text:
Articles & Reviews
News Archives
Display Drivers
Graphics Cards
Operating Systems
Phoronix Premium
Ad-Free Browsing
Support Phoronix
Single-Page Article Viewing
Robocraft Is Rolling Over To Linux
Published on 22 August 2014 03:40 PM EDT
Written by
Michael Larabel
5 Comments
Robocraft, a game described as "Minecraft Meets World of Tanks", will soon be released on Steam for Linux.
It was announced today via
this Steam Community posting
that Robocraft has been ported to Linux and OS X and that these ports will soon be released on Steam. "We have completed the work to get the Mac and Linux versions up and running and they are in test now and will launch very soon via Steam." Robocraft is powered by the Unity Game Engine.
Robocraft is self-described as "If Minecraft smashed straight into World of Tanks you'd get Robocraft. BUILD, DRIVE & FIGHT in this Free-To-Play MMO Robot vehicle shooter. Build a robot battle vehicle, add wheels, jets, guns, wings and take it into intense online team battles."
Those wishing to learn more can stop by
the Steam Store
(it's free to play) or watch the game trailer embedded below.
5 Comments
News Archives
Recent Gaming News
Cube2-Forked Red Eclipse 1.5 Sees Huge Update
BioShock Infinite Linux Gaming Benchmarks
BioShock Infinite Now Available For Linux, Demands Binary Graphics Drivers
Divinity: Original Sin Is Still En Route To Linux Gaming Systems
0 A.D. Alpha 18 Brings Nomad Game Mode
Popular Phoronix News
Chrome OS Switches To "Freon" Graphics Stack To Replace X11
"Why Linux Is Still Not Ready For The Desktop"
Introducing The Library Operating System For Linux
Google To Shutdown Google Code, Concedes To GitHub
There's Now More Than 1,000 Games On Steam For Linux
About The Author
Michael Larabel is the principal author of Phoronix.com and founded the web-site in 2004 with a focus on enriching the Linux hardware experience and being the largest web-site devoted to Linux hardware reviews, particularly for products relevant to Linux gamers and enthusiasts but also commonly reviewing servers/workstations and embedded Linux devices. Michael has written more than 10,000 articles covering the state of Linux hardware support, Linux performance, graphics hardware drivers, and other topics. Michael is also the lead developer of the
Phoronix Test Suite
, and
automated testing software. He can be followed via
or contacted via
Latest Articles & Reviews
OS X 10.10 vs. Ubuntu 15.04 vs. Fedora 21 Tests: Linux Sweeps The Board
The New Place Where Linux Code Is Constantly Being Benchmarked
18-GPU NVIDIA/AMD Linux Comparison Of BioShock: Infinite
Phoronix Test Suite 5.6 Adds New Phoromatic Enterprise Benchmarking Features
OpenGL Threaded Optimizations Responsible For NVIDIA's Faster Performance?
Big Graphics Card Comparison Of Metro Redux Games On Linux
Latest Linux News
Git 2.4.0-rc0 Does A Ton Of Polishing
The Most Common, Annoying Issue When Benchmarking Ubuntu On Many Systems
Mesa Is At Nearly 1,500 Commits This Year
Gestures & Other GTK3 Features For LibreOffice
It's Now Easier To Try PHP 7 On Fedora & RHEL
BQ Is Cleaning Up Their Aquaris E4.5 Ubuntu Kernel
Allwinner Continues Jerking Around The Open-Source Community
NVIDIA Linux 349.12 Beta Has Improved G-SYNC & VDPAU Features
Canonical Just Made It Even Easier To Benchmark Ubuntu Linux In The Cloud
NVIDIA GeForce GTX TITAN X Linux Testing Time
Most Viewed News This Week
Introducing The Library Operating System For Linux
AMD Is Hiring Two More Open-Source Linux GPU Driver Developers
New SecureBoot Concerns Arise With Windows 10
GNU Nano 2.4.0 Brings Complete Undo System, Linter Support & More
GNOME Shell & Mutter 3.16.0 Released
Systemd Change Allows For Stateless Systems With Tmpfs
GCC 5 Compiler Is Getting Close To Being Released
Chromebooks Powered By The MIPS Pistachio, Linux Support Evolving
Legal Disclaimer & Disclosure
Contact Us
Copyright © 2004 - 2015 by
Phoronix Media
All trademarks used are properties of their respective owners. All rights reserved.
Powered by the Phoronix Content Management System (PHXCMS-9).


text:
When two people have a special connection. Kindred Spirits is a painting I've created of two little friends enjoying a warm day out in the garden at a garden party, the birds are singing and everyone is peaceful, content and happy. One of the girls wears a daisy chain around her neck, they have been collecting the little flowers to give and share and handing out balloons to everyone. Little bird is nesting in one girls' hair….white puffy clouds in the background and I've given this a vintage look, which I hope you'll really like too. Thanks so much for dropping by to look at my art today!
Adorable, you have amazing talent! -Sherry
thank you so much for your lovely response Sherry! :)
– © Karin Taylor
Gorgeous Karin. I am so pleased to see you back here. I was thinking you had left RB or something was wrong, so I am glad to see you back in full swing again.
Thanks so much Christina, I had taken time away from RB and only really decided to return today, it's lovely to see you again, thank you for your lovely response, so glad you like it! ;)
– © Karin Taylor
Best buds. Gorgeous creation!
Thank you so much Bunny, indeed I think they are :) (best buds that is)
– © Karin Taylor
Beautiful, and I love those translucent balloons. You add the most wonderful special touches: the wind-blown daisy chain necklace, the single heart balloon, the bird sitting on the end of a twig decorating her hair, and the dresses— as always, darling, Karin.
Frannie, thank you so much, I'm so happy you noticed the balloons, as I was really pleased with how they turned out in particular, and thanks for noticing those little additional touches and details! I am so proud of this one :) x
– © Karin Taylor
This is so sweet; I love how you softly blended your colours together, and how even with this overal vintage look, there are hints of colourful subjects like the butterflies and the birds.
Thank you so much Mui-Ling, I've developed a new technique here and that's really helping to pull it all together and give it a more sophisticated look I'm hoping, so glad you like the additional hints of colour in the little birdies, etc :)
– © Karin Taylor
Marvelous Karin!! :)
Thank you so much Keith, I am so proud of this one, so happy you like it! :)
– © Karin Taylor
Feels like a great birthday party coming up. Soft and adorable.
Many thanks Sandra, so happy you like it :)
– © Karin Taylor
They're so pretty with wonderful dresses. I also love the balloon and muted colours.
thank you so much Josette, very pleased you like those elements and details, this is my most favourite piece, I feel as though it's the best one I've done so far :)
– © Karin Taylor
Wonderful work, lovely tones and pastel palette!

subset: commoncrawl


text:
EventInfo.GetRaiseMethod Method ()
.NET Framework 2.0
Returns the method that is called when the event is raised. Namespace: System.Reflection
Assembly: mscorlib (in mscorlib.dll)
Assembly: mscorlib (in mscorlib.dll)
- ReflectionPermission when invoked late-bound through mechanisms such as Type.InvokeMember. Associated enumeration: ReflectionPermissionFlag.MemberAccess.
Windows 98, Windows 2000 SP4, Windows CE, Windows Millennium Edition, Windows Mobile for Pocket PC, Windows Mobile for Smartphone, Windows Server 2003, Windows XP Media Center Edition, Windows XP Professional x64 Edition, Windows XP SP2, Windows XP Starter Edition
The .NET Framework does not support all versions of every platform. For a list of the supported versions, see System Requirements.


text:
It's getting to be that time of year again. On October 2nd, Wizards of the Coast is proud to release its newest set to Magic the Gathering. Zendikar features new mechanics and nearly 249 new cards to add to the set. But, on September 26th, there is plenty of places to get an inside look at the new set.
Like every new set, the DCI will host a tournament at many participating locations. www.examiner.com/x-22992-Baltimore-Magic-the-Gathering-Examiner~y2009m9d11-Looking-for-More-Magic-The-Gathering-in-Baltimore Many of these places will hold their own prerelease tournaments and will hand out special prizes to the participants.


text: Web exclusive! One of the original MAD men, this spy has only one mission… destroy the other spy! Price: Price pending This Halloween, only one spy will make it out on top… the other will have to go home with nothing. Choose your side and get ready for some booby-trapping madness with the Spy Vs. Spy White Spy Adult Costume. It includes a white hat, a white character mask, a head sock, a white long sleeve shirt, and matching pants. Featured in white Includes: hat, mask, head sock, shirt, and pants Officially licensed Spy vs. Spy costume Care Instructions: spot clean Material: polyester Gloves and shoe covers not included Imported More


text:
Page 1
Dopamine and α-synuclein dysfunction in Smad3
null mice
Tapia-González et al.
Tapia-González et al. Molecular Neurodegeneration 2011, 6:72
http://www.molecularneurodegeneration.com/content/6/1/72 (13 October 2011)
Page 2
Dopamine and a-synuclein dysfunction in Smad3
null mice
Open Access
Silvia Tapia-González1,3, Rosa M Giráldez-Pérez1,3, M Isabel Cuartero1,3, M José Casarejos1,3, M Ángeles Mena1,3,
Xiao-Fan Wang2and Amelia Sánchez-Capelo1,3*
Background: Parkinson's disease (PD) is characterized by dopaminergic neurodegeneration in the substantia nigra
(SN). Transforming growth factor-b1 (TGF-b1) levels increase in patients with PD, although the effects of this
increment remain unclear. We have examined the mesostriatal system in adult mice deficient in Smad3, a molecule
involved in the intracellular TGF-b1 signalling cascade.
Results: Striatal monoamine oxidase (MAO)-mediated dopamine (DA) catabolism to 3,4-dihydroxyphenylacetic acid
(DOPAC) is strongly increased, promoting oxidative stress that is reflected by an increase in glutathione levels.
Fewer astrocytes are detected in the ventral midbrain (VM) and striatal matrix, suggesting decreased trophic
support to dopaminergic neurons. The SN of these mice has dopaminergic neuronal degeneration in its rostral
portion, and the pro-survival Erk1/2 signalling is diminished in nigra dopaminergic neurons, not associated with
alterations to p-JNK or p-p38. Furthermore, inclusions of a-synuclein are evident in selected brain areas, both in the
perikaryon (SN and paralemniscal nucleus) or neurites (motor and cingulate cortices, striatum and spinal cord).
Interestingly, these a-synuclein deposits are detected with ubiquitin and PS129-a-synuclein in a core/halo cellular
distribution, which resemble those observed in human Lewy bodies (LB).
Conclusions: Smad3 deficiency promotes strong catabolism of DA in the striatum (ST), decrease trophic and
astrocytic support to dopaminergic neurons and may induce a-synuclein aggregation, which may be related to
early parkinsonism. These data underline a role for Smad3 in a-synuclein and DA homeostasis, and suggest that
modulatory molecules of this signalling pathway should be evaluated as possible neuroprotective agents.
Keywords: Smad3, Parkinson's disease, Synucleinopathy, Dopamine, Astrocytes, TGF-β, α-Synuclein, MAO-B, MAPK
PD is characterized by the progressive degeneration
of dopaminergic neurons in the SN pars compacta. These
neurons project to the ST and secrete DA in order
to control voluntary movements and rewarding events [1].
Another significant feature of PD is the presence of cyto-
plasmic and neuritic inclusions of a-synuclein, known
as LB and neurites. The aetiology of idiopathic PD is
unknown, although current hypotheses focus on increased
oxidative stress, aberrant protein folding, defective protea-
some degradation or mitochondrial dysfunction. Given this
diversity in the underlying molecular mechanisms, it has
been suggested that multiple factors may cause the disease
[2]. However, the central mechanisms that induce dopami-
nergic neuron loss and a-synuclein aggregation remain
TGF-b1 is a cytokine that might mediate some molecu-
lar mechanisms of the disease. This molecule is expressed
at low levels in uninjured brains, while it is up-regulated
in the brain in association with diseases such as Parkin-
son's and Alzheimer's disease, Down syndrome, ischemic
lesion, hydrocephalus or spinal cord injury [3-10].
TGF-b1 is a highly pleiotropic molecule that can regu-
late cell proliferation, migration, differentiation and apop-
tosis. Its effect is cell type and context-dependent, and it
may provide signals for both cell survival and apoptosis
[11]. Moreover, a role for TGF-b in neuronal plasticity is
becoming evident. In Drosophila, the maintenance and
* Correspondence: firstname.lastname@example.com
1Departamento de Neurobiología-Investigación, Hospital Ramón y Cajal,
IRYCIS, Madrid, Spain
Full list of author information is available at the end of the article
Tapia-González et al. Molecular Neurodegeneration 2011, 6:72
© 2011 Tapia-González et al; licensee BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative
Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and
reproduction in any medium, provided the original work is properly cited.
Page 3
specialization of synapses at the neuromuscular junction
requires TGF-b-mediated transcriptional regulation
[12,13]. In Aplysia, TGF-b1 induces long-term facilitation
in sensory-motor synapses [14].
During mammalian embryonic development, TGF-b3
(but not TGF-b1) is necessary for the survival of midbrain
dopaminergic neurons at perinatal stages [15]. Hence,
while TGF-b3 appears to exert its effects on newborns
neurons, TGF-b1 might have pathological effects in adults.
The context-dependent effects of these factors are clearly
observed in sensory neurons, where ontogenic neuronal
death is increased by TGF-b [16]. In the nigrostriatal sys-
tem of MPTP-treated mice, adenoviral overexpression of
active TGF-b1 produces a decrease in the survival of dopa-
minergic neurons, accompanied by higher levels of striatal
DA depletion [17,18]. Despite these studies, the role of
TGF-b in the adult mesostriatal system is not well
The TGF-b subfamily of cytokines exert their effect by
binding to heteromeric receptor complexes at the cell sur-
face that contain a type I (primarily ALK5, but also ALK2
or ALK1) and a type II receptor (TbR-II or ActR-IIB).
Ligand binding to the receptor complex promotes the
recruitment and phosphorylation of Smad3 and/or Smad2.
Activated Smad3/2 interacts with Smad4 inducing their
translocation to the nucleus, where they bind to DNA
through specific Smad-binding elements, thereby promot-
ing targeted gene transcription. Some of the ligands that
can activate Smad3 and/or Smad2 are TGF-b1, -b2, -b3,
Inhibin, Activin, Nodal, GDF1, Vg1, and Lefty. This cano-
nical TGF-b pathway can cross-talk at multiple levels with
other signalling pathways such as those mediated by
MAPK, Fas, CaMKII, PKC, etc [11].
To address the role of TGF-b in the adult nigrostriatal
system, we have analyzed Smad3 null mice. We provide
evidence that Smad3 deficiency diminishes the trophic
and astrocytic support to nigral dopaminergic neurons, as
well as strongly altering DA metabolism. In addition a-
synuclein over-expression is produced in these mice, with
the formation of aggregates in different telencephalic,
mesencephalic and romboencephalic regions.
Smad3 is expressed in dopaminergic neurons of the SN
To determine whether molecules related to TGF-b sig-
nalling are expressed in the adult murine nigrostriatal
system, we examined the expression of mRNA transcripts
encoding: TGF-b1, -b2 and -b3 ligands; type I receptors
ALK1, ALK2 and ALK5; the type II receptor TbR-II; the
Smad signalling molecules Smad2, Smad3, Smad4; and
the inhibitor Smad7. RT-PCR analysis showed that all
these molecules are expressed in the VM and ST of adult
mice, with the exception of TGF-b3 and AKL1 in the
VM (Figure 1A). The absence of TGF-b3 expression in
the VM suggests that this molecule is not critical for
Smad3 signalling in adults, in contrast with perinatal
stages [15]. We further assessed whether TGF-b signal-
ling might be present in dopaminergic neurons by histo-
logical analysis of Smad3 on tyrosine hydroxylase (TH)
positive neurons. Dopaminergic neurons in the SN
express Smad3, primarily in the cytoplasm, although
some neurons with nuclear localization were also
detected (Figure 1B, 2F, arrows). We also observed a
population of TH+/Smad3-and Smad3+/TH-cells in the
SN. The presence of Smad3 in TH-ir cells indicates that
TGF-b ligands can act directly on dopaminergic neurons.
We further assessed Smad3 expression in other brain
areas related to parkinsonism, such as the ST and the
motor cortex. Confocal analysis suggested that Smad3
was present in the nucleus of cells of the striatal matrix
compartment (the TH-fibre rich region). On the other
hand, TH-fibre innervation was detected throughout the
motor cortex, with Smad3 mainly expressed in the
nucleus of cells of the third cortical layer (Figure 1B).
Smad3 depletion leads to fewer dopaminergic neurons in
the rostral SN
Because TGF-b1 can provide both pro-survival or death
signals to cells, we determined whether Smad3 deficiency
altered the number of dopaminergic neurons in the VM.
We analyzed the nigrostriatal system in 2- to 3 -month-
old Smad3 knock-out male mice in the basal state. These
mice develop to term and live to adulthood, surviving to
2 to 4 months of age. They exhibited subtle phenotypic
abnormalities, such as osteopenia, while 1/3 of the mice
developed malformations of the forelimbs and rib cage
[19], curvature of the spine (kyphosis) and resting tremor
(data not shown). The brains of Smad3-/-mice exhibited
a grossly normal morphology. Indeed, when we used
unbiased stereological methods to quantify the number
of dopaminergic neurons along the rostro-caudal axis of
the A9, A10 and A8 areas of Smad3-/-and Smad3+/+
mice, the numbers were similar in the null and wild-type
mice genotypes in both the A10 and A8 dopaminergic
areas (Figure 2A, Additional file 1A). However, Smad3
depletion led to a 34.8% decrease in the number of TH-ir
neurons in the rostral SN (P = 0.004, Figure 2B-C). We
could not detect stronger Smad3 expression in the rostral
portion of the SN (data not shown), suggesting a func-
tional spatial differentiation of this signalling molecule.
Although the number of dopaminergic neurons
decreased in the rostral portion of SN in Smad3 null
mice, TH protein levels in the VM were similar to those
found in control mice (P = 0.566, data not shown). We
assessed apoptosis in the SN of these mice using TUNEL
assay (to measure nuclear DNA fragmentation) and mor-
phological criteria (shrunken, smooth-appearing nuclei
that have lost the granularity, associated with nuclear
Tapia-González et al. Molecular Neurodegeneration 2011, 6:72
Page 2 of 22
Page 4
Figure 1 Smad3 is expressed in dopaminergic neurons of the SN. (A) Expression of TGFb ligands, receptors and Smad3 signalling molecules
in the VM and ST of adult mice. RT-PCR from three adult mice are shown under basal conditions for the expression of TGF-b1, -b2, and -b3
ligands, ALK1, ALK2, ALK5 type I receptors, TbR-II type II receptor, Smad2 and Smad3 receptor activated Smads, the common Smad4, and the
inhibitory Smad7. Every PCR reaction was performed using 1 μl of the cDNA pool, except for TGF-b3 and ALK1 in the VM where 1, 2 and 3 μl
were tested. (B) Coronal sections were double immunostained for TH and Smad3. SN dopaminergic neurons express Smad3 in the cytoplasm,
although some neurons have Smad3 in the nucleus (arrows). Quantitative estimate of the values representing colocalized pixels in the dual-color
image shows the co-localization of both signals in the SN (Pearson correlation coefficient = 0.62). Confocal images of TH/Smad3 double-labelling
in the ST and the motor cortex (Cx-M) suggested region co-localization, but not at the cellular level. Scale bar 50 μm.
Tapia-González et al. Molecular Neurodegeneration 2011, 6:72
Page 3 of 22
Page 5
chromatin condensation) [20]. We could not detect a
clear increase in apoptotic cells in the SN of Smad3-/-
mice (Additional file 1B). Considering that the criteria
used to measure apoptosis last for a few hours (both
nuclear condensation and fragmentation), it seemed pos-
sible that neuronal death might be greater at older ages.
Unfortunately, these knockout mice did not survive
beyond 4 months of age and thus, an analysis of older
animals was not possible.
TGF-b3, but not TGF-b1, has been shown to be
required for the survival of dopaminergic neurons in
perinatal stages [15]. In order to discriminate between a
developmental defect and a degenerative process in the
SN of adult Smad3 null mice, we quantified the number
of rostral TH-ir neurons in day 0 of postnatal age (P0).
Similar number of dopaminergic neurons was found in
Smad3 null mice and control mice (Figure 2E), indicating
a postnatal degenerative process in Smad3 deficiency
(Figure 2C).
We evaluated whether Smad3 was necessary to maintain
dopaminergic striatal innervation from nigral dopaminer-
gic neurons, and whether the rostral deficit in DA neurons
might have a positional effect in the ST. We quantified the
optical density of TH-ir fibres along the rostro-caudal axis,
as well as in the dorsal and ventral ST. No changes were
observed between the wild type and mutant mice (Addi-
tional file 2A). Furthermore, there was no alteration in
striatal TH protein content (Additional file 2B). Dopamine
transporter (DAT) immunoreactivity was also assayed in
the ST of these mice. Smad3 deficiency resulted in a not-
statistically significant trend to decrease DAT expression
in the rostral ST (P = 0.095, Additional file 3A-C). This
trend is observed in the rostral ST, similar to the rostral
decrease on TH-ir neurons in the SN, although in a wider
Figure 2 Degeneration of dopaminergic neurons in the rostral SN of Smad3 null mice. (A) Quantification of TH-ir neurons was determined
using unbiased stereological methods in the midbrain A9, A10 and A8 dopaminergic areas (which include the SN, ventral tegmental area and
retrorubral field, respectively). No clear differences were observed between mice when the entire pool of TH-ir cells was counted in these areas.
The stereological counting method had a CE < 0.07 for each mouse. (B-C) Representation of the SN (A9) along the rostro-caudal axis showed
that Smad3-/-mutant mice had fewer TH-ir neurons than Smad3+/+in the initial rostral 500 μm of the SN (*P < 0.05, **P < 0.01, Mann-Whitney
Rank Sum test). Similar values were detected between mice in the middle and caudal regions. Unpaired t-test, n = 5 for Smad3+/+and n = 6 for
Smad3-/-mice. (D) Unbiased stereological quantification of Nissl(+) neurons in the ST of Smad3+/+and Smad3-/-mice (Unpaired t-test, n = 4
mice per genotype). CE ≤ 0.05. (E) Quantification of TH-ir neurons in the rostral portion of the midbrain at perinatal stage (P0). Similar values
were found between Smad3+/+and Smad3-/-mice, indicating no developmental defect, but a postnatal degenerative process in Smad3
deficiency (Student's t-test, n = 3-4 per genotype). (F) Confocal microscopic images of Smad3/TH double-labelled neurons in the SN of Smad3+/+
and Smad3-/-mice. In control tissues, some neurons show Smad3 translocated to the nucleus (arrow). Smad3 null mutant mice have no
expression of the protein.
Tapia-González et al. Molecular Neurodegeneration 2011, 6:72
Page 4 of 22
Page 6
area. We further analyzed whether there are any neuronal
loss in the ST. Unbiased stereological quantification of
Nissl(+) neurons showed that Smad3 deficiency did not
alter the number of striatal neurons (Figure 2D), suggest-
ing that the pro-survival effect of Smad3 may be specific
to the dopaminergic neurons of the SN in the mesostriatal
system. Together, the loss of TH-ir neurons in the SN and
the trend to decrease the presynaptic DA marker in the
ST (DAT) seems to suggest an early loss of dopaminergic
neurons in Smad3 null mice.
Smad3 null mice have less GFAP expressing astrocytes
The effect of the absence of Smad3 on astrocytes -expres-
sing glial fibrillary acidic protein (GFAP)- and microglia
-detected by CD11b-ir, which is upregulated on activated
microglia- was explored in the nigrostriatal system of
these mice. Microglial cells were not clearly altered in
Smad3 deficient mice (data not shown). However, the
number of astrocytes in the SN apparently decreased even
when assessed at low magnification (Figure 3A-B). Quanti-
fication of the ventral, middle and lateral SN (Additional
file 4A-C) showed that Smad3-/-mice had 38.0% less
GFAP-ir cells than Smad3+/+mice (P = 0.004) (Figure 3G).
Indeed, in the ST, Smad3 deficiency induces a 32.1%
decrease in GFAP-ir cells in the matrix compartment
of the dorsal zone (the TH-fibre rich region; P = 0.014,
Figure 3H). To further explore this effect we examined the
levels of GFAP protein expressed in the VM and ST of
these animals. The GFAP protein content is decreased in
null mutant mice by 63.9% in the VM (P < 0.001) and by
55.3% in the ST (P = 0.004, Figure 3I). Moreover, while
morphological changes were not apparent in astrocytes
from the SN (Figure 3A-D), some atrophy in the arboriza-
tion of astrocytes, with thin branching, is exhibited in the
ST of Smad3 null mice (Figure 3E-F).
We searched for Smad3 immunoreactivity in GFAP(+)
astrocytes within the SN of wild-type mice. Co-localization
of Smad3 and GFAP was evident (Figure 3J), suggesting
that Smad3 could provide a survival signalling for astro-
cytes. Astrocytes are the main element of brain homeo-
static system and loss of astrocytes in Smad3 deficient
mice may decrease trophic and metabolic support to sur-
rounding neurons.
Striatal DA catabolism and oxidative stress increase when
Smad3 is depleted
We further tested whether striatal DA levels is altered.
Similar levels of DA were found in Smad3 null mice and
control mice, although a strong increase in DA catabo-
lism was detected in the former (Figure 4A). Thus, in
Smad3 deficient mice the striatal levels of DOPAC,
homovanillic acid (HVA), and 3-methoxy-tyramine (3-
MT) increased by 94.5% (P = 0.001), 59.8% (P < 0.001),
and 28.1% (P = 0.007), respectively. The increase in DA
oxidation was primarily mediated by MAO since the
DOPAC/DA ratio augmented 98.8% (P = 0.004, Figure
4B). Catechol-O-methyl transferase (COMT) activity,
quantified by 3-MT/DA ratio, did not increase signifi-
cantly (33.9%, P = 0.101). This Smad3-dependent effect
might be relatively specific for DA because striatal sero-
tonin was not clearly altered (Additional file 5). It is
known that the oxidation of DA to DOPAC is accompa-
nied by the stoichiometric reduction of oxygen to H2O2
[21]. Hence, the 2-fold increment in MAO-mediated DA
oxidation must increase striatal oxidative stress. The nor-
mal detoxification pathway for H2O2is the glutathione
system, supplied by the mitochondria. We measured
striatal glutathione content in its reduced (GSH) and oxi-
dized (GSSG) forms (Figure 4C). Smad3-/-mice had
35.9% more striatal GSH than Smad3+/+mice (P =
0.043). We could speculate on a compensatory mechan-
ism by anti-oxidant GSH defence in order to detoxify
excess of H2O2produced by increased DA turnover in
Smad3 deficiency.
MAO-B increases in the ventral midbrain of Smad3
null mice
The increment in DA catabolism without a modification
in DA levels may be related to alterations in different
mechanisms, such as increased MAO levels or decreased
vesicular DA storage. Once re-uptaked from the presynap-
tic terminals, DA can either be repackaged into synaptic
vesicles by the vesicular monoamine transporter (VMAT-
2) or catabolyzed by MAO. We analyzed the striatal
VMAT-2 content and we observed that Smad3 deficiency
did not alter their protein levels (Figure 5B). However,
Smad3 ablation increased the MAO-B content in the VM
by 38.9% (P = 0.010, Figure 5A). In the ST, the change in
MAO-mediated DA catabolism (Figure 4B) does not seem
to correlate with protein content alteration, suggesting
changes in MAO-B activity for the 2-fold increment in
DOPAC/DA ratio. On the other hand, dopaminergic
somatodendrites in SN can synthesize, store and release
DA. The increment in MAO-B protein levels could parti-
cipate in the catabolism of DA in somatodendrites. More-
over, the SN contains high levels of MAO-B positive
astrocytes and its elevation in mouse brain results in par-
kinsonism [22]. Moreover, MAO-B activity levels have
been found to be doubled in the SN of PD [23].
Smad3 deficiency decreases the phosphorylation of the
Smad2 linker region
Smad3-deficiency may induce an increment in Smad2 sig-
nalling in an attempt to compensate for the TGF-b signal-
ling deficit. In order to evaluate this possibility we
measured the phosphorylation of Smad2 at two different
locations (Figure 6A). The carboxyl terminus is phos-
phorylated by TGF-b activated type I receptors, while the
Tapia-González et al. Molecular Neurodegeneration 2011, 6:72
Page 5 of 22
Page 7
Figure 3 Smad3-/-mice have fewer nigrostriatal astrocytes. In the SN the number of GFAP-ir astrocytes decreased in Smad3 deficiency (A-B,
G). Illustrations depict non-activated astrocytes as verified by the labelling of thin, elongated processes in the wild-type (C) and null (D) mice. In
the ST, atrophic astrocytes could be detected in Smad3-/-null mice (F, arrow) when compared to Smad3+/+mice (E). (H) Number of GFAP-ir
cells in the striatal matrix and striosomes. The figure represents the GFAP-ir cells per mm2in one striatal hemisphere. (I) The GFAP protein
content is decreased in both the VM and ST of Smad3 null mice (Student's t-test, n = 4-6 per genotype) (J) Confocal images of Smad3 and
GFAP double-labelled cells in the SN of wild-type mice showed co-localization of Smad3 in astrocytes. *P < 0.05, **P < 0.01, ***P < 0.001
Tapia-González et al. Molecular Neurodegeneration 2011, 6:72
Page 6 of 22
Page 8
linker region is phosphorylated by regulatory intracellular
signals such as MAPKs, CaMKII or CDKs [24]. In immu-
noblots, phosphorylation of the Smad2 carboxy-terminal
was not altered in the VM or the ST of Smad3-/-mice
(Figure 6B). However, the phosphorylation of the linker
region decreased by 48.4% in the VM (P = 0.028) and
20.8% in the ST (P = 0.037, Figure 6C). Decreased phos-
phorylation in the linker region might be indicative of
weaker modulatory intracellular signalling. However, there
is no indication of any significant compensatory up-regula-
tion in activity at the Smad2 carboxy-terminal.
Cross-talk between Smad3 and Erk1/2, but not JNK or
p38 MAPKs
The decreased activation in the linker region of Smad2
may imply a deficiency in MAPK signalling. Extensive
cross-talk between TGF-b signalling and JNK, Erk or
p38 occurs at different levels to promote or inhibit
Figure 4 Smad3 ablation induces an increase in MAO-mediated DA catabolism in the ST. (A) Loss of Smad3 leads to increased catabolism
of striatal DA, with no alterations in total neurotransmitter content. (B) The catabolism of DA in Smad3 deficient mice is primarily MAO-mediated
(DOPAC/DA ratio) as compared to COMT (3-MT/DA). (C) The striatal GSH content of Smad3-/-mice is also higher than in Smad3+/+mice. (*P <
0.05, **P < 0.01, ***P < 0.001, Student's t-test, n = 6 per genotype)
Tapia-González et al. Molecular Neurodegeneration 2011, 6:72
Page 7 of 22
Page 9
apoptosis [11]. Furthermore, JNK-cJun activation has
been implicated in the pathogenesis of PD [25]. In order
to determine whether an interaction between Smad3
and MAPKs exists in the nigrostriatal system, we evalu-
ated the endogenous levels of phosphorylated JNK, p38
and Erk1/2 in Smad3 null mice. The phosphorylation of
JNK and p38 in the VM and ST was similar in Smad3
+/+and Smad3-/-mice (Additional file 6A-B), as were
the striatal levels of p-Erk1/2 (P = 0.537, for p-ERK2p42,
Figure 7A). However, Smad3 ablation led to a significant
decrease in p-Erk1p44(38.7%, P = 0.032) and p-Erk2p42
(20.3%, P = 0.038) in the VM (Figure 7A). Immunohis-
tochemistry for p-Erk1/2 showed a granular distribution
in dopaminergic neurons of the SN. Interestingly, a gen-
eral decrease in the number of these cytoplasmic gran-
ules labelled for p-Erk1/2 was detected in Smad3 null
mice (Figure 7B, data not shown). In summary, two
MAPKs implicated in the induction of cell death, such
as JNK and p38, are not regulated by Smad3 under
basal conditions. However, activation of the pro-survival
MAPK Erk1/2 was diminished in nigral dopaminergic
neurons after Smad3 depletion. These results suggest
that Smad3 activates Erk1/2 in dopaminergic neurons
and that Smad3 deficiency may decrease the trophic
support provided by Erk1/2 signalling.
Increased expression and aggregated a-synuclein in
The molecular events detected in Smad3-/-mice were
highly suggestive of early signs of parkinsonism. One of
the pathological hallmarks in PD is abnormal a-synuclein
aggregation and inclusion body formation. When we
examined the expression of a-synuclein in immunoblots
of the VM and ST of these mice, similar protein levels
were found in the VM (Figure 8A). However, Smad3 defi-
ciency increased the expression of a-synuclein in the ST
by 20% (P = 0.028). To further assess this response, we
performed immunohistochemistry in the ST and we
quantified the optical density of a-synuclein fibres in this
area (Figure 9H-J). Smad3-/-mice displayed a 16% trend
to increase (P = 0.024) in the density of a-synuclein
along the rostrocaudal ST when compared to wild-type
mice (Figure 8B).
Biochemically, a-synuclein inclusions are resistant to
extraction with non-ionic detergents like Triton X-100.
Conversely, extraction with ionic detergents such as SDS
disrupts a-synuclein aggregation into monomers and
SDS-stable oligomers [26]. We detected aggregated forms
of a-synuclein by immunohistochemistry (see below,
Figures 9K, N, R and 10). A range of aggregates is
observed, from strongly- to mildly-detected, in male
Smad3 deficient mice at this age (2-3 months old). We
further characterized these aggregates by sequentially
extracting tissue from the VM, ST and M1 motor cortex
in detergents of increasing strength [27]. To verify the
specificity of the signal detected, a-synuclein null mice
were used as a control. In Western blots, a specific 12-14
kDa band that corresponded to a-synuclein monomers
was detected, which was not detected in the a-synu-
clein-/-mice. Indeed, most a-synuclein monomers were
extracted in the Triton X-100 and SDS soluble fractions
(Figure 8C). It should be noted that the antibody reacts
with non-specific bands in the a-synuclein-/-mice that
differ from those observed in mice of the Smad3 colony.
Interestingly, soluble oligomers of 45-50 kDa could be
detected in the Triton X-100 phase of VM from Smad3
null mice and a faint 22-24 KD band was detected in
Smad3-/-mice, mainly present in the primary motor cor-
tex, suggestive of soluble dimers. High molecular weight
SDS-insoluble aggregates were detected in the VM and
motor cortex of Smad3 null mice (Figure 8D). Quantifi-
cation of insoluble monomeric a-synuclein highlighted a
2.1-fold induction (p = 0.010) in the motor cortex in
Smad3 null mice (Figure 8E). Interestingly, there was a
clear increment of SDS-insoluble monomers and oligo-
mers in the spinal cord of Smad3 null mice, primarily in
the lumbar region (Figure 8F), suggesting the appearance
of inclusions in these regions.
We further characterized a-synuclein by immunohisto-
chemistry along the rostrocaudal brain axis, and we
Figure 5 Smad3 deficiency leads to increased MAO-B levels in the ventral midbrain. (A) Smad3-/-mice had higher MAO-B levels in the VM
than Smad3+/+mice. (B) VMAT-2 protein contents are not altered in Smad3-/-mice. Bars represent values normalized to b-tubulin from 6 mice
per group, each measured two to three times. **P < 0.01, Student's t- test.
Tapia-González et al. Molecular Neurodegeneration 2011, 6:72
Page 8 of 22
Page 10
detected a-synuclein aggregates in neurites and cell bodies
of different telencephalic, mesencephalic and romboence-
phalic regions (Figure 9 A-Q). Indeed, the specificity of
this staining was verified in a-synuclein null mice (data
not shown). To further study the distribution of a-synu-
clein, we compared old mice (21 month-old wild-type and
19 month-old heterozygous for Smad3), as well as young
mice (3 month-old wild-type and Smad3 null mice). There
was no difference in the distribution of a-synuclein
between young Smad3 heterozygous and wild-type mice.
However, a-synuclein aggregates where detected in young
Smad3 null mice and in old Smad3 heterozygous mice,
Figure 6 Decreased Smad2 phosphorylation in Smad3 null mice. (A) The MH1 and MH2 domains in Smad2, conserved in other Smads, are
connected by a less-conserved linker region. TGF-b and MAPKs signalling induces phosphorylation of certain sites in Smad2. (B) The level of
activated Smad2 (carboxy-terminal phosphorylation) is not modified by Smad3 ablation. (C) However, in Smad3 null mice the linker region is less
phosphorylated, suggesting a decrease in an intracellular regulatory signal of Smad2 (*P < 0.05, Student's t-test, n = 6 per genotype, each
measured two to three times).
Tapia-González et al. Molecular Neurodegeneration 2011, 6:72
Page 9 of 22
Page 11
suggesting a gene dosage and age dependent progressive
effect. Interestingly a-synuclein aggregates were detected
in fibres of the primary and secondary motor cortex, cin-
gulate cortex, ST and corpus callosum (Figure 9A-D, H-
K). Moreover, perinuclear a-synuclein aggregates were
evident in the cell bodies of the SN, paralemniscal nucleus,
and pontine nuclei of old heterozygous and primarily
young Smad3 null mice (Figure 9L-N). Furthermore,
Smad3 deficient mice manifested a clear increase in stain-
ing, associated with an irregular morphology and neurite
thickness, in fibres from the internal capsule, corpus callo-
sum, lateral lemniscus, cerebral peduncle and spinal
trigeminal tract (Figure 9A-E, O-P). At the level of the 2nd
and 3rdcerebellar lobe (Figure 9E), and of the anterior
white matter of the spinal cord (Figure 9G, Q) positively
labelled cells resembling glia were detected (star shape
appearance, arborizing processes, and a mean diameter of
4.3 ± 0.5 μm) [28], together with positive fibres. In the
lateral lemniscus, a broad dentritic arbor was detected
(Figure 9P).
In order to verify that the a-synuclein aggregates
detected in the SN of Smad3 deficient mice are degener-
ating dopaminergic neurons, we performed confocal
imaging of a-synuclein and TH double-labelled neurons
Figure 7 Decreased Erk1/2 signalling in dopaminergic neurons of Smad3-/-mutants. (A) Analysis of the phosphorylation state of Erk1/2
MAPK in the VM and ST of Smad3 null mice identifies a specific decrease in Erk1/2 activation in the VM (*P < 0.05, Student's t-test, n = 6 per
genotype, each measured two to three times). (B) Immunolabelling for p-Erk1/2 defines granular cytoplasmic structures in dopaminergic neurons
that are fewer when Smad3 is depleted.
Tapia-González et al. Molecular Neurodegeneration 2011, 6:72
Page 10 of 22
Page 12
(Figure 9R). Co-localization of TH and a-synuclein is
observed in the cytoplasm of dopaminergic neurons of
the SN, both in control and Smad3 deficient mice.
Furthermore, a-synuclein aggregates present in Smad3
deficiency co-localize with TH in this brain area, further
suggesting a relationship between the dopaminergic
alterations detected in Smad3 deficiency and the pre-
sence of a-synuclein aggregates.
Figure 8 Increased expression and aggregated a-synuclein in Smad3-/-mice. (A) Immunoblots of ST and VM tissue show that Smad3
deficiency increases the striatal a-synuclein content (*P < 0.05, Student's t-test with n = 6 mice per genotype, measured in triplicate). (B) Further
evaluation of a-synuclein expression by immunohistochemistry along the rostro-caudal axis of the ST. Twenty brain sections per mice were
counted from Smad3+/+(n = 5) and Smad3-/-(n = 6). Figure indicates values for the rostral 1000 μm. O.D. quantification shows an increase in
the ST of Smad3 null mice (*P < 0.05, Student's t-test). (C-F) Several nervous system regions from Smad3 wild-type and null mice were subjected
to graded detergent extraction (Triton-soluble, SDS-soluble, and SDS-insoluble fractions) and analyzed in Western blots. The molecular weight
markers on the left indicate the protein size in kilodaltons (**P < 0.01, Student t-test, n = 3 for Smad3+/+, n = 5 for Smad3-/-, n = 2 for a-
Tapia-González et al. Molecular Neurodegeneration 2011, 6:72
Page 11 of 22
Page 13
Figure 9 Aggregation of a-synuclein in specific brain areas. (A-G) Rostrocaudal distribution of a-synuclein in Smad3 wild-tupe and deficient
mice. Red points represent positive a-synuclein staining in wild-type and null mutant. Blue asterisks illustrate cellular and neuritic aggregates
present in old heterozygous and young Smad3 null mice. (H-Q) Photomicrography showing a-synuclein positive structures. (H-J) ST striatum, (K)
M1 primary motor cortes, cc corpus callosum in Smad3-/-, (L-N) SN substantia nigra, (O) cp cerebral peduncle, (P) ll lateral lemniscus, and (Q) fa,
anterior funiculus of the spinal cord. The double arrows show positive fibres detected in Smad3 deficient mice, asterisks point to synaptic
terminals with a punctuate morphology, and the arrowheads indicate perykarial inclusions. (R) Confocal microscopic images of TH/a-synuclein/
Dapi triple labelling in the SN of Smad3+/+and Smad3-/-mice. In control tissues, a-synuclein is present in the cytoplasm of dopaminergic
neurons. The aggregates observed in Smad3 deficiency are double-labelled for both molecules (arrow). The window is further magnified to get a
better appraisal of the accumulation of a-synuclein in a TH(+) neuron.
Tapia-González et al. Molecular Neurodegeneration 2011, 6:72
Page 12 of 22
Page 14
a-Synuclein deposits in Smad3 deficiency accumulates
ubiquitin and PS129-a-synuclein
Phosphorylation of serine 129 is a widespread a-synuclein
post-translational modification within LB and inclusions
isolated from human brain with PD, Multiple System
Atrophy (MSA), dementia with Lewy bodies (DLB) and
other synucleinopathies. Furthermore, the majority of a-
synuclein found in LB are mono- to tri-ubiquitinated [29].
In order to evaluate whether these post-translational mod-
ifications of a-synuclein are present in the aggregated
forms detected in Smad3 deficiency, we performed dou-
ble-immunostaining using anti-PS129-a-synuclein and
anti-ubiquitin antibodies in the SN of Smad3 deficient
mice (Figure 10A). In the wild-type mice ubiquitin could
be found in the cytoplasm of neurons and PS129-a-synu-
clein in neurites and dendrites. However, in Smad3 defi-
cient mice we could detect aggregates with both proteins.
Interestingly, in mice with less clear deposits of a-synu-
clein the distribution of ubiquitin and PS129-a-synuclein
revealed a core of ubiquitin, surrounded by a halo of
PS129-a-synuclein (Figure 10A, arrows). It is of note that
accumulation of ubiquitin could also be observed within
these halos. On the other hand, mice with more clear
inclusions detected by anti-a-synuclein antibody show
stronger accumulation of ubiquitin, surrounded by a
strong rim of PS129-a-synuclein. These ubiquitin/PS129-a-
synuclein double-labelled aggregates were found isolated
(arrowhead) or in small groups (double arrowhead). Dou-
ble-labelling with a-synuclein antibody shows co-localiza-
tion with ubiquitin within the core of these deposits
(Figure 10B).
This core/rim cellular distribution may resemble those
observed in human patients with LB diseases [29] and
further suggest a key role of Smad3 signalling in the
homeostasis of a-synuclein.
The presence of a-synuclein inclusions, with post-
translational modifications observed in the human LB, in
specific brain areas of Smad3 deficient mice suggests a
neuropathological status in these mice, which together
with the strong DA catabolism, the loss of dopaminergic
neurons and decreased trophic and astroglial support
detected, may resemble early symptoms of parkinsonism
in these mice.
The strong catabolism of striatal DA associated with the
increment of a-synuclein expression and inclusions in
selected brain areas, which are phosphorylated at Ser129
and ubiquinated, might represent an interesting model
of parkinsonism and/or a-synucleinopathies. Moreover,
Smad3 knockout mice have fewer astrocytes in the SN,
less nigral dopaminergic neurons, diminished trophic
support mediated by Erk1/2 signalling, and increased
oxidative stress (Figure 11).
In the mouse brain, TGF-b ligands, receptors and Smad
signalling molecules are expressed in the VM and ST.
Indeed, the dopaminergic neurons of the SN express
Smad3, suggesting that TGF-b ligands can act directly on
these neurons. The loss of dopaminergic neurons in
Smad3-/-mice from the rostral SN might result from an
alteration during the perinatal stage due to TGF-b3 signal-
ling [15]. However, adult Smad3-/-mice do not manifest
the overt DA neuron loss observed in TGF-b3 null mice,
resembling the effect of TGF-b1 deficiency. Moreover, the
decrease in the number of TH-ir neurons in the rostral
SN is not observed at P0, suggesting no developmental
defect but a postnatal degenerative process. An alternative
pathway could account for TGF-b3 during perinatal stage,
such as those involving Smad2. Moreover, mRNA TGF-b3
is not present in the adult VM, and several ligands should
be considered to activate Smad3 in the adulthood, such as
TGF-b1, -b2, Inhibin, Activin, or GDF1. Our results sug-
gest that Smad3 deficiency may be particularly important
for adult midbrain pathology since we detect a progressive
loss of dopaminergic neurons and a progressive onset of
a-synuclein aggregation, with a late onset in the heterozy-
gous mice (19-20 months), when compared with the early
onset in the null mice (2-3 months). These neurons may
be compromised by decreased Erk1/2 trophic support and
increased striatal oxidative stress. It has already been
established that Erk1/2 can phosphorylate Smad3 in the
linker region, and Erk1/2 and Smad3 converge on the pro-
moters of TGF-b targeted genes [30]. On the other hand,
TGF-b can activate Ras-Erk1/2 signalling in a Smad-inde-
pendent manner [31]. However, our studies show another
level of interaction, because Erk1/2 activity is dependent
on Smad3 in dopaminergic neurons. Erk1/2 signalling has
been implicated in numerous biological processes such as
cell survival and neurite outgrowth. Erk1/2 can be com-
partmentalized to different subcellular structures, such as
the nucleus, Golgi, endosomes or plasma membrane.
Notably, phosphorylated Erk1/2 has been detected in PD
within SN neurons, and with a granular distribution in
mitochondria and autophagosomes [32].
Primary cultures of astrocytes derived from human and
mouse brain are known to secrete TGFb molecules and to
express the Smad3 signalling pathway. In vitro studies have
shown the role of TGFb/Smad3 in the secretion of TGF-
b2 for the increment of extracellular matrix proteins in
human optic nerve head astrocytes, in the context of glau-
coma [33], the regulation of iNOS production by TGF-b1/
Smad3 after inflammation [34], or the induction of
Jagged1-Notck signalling for oligodendrocyte progenitor
proliferation and differentiation [35]. Interestingly, TGF-b1
arrests astrocyte proliferation in G1 phase, through
the induction of p15INK4B[36]. Hence, we could expect
an increase in the number of astrocytes in Smad3 defi-
ciency. However, we observe a decrease in the number of
Tapia-González et al. Molecular Neurodegeneration 2011, 6:72
Page 13 of 22
Page 15
Figure 10 a-synuclein deposits in Smad3 deficiency are ubiquitinated and S129 phosphorylated. (A) Confocal images of aggregates
double-labelled with ubiquitin and PS129-a-synuclein in the ventral midbrain of Smad3 deficient mice. Different morphologies could be
detected, from a clear core of ubiquitin surrounded by a halo of PS129-a-synuclein (arrows), to strong accumulation of ubiquitin and PS129-a-
synuclein in isolated (arrowhead) or groups (double arrowhead) of deposits. (B) a-synuclein staining co-localize with ubiquitin within the core of
the aggregates.
Tapia-González et al. Molecular Neurodegeneration 2011, 6:72
Page 14 of 22


text:
George Overstreet
Published: Wednesday, August 29, 2012 at 12:42 p.m.
Last Modified: Wednesday, August 29, 2012 at 12:42 p.m.
For many years George Overstree has been a driving force behind the Polk County Toys For Tots program.
Occupation: Retired
Years in Polk: 85
Spouse: Alice Ray-Overstreet
Children: Cindy Connors
Civic Activities: Mr. Overstreet is also active with his church, the First United Methodist Church on Lake Morton in Lakeland.
Reader comments posted to this article may be published in our print edition. All rights reserved. This copyrighted material may not be re-published without permission. Links are encouraged.


text:
Cantillon: Will Suntory swoop get lost in translation?
- Agribusiness & Food
- January 16, 2014, 01:00
Like most surprises, it turns out the Suntory bid for US bourbon giant Beam to create a global heavyweight was not that much of a shock after all.Sunt(...)
Suntory acquires Cooley parent Beam in $16bn deal
- Agribusiness & Food
- January 14, 2014, 01:00
Beam, one of the biggest names in US whiskey and parent of Cooley Distillery, is to be acquired by Suntory Holdings in a deal valued at $16 billion(...)
A whiskey story
- Drink
- December 7, 2013, 01:00
The space given over to Irish whiskey in the Celtic Whiskey Shop, Dublin's premium spirit shop, should be expanding. After all, Irish whiskey is going(...)
Distillers in high spirits as the whiskey sector enters golden era
- Agribusiness & Food
- November 8, 2013, 01:00
Irish whiskey's renaissance has matured to the point where multinationals and domestic players have committed to spending almost €400 million on build(...)
Beam reports sales of Kilbeggan down 26%
- Agribusiness & Food
- August 9, 2013, 01:00
US drinks giant Beam said yesterday that sales of its Irish brand, Kilbeggan Whiskey, were down 26 per cent in the first half of the year.Kilbeggan is(...)
New €12m distillery for Slane Castle
- Agribusiness & Food
- July 12, 2013, 01:00
The Mount Charles family is committing some €2.75 million of its own funds to a new €12-million craft whiskey distillery and visitor centre at Slane C(...)
A spirit of adventure, distilled over 20 years
- Agribusiness & Food
- June 28, 2013, 01:00
Donard Gaynor's knowledge of the global spirits market, particularly the buying habits of whiskey drinkers, is older than some of the rare malts resti(...)


text:
Cobbtown is a populated place located in Jefferson County at latitude 43.726 and longitude -76.044.
The elevation is 722 feet. Cobbtown appears on the Sandy Creek U.S. Geological Survey Map. Jefferson County is in the Eastern Time Zone (UTC -5 hours).
Jefferson County: Cities & Towns | ZIP Codes | Schools | Features (airports, churches, hospitals, etc.)
BIG Map |
Driving Directions |
Data Source - USGS GNIS FID: 972329
BOUNDARY MAPS: Town of Ellisburg | Jefferson County
Cobbtown, NY
(< 0.1 mile)
Mannsville, NY (1.4 miles SW)
Pierrepont Manor, NY (1.4 miles NW)
Winona, NY (3.1 miles E)
Wardwell, NY (3.1 miles NNW)
North Boylston, NY
(3.4 miles SSE)
Giddingsville, NY (3.6 miles N)
Allendale, NY (4.1 miles NNE)
Taylor Settlement, NY (4.3 miles NNW)
Ellisburg, NY (4.5 miles W)
Saxe Corner, NY
(3.9 miles NW)
Community Park, NY (4.3 miles SSW)
Hemlock District, NY (5.3 miles SSE)
Green Haven Mobile Home Community, NY (5.5 miles SSW)
North Landing, NY (7.1 miles W)
Jefferson Courts Mobile Home Park, NY
(8.7 miles N)
Lyon Corner, NY (8.8 miles N)
Sandy Acres Mobile Home Park, NY (10.1 miles N)
Pine Ridge Mobile Home Park, NY (10.2 miles N)
Babbitt Corner, NY (10.9 miles NNW)