~]$ sudo curl  10.254.146.189:8080
^C[osboxes@osboxes ~]$ sudo curl  10.254.146.189:27017

It looks like you are trying to access MongoDB over HTTP on the native driver port.

root@mongodb:/# netstat -an
Active Internet connections (servers and established)
Proto Recv-Q Send-Q Local Address           Foreign Address         State
tcp        0      0 0.0.0.0:27017           0.0.0.0:*               LISTEN
tcp        0      0 172.17.99.2:60724       151.101.128.204:80      TIME_WAIT
tcp        0      0 172.17.99.2:60728       151.101.128.204:80      TIME_WAIT

mongodb container has no errors on startup.
Trying to follow steps in https://kubernetes.io/docs/user-guide/debugging-services/#iptables, stuck in the part where it says " try restarting kube-proxy with the -V flag set to 4" since I dont know how to do it.
I'm not a networking person, so dont know how and what needs to be analyzed in this. Any kind of tips to debug would be of great help.
Thanks.

A: :)
As a side note, have in mind that curl performs HTTP requests by default, but the port 27017 in the host you are trying to reach is not binded to an application that understands such protocol. Typically, what you would you in these scenarios is to use netcat:
nc -zv mongoservice 27017
This reports whether the port 27017 from such host is open or not.

*

*nc = netcat

*-z scan for listening daemons without sending data

*-v adds verbosity

Regarding your MongoDB file, you must remember to set the targetPort directive. As explained in Kubernetes docs regarding targetPort:
This specification will create a Service which targets TCP port 80 on any Pod with the run: my-nginx label, and expose it on an abstracted Service port (targetPort: is the port the container accepts traffic on, port: is the abstracted Service port, which can be any port other pods use to access the Service). View service API object to see the list of supported fields in service definition.
Therefore, just set it to 27017 for consistency.
You should not run into issues after following these advices. Keep the good work and learn as much as you can!

A: The iptables rules looks ok, but it is not sure what network solution (flannel/calico) used in your kubernetes. You may check whether you can access kube dns pod IP from your web pod.

A: thanks. I had got a clue on this and since I was using the flannel network, there was an issue with the communication between the pods in the flannel network.
Particularly this part, FLANNEL_OPTIONS="--iface=eth1" as mentioned in the link http://jayunit100.blogspot.com/2015/06/flannel-and-vagrant-heads-up.html
Thanks.

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text:
Q: Edit windows HOST file using Ruby I would like to copy content from another file into my windows host file.
I have tried running it as a administrator but it still does not work.
Any help or alternative methods to do this would be appreciated!
Filetocopy = File.read('assets/backup/copy_host')
File.open('C:/Windows/System32/drivers/etc/hosts', 'w+') do |copy|
   copy.puts Filetocopy.to_s
end

The error I get is - Permission denied @ rb_sysopen - C:/Windows/System32/drivers/etc/hosts


Damien Seguin will carry the French flag during the London 2012 Paralympic Games Opening Ceremony.
The French 2.4mR sailor has been one of the standout sailors in the class having won gold at Beijing 2008, silver at Athens 2004 and three World Championships.
Seguin said, "It is an honour and a pride in having been chosen by the French Paralympic Committee and Sports to carry the colours of France in the Paralympics. Thanks to the staff but also for people who helped me get to that level, to be what I am today. It is also an award for my discipline because it is the first time a marine athlete has been assigned this responsibility."
Jean-Pierre Champion, President of FFV, added, "Damien Seguin is a great competitor and a great champion. He has the desire to win and to be a great captain."


Ample, a technology company focused on solving the energy delivery challenge for electric cars, co-founded by Khaled Hassounah and John de Souza, announced today that it has closed a $31 million Series A round of financing. Shell Ventures led the investment round with participation from Moore Strategic Ventures, Repsol Energy Ventures, Hemi Ventures, and TRIREC. The financing will be used to deploy Ample's technology in multiple locations over the coming months. Steve McGrath of Shell Ventures and James McIntyre of Moore Strategic Ventures will be joining Ample's Board of Directors.
"Building a scalable solution for charging electric vehicles requires a new approach to infrastructure, and partners who are committed to developing electrification as the new energy downstream," said Khaled Hassounah, CEO and co-founder of Ample.
Ample has invented an economical, rapidly deployable and widely accessible platform that delivers a full charge to any electric car in minutes. An alternative to traditional charging, Ample uses autonomous robotics and smart-battery technology, making it feasible for anyone to own an electric car regardless of driving needs, economic means, or geographic location.
Moore Strategic Ventures, LLC is the privately held investment company for Louis M. Bacon, Founder and CEO of Moore Capital Management, LP.


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For additional information, visit www.businessdevelopment.org/eerl.
Answers to commonly asked questions are listed below.
Who is eligible to receive an EERL Loan?
Does the interest rate change over the life of the loan?
Is there a penalty for paying off the loan more quickly than planned?
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text:
Labor Relations Director Salaries in Canada
Alternate Job Titles: Director of Industrial Relations, Director of Labor Relations
What is the average Labor Relations Director salary in Canada?
How much does a Labor Relations Director in Canada make? The median annual Labor Relations Director salary in Canada is C$125,601, as of December 28, 2020, with a range usually between C$113,050-C$132,995 not including bonus and benefit information and other factors that impact base pay. However, the salary for someone with the title Labor Relations Director may vary depending on a number of factors including industry, company size, location, years of experience and level of education. Our team of Certified Compensation Professionals has analyzed survey data collected from thousands of HR departments at companies of all sizes and industries to present this range of annual salaries for people with the job title Labor Relations Director in Canada.
This chart describes the expected percentage of people who perform the job of Labor Relations Director that make less than that annual salary. For example the median expected annual pay for a typical Labor Relations Director in Canada is C$125,601, so 50% of the people who perform the job of Labor Relations Director in Canada are expected to make less than C$125,601.
Labor Relations Director Salaries by Percentile
10th Percentile Labor Relations Director Salary C$101,624 Canada December 28, 2020
50%(Median) C$125,601
10%C$101,624
This chart describes the expected percentage of people who perform the job of Labor Relations Director in , that make less than that salary. For example 50% of the people who perform the job of Labor Relations Director in are expected to make less than the median.
Job Description for Labor Relations Director
Directs and oversees a company's labor relations programs, policies, and procedures. Establishes and maintains satisfactory labor-management relations, interprets the collective bargaining agreements, administers grievance procedures including arbitrations, and assists all levels of management on labor matters. Requires a bachelor's degree in a related area. Demonstrates expertise in a variety of the field's concepts, practices, and procedures. Typically reports to top management. Manages a departmental sub-function within a broader departmental function. Creates functional strategies and specific objectives for the sub-function and develops budgets/policies/procedures to support the functional infrastructure. Deep knowledge of the managed sub-function and solid knowledge of the overall departmental function. Typically requires 5+ years of managerial experience. View full job description
View Human Resources Jobs by Salary Range: <$30K, $30K–$50K, $50K–$80K, $80K–$100K, >$100K
Categories: Human Resources , Executive and Management
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text:
Polska na zawodach Pucharu Europy w Lekkoatletyce 1994 – wyniki reprezentacji Polski w 15. edycji Pucharu Europy w 1994.

Zarówno reprezentacja żeńska, jak i męska, wystąpiły w zawodach tzw. I ligi (II klasie rozgrywek), które odbyły się w dniach 11–12 czerwca 1994 w Walencji.

Mężczyźni
Polska zajęła 2. miejsce wśród ośmiu drużyn, zdobywając 112,5 punktów i awansowała do tzw. superligi. Zwycięstwa pozbawiła ją dyskwalifikacja za doping Piotra Perżyły.

 100 m: Marek Zalewski – 4 m. (10,49)
 200 m: Robert Maćkowiak – 2 m. (21,14)
 400 m: Tomasz Jędrusik – 3 m. (47,28)
 800 m: Ryszard Ostrowski – 2 m. (1:51,13)
 1500 m: Michał Bartoszak – 4 m. (3:41,78)
 5000 m: Piotr Gładki – 6 m. (13:56,52)
 10000 m: Sławomir Kąpiński – 2 m. (28:30,15)
 110 m ppł: Ronald Mehlich – 2 m. (13,87)
 400 m ppł: Piotr Kotlarski – 1 m. (50,36)
 3000 m z przeszkodami: Rafał Wójcik – 4 m. (8:43,81)
 skok wzwyż: Jarosław Kotewicz – 1 m. (2,24)
 skok o tyczce: Bogusław Kopkowski – 5 m. (5.20)
 skok w dal: Dariusz Bontruk – 5 m. (7,64)
 trójskok: Krystian Ciemała – 6 m. (16,38, wiatr +3,0)
 pchnięcie kulą: Piotr Perżyło – zdyskwalifikowany za doping (pierwotnie 2 m. 18,85)
 rzut dyskiem: Marek Majkrzak – 6 m. (56,02)
 rzut młotem: Lech Kowalski – 4 m. (71,98)
 rzut oszczepem: Mirosław Witek – 2 m. (78,84)
 sztafeta 4 × 100 m: Tomasz Pokora, Robert Maćkowiak, Marek Zalewski, Marcin Krzywański – 2 m. (39,93)
 sztafeta 4 × 400 m: Piotr Rysiukiewicz, Tomasz Czubak, Piotr Kotlarski, Tomasz Jędrusik – 1 m. (3:06,52)

Kobiety
Polska zajęła 1. miejsce wśród ośmiu drużyn, zdobywając 106 punktów i awansowała do tzw. superligi.

 100 m: Izabela Czajko – 7 m. (11,62)
 200 m: Monika Borejza – 6 m. (24,13)
 400 m: Elżbieta Kilińska – 1 m. (52,98)
 800 m: Małgorzata Rydz – 1 m. (2:00,40)
 1500 m: Małgorzata Rydz – 2 m. (4:18,23)
 3000 m: Anna Brzezińska – 1 m. (8:57,50)
 10000 m: Małgorzata Sobańska – 7 m. (34:04,99)
 100 m ppł: Urszula Włodarczyk – 3 m. (13,45)
 400 m ppł: Sylwia Pachut – 1 m. (57,45)
 skok wzwyż: Katarzyna Majchrzak – 4 m. (1,90)
 skok w dal: Agata Karczmarek – 1 m. (6,89)
 trójskok: Urszula Włodarczyk – 2 m. (13,61, wiatr +3,4)
 pchnięcie kulą: Krystyna Danilczyk – 2 m. (18,29)
 rzut dyskiem: Renata Katewicz – 2 m. (60,78)
 rzut oszczepem: Genowefa Patla – 1 m. (61,60)
 sztafeta 4 × 100 m: Izabela Czajko, Sylwia Pachut, Monika Borejza, Dorota Brodowska – 4 m. (45,05)
 sztafeta 4 × 400 m: Barbara Grzywocz, Monika Warnicka, Sylwia Pachut, Elżbieta Kilińska – 2 m. (3:33,71)

Bibliografia
 Francisco Ascorbe, Félix Capilla, José Luis Hernández History of the European Cup. Statistics handbook, wyd. Malaga 2006

1994 w lekkoatletyce
Polska na zawodach Pucharu Europy w lekkoatletyce
1994 w polskim sporcie

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Warner Music Group has become the second major record label to sign with online video site Hulu.
Hulu.com , which is mostly known for its full length movies and TV episodes, has just signed a music deal with EMI music to deliver high quality music and videos.
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Music labels are more willing to work with Hulu over YouTube since Hulu provides copyrighted and professional content only.


Professor Elizabeth Anderson has been awarded an American Council of Learned Societies Fellowship. ACLS Fellowships support a faculty member through a year of continuous full-time research and writing. The criteria used in judging ACLS fellowship applications include the potential of the project to make an original and significant contribution to knowledge, and the scholarly record and career trajectory of the applicant. There were 1121 applicants for 65 different ACLS Fellowships this year. Liz received one of only 20 fellowships available to full professors. Congratulations, Liz!
Project Abstract- I aim to develop a theory of how people can improve their moral beliefs through experiments in living and knowledge-enhancing practices of contention or interpersonal claim-making. I shall develop this theory through case studies in the history of contention over slavery and emancipation, mostly in the U.S., using work in social and moral psychology on cognitive biases and the influence of social situations on value judgments. The result will be a pragmatist, naturalized moral epistemology in the social contract tradition.


text:
Chloroquine treatment of ARPE-19 cells leads to lysosome dilation and intracellular lipid accumulation: possible implications of lysosomal dysfunction in macular degeneration
Patrick M Chen1,2,
Zoë J Gombart1 &
Jeff W Chen1
Age-related macular degeneration (AMD) is the leading cause of vision loss in elderly people over 60. The pathogenesis is still unclear. It has been suggested that lysosomal stress may lead to drusen formation, a biomarker of AMD. In this study, ARPE-19 cells were treated with chloroquine to inhibit lysosomal function.
Chloroquine-treated ARPE-19 cells demonstrate a marked increase in vacuolation and dense intracellular debris. These are identified as chloroquine-dilated lysosomes and lipid bodies with LAMP-2 and LipidTOX co-localization, respectively. Dilation is an indicator of lysosomal dysfunction. Chloroquine disrupts uptake of exogenously applied rhodamine-labeled dextran by these cells. This suggests a disruption in the phagocytic pathway. The increase in LAMP protein levels, as assessed by Western blots, suggests the possible involvement in autophagy. Oxidative stress with H2O2 does not induce vacuolation or lipid accumulation.
These findings suggest a possible role for lysosomes in AMD. Chloroquine treatment of RPE cells may provide insights into the cellular mechanisms underlying AMD.
Age-Related Macular Degeneration (AMD) is the leading cause of progressive central vision loss in elderly people over the age of 60 [1–3]. The clinical hallmarks of "dry" AMD, which accounts for 85-90% of AMD patients, is the appearance of yellow pigments known as drusen and marked photoreceptor death within the macula [1, 4]. While it has been established that smoking, light exposure and genetics are risk factors for AMD, its cellular-molecular pathogenesis remains unclear [4].
Retinal pigment epithelium (RPE) metabolism is an important factor in drusen buildup along the Bruch's membrane, located strategically between the choroid and RPE [4]. The RPE, a highly specialized monolayer epithelium that forms the outermost layer of the retina, is among the most active phagocytic systems in the body [5, 6]. On a daily basis, the outer segment tips of photoreceptors are phagocytosed into the RPE, and digested in phago-lysosomes within the RPE [7]. Autophagy also contributes to the heavy load of material the RPE digests [8]. In theory, lysosomal overload may thus lead to a buildup of biological "waste products", reducing RPE efficiency and contributing to extracellular protein-lipid deposits along Bruch's membrane [4, 8–10].
Lysosomal overload and dysfunction in RPE is suspected to be a critical and early cause of AMD [4, 11]. It is well established that lipofuscin, a pigmented aggregate of proteins and lipids, a primary component of drusen, and an AMD biomarker, is sequestered by lysosomes in RPE [12, 13]. At critical concentrations, N-retinylidene-N- retinylethanolamine (A2-E), a fluorescent pigment of lipofuscin, inhibits lysosomal ATPase proton pumps, inhibits critical enzymes and causes lysosomal compartment leakage into RPE cytoplasm [4, 14, 15]. Recently, it has been shown that the variant B mutation in cystatin C, a widely expressed lysosomal protease inhibitor, inhibits proteolytic regulator secretion, mistargets signaling, causes inappropriate cell protein retention and is associated with AMD and Alzheimer's [16]. Furthermore, proteins modified by lipid peroxidation similar to those found in lipofuscin have been shown to reduce proteolytic activity of lysosomes of RPE cells [17]. Finally, studies have begun to uncover a link between retinal degeneration and Niemann-Pick type C, a known lysosomal storage disease. A recent study observed that mice with mutations in the Npc1 and Npc2 gene, which transcribe proteins that mediate the exit of lipoproteins from lysosomes, demonstrate striking retinal degeneration, upregulation of autophagy and marked lipofuscin accumulation within the RPE [18]. These aforementioned studies suggest that abnormalities in the structural integrity and enzymatic activity of the lysosomes of RPE cells may play a role in the pathogenesis of AMD.
In this study, we investigate the possible role of lysosomes in AMD by treating in vitro human adult retinal pigmented epithelium-19 (ARPE-19) cells, which have previously been used as a model for the study of the etiology and development of AMD [19, 20], with chloroquine, a known lysosomotropic agent. The effects of chloroquine as a retinopathic agent, as observed by lysosomal dysfunction and RPE degradation, have been demonstrated in various animal models [21–24]. We use the ability of chloroquine to increase pH [25] to both understand the general effects of chloroquine on ARPE-19, and as a model for lysosomal inhibition. The results demonstrate that chloroquine induces vacuole formation, cell death, cytosolic lipid buildup and decreased exogenous dextran uptake in ARPE-19.
ARPE-19 Lysosomal Inhibition with Chloroquine Treatment
Chloroquine is a known lysosomotropic agent that increases lysosomal pH by accumulating within lysosomes as a deprotonated weak base. To study the effects of lysosomal dysfunction in ARPE-19, it was necessary to establish an in vitro model utilizing chloroquine. We determined the concentration of chloroquine that substantially changed lysosomal activity, but did not result in cell necrosis.
To find an optimal concentration of chloroquine that did not affect ARPE-19 cell viability, we utilized both DAPI nuclei staining and the MTT assay (Figure 1). For DAPI cell quantification, the nuclei of ten random areas of the coverslip were counted. The results (not shown) of these counts were averaged, expressed as the percentage of the control, and analyzed by student t-test and one-way ANOVA with a TI-89 Texas Instruments graphing calculator. The DAPI analysis showed cell viability was time and dosage dependent, with concentrations of 10-20 μg/ml at 24 hour incubation periods not significantly (p < 0.05, n = 6) affected.
Cell Viability Assay. MTT assay shows chloroquine toxicity is both time and dose dependent. Chloroquine concentrations of 10-30 μg/ml (p < 0.05) do not significantly affect cell viability.
The MTT assay, which evaluated cell proliferation by measuring metabolic activity, showed similar results. Figure 1 demonstrates that cell viability and metabolism is relatively unaffected from 10-30 μg/ml and is dosage dependent. Student t-test shows no significant difference from 10-20 μg/ml (p < 0.05, n = 6) with significant difference in cell viability between 10 and 40 μg/ml (p = 0.033, <0.05, n = 6).
Chloroquine Induces Cytosolic Vacuolation and Dense Body Formation
Having determined that low concentrations (10-20 μg/ml) at 24-hour incubation did not significantly reduce viability, we evaluated the cytoplasmic cellular changes in ARPE-19 with phase contrast microscopy. Figure 2 shows the progression of cytoplasmic changes of ARPE-19 treated with chloroquine under phase contrast microscopy. We found that 10-20 μg/ml of chloroquine induced marked vacuolation within the cytoplasm of ARPE-19, as can be seen in Figure 3. In addition to increased vacuole size and dispersion, we note the appearance of dense, black, circular formations interspersed throughout the cytoplasm and around the large vacuoles. Vacuole and dense body size increased with chloroquine dosage. In a parallel study performed on NIH 3T3 cells, we noted the dilation of lysosomes at similar concentrations of chloroquine treatment (Figure 4). However, there was no concurrent development of the dense, black formations.
Cytoplasmic Changes with Chloroquine in ARPE-19. Phase contrast microscopy of ARPE-19 with 0 (A), 10 (B), 20 (C), 40 (D), 80 (E) μg/ml chloroquine. Chloroquine treatment causes membrane-enclosed vacuoles (red arrows) and membranous dense bodies (blue arrows). ARPE treated with high chloroquine dosage show signs of cell death. Bar size = 25 microns.
Co-localization of LAMP-2 and LipidTOX. (A-D) Control ARPE-19. (E-H) 20 μg/ml chloroquine-treated ARPE-19. Control phase contrast (A) shows no vacuolation and LAMP-2 (B, green), and LipidTOX(C, red) staining is observed throughout cell. D is fluorescent overlay. Chloroquine-treated cell in phase contrast (E) shows intense vacuolation (white arrows). These vacuoles co-localize with LAMP-2 staining (F, corresponding white arrows). Chloroquine treatment also induces dense body formation (E, yellow arrows). Dense formations co-localize with neutral lipid (G, corresponding yellow arrows). Overlay (H) shows no overlap between lipids and vacuoles. Bar size = 25 microns.
Co-localization of LAMP-1 in NIH/3T3. NIH/3T3 cells show a similar pattern of vacuole formation with treatment of chloroquine. Phase contrast and corresponding LAMP-1 co-localization of 25 μg/ml chloroquine-treated cells (C, D) show vacuolation of LAMP-1, when compared with control (A, B). The arrows in C and D note that the borders of the vacuoles correspond with LAMP-1 protein. Bar size = 25 microns.
Lamp-2 Co-localizes with Dilated Vacuoles, LipidTOX Co-localizes with Dense Formations
To determine if the abnormal vacuoles were dilated lysosomes, as documented in a previous study using calf RPE [26], and if the dense bodies contained lipid, ARPE-19 cells were doubled stained with LAMP-2 antibody and LipidTOX neutral lipid. Inhibition of the essential lysosomal protease cathepsin B and other mucopolysaccharides is characteristic of chloroquine-dilated lysosomes [25]. Moreover, lipid co-localization would suggest a form of intracellular "biological debris" accumulation within the cell in response to lysosomal enzymatic inhibition.
LAMP-2 antibody staining of chloroquine-treated cells was used to co-localize dilated vacuoles with lysosomes and phago-lysosomal complexes. As shown in Figure 3 LAMP-2 antibody staining co-localizes with membranes of dilated vacuoles. We did not observe noticeable difference in the strength of antibody staining between control and chloroquine-treated cells. LipidTOX staining localized in the perinuclear and peripheral area of the cell. Figure 3 shows co-localization of dense bodies with LipidTOX, especially in the periphery of cells. As a control for non-specific staining, cells washed with Triton × showed no LipidTOX staining. Chloroquine caused no morphological changes in the Golgi apparatus or mitochondria (Figure 5). Figure 5 shows no co-localization, and there was no marked decrease in mitochondrial staining between control and 10-20 μg/ml chloroquine treatment. It appears that co-localized lipids may represent intracellular debris, the consequence of the dysfunctional lysosomes.
Chloroquine Effect on ARPE-19 Golgi and Mitochondria. (A-D) Mitochondria co-localization in (A-B) control and (C-D) 20 μg/ml-treated ARPE-19. There is no difference in MitoTracker staining (red) intensity, shape or localization between control (B) and treatment (D). Similarly, there were no noticeable changes in Golgi apparatus (E-H). Golgi co-localization of control (E-F) and 20 μg/ml chloroquine treatments. Golgi is stained with Golgin-97 antibody staining (green). Bar size = 25 microns.
ARPE-19 Phagocytic Pathway is Disrupted by Chloroquine
We use Western blotting of LAMP-1 and 2 (Figure 6A) and exogenous rhodamine-labeled dextran (Figure 6B) to observe disruption of chloroquine-treated autophagic and phagocytic pathways, respectively. If phagocytic function is normal, LAMP-1 and LAMP-2 protein levels should remain relatively the same despite chloroquine treatment. Quantitative examination of the Western blots was performed by densitometry [27] (ImageJ Software, available at http://rsb.info.nih.gov/ij/). Western blots in Figure 6 show marked increase of LAMP-1 in chloroquine-treated cells, especially at 10 μg/ml. LAMP-2 shows a similar, but less drastic, upregulation trend.
Phagocytic Activity and LAMP Protein. (A) Comparisons of LAMP-1 and LAMP-2 protein levels at different concentrations of chloroquine. Protein levels measured in ARPE-19 by Western blot with beta-actin as loading control. Western shows marked qualitative increase in LAMP 1-2 band size between control and 10 μg/ml chloroquine treatment, while loaded control is constant. (B) There is a significant increase in LAMP-1 and 2 (p < 0.05) at 10 μg/ml with overall upregulation trend response to chloroquine treatment. Immunoblots (n = 6) were scanned, and densitometry was performed on bands with ImageJ (NIH, Bethesda MD). Relative band density measurement was repeated several times to ensure maximum and minimum values.
Exogenous rhodamine-labeled dextran was applied to the chloroquine-treated cells. We observe a striking decrease in intracellular dextran in chloroquine-treated cells (Figure 7A). Dextran retention and uptake was quantified by comparing relative maximas with ImageJ. There is a significant decrease in intracellular dextran between the control and 10 μg/ml treatment (Figure 7B). These findings suggest either increased exocytosis or decreased phagocytic uptake.
Phagocytic Activity Measured by Dextran Uptake. (A-B) Immunofluorescence of exogenous rhodamine-labeled dextran (red) uptake with ImageJ markings (crosshairs) of counted dextran maximas. 20 μg/ml chloroquine-treated cells at 24 hours (B) showed striking decrease in fluorescent dextran uptake when compared to control (A). (C) Quantization of dextran uptake between control and chloroquine-treated ARPE-19. There is a significant (p < 0.05) decrease in relative dextran uptake in individual sampled ARPE-19 cells between control and chloroquine-treated cells. Quantitation was performed by using ImageJ, isolating cells of relative same size (~20-25 microns), and then calculating florescent maximas (tolerance = 20, based on control). Maximas were averaged and standard t-test was performed (n = 10).
H2O2 Oxidative Stress Test
Oxidative stress is one of the primary causes of AMD, and is suspected as a primary catalyst of the disease [28]. We investigated whether intracellular lipid accumulation occurs to the same extent in ARPE cells treated with H2O2. No cytoplasmic dense aggregate formation or lysosomal dilation occurred in H2O2-treated cells under phase contrast (Figure 8). There were no striking differences in LAMP-2 and LipidTOX co-localization staining patterns in control and H2O2-treated ARPE-19 cells. This suggests that aforementioned lipid buildup in ARPE-19 observed with chloroquine treatment is accelerated by induced alteration in lysosomal function, and not secondary to oxidative stress.
Oxidative Stress Test. (A) phase contrast, (B) LAMP-2 and (C) LipidTOX staining of 10 mM hydrogen peroxide-treated ARPE-19. (D) Overlay showing co-localization of neutral lipid and LAMP-2. There are no differences in cell cytoplasm (vacuoles, lipids) in phase or fluorescence between control (Fig 2 A-D) and hydrogen peroxide treatment.
The molecular mechanisms by which drusen and AMD develop are not clearly understood at this time. Drusen has been postulated to be the combination of oxidative stress, lipofuscin and POS overload lysosomal capacity [4]. Here, we investigate the possible role of lysosomal malfunction in AMD by examining the effects of chloroquine, a lysosomotropic agent, on ARPE-19 cells. Our findings detail a time-dosage toxicity curve, marked vacuolation in the cytoplasm, intracellular debris accumulation, and altered phagocytic pathways in chloroquine treated ARPE-19.
Chloroquine proved to be an apt in vitro model for observing the buildup of intracellular lipid in ARPE-19 cells. Chloroquine induced intense vacuolation of cells. A similar observation has recently been observed under similar conditions [29] and in bovine RPE under slightly hypoxic conditions [23]. Vacuolation has been shown as an indicator of altered acidification through a proton "trapping" mechanism [25] and inhibition of key lysosomal enzymes, including cathepsin D [30]. This is consistent with in vivo findings that have demonstrated that retinal aging has been associated with a decrease in lysosomal glycosidases and downregulation of cathepsin D [31]. We further confirm the validity of dilation, and thus induced inhibition of phago-lysosomes, by successful co-localization with LAMP-2. Perhaps the most striking finding was the co-localization of dense formation buildup with lipid staining. The lack of dense bodies in 3T3 cells (Figure 4) suggests that this phenomenon may be unique to cells of the retinal epithelium. RPE cells by the nature of their function to phagocytose POS are likely to be more dependent upon lysosomal activity. Aberrations in this activity may lead to the accumulation of the lipid bodies that ultimately may contribute to drusen formation. Finally, in contrast to the well-established mitochondrial-axis theory [32], we found that the accumulation of lipid, induced by chloroquine, is not seen with an oxidative stress model (Figure 8), which is possibly only a secondary factor.
Interestingly, increased LAMP-1 and 2 protein levels, as well as decreased retention/uptake of exogenous dextran, suggest that alterations in the phagocytic pathway occur after lysosomal inhibition by chloroquine. Upregulation of LAMP could indicate a number of intracellular changes. It could be a reflection of a general increase in lysosomal surface membrane size due to dilation, or an overall increase in lysosomes. Furthermore, it may indicate a compensatory upregulation in phagosome-lysosomal fusion and movement [33]. It is possible that the striking increase in LAMP-1 is an indication of the onset of apoptosis, an observation noted in glioblastoma cells in culture [34]. In contrast, chloroquine at low dosages does not seem to induce increased autophagy, as autophagic-associated vacuolation is correlated with decreased LAMP-2 levels [35]. At the time of publication, we have initiated studies on siRNA knockdown of LAMP protein to eludicate the mechanism of these protein changes in light of vacuolation.
Our exogenous dextran findings suggest either a lack of intracellular retention or increased exocytosis due to lysosomal inhibition. There is much debate on the mechanisms that occurs in AMD. There is a belief that the combination of oxidative stress and a decrease in ATP drives the exocytosis of partially digested intracellular proteins into the Bruch's membrane [8]. However, recent studies have investigated the role of ABCA4 transporter mutations in reducing movement of oxidized lipids in Stargardt's AMD [36]. Similarly, it has been shown that oxidized lipid-proteins can block phagosome formation, and thus deter internalization into RPE [37].
Another candidate for the cause of lysosomal dysfunction in cells is the degradation of the glycocalyx, which protects the vital LAMP-2 membrane protein. While the exact role of LAMP-2 is still unclear, it has been proposed that it is responsible for equilibrating proton levels in lysosomes. Moreover, recent studies suggest LAMP-2 may be critical in communication between phagosomes and lysosomes, and in receptor-targeting for lysosome maturation [38–40]. We therefore postulate a "LAMP-2/Lysosomal Inhibition" AMD biogenesis model (Figure 9). In this postulated model, lysosomal inhibition begins with the modification or degradation of the asparagine-linked glycocalyx of LAMP-2. Removal of the glycocalyx results in rapid proteolysis of LAMP-2 [41]. The glycocalyx shield could be slowly modified or degraded over time by the powerful proteases within the lysosomal lumen, perhaps as a result of aging. Further investigation will be required to understand the causes of the breakdown of the glycocalyx. Nevertheless, after LAMP-2 has been degraded, a pH shift occurs, dynein is no longer able to pull mature phagosomes to lysosomes or mistargeting of M6PR/Rab7 occurs [42]. Consequently, the lysosome is incapable of degrading extra- and intracellular material by phagolysosomal fusion. This results in the accumulation of oxidized lipofuscin. The combination of mitochondrial oxidative stress and amassing of senile, inefficient mitochondria and iron in lysosomes further expedites lipofuscin buildup.
Possible LAMP-2/Lysosomal Inhibition Model of Pathogenesis of AMD. An unknown cause results in loss of the protective glycocalyx of LAMP-2. Proteolysis of LAMP-2 occurs. Loss of LAMP-2 results in either 1) A pH shift and loss of acidity of lysosome, 2) dynein no longer moving late phagosome to microtubule sorting center near Golgi for fusion with lysosome or 3) perturbation of M6PR/Rab recycling such that M6PR/Rab7 does not tag late endosomes. The loss of functionality of M6PR/Rab7 results in a lack of phagosome lysosome-fusion. Any of these results in loss of the lysosome's ability to degrade intra- and extracellular material. Subsequently, undegraded material is oxidized, turning to lipofuscin. Ultimately, the combination of oxidative stress, Fe+ accumulation, senile mitochondria and decrease in ATP results in inefficient turnover of organelles and increased inhibition of lysosomes. RPE death triggers photoreceptor and macula death.
In summary, these studies demonstrate two new findings. First, we show the morphological changes in ARPE-19 due to chloroquine. Also, we demonstrate that lysosomal dysfunction leads to the accumulation of lipids and their byproducts in ARPE-19 cells which may simulate the situation in AMD. These findings seem to be specific for this particular cell line, and we demonstrate that despite lysosome dilation with chloroquine treatment, this profuse lipid accumulation is not seen with NIH 3T3 cells (Figure 4), neuroblastoma or glioblastoma cell lines that we have tested. This is perhaps a reflection of the intense lysosomal/autophagocytic activity of the RPE cells. These cells may be more susceptible to perturbations of the lysosomal degradation systems and are thus a good model for studying potential mechanisms of AMD. There is a piqued interest in the relationship between lysosomal inhibition and AMD as seen in the recent literature on the correlation between iron overload and reduced Cat-D activity [43], the conversion of protease substrates to lysosomal enzyme substrate inhibitors [44] and the possible usage of toxins to elevate low pH in ARPE lysosomes [45].
These studies demonstrate the dose and time dependent response of ARPE-19 to chloroquine. Moreover, we have demonstrated that chloroquine induces intracellular vacuolation and lipid accumulation, and disrupts the phagocytic pathway. Our observations appear to be independent of oxidative stress. Lysosomal malfunction may thus have a pivotal role in the pathogenesis of AMD.
Cell Culture Growth Technique
Human ARPE-19 tissue culture cells were obtained from American Tissue Culture Center, Rockville, MD (ATCC, CRL-2502). Cells were grown in Ham's F12 Media: DMEM (1:1) (ATCC, 30-2006) with 10% fetal bovine serum (FBS) (ATCC, 30-2030) as monolayers at 37°C with 5% CO2 in 100 × 20 mm round culture dishes (Falcon, 35-3003). NIH/3T3 cells, embryonic mouse fibroblast cells, were also obtained from ATCC (CRL-1658) and grown in DMEM (ATCC, 30-2002) with 10% calf bovine serum (ATCC, 30-2030) at the same conditions as ARPE cells. The cells were propagated by splitting at a ratio of 1:5 with a trypsin-EDTA solution. Studies were done on both ARPE-19 and 3T3 cells during passages 5-15.
Chloroquine was obtained from Sigma (C6628, St. Louis, MO). Chloroquine stock solutions were made at 10 mg/ml in double deionized water, and sterilized by filtration with an Acrodisc syringe filter with a 0.2 μm Tuffryn membrane (Pall Life Sciences, New York). Aliquots of the sterile stock solutions were stored at -20°C until use. The appropriate amounts of stock solution were added to the media to achieve the desired concentration. To test the effect of the carrier, equivalent volumes of sterile water were also added to a set of ARPE-19 cells.
Response of ARPE-19 Cells to Chloroquine
Approximately 50,000 cells were seeded into each well of a 12-well plate (Falcon 35-3043) and grown to 50% confluency on sterile glass coverslips. These were incubated with chloroquine at concentrations of 0, 10, 20, 40, 80 and 100 μg/ml for 24 hours. In a separate set of experiments, cells were incubated at 0, 40 and 80 μg/ml for 2 hours and 6 hours. To demonstrate that the carrier did not affect the ARPE-19 cells, sterile water, in a quantity equivalent to that of the carrier, was added to a triplicate set of RPE cells. Each point on the graph in Figure 1 represents seven independent experiments. The loss of cells after the different chloroquine treatments was determined by counting the cells on the coverslip. To visualize the cells, 4'-6-Diamidino-2-phenylindol (DAPI) was used to stain the cell nuclei. After the designated treatment, coverslips were fixed in situ in 3.7% paraformaldehyde. Coverslips were washed three times with 1X phosphate buffered saline (PBS), and then once with PBS containing 1 μg/ml DAPI and 0.1% saponin. The stained cells were visualized through a DAPI filter with a 20× panfluorochrome objective with a BX50 Olympus microscope. Ten random fields from each coverslip were photographed and counted.
An MTT assay kit was obtained from Roche (11465007001). Approximately 5,000 cells were seeded into each well of a 96-well plate, and were grown to ~50% confluency. Chloroquine treatments of 20, 30, 40, 50, 60, and 80 μg/ml were administered in 100 μl of media/well, and cells were incubated for 24 hours at 37°C. After this first incubation period, 10 μl of MTT labeling reagent was added to each well, bringing the final concentration of MTT labeling reagent to 10 mg/ml. The cells were incubated for 4 hours. At this point, 100 μl of the solubilization solution was added to each well, and the plate was incubated overnight. After this last incubation period, the spectrophotometric absorbance of the samples was measured using an ELISA reader.
LAMP-2 and Cy2 Secondary Antibodies
Anti-human LAMP-2 monoclonal antibody (anti-mouse IgG1) was obtained as stock hybridoma tissue supernatants from The Developmental Studies Hybridoma Bank (University of Iowa) and diluted (1:50) in a 0.1% Bovine Albumen Serum (BSA)/0.1% saponin/PBS solution. The secondary goat-anti-mouse cy2 or rhodamine antibody (Kirkegaard and Perry, Gaithersburg, MD) was diluted (1:60) with 0.1% BSA/0.1% saponin/PBS.
LAMP-2 Immunofluorescent Staining
RPE and 3T3 cells were stained for LAMP-2 as previously described [46].
Neutral Lipid Stain
Stock HCS LipidTOX Red Neutral Lipid Stain (Invitrogen, H34475) was diluted 1:4000 in PBS. Coverslips with LipidTOX were incubated for 15-20 minutes at room temperature. LipidTOX was then removed and cells were washed once with 1X PBS. To test LipidTOX specificity, cells were washed with 0.1% Triton X-100 in PBS prior to staining with LipidTOX. Triton X-100-washed cells functioned as a control, demonstrating no staining with the LipidTOX.
Epifluorescence and Phase Contrast Microscopy
An Olympus BX50 microscope with phase contrast and epifluorescent capabilities, attached to an Olympus DP11 or DP20 digital camera, was used for DAPI (20×) images. This was also used for phase contrast and fluorescent-labeled studies (60×, oil immersion).
Mitochondria Staining
MitoTracker CMX Ros Red (Invitrogen, M7512) was dissolved in dimethyl sulphoxide (DMSO [Sigma]) to achieve a final stock concentration of 1 mM. Cells were grown to 50% confluency and treated with chloroquine concentrations of 10, 20 and 40μg/ml. To stain the mitochondria, cells were incubated for 15 minutes at 37°C in fresh media with MitoTracker at a concentration of 300 nM, and then fixed with 3.7% paraformaldehyde. Cells were washed in 1× PBS three times, viewed with epifluorescence, and photographed as previously described.
Golgi Stain
Anti-Golgin 97 mouse IgG1 (Invitrogen, 49363A) was dissolved in 1× PBS to create a final concentration of 200 μg/ml. The cells were incubated with anti-golgin 97 and the LAMP-2 antibodies, both diluted 1:50 in 0.1% BSA/0.1% saponin/PBS. The Golgi stain was visualized with rhodamine-labeled goat anti-mouse secondary antibody (Kirkegaard and Perry, Gaithersburg, MD).
Western blots were performed with modifications for LAMP-2 [39]. LAMP-2 was administered at a concentration of 1:2000, followed by incubation with goat anti-mouse peroxidase-conjugated secondary antibodies (BioRad, 1:3000). Antigen-antibody complexes were detected using chemiluminescence reagent kit (Perkin-Elmer ECL). The blots were stripped and then probed with actin to function as a control.
H202 Oxidative Stress Study
A stock concentration of 3% hydrogen peroxide (H2O2, 0.88 M) was diluted in the growth media to attain final concentrations ranging from 0.2 to 10 mM [28]. The treated cells were incubated for 24 hours. Following incubation, cells were visualized by phase contrast microscopy and stained with LAMP-2, LipidTOX, Anti-Golgin 97, and MitoTracker.
Dextran Uptake Studies
Rhodamine-conjugated dextran, avg. 10,000 MW, was obtained from Invitrogen. A stock solution of dextran diluted in sterile, deionized water was prepared. Cells were grown in different concentrations of chloroquine on round glass coverslips in 24-well plates for 24 hours. After 24 hours of incubation, the media was removed and replaced with new media containing the same amount of chloroquine, as well as an additional 0.05 mg/ml of rhodamine-conjugated dextran. At this point, the cells were incubated for 6 hours, and then fixed in 3.7% paraformaldehyde in PBS. Cells were mounted in a 10% glycerol-PBS solution, visualized, and photographed immediately.
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Eskelinen EL: Roles of LAMP-1 and LAMP-2 in lysosome biogenesis and autophagy. Mol Aspects Med. 2006, 27: 495-502. 10.1016/j.mam.2006.08.005
Huynh K, Eskelinen E, Scott C, Malevanets A, Saftig P, Grinstein S: LAMP proteins are required for fusion of lysosomes with phagosomes. Embo J. 2007, 26: 313-324. 10.1038/sj.emboj.7601511
Saftig P, Klumperman J: Lysosome biogenesis and lysosomal membrane proteins: trafficking meets function. Nature. 2009, 10: 623-635.
Kundra R, Kornfield S: Asparagine-linked oligosaccharides protect Lamp-1 and Lamp-2 from intracellular proteolysis. J Biol Chem. 1999, 274: 31039-31046. 10.1074/jbc.274.43.31039
Luzo J, Pryor P, Bright N: Lysosomes: fusion and function. Nature. 2007, 8: 622-632.
Chen H, Lukas T, Du N, Suyeoka G, Neufield A: Dysfunction of retinal pigment epithelium with age: increased iron decreases phagocytosis and lysosomal activity. Invest Ophthalmol Vis Sci. 2009, 50: 1895-1902. 10.1167/iovs.08-2850
Krohne T, Kaemmerer E, Holz F, Kopitz J: Lipid peroxidation products reduce lysosomal protease activities in human retinal pigment epithelial cells via two different mechanisms of action. Exp Eye Res. 2009, 90: 261-266. 10.1016/j.exer.2009.10.014
Liu J, Lu W, Reigada D, Nguyen J, Laties AM, Mitchell CH: Restoration of lysosomal pH in RPE cells from cultured human and ABCA4-/- mice: pharmacologic approaches and functional recovery. Invest Opthalmol Vis Sci. 2009, 49: 772-780. 10.1167/iovs.07-0675. 10.1167/iovs.07-0675
Goldenthal KL, Hedman K, Chen J, August J, Willingham M: Postfixation detergent treatment for immunofluorescence suppresses localization of some integral membrane proteins. J Histochem Cytochem. 1985, 33: 813-820.
We thank Dr. Zhi-Gang Xiong (Dow Neurobiology laboratory, Legacy Holladay Park Research Center, Portland, Oregon) for helpful discussions and technical assistance with instrumentation and Mr. Jeff Gadette (Oregon Episcopal School (OES), Portland, Oregon) for his advice on statistics. Furthermore, we thank Dr. Robert Maue (Dartmouth College, Hanover, NH), Dr. Bill Lamb (OES), Mr. Peter Langley (OES), and Dr. Tanja Horvat (OES) for critiquing this manuscript. Funding for this study was obtained in part from grants from the Oregon Research and Education Foundation and The Legacy Foundation Research Fund.
Department of Neurological Surgery, Legacy Clinical Research and Technology Center, 1225 NE 2nd Ave., Portland, OR, 97232, USA
Patrick M Chen
, Zoë J Gombart
& Jeff W Chen
Department of Biological Sciences, Dartmouth College, 103 Gilman Hall, Hanover, NH, 03755, USA
Search for Patrick M Chen in:
Search for Zoë J Gombart in:
Search for Jeff W Chen in:
Correspondence to Jeff W Chen.
PC and JC conceived, designed and carried out these experiments. PC performed analysis and drafted manuscript. ZG carried out experiments and performed statistical analysis. All authors read and approved the final manuscript.
Chen, P.M., Gombart, Z.J. & Chen, J.W. Chloroquine treatment of ARPE-19 cells leads to lysosome dilation and intracellular lipid accumulation: possible implications of lysosomal dysfunction in macular degeneration. Cell Biosci 1, 10 (2011) doi:10.1186/2045-3701-1-10
Retinal Pigment Epithelium
Retinal Pigment Epithelium Cell
Chloroquine Treatment
Lysosomal Dysfunction


text:
Elon Musk unveils his test car tunnel as a fix for traffic in L.A.
Dennis Romero
Dec 19th 2018 5:10AM
LOS ANGELES — Billionaire Elon Musk debuted a long-awaited test tunnel on Tuesday to demonstrate technology that he said could revolutionize transportation in what is often called the most freeway-congested city in the nation.
The less than 2-mile tunnel took VIPs and journalists on rides in a Tesla electric vehicle with horizontal wheels added to the chassis in order to keep the vehicle aligned on a track. While riders were taken at speeds of about 50 to 60 miles per hour, Musk said the tunnel would ultimately be good for speeds of 150.
The idea would be to use driverless vehicles to make the trip in the quiet and comfort of the underworld.
The tunnel has been nearly two years in the making — it was initially approved by state officials as a subterranean walkway — and was the brainchild of Musk after he complained in 2016 about his "soul-crushing" commute from Bel Air to his rocket company SpaceX in Hawthorne.
The Loop system from his adjacent Boring Company startup, once up and running, would take vehicles underground on an elevator and then send them zipping along a track to their destination.
An earlier concept of public-transportation-like "skates" based on Tesla chassis was abandoned, apparently, in favor of regular Teslas with the enhanced wheels. Those who tried the tunnel described a very bumpy ride at less than optimum speeds.
Elon Musk's tunneling company unveiled its 1.14-mile tunnel in Los Angeles County tonight, and I rode through it in a Tesla SUV. It was a bumpy ride! More here, check back for updates: https://t.co/JBEy6e8NcZpic.twitter.com/8sSUGGwDM5
— Laura J. Nelson 🦅 (@laura_nelson) December 19, 2018
During a presentation, Musk called his project "a little rough around the edges."
But he continued to make big promises, even as another planned tunnel near the dreaded 405 freeway was recently nixed as part of a settlement with neighborhood groups that sued the city for fast-tracking its environmental review process.
Musk said Boring's technology will eventually be able to dig tunnels 15 times faster than contemporary methods.
Musk's demonstration Tuesday included the entrepreneur's usual trappings for an unveiling, like nightclub-style lighting and music, but it left many questions remaining, including how a one- or even two-way tunnel of that size would alleviate much traffic in Los Angeles, where about 20,000 vehicles per hour pass the Boring Company in Hawthorne along the 405 freeway, according to California Department of Transportation data.
Also, the tunnel's width, and the Tesla's use of horizontal wheels that tuck under the vehicle, appeared specific to Musk's own automaker. And the elevators used could create choke points that simply mean more traffic, experts have said.
"New capacity has never helped with congestion and never will," said James Elliott Moore II, director of the Transportation Engineering Program at the University of Southern California.
Elon Musk unveils rockets to fly people around Earth
TOPSHOT - Billionaire entrepreneur and founder of SpaceX Elon Musk speaks below a computer generated illustration of his new rocket at the 68th International Astronautical Congress 2017 in Adelaide on September 29, 2017. Musk said his company SpaceX has begun serious work on the BFR Rocket as he plans an Interplanetary Transport System. / AFP PHOTO / PETER PARKS (Photo credit should read PETER PARKS/AFP/Getty Images)
ADELAIDE, AUSTRALIA - SEPTEMBER 29: Elon Musk during his presenation at the Tesla Powerpack Launch Event at Hornsdale Wind Farm on September 29, 2017 in Adelaide, Australia. Tesla will build the world's largest lithium ion battery after coming to an agreement with the South Australian government. The Powerpack project will be capable of an output of 100 megawatts (MW) of power at a time and the huge battery will be able to store 129 megawatt hours (MWh) of energy. Tesla CEO Elon Musk has promised to build the Powerpack in 100 days, or he will deliver it for free. (Photo by Mark Brake/Getty Images)
Billionaire entrepreneur and founder of SpaceX Elon Musk speaks in below a computer generated illustration of his new rocket at the 68th International Astronautical Congress 2017 in Adelaide on September 29, 2017. Musk said his company SpaceX has begun serious work on the BFR Rocket as he plans an Interplanetary Transport System. / AFP PHOTO / PETER PARKS (Photo credit should read PETER PARKS/AFP/Getty Images)
Billionaire entrepreneur and founder of SpaceX Elon Musk speaks at the 68th International Astronautical Congress 2017 in Adelaide on September 29, 2017. Musk said his company SpaceX has begun serious work on the BFR Rocket as he plans an Interplanetary Transport System. / AFP PHOTO / PETER PARKS (Photo credit should read PETER PARKS/AFP/Getty Images)
Billionaire entrepreneur and founder of SpaceX Elon Musk speaks below a computer generated illustration of his new rocket at the 68th International Astronautical Congress 2017 in Adelaide on September 29, 2017. Musk said his company SpaceX has begun serious work on the BFR Rocket as he plans an Interplanetary Transport System. / AFP PHOTO / PETER PARKS (Photo credit should read PETER PARKS/AFP/Getty Images)
ADELAIDE, AUSTRALIA - SEPTEMBER 29: SpaceX CEO Elon Musk speaks at the International Astronautical Congress on September 29, 2017 in Adelaide, Australia. Musk detailed the long-term technical challenges that need to be solved in order to support the creation of a permanent, self-sustaining human presence on Mars. (Photo by Mark Brake/Getty Images)
ADELAIDE, AUSTRALIA - SEPTEMBER 29: Sunsets on a wind turbine on the way to Tesla Powerpack Launch Event at Hornsdale Wind Farm on September 29, 2017 in Adelaide, Australia. Tesla will build the world's largest lithium ion battery after coming to an agreement with the South Australian government. The Powerpack project will be capable of an output of 100 megawatts (MW) of power at a time and the huge battery will be able to store 129 megawatt hours (MWh) of energy. Tesla CEO Elon Musk has promised to build the Powerpack in 100 days, or he will deliver it for free. (Photo by Mark Brake/Getty Images)
ADELAIDE, AUSTRALIA - SEPTEMBER 29: Tesla Powerpack batteries during Tesla Powerpack Launch Event at Hornsdale Wind Farm on September 29, 2017 in Adelaide, Australia. Tesla will build the world's largest lithium ion battery after coming to an agreement with the South Australian government. The Powerpack project will be capable of an output of 100 megawatts (MW) of power at a time and the huge battery will be able to store 129 megawatt hours (MWh) of energy. Tesla CEO Elon Musk has promised to build the Powerpack in 100 days, or he will deliver it for free. (Photo by Mark Brake/Getty Images)
The professor said congestion pricing — charging commuters according to the density of traffic — is the only way to solve the problem Musk is tackling.
"If his real goal is to reduce congestion, I would say focusing on a way to make it simple to assess congestion tolls would move ahead further than boring tunnels," he said.
Boring Company has loosely proposed a network of tunnels beneath Los Angeles — it's not clear if public money would be involved — and his Dugout Loop plan to bring baseball fans to Dodger Stadium via a tunnel funded privately is in the environmental review stage.
More from NBC News:
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When I was in school, writing was not my strong points. I get lazy sometimes that I asked classmates to either copy something for me or I borrow their notes to copy. Until the scanner software was made available for public use. Oh, how I loved the fact that all I needed to do was have a few cents so I could photocopy notes and pages off a book for me to study. No more writing unless it is something that is imparted by the teacher from her own research. It was cool! To this day, I still have said photocopies that I accumulated through the years. I did have books but there were reference materials we could only get from the library and that was what I had copied and saved after all these time. I actually have boxes and boxes of them stacked in my room at my parents house. They do not want to get rid of any items I left behind which is sweet. Right now, the husband and I have our own printer/scanner we use for documents. It is handy and is good for when we need copies of documents for our own personal records.


One of a monthly promotional series put out by Capitol Records. Each promotes various Capitol albums that are new for the month. Each side carries its own promo number.
The jackets for this series include thumbnails for each album being promoted, plus extensive narrative. The catalog # for each album is also listed.
The May, 1959 issue may be most known for track B6 "Hey Miss Lucy" by Esquerita.


Motorcycle Santa Wins the Photo Contest!
Congrats to Phil Thompson (a.k.a. Motorcycle Santa) for winning our Holiday Photo Contest with 303 votes! On GigMasters, he's listed as Phil Thompson Pianist & Vocalist, Duos, Trios & DJ and now we know he's a fast-riding mobile Santa as well.
To view the results (and to see all of the great pics) visit our contest page on Facebook. There were 906 votes total and it was a close match all the way to the end. And a special shout-out to The Hanson Family Singers for their strong finish in second place with 263 votes.
Thanks to everyone who entered and voted! Stay tuned to Inside the Gig for more contests.


Victor AD & Erigga teams up to deliver a new track tagged Why stream and download mp3 below.
Victor AD makes a new song featuring Erigga tagged Why.
The jam came after there dropped "Motivation" last year.
Cop the jam below. Download mp3 Victor AD ft Erigga – Why.
Stream and download Victor AD ft Erigga – Why.


Results of the photographic competition Theatre night, Month of photography, contest lasted from 9th till 20th november 2011.
Cultural centre Dubrava second year in a row successfully hosted the project Month of Photography, which lasted throughout the month of november and includes a number of forums and photo workshops, photo exhibitions and fair od fotoequipment and photography.
During this project we organized a foto competition named Theatre night that lasted from 9th till 20th november and brought together 45 participants from different countries.
Five-member committee has decided that this year's winning photo contests Theatre Night belongs to the author Julien Duval. It is interesting that all the committee members, the maximum amount of points, assigned to the same author but different photos, so we can boast Julien Duval as a complete and very good photographer. Equal praise, with some minor points, go to the authors Christoph Ullman, Sonya Lukas and Martina Mijić.
Also, we thank all the applicants and hope to see you again at Month of photography!


text:
BlackBerry Limited, tidligere kendt som Research In Motion (RIM), (, ) er en canadisk multinational telekommunikationsvirksomhed med hovedsæde i Waterloo, Ontario, Canada som designer, producerer og sælger trådløse løsninger på mobiltelefonimarkedet. BlackBerry tilbyder platforme og løsninger for adgang til informationer via e-mail, tale, instant messaging, SMS, Internet- og intranetbaserede applikationer og browsing. RIM's portefølge inkluderer BlackBerry, håndholdte enheder, softwareudviklingsværktøjer og øvrig soft- og hardware. Virksomheden blev etableret af Mike Lazaridis, som i dag fungerer som co-CEO sammen med Jim Balsillie. Virksomheden er i Danmark mest kendt for smartphonen BlackBerry, der kom på markedet i 2002.

Historie
Før produktionen af BlackBerry arbejde RIM sammen med RAM Mobile Data og Ericsson på at lave det Ericsson-udviklede Mobitex trådløse netværk til en tovejs personsøger og trådlås e-mail computernetværk. Omdrejningspunktet for denne udvikling var udgivelsen af Inter@ctive pager 950, som kom på markedet i 1998. Personsøgeren var omtrent på samme størrelse som et stykke håndsæbe og konkurrerende imod det af Motorola udviklede SkyTel tovejs personsøgernetværk.

RIM's tidlige udvikling blev finansieret af institutionelle investorer og venturekapitalister fra Canada. Working Ventures Canadian Fund Inc. led the first venture round. Virksomheden blev noteret på Toronto Stock Exchange i Januar 1998 med symbolet RIM.

Vækst
RIM var været igennem en stor vækst efter årtusindskiftet og har i dag omkring 10 % af smartphonemarkedet (maj 2010)Desuden er virksomheden med en andel på 3% verdens sjettestørste fabrikant på markedet blandt samtlige mobiltelefonproducenter.

Referencer

Eksterne henvisninger

Research In Motion website
ValueWiki Research in Motion Profile
RIM profile at Google Finance

Etableret i 1984
Ontario
Elektronikvirksomheder fra Canada
Mobiltelefonproducenter
Selskaber noteret på NASDAQ
Telekommunikationsvirksomheder fra Canada
Blackberry


text:
PS5 VS Xbox Series X: Which One Is Better?
by Geralt of Sanctuary
The PlayStation 5. The Xbox Series X.
The next generation of console gaming is upon us. As is our sacred duty, we must now compare the two in a variety of very important scientific categories: Size. Design. Molecular density. Usefulness at a picnic.
And of course, the dreaded essay section. Which next generation console reigns supreme? Let's find out.
Xbox Series X Vs. PS5 Round One: Design
First impressions are everything. Before the power goes on and the games are downloaded and we wait for the day one patches, we see the fruits of months of hard work from designers and engineers. A game console is something that sits on our shelves for years, gathering dust and harboring cockroaches until smaller, more sensible models are released.
Xbox Series X: Sometimes the design process can be pretty simple. For the original Xbox, Microsoft made a box shaped like an X. Over the years the company has slowly evolved that design into the Series X, a box with an X on it. One has to admire the simplicity. This dark monolith for ants commands respect, fealty, and perhaps regular offerings of fruit and fresh meat.
Spaceship.
Photo: Sony
PlayStation 5: After decades of PlayStation consoles that look nothing like any sort of station, Sony finally gives us a game console design worth of the starship Enterprise, or at least one of the ships from Babylon 5. If we're being honest, it's as if Sony's engineers attempted to out-prototype those wacky internet kids who mock up their dream PlayStation models every console generation.
Winner: Xbox Series X
Vertical or horizontal, the Xbox Series X is ready to do some serious gaming business. The PS5 is drunk. It thinks it looks cool, but I keep looking over at it, wincing, and shaking my head. Also it makes me crave Oreo tacos, which are not a thing. Also it's too damn big.
Xbox Series X Vs. PS5 Round Two: Price
How much should a powerful new video game console cost? Spurious science says the answer is somewhere in the $300 to $500 range.
Xbox Series X: $500
PS5 (with optical disc): $500
Total: $1,000
Winner: People With $500 Who Were Able To Secure A Preorder
When it comes to buying things, the true winners are the money-havers. Note that these results do not take into account the sub-models of the Xbox Series X and PS5.
Xbox Series X Vs. PS5 Round Three: Sub-Models
Photo: Sony / Microsoft
While evaluating the Xbox Series X and PS5 it's important we don't lose sight of both consoles' optical drive-free counterparts: the smaller, cheaper, less powerful Xbox Series S, and the slightly skinnier digital PS5.
Xbox Series S: It's small, it's white, its profile's pretty tight. For the first time ever in a console launch, customers can opt to spend $200 less than they would for the $500 Xbox Series X for a less powerful version of the hardware. It's such an iPhone move. While I've not had the pleasure of playing with the Series S personally, I am not averse to the idea. So what that it doesn't do 4K games.
PS5 Without An Optical Drive: This system features the full power of the PS5 without the optical drive for $100 less than the PS5's $500 asking price. This teaches us that a 4K Blu-ray drive costs around $100, while making us ask ourselves how much we really need such a drive.
Winner: Xbox Series S
As strange as it is to have two console models with such a large power discrepancy at launch, the value proposition of the Xbox Series S is clear, and its $300 pricing makes it a much more attractive option than the missing slit PS5.
Xbox Series X Vs. PS5 Round Four: Power
Unless you are attempting to build a sturdy, load-bearing wall, the exterior design of a video game console is nowhere as important as what's on the inside, not including living and/or dead insects.
Xbox Series X: Powered by a custom eight-core AMD Zen 2 CPU, 16GB of GDDR6 memory, and a custom AMD RDNA 2 GPU, the Xbox Series X is on the cutting edge of console gaming technology. Add exciting new super-speedy SSD storage and you've got a recipe for one hell of a gaming device.
PS5: Powered by a custom eight-core AMD Zen 2 CPU, 16GB of GDDR6 memory, and a custom AMD RDNA 2 GPU, the PS5 is on the cutting edge of console gaming technology. Add exciting new super-speedy SSD storage and you've got a recipe for one hell of a gaming device.
While the Xbox Series X and PlayStation 5 share a lot of similar components, Microsoft's machine edges out Sony's in terms of pure power. The Xbox's Zen 2 CPU runs at 3.8GHz to the PS5's 3.5GHz. The PS5 graphics processor sports 10.28 teraflops of power to the Xbox's 12 teraflops. The Xbox Series X and PS5 have the same amount of memory, but the Xbox accesses its memory much faster with more bandwidth. Sony's machine is more versatile, adjusting CPU and GPU frequency as needed, but in terms of raw power, Microsoft comes out on top.
Xbox Series X Vs. PS5 Round Five: Taste
"How does my game console taste?" is one of the most important questions a person can ask, as it often leads to console licking, which is the best. Go on, lick it.
Xbox Series X: Tastes like plastic.
PS5: Also tastes like plastic. Weird.
Winner: PlayStation 5
With both plastic consoles tasting strangely of plastic, this particular criteria comes down to mouthfeel, as with many contests in life. While the Xbox Series X does some interesting things with textures, treating the tongue to the pure delight of all the little vent holes, it's largely uniform. The PS5's grayish-white wings have a similarly rough plastic feel, but the black bits in the middle are shiny, polished plastic. It's like a playground for the tongue.
Xbox Series X Vs. PS5 Round Six: Load Times
Determining the best-tasting console is easy; which console has the best loading times is a bit more complex. Both the PlayStation 5 and Xbox Series X utilize a combination of incredibly fast solid-state drives and creative data and compression management to speed up game and game asset loading to speeds never-before-seen from a gaming console. Both consoles rely on games specifically optimized to utilize these new technologies in order to fully benefit from them, but older, backward-compatible PS4, Xbox One, Xbox 360, and Xbox games should see some sort of benefit as well.
It's a bit hard to compare new games, as the games benefiting most from the load time reductions are platform or publisher exclusives. Forza Horizon 4 loads in under 30 seconds now on the Xbox Series X, but that means nothing to a PS5 player. Likewise, new PS5 owners are sure to be impressed by how Insomniac's Miles Morales transitions from start screen to gorgeous open-world New York City in a few seconds, but Xbox Series X owners can't play that.
Instead, we have to look at how fast legacy games load. Or, to be more precise. We did test out The Witcher 3 on both the PS5 and Xbox Series X, with the Sony console several seconds faster, but GameSpot tested a bunch more. They loaded a number of older games, including Final Fantasy XV, Red Dead Redemption 2, and Destiny 2, none of which have received next-gen performance patches as of yet. In almost all cases, the Xbox Series X hard drive performed faster than the PS5's. Sometimes the differences are miniscule. The Xbox Series X makes it to the Batman: Arkham Knight menu one second faster than the PS5. Not a big deal. Other times the gap is bigger, like Monster Hunter: World taking 23 seconds to load on Xbox to the PS5's 32 seconds.
Both consoles' loading speeds are impressive, especially with new or newly-enhanced games. The Xbox Series X's are just slightly more impressive.
Xbox Series X Vs. PS5 Round Seven: New Games
Screenshot: Sony
This one's going to be pretty easy, not going to lie. Feel free to skip down to the PlayStation 5 winning part.
Xbox Series X: There are no new, console-exclusive games on the Xbox Series X. There are a couple of next-generation exclusives: Yakuza: Like a Dragon won't be on PS5 until March. Indie shooter Bright Memory is still on track to make its console debut on Xbox. And Gears Tactics counts, if you're into that sort of thing. However, the Xbox's biggest potential system-seller, Halo Infinite
, slipped to 2021 earlier this year, while horror exclusive The Medium was pushed to next year just this morning.
PS5: Marvel's Spider-Man: Miles Morales. Also, Bugsnax. And Demon's Souls.
Oh hey, and there's that new Sackboy game.
Xbox Series X Vs. PS5 Round Eight: Usefulness At A Picnic
Surprised? I said it right in the lede. One day you'll be at a picnic with a game console and you'll thank me for this.
Right off the bat let's eliminate points for holding down the picnic blanket on a windy day, as that's something both consoles can do. Also we've already covered sitting on either console. The PS5 and Xbox Series X are both equally effective in warding off rabid bears, which is to say not at all (apologies to the families of our test subjects).
Xbox Series X: Our most useful imagined use for an Xbox Series X at a picnic would involve having the console connected to power, preferably while running a demanding game. The heat that rises from the system's top vents is not enough to cook food, but is certainly enough to keep it warm. Plug the Series X into a long extension cord, fire up a purely hypothetical (until embargo) Assassin's Creed Valhalla, and you've got the most powerful burger and dog warmer this side of an exploding Samsung Galaxy Note 7.
PS5: You're never lacking serving platters if there's a PS5 at your picnic. Pry off those side panels and fill them with chips, your favorite dip, macaroni or potato salad, or a delicious, fluffy pile of marshmallowy ambrosia. For a non-food use, stand the console up and let the kids use it as a slide.
Beneath the trees, where nobody sees, they'll trace some rays, and load games with ease, and that's the way the PlayStations have their picnic.
Xbox Series X Vs. PS5 Round Nine: Innovation
The Xbox Series X and PlayStation 5 are both incredibly powerful video game playing devices, but what else do they do? Being more powerful than the older one is nice, but what do these consoles bring to the table that's new, besides faster loading screens?
Xbox Series X: The Xbox Series X does everything an Xbox One X does, only better and faster, but it doesn't do a lot new. The controllers are slightly redesigned, but overall feel familiar. The games run a bit faster and at higher resolutions, but nothing completely transformative. The closest thing the Series X and S have to a game-changing feature is Quick Resume, literally a game-changing feature that allows players to pause multiple games at the same time, picking up right where they left off upon returning. When it works, it's amazing, but it's not exactly a must-have next-generation feature.
PS5: If all the PlayStation 5 had in terms of innovation was the new DualSense controller, with its powerful haptic feedback and triggers capable of providing degrees of resistance against players' thumbs, that would be enough. I can't wait to see reactions on the internet once new PS5 owners get their hands on Astro's Playroom, the free pack-in game built to highlight the new controller tech. I had a smile on my face and a tingling in my hands the entire time I played.
The PlayStation 5 is also doing something new with its dashboard, integrating game tips and challenges into new titles. In supported games players will be able to hit the PlayStation button on their controller to bring up a list of tasks or unattained trophies. In some cases clicking on them will transport the player to exactly the spot in the game they need to complete said challenges or achievements. We've not had much experience with this feature during our time with Sony's review units, but I'm excited to see what comes of it.
The Xbox Series X makes games play better. The PlayStation 5 has the potential to make them feel better.
Xbox Series X Vs. PS5 Round Ten: Store And Media Support
In these days of always-connected consoles with options for foregoing optical media entirely in favor of downloading games and streaming movies directly from the internet, talking about how games are sold and media is streamed is very important.
Xbox Series X: The Series X uses the same store and media apps as the Xbox One. They're fine. It can get a little confusing knowing which versions of games you're looking at now that there is a new Series X layer added, but we'll figure it out.
Not pictured: The PS5 store.
Screenshot: Microsoft / Kotaku
PS5: We're not allowed to talk about any of this yet. I'd explain why, but we're not allowed to talk about that either.
Winner: We Can't Say
We'd love to, but we can't.
Xbox Series X Vs. PS5 Round Eleven: Backward Compatibility
How are we on eleven? Science said we'd only have to do five tops, followed by fruit juice and Nutter Butters, dammit. Fine. Let's talk about backward compatibility, AKA the vast majority of the PlayStation 5 and Xbox Series X launch libraries.
Xbox Series X: Every game that runs on the Xbox One will run on the Xbox Series X, except for Kinect games and face it, if you're still playing Kinect games the government is probably watching you play them. That includes just about every Xbox One game, plus all of the Xbox 360 and original Xbox games that are playable on the Xbox One via its backward compatibility. That also includes all games on the Xbox Game Pass subscription service. A new Xbox Series X owner can, without regrets, toss their Xbox One in the garbage. Or donate it to a children's hospital. In fact, forget I mentioned garbage.
PS5: Most PlayStation 4 games can be downloaded and played on the PlayStation 5 at launch. I've had no problem downloading and installing the games I was playing on my PS4 before the PS5 arrived, picking up where I left off via cloud saves. I cannot hook up PlayStation VR right now, as I lack the free camera adapter Sony is distributing, but they are playable with that hardware. It's a nice library of games for a console launch, but it would be much nicer with a nice selection of PlayStation, PS2, and PS3 games to play.
Not only does the Xbox Series X have four generations of playable games at launch, it also has Xbox Game Pass, a relatively inexpensive monthly service that gives subscribers instant access to hundreds of them.
Xbox Series X Vs. PS5 Round Twelve: The Future
The battle between the Xbox Series X and PlayStation 5 is just beginning, and by battle I mean friendly scuffle, because who needs console wars anymore. As decades of gaming consoles has proven, two competitive consoles and whatever underpowered but lovable bullshit Nintendo's put out can exist together in complete harmony. Can anyone predict where the Xbox Series X and PlayStation 5 will be in five years? Sure, I'll give it a shot right now.
Xbox Series X: In five years' time the Xbox Series X will be the tired old man of the Xbox Series series of consoles. Microsoft console gamers will be eagerly awaiting news of the upcoming Xbox Series X2 while playing games on their Xbox Series XYZ consoles, which will be adorable little cubes with HDMI outputs and tiny little Xs on the front.
PS5: In grand PlayStation tradition, Sony will release a smaller, sleeker version of the PS5 hardware that's only 12 inches tall instead of 15. Fans will call it adorable. I'll hold off on buying one but cave at the last minute.
Overall Winner: Console Gamers And Science
Science never loses, so the title of "Best New Console" goes to both science, for being right all the time, and console gamers, who have two shiny new game consoles to play with. Turns out the "Best New Console" was inside us all along.
Tags: PS5seriesXbox
Lock and unlock iPhone and iPad flip screen
Ms. Marvel: news, rumors, broadcast date and release
Geralt of Sanctuary
Since I had my first console when I was 10, I've never stopped playing. I'm a multiplatform player and my favorite genre is RPG.
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Attention Destiny 2 players – start using your Gunsmith parts today!
Shenmue is getting an anime series from Crunchyroll and Adult Swim
Warzone cheaters have started roaming around in flying cars
Sony asks Microsoft to honor multiplatform agreements with Activision games
The OPPO Find X5 will finally be presented at the end of February
The Lord of the Rings (series): Premiere, platform, cast, plot…
XCOM and Civilization veterans form a studio
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