text:
HMG-1 as a late mediator of endotoxin lethality in mice.
ABSTRACT Endotoxin, a constituent of Gram-negative bacteria, stimulates macrophages to release large quantities of tumor necrosis factor (TNF) and interleukin-1 (IL-1), which can precipitate tissue injury and lethal shock (endotoxemia). Antagonists of TNF and IL-1 have shown limited efficacy in clinical trials, possibly because these cytokines are early mediators in pathogenesis. Here a potential late mediator of lethality is identified and characterized in a mouse model. High mobility group-1 (HMG-1) protein was found to be released by cultured macrophages more than 8 hours after stimulation with endotoxin, TNF, or IL-1. Mice showed increased serum levels of HMG-1 from 8 to 32 hours after endotoxin exposure. Delayed administration of antibodies to HMG-1 attenuated endotoxin lethality in mice, and administration of HMG-1 itself was lethal. Septic patients who succumbed to infection had increased serum HMG-1 levels, suggesting that this protein warrants investigation as a therapeutic target.
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Article: No Signs of Inflammation during Knee Surgery with Ischemia: A Study Involving Inhaled Nitric Oxide.[Show abstract] [Hide abstract]
ABSTRACT: Nitric oxide donors and inhaled nitric oxide (iNO) may decrease ischemia/reperfusion injury as reported in animal and human models. We investigated whether the attenuation of reperfusion injury, seen by others, in patients undergoing knee arthroplasty could be reproduced when patients had spinal anesthesia. 45 consecutive patients were randomized into three groups (n = 15). Groups 1 and 3 were receiving iNO 80 ppm or placebo (nitrogen, N2) throughout the entire operation, and group 2 only received iNO in the beginning and at the end of the operation. Blood samples were collected before surgery, at the end of the surgery, and 2 hours postoperatively. Muscle biopsies were taken from quadriceps femoris muscle before and after ischemia. There were no increases in plasma levels of soluble adhesion molecules: ICAM, VCAM, P-selectin, E-selectin, or of HMGB1, in any of the groups. There were low numbers of CD68+ macrophages and of endothelial cells expression of ICAM, VCAM, and P-selectin in the muscle analyzed by immunohistochemistry, prior to and after ischemia. No signs of endothelial cell activation or inflammatory response neither systemically nor locally could be detected. The absence of inflammatory response questions this model of ischemia/reperfusion, but may also be related to the choice of anesthetic method EudraCTnr.Mediators of Inflammation 01/2014; 2014:620281. · 3.88 Impact Factor
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tilled water, 60 to 95% ammonium sulfate precipi-
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man degradation of proteolytically derived internal
peptides of the protein yielded two unambiguous
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revealed that PEDF contains identical sequences.
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histidine tag, cloned into pCEP4 (Invitrogen), and
transfected into human embryonic kidney cells.
Recombinant PEDF was purified from the condi-
tioned media with the Xpress Protein Purification
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freed of associated epithelium and as much of the
endothelium as possible, washed extensively in ice-
cold phosphate-buffered saline (PBS, pH 7.4), and
minced into small fragments that were incubated for
24 hours in PBS containing 0.5 mM phenylmethane-
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final concentration of 10 ?g of protein per milliliter.
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cells or human dermal microvascular endothelial cells
(Clonetics, San Diego, CA) as described (28). To com-
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ground migration (Bkgd) toward vehicle (0.1% bovine
serum albumin) and then normalized data by setting
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All experiments were repeated two to five times.
Statistics were performed on raw data before nor-
malization with the Student's t test. Standard errors
were converted to percentages.
27. Single-letter abbreviations for the amino acid resi-
dues are as follows: A, Ala; D, Asp; E, Glu; F, Phe; G,
Gly; I, Ile; L, Leu; M, Met; P, Pro; Q, Gln; R, Arg; S, Ser;
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29. PEDF antipeptide antibody (anti-PEDF) was raised in
rabbits against a peptide containing PEDF amino acids
327 to 343, conjugated to Keyhole-limpet hemocyanin,
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antisera against bacterial recombinant PEDF/EPC-1 (an-
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bodies to TGF?, and all angiogenic inducers (R & D
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31. We thank A. Mountz for VEGF measurements; B.
Kennedy and the Midwest Eye Banks and Transplanta-
tion Center for human eye tissue; M. K. Francis and V.
Cristofalo for anti–EPC-1; M. O'Reilly and J. Folkman for
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Hawkins, R. O'Grady, and Y. Mu for assistance with
retinoblastomas. Supported by the National Eye Insti-
cer Institute, and the Chicago Baseball Charities.
15 March 1999; accepted 3 June 1999
HMG-1 as a Late Mediator of
Endotoxin Lethality in Mice
Haichao Wang,1,3* Ona Bloom,3Minghuang Zhang,3
Jaideep M. Vishnubhakat,3Michael Ombrellino,2,3Jiantu Che,3
Asia Frazier,2,3Huan Yang,3Svetlana Ivanova,3
Lyudmila Borovikova,3Kirk R. Manogue,3Eugen Faist,4
Edward Abraham,5Jan Andersson,6Ulf Andersson,7
Patricia E. Molina,2Naji N. Abumrad,2Andrew Sama,1
Kevin J. Tracey2,3
release large quantities of tumor necrosis factor (TNF) and interleukin-1 (IL-1),
of TNF and IL-1 have shown limited efficacy in clinical trials, possibly because
these cytokines are early mediators in pathogenesis. Here a potential late
mediator of lethality is identified and characterized in a mouse model. High
mobility group–1 (HMG-1) protein was found to be released by cultured mac-
showed increased serum levels of HMG-1 from 8 to 32 hours after endotoxin
exposure. Delayed administration of antibodies to HMG-1 attenuated endo-
toxin lethality in mice, and administration of HMG-1 itself was lethal. Septic
patients who succumbed to infection had increased serum HMG-1 levels, sug-
gesting that this protein warrants investigation as a therapeutic target.
Mortality rates for systemic bacterial infec-
tion have not declined significantly, despite
advances in antibiotic therapy and intensive
care. Bacteria do not directly cause lethal
shock and tissue injury. Rather, bacterial en-
dotoxin (lipopolysaccharide, LPS) stimulates
the acute, early release of cytokines such as
TNF and IL-1? from macrophages, and it is
these host products that mediate damage (1).
Macrophages from C3H/HeJ mice do not re-
lease TNF and IL-1 when stimulated by LPS;
these animals are resistant to LPS lethality
(2). Normal, LPS-responsive mice can be
protected from lethal endotoxemia by thera-
peutic agents that selectively inhibit cytokine
action or prevent cytokine release (3).
Translating these pathogenic insights into
clinical therapy has proved difficult, in part
because these "early" mediators (TNF and
IL-1) are released within minutes after LPS
exposure (4). Thus, even a minimal delay in
treatment directed against TNF or IL-1 is
ineffective (3, 5). Paradoxically, LPS-respon-
sive mice treated with lethal doses of LPS
succumb at latencies of up to 5 days, long
1Department of Emergency Medicine and
ment of Surgery, North Shore University Hospital–
New York University School of Medicine, Manhasset,
NY 11030, USA.3The Picower Institute for Medical
Research, Manhasset, NY 11030, USA.4Department
of Surgery, Klinicum Grosshadern, Ludwig-Maximil-
ians University, Munich, Germany.5Division of Pul-
monary Sciences and Critical Care Medicine, Univer-
sity of Colorado Health Sciences Center, Denver, CO
80262, USA.6Department of Infectious Disease, Karo-
linska Institute, Huddinge University Hospital, Stock-
holm, Sweden.7Department of Rheumatology, Astrid
Lindgren's Children's Hospital, Karolinska Institute,
Stockholm, Sweden.
*To whom correspondence should be addressed. E-
mail: email@example.com
9 JULY 1999VOL 285SCIENCE www.sciencemag.org
Page 2
after serum TNF and IL-1 have returned to
basal levels. Moreover, mice deficient in
TNF die within several days of LPS admin-
istration (6), suggesting that mediators other
than TNF might contribute causally to endo-
toxin-induced death.
To identify potential "late" mediators of
endotoxemia, we stimulated murine macro-
phage-like RAW 264.7 cells with LPS and
analyzed the conditioned culture medium
by SDS–polyacrylamide gel electrophore-
sis (PAGE). LPS stimulation for 18 hours
induced the appearance of a 30-kD protein
that was not apparent at earlier time points.
The NH2-terminal sequence of this late-ap-
pearing factor (Gly-Lys-Gly-Asp-Pro-Lys-
Lys-Pro-Arg-Gly-Lys-Met-Ser-Ser) was
identical to murine HMG-1, a 30-kD mem-
ber of the high mobility group (HMG)
nonhistone chromosomal protein family (7,
8). Based on the HMG-1 sequence in Gen-
Bank (accession no. M64986), we designed
primers and isolated HMG-1 cDNA after
polymerase chain reaction (PCR) amplifi-
cation. Recombinant HMG-1 (rHMG-1)
protein was expressed in Escherichia coli,
purified to homogeneity, and used to gen-
erate polyclonal antibodies (9).
Immunoblot analysis revealed that large
amounts of HMG-1 were released from RAW
264.7 cells in a time-dependent manner (Fig.
1A), beginning 6 to 8 hours after stimulation
with LPS. Cell viability, as judged by trypan
blue exclusion and lactate dehydrogenase re-
lease, was unaffected by LPS concentrations
that induced the release of HMG-1, indicat-
ing that HMG-1 release was not due to cell
death. HMG-1 mRNA levels were unaffected
by LPS treatment (Fig. 1B), indicating that
HMG-1 release is unlikely to be linked to
increased transcription of the gene. Stimula-
tion of RAW 264.7 cells for 18 hours with
TNF (5 to 100 ng/ml) or IL-1? (5 to 100
ng/ml) also induced HMG-1 release in a cy-
tokine dose-dependent manner. In contrast,
stimulation with interferon-? (IFN-?) alone
did not induce HMG-1 release, even at con-
centrations up to 100 U/ml; however, IFN-?
increased by three- to fivefold the amount of
HMG-1 released by stimulation with either
TNF or IL-1 (10, 11). Pulse labeling experi-
ments with
most of the HMG-1 released during the first
12 hours after TNF and IFN-? stimulation
was derived from a preformed protein pool.
Radioactivity was incorporated into newly
synthesized HMG-1 from 12 to 36 hours after
macrophage stimulation (10, 11).
We next examined the inducible release of
HMG-1 from other cell types. LPS triggered
HMG-1 release from human primary peripheral
blood mononuclear cells and primary macro-
phages from LPS-sensitive mice (C3H/HeN),
but not from macrophages from LPS-resistant
C3H/HeJ mice (11, 12). Human primary T
35S-methionine revealed that
cells, rat adrenal (PC-12) cells, and rat primary
kidney cells did not release HMG-1 after stim-
ulation with LPS, TNF, or IL-1?. Like other
macrophage products (for example, TNF, IL-
1?, and macrophage migration inhibitory fac-
tor), HMG-1 lacks a classical secretion signal
sequence, so the mechanism of release remains
to be determined.
To determine if HMG-1 was released sys-
temically during endotoxemia in mice, we
measured serum HMG-1 levels after LPS
administration. Serum HMG-1 was readily
detectable 8 hours after administration of a
median lethal dose (LD50) of LPS and was
maintained at peak, plateau levels from 16 to
32 hours after LPS treatment (Fig. 1C).
About 20 to 50 ?g of HMG-1 was released
into the murine circulation within 24 hours
after endotoxin administration [assuming a
distribution half-life (t1/2) of 3 min and an
elimination t1/2of 20 min]; this is compara-
ble to the quantity of TNF and IL-1 released
by LPS treatment. The kinetics of HMG-1
appearance in the blood of LPS-treated mice
differs from that of previously described le-
thal LPS-induced mediators.
Passive immunization of unanesthetized
(anti–HMG-1) 30 min before a lethal dose
(LD100) of LPS did not prevent LPS-induced
death (Fig. 2A). Based on the kinetics of
HMG-1 accumulation in serum (Fig. 1C), and
the relatively short biological half-life of anti-
bodies to cytokines (3, 13), we reasoned that
complete neutralization of a late-appearing me-
diator might require repeated dosing. Adminis-
tration of anti–HMG-1 in two doses (one 30
min before LPS and one 12 hours after LPS)
increased the survival rate of the mice to 30%.
With three doses of antiserum (?30 min, ?12
hours, ?36 hours), 70% of the treated mice
survived, as compared with 0% survival in
controls treated with three matched doses of
preimmune serum (P ? 0.05). No late death
occurred over 2 weeks, indicating that anti–
HMG-1 did not merely delay the onset of LPS
lethality, but provided lasting protection.
To investigate whether antibody treatment
could be delayed until after administration of
LPS, we injected anti–HMG-1 beginning 2
hours after LPS (followed by additional doses
at 12 and 36 hours after LPS). This delayed
treatment conferred
against an LD100of LPS (Fig. 2B). Preimmune
Fig. 1. (A) Release of HMG-1 from cultured
macrophages after stimulation with LPS. Mu-
rine macrophage-like RAW 264.7 cells (Ameri-
can Type Culture Collection, Rockville, Mary-
land) were cultured in RPMI 1640 medium,
10% FBS, and 1% glutamine. When 70 to 80%
confluence was reached, cells were resuspend-
ed in serum-free OPTI-MEM I medium and
seeded onto tissue culture plates (5 ? 106cells
per well). After 2 hours, RAW 264.7 cells were
treated with LPS (E. coli 0111:B4, 100 ng/ml)
and proteins in the cell-conditioned medium
were fractionated by SDS-PAGE, excised from
Coomassie- stained SDS-PAGE gels, and subject-
ed to NH2-terminal sequencing analysis (Com-
monwealth Biotechnologies, Richmond, Virginia).
Polyclonal antisera against purified recombinant
HMG-1 were generated in rabbits (Biosynthesis,
Lewisville, Texas); immunoblotting showed that
antiserum reacted with native HMG-1 released
by RAW cells (inset). HMG-1 levels were mea-
sured by optical intensity of bands on immuno-
blots with NIH 1.59 image software, with refer-
ence to standard curves generated with purified
rHMG-1. Data are shown as the mean ? SE (n ?
3). (B) Expression of HMG-1 mRNA in macro-
phages. Murine macrophage-like RAW 264.7 cells
were cultured in RPMI 1640, 10% FBS, and 1%
glutamine, and stimulated with LPS (1 ?g/ml) for
isolated with the SV Total RNA Isolation System
(Promega) and levels of HMG-1 mRNA were de-
termined by reverse transcriptase (RT)–PCR with
the Access RT-PCR System (Promega; ?-actin
T-3? and 5?-CCTAGAAGCATTTGCGGTGCAC-
GATG-3?; and HMG-1 primers, 5?-ATGGGCA-
AAGGAGATCCTA-3? and 5?-ATTCATCATCAT-
CATCTTCT-3?). (C) Accumulation of HMG-1 in
serum of LPS-treated mice. Male Balb/C mice (20 to 23 g) were treated with LPS [10 mg/kg,
intraperitoneally (ip)]. Serum was assayed for HMG-1 by immunoblotting; the detection limit is?50 pg.
Data are shown as the mean ? SE (n ? 3).
www.sciencemag.org SCIENCEVOL 285 9 JULY 1999
Page 3
serum–treated controls all developed lethargy,
groomed and active, had no diarrhea, and were
viable. To clarify that anti–HMG-1 protected
mice from LPS lethality, we purified the immu-
noglobulin G (IgG) fraction from anti–HMG-1
and administered it to mice exposed to an
LD100of LPS (11, 14). The highest dose of
anti–HMG-1 IgG tested, 5 mg per mouse, con-
ferred complete protection against an LD100of
LPS, whereas all control mice given compara-
ble doses of rabbit IgG died (Table 1). Treat-
ment with anti–HMG-1 IgG (2 mg per mouse)
significantly reduced serum HMG-1 levels,
whereas no reduction was observed after treat-
ment with a lower dose of antibodies (0.5 mg
per mouse) or with control IgG (5 mg per
mouse). Antiserum against a chemically syn-
thesized peptide corresponding to the first 12
amino acids of HMG-1 also significantly atten-
uated the lethality of endotoxemia in mice (15).
To determine if HMG-1 was toxic, we
administered highly purified rHMG-1 to un-
anesthetized Balb/C mice (10 to 50 ?g per
mouse). Within 2 hours, the mice developed
signs of endotoxemia, including lethargy, pi-
loerection, and diarrhea. At higher doses (500
?g per mouse), three of five mice died at 18,
30, and 36 hours after rHMG-1 administra-
tion. Toxicity and lethality were not observed
in control mice treated with a protein fraction
purified from E. coli transformed with a plas-
mid devoid of HMG-1 cDNA (9), indicating
that the toxicity we observed was specific to
HMG-1. To exclude the possibility that en-
dotoxin contamination of HMG-1 prepara-
tions mediated lethality, we injected rHMG-1
into LPS-resistant mice. rHMG-1 (500 ?g
per mouse) was lethal within 16 hours both to
C3H/HeJ (n ? 4) and C3H/HeN (n ? 3)
mice, indicating that HMG-1 itself is toxic
even in the absence of LPS signal transduc-
tion. When sublethal doses of rHMG-1 were
injected into Balb/C mice together with sub-
lethal doses of LPS, the combined challenge
was lethal to 90% of the mice, as compared
with 0% lethality in mice exposed to LPS or
HMG-1 alone (Fig. 2C). Thus, HMG-1 itself
mediates lethality in both LPS-sensitive and
LPS-resistant mice.
Animal models of human sepsis, including
the murine endotoxemia model used here, have
inherent limitations (16). As an initial step in
determining whether HMG-1 participates in the
pathogenesis of human sepsis, we studied 8
Table 1. Protection against LPS lethality by anti–
HMG-1 IgG. Balb/C mice (male, 20 to 23 g, three
to six mice per group) were injected intraperito-
neally with IgG purified from anti–HMG-1 or con-
trol rabbit IgG 30 min before injection of an LD100
of LPS. All mice were then treated with additional
doses of anti–HMG-1 IgG or control IgG at 12 and
24 hours after LPS. Serum HMG-1 levels were
determined by immunoblots (under denaturing
conditions) 14 hours after LPS challenge (n ? 3
per group). ND, not determined.
Dose of HMG-1–
IgG (mg/mouse)
2 weeks
after LPS
1003 ? 180
1070 ? 20
415 ? 240
0 (0/6)
17 (1/6)
50 (3/6)*
100 (3/3)*
*P ? 0.05 versus treatment with control rabbit IgG.
Fig. 2. (A) Anti–HMG-1 protect against
LPS lethality in mice. Polyclonal anti-
bodies against rHMG-1 were generated
in rabbits, and antiserum was assayed
for specificity and titer by enzyme-
linked immunosorbent assay and im-
munoblotting. Antibodies reacted spe-
cifically with HMG-1 and did not cross-
react with LPS, other bacterial proteins,
TNF, or IL-1?. Immunoblots of lysates
of macrophages or E. coli transformed
with plasmid containing HMG-1 cDNA
revealed only one band of immunore-
activity. Male Balb/C mice (20 to 23 g)
were randomly grouped (10 mice per
group) and treated with an LD100of LPS
(25 mg/kg). Anti–HMG-1 (Ab) or preim-
mune serum (0.2 ml per mouse, ip) was
administered 30 min before LPS. Additional doses of preimmune (0.4 ml, ip) or anti–HMG-1 (0.4
ml, ip) were administered at 12 and 36 hours after LPS as indicated. (B) Delayed administration of
anti–HMG-1 protects against LPS lethality in mice. Male Balb/C mice (20 to 23 g) were randomly
grouped (seven mice per group) and treated with an LD100of LPS. Anti–HMG-1 or preimmune
serum (0.4 ml per mouse) was administered at 2, 24, and 36 hours after LPS. (C) Administration of
rHMG-1 is lethal to mice. Recombinant HMG-1 was purified and LPS content determined by the
Limulus Amoebocyte Lysate Test (Bio-Whittaker, Walkersville, Maryland). Purified rHMG-1 protein
contained ?2.5 ng of LPS per microgram of rHMG-1. Male Balb/C mice (20 to 23, 10 animals per
group) were injected with a nonlethal dose of LPS (3.1 mg/kg, ip). Purified rHMG-1 protein was
administered intraperitoneally in the doses indicated at 2, 16, 28, and 40 hours after LPS.
Fig. 3. Increased serum HMG-1
levels in human sepsis. Serum was
obtained from 8 healthy subjects
and 25 septic patients infected
with Gram-positive [Bacillus fragi-
lis (one patient), Enterococcus fae-
calis (one patient), Streptococcus
pneumoniae (four patients), Liste-
ria monocytogenes (one patient),
or Staphylococcus aureus (two pa-
tients)], Gram-negative [E. coli
(seven patients), Klebsiella pneu-
moniae (one patient), Acineto-
bacter calcoaceticus (one patient),
Pseudomonas aeroginosa (one pa-
tient), Fusobacterium nucleatum
(one patient), Citrobacter freundii
(one patient)], or unidentified
pathogens (five patients). Serum
was fractionated by SDS-PAGE, and HMG-1 levels were determined by immunoblotting analysis with
reference to standard curves of purified rHMG-1 diluted in normal human serum; the detection limit is
?50 pg. *P ? 0.05 versus normal. **P ? 0.05 versus survivors.
9 JULY 1999VOL 285SCIENCEwww.sciencemag.org
Page 4
normal subjects and 25 critically ill septic pa-
dysfunction. HMG-1 was not detectable in the
serum of normal subjects, but significant levels
were observed in critically ill patients with sep-
sis (Fig. 3), and these levels were higher in
patients who succumbed as compared to pa-
tients with nonlethal infection.
HMG-1 is a highly conserved protein with
?95% amino acid identity between rodent and
human (17–20). It has previously been charac-
terized as a nuclear protein that binds to cruci-
form DNA (21), and as a membrane-associated
protein termed "amphoterin" that mediates neu-
rite outgrowth (19, 20). Extracellular HMG-1
interacts directly with plasminogen and tissue
type plasminogen activator (tPA), which en-
hances plasmin generation at the cell surface;
this system plays a role in extracellular prote-
olysis during cell invasion and tissue injury
(19). In addition, HMG-1 has been suggested to
bind to the receptor for advanced glycation end
products (RAGE) (22).
As with other inflammatory mediators
such as TNF and IL-1, there may be protec-
tive advantages of extracellular HMG-1 when
released in nontoxic amounts. Macrophages
release HMG-1 when exposed to the early,
acute cytokines, indicating that HMG-1 is
also positioned as a mediator of other inflam-
matory conditions associated with increased
levels of TNF and IL-1 (for example, rheu-
matoid arthritis and inflammatory bowel dis-
ease). Indeed, in most inflammatory scenari-
os, LPS is probably not the primary stimulus
for HMG-1 release; it seems more likely that
TNF and IL-1 function as upstream regula-
tors of HMG-1 release. The delayed kinetics
of HMG-1 release suggest that serum HMG-1
levels may be a convenient marker of disease
severity. Moreover, the observations that
HMG-1 itself is toxic, and that anti–HMG-1
prevents LPS lethality, point to HMG-1 as a
potential target for therapeutic intervention.
References and Notes
1. K. J. Tracey et al., Science 234, 470 (1986); C. A.
Dinarello, FASEB J. 8, 1314 (1994); D. C. Morrison and
J. L. Ryan, Annu. Rev. Med. 38, 417 (1987); L. L.
Moldawer, Crit. Care Med. 22, S3 (1994).
2. B. Beutler, N. Krochin, I. W. Milsark, C. Luedke, A.
Cerami, Science 232, 977 (1986); J. W. Killion and
D. C. Morrison, Infect. Immun. 54, 1 (1986); A. Pol-
torak et al., Science 282, 2085 (1998).
3. K. J. Tracey et al., Nature 330, 662 (1987); M. Bianchi
et al., J. Exp. Med. 83, 927 (1996); H. R. Alexander,
G. M. Doherty, C. M. Buresh, D. J. Venzon, J. A.
Norton, J. Exp. Med 173, 1029 (1991); G. Wakaba-
yashi, J. A. Gelfand, J. F. Burke, R. C. Thompson, C. A.
Dinarello, FASEB J. 5, 338 (1991).
4. D. G. Hesse et al., Surg. Gynecol. Obstet. 166, 147
(1988); H. R. Michie et al., N. Engl. J. Med. 318, 1481
5. M. P. Fink, Crit. Care Med. 23, 989 (1995); E. Abra-
ham, Chest 113 (suppl.), 224S (1998).
6. M. W. Marino et al., Proc. Natl. Acad. Sci. U.S.A. 94,
8093 (1997); F. Amiot, C. Fitting, K. J. Tracey, J. M.
Cavaillon, F. Dautry, Mol. Med. 3, 864 (1997).
7. M. E. Bianchi, M. Beltrame, G. Paonessa, Science 243,
1056 (1989); J. M. Walker, K. Gooderham, J. R. Hastings,
E. Mayes, E. W. Johns, FEBS Lett. 122, 264 (1980); L.
Einck and M. Bustin, Exp. Cell Res. 156, 295 (1985).
8. HMG-1 has also been termed "amphoterin" [J. Park-
kinen et al., J. Biol. Chem. 268, 19726 (1993)].
HMG-1 and amphoterin are the same protein; we use
the name "HMG-1" to reflect the original description
of this protein as the first member of the HMG
protein family.
9. HMG-1 was cloned by DNA amplification of the
648–base pair (bp) open reading frame from Rat
Brain Quick-Clone cDNA (5 ng; Clontech, Palo Alto,
CA) with the following primers: 5?-CCCGCGGATC-
and 5?-CCCGCAAGCTTATTCATCATCATCATCTTCT-
3? (PCR at 94°C for 1 min, 56°C for 2 min, 72°C for
45 s; 30 cycles). The 680-bp PCR product was
digested with Bam HI and Hind III and subcloned into
the Bam HI–Hind III cloning sites of the pCAL-n
vector (Stratagene, La Jolla, CA). The recombinant
plasmid was transformed into E. coli BL21(DE3)pLysS
(Novagen, Madison, WI), and positive clones were
confirmed by DNA sequencing of both strands. Trans-
formed cells were induced with isopropyl-D-
thiogalactopyranoside, and rHMG-1 protein was pu-
rified with a calmodulin-binding resin column (Strat-
agene). As controls for experiments involving admin-
istration of rHMG-1 to mice, we purified proteins
from E. coli BL21(DE3)pLysS that had been trans-
formed with a plasmid that lacks the HMG-1 cDNA
insert (pCAL-n). The amount of control material
administered to mice was normalized to the number
of E. coli that produce 0.5 mg of rHMG-1.
10. H. Wang and K. J. Tracey, unpublished observations.
11. Supplementary data can be found on Science Online
at www.sciencemag.org/feature/data/1037699.shl.
12. Macrophages were obtained from the peritoneal cavity
of LPS-sensitive (C3H/HeN and Balb/C) or LPS-resistant
(C3H/HeJ) mice 4 days after intraperitoneal injection
with 2.0 ml of thioglycollate broth (4%; Difco, Detroit,
MI). Macrophages were pooled from four mice, resus-
pended into RPMI 1640, 10% fetal bovine solution
(FBS), and 1% glutamine, and plated at a density of 4 ?
106cells per well in six-well Falcon Primaria tissue
culture plates. After 24 hours, the culture medium was
replaced with serum-free OPTI-MEM-I medium, and LPS
(1 ?g/ml) was added. The level of HMG-1 in the
culture medium was determined 18 hours later by
immunoblotting. HMG-1 was not detectable in
culture medium of LPS-stimulated C3H/HeJ murine
macrophages; HMG-1 levels reached 1 ?g/106cells
in the culture medium of LPS-stimulated C3H/HeN
murine macrophages.
13. L. B. Hinshaw et al., Circ. Shock 30, 279 (1990); C.
Nathan and M. Sporn, J. Cell Biol. 113, 981 (1991);
D. G. Remick et al., Am. J. Pathol. 136, 49 (1990); R. E.
Walker et al., J. Infect. Dis. 174, 63 (1996).
14. IgG was purified from anti–HMG-1 by Protein G–
Sepharose HiTrap affinity chromatography (Phar-
macia Biotech). Purified IgG fractions were de-
salted by ultrafiltration through Centricon-10 (Mil-
lipore), followed by two washes with 1? phos-
phate-buffered saline. The specificity of IgG was
confirmed by immunoblot analysis of macrophage
lysates, which revealed one band of 30 kD. Anti–
HMG-1 IgG did not cross-react with LPS, TNF, IL-1,
or bacterial proteins on immunoblots.
15. H. Wang and K. J. Tracey, unpublished observations.
16. M. P. Fink and S. O. Heard, J. Surg. Res. 49, 186
(1990); K. J. Tracey, and E. Abraham, SHOCK 11, 224
(1999); E. A. Deitch, ibid. 9, 1 (1998).
17. K. L. Lee et al., Nucleic. Acids Res. 15, 5051 (1987); S.
Ferrari, L. Ronfani, S. Calogero, M. E. Bianchi, J. Biol.
Chem. 269, 28803 (1994); S. Ferrari, P. Finelli, M.
Rocchi, M. E. Bianchi, Genomics 35, 367 (1996).
18. J. F. Maher and D. Nathans, Proc. Natl. Acad. Sci.
19. J. Parkkinen et al., J. Biol. Chem. 268, 19726 (1993).
20. M. Salmivirta, H. Rauvala, K. Elenius, M. Jalkanen, Exp.
Cell Res. 200, 444 (1992).
21. M. E. Bianchi and M. Beltrame, Am. J. Hum. Genet. 63,
1573 (1998); D. Landsman and M. Bustin, Bioessays
15, 539 (1993); M. Bustin and N. K. Neihart, Cell 16,
181 (1979).
22. O. Hori et al., J. Biol. Chem. 270, 25752 (1995); A. M.
Schmidt et al., Diabetes 45, S77 (1996).
23. We thank C. Dang for technical assistance; J. Eaton, J.
Roth, B. Sherry, M. Bukrinsky, and M. Symons for
critical reading of the manuscript; and D. Prieto for
administrative assistance.
11 December 1998; accepted 3 May 1999
Ploidy Regulation of Gene
Timothy Galitski,1Alok J. Saldanha,1,2Cora A. Styles,1
Eric S. Lander,1,2Gerald R. Fink1,2*
Microarray-based gene expression analysis identified genes showing ploidy-
dependent expression in isogenic Saccharomyces cerevisiae strains that varied
in ploidy from haploid to tetraploid. These genes were induced or repressed in
proportion to the number of chromosome sets, regardless of the mating type.
Ploidy-dependent repression of some G1cyclins can explain the greater cell size
associated with higher ploidies, and suggests ploidy-dependent modifications
of cell cycle progression. Moreover, ploidy regulation of the FLO11 gene had
direct consequences for yeast development.
Changes in the number of chromosome sets
occur during the sexual cycle, during meta-
zoan development, and during tumor progres-
sion. Organisms with a sexual cycle double
their ploidy upon fertilization and reduce
their ploidy by half at meiosis. In the devel-
opment of almost all plants and animals, spe-
cialized polyploid and polytene cell types
arise though endocycles, cell cycles lacking
cell division (1). Aberrant cell cycle control
during tumor progression is thought to result
in polyploidy and altered cell behavior (2).
Cells of different ploidy typically show very
different developmental, morphological, and
physiological characteristics. However, a lack
1Whitehead Institute for Biomedical Research, 9 Cam-
bridge Center, Cambridge, MA 02142, USA.2Depart-
ment of Biology, Massachusetts Institute of Technol-
ogy, Cambridge, MA 02139, USA.
*To whom correspondence should be addressed. E-
mail: email@example.com
www.sciencemag.orgSCIENCEVOL 2859 JULY 1999


text:
Post Content
Following up on its excellent guides for businesses and higher education, the CDC recently released new guidance for child care providers to help them prepare for the upcoming flu season. Child care providers, both center-based and home-based, are critically important partners in helping to limit the spread of flu. H1N1 flu disproportionately impacted young people during the outbreak in the spring. The expected resurgence of the virus this fall will likely continue this trend.
The toolkit for child care providers includes:
- Question & Answers on H1N1
- Fact sheets for child care providers and for parents
- Prevention posters informing parents to keep sick kids home
- Template letters for parents
Tags: Child Care Providers, DPH, flu, H1N1, influenza, Massachusetts, Prevention
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This week's flu report shows a slight dip in rates of flu-like illness since last week's report – which is entirely in keeping with the unpredictable nature of flu season. One thing we know for sure is that no matter what, the single best way to …Continue Reading Weekly Flu Report, December 12, 2014
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The December monthly meeting of the Public Health Council featured the consideration of one Determination of Need (DoN) request, two votes on final amendments to existing regulations, and an informational presentation to the Council on a key DPH community initiative. First, the Council took up …Continue Reading Highlights of the Public Health Council Meeting, December 10, 2014
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subset: commoncrawl


text:
Winnipeg Free Press - ONLINE EDITION
Provinces being asked to fund Experimental Lakes Area
Environmental groups are calling on Ontario and Manitoba to split the cost of funding to keep the Experimental Lakes Area open.
Each province is being asked to pitch in $1 million a year for the next three years. Ottawa announced last winter it's cutting funding this April.
For decades the ELA has been one of the world's biggest natural laboratories and the source of pioneering work on pollution, like identifying phosphorus as a pollutant, leading to its removal from laundry detergent plus dozens of other practical measures to protect the environment.
Environmentalists and scientists in Canada strenuously lobbied the federal government for the funding to resume. And the decision triggered criticism from around the world, with scientists sending open letters calling for Canada to not abandon the ELA research.
Now the Boreal Forest Network and the Council of Canadians is calling upon the Ontario and Manitoba governments to fund the lakes in an open letter Thursday.
"Sadly our federal government is failing in its responsibility to foster credible scientific research to base decisions affecting our water, environment and climate. For this reason we look to your governments to fulfill this critical role," an announcement from the groups stated.
The lakes are located in Ontario and the Freshwater Institute is located in Manitoba. Each province would need to contribute $1 million annually until federal funding is re-instated." The groups are asking for a three year minimum commitment.
Earlier this week, Diane Orihel, founder of the Coalition to Save ELA, was presented with the prestigious Diamond Jubilee Medal. This award recognizes Canadians who have made significant contributions to their communities and who have brought credit to Canada.
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UFC fighter Tim Kennedy joins Cam & Co. for MMA Week, April 22- 26. Kennedy discusses how his time in the armed forces shaped who he is today, as well as the hog overpopulation problem in his home state of Texas. Originally aired on Sportsman Channel Cam & Co. 4/26/13.
Kennedy discusses how his time in the armed forces shaped who he is today.
December 19th, 2014Steve Chapman: Targeting 'Assault Weapons' Again
The journalist contends that the Newtown families' lawsuit ignores the fact that the vast majority of AR-15s are used responsibly.
View the latest Cam & Co shows.
Have a Second Amendment or firearm-related question? Ask Cam!
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Winnipeg Free Press - PRINT EDITION
The Buzz
East Meets Western
A Fistful of Dollars, the 1964 movie that arguably launched Clint Eastwood's movie career, was a western-ized remake of Akira Kurosawa's 1961 samurai movie Yojimbo. Proving that the cinematic cultural exchange goes both ways is the trailer for Yurusarezaru mono, a Japanese remake of Eastwood's Oscar winning 1992 film Unforgiven, starring Ken Watanabe (Inception) as a retired samurai tempted to get back in the violence biz by a large bounty. The trailer is at http://wfp.to/V4s.
Get behind the wheel
If people you know are hard to reach on Sept. 17, it's probably because they have immersed themselves in the latest Grand Theft Audio video game (bit.ly/GTA5_video). With a reimagined open-world concept in which players can drive any vehicle they want, go scuba diving, fly planes, go golfing, head out on rampages, rob banks and generally cause mayhem, the newest edition of the popular and controversial series will be flying off shelves this fall. Early trailers and sneak peeks of the most anticipated game of the year suggest that Rockstar Games has gone all-out in this new edition, tying together addictive and expansive online play with a new multi-character approach to the main story that will let you play from any of the three main characters' perspectives at any time in the game. This seemingly small change will not only extend the game play time, but gives the series a whole new twist. bit.ly/GTA5_video
Sharks take a bite out of summer
Remember those wide-eyed, innocent times, before Sharknado rained fin-flashing, razor-toothed doom from above, when you thought being a Prairie dweller meant safety from shark attacks? Well, fellow flatlander, those days are over. And if Space's recent broadcast of the instant camp classic wasn't enough to scare you silly, wait til 2010's Sharktopus (Tuesday, July 23 at 8 p.m. on Space) brings its terrifying tandem of teeth and tentacles to your TV screen. And then ... be very afraid.
Five days more ...
To the barricades as Cameron Mackintosh's 25th anniversary production of Les Miserables arrives at the Centennial Concert Hall July 23 for an eight-performance run. Come for the fabulous score -- think I Dreamed a Dream, On My Own and Master of the House -- and stay for the uplifting, epic Victor Hugo story about an unjustly hounded fugitive Jean Valjean. The new staging was inspired by Hugo's paintings.
Perhaps Lord Voldemort had a hand in it
Even an amateur sleuth might think it suspicious that Harry Potter author J.K. Rowling is unmasked as the real writer of an obscure mystery novel, The Cuckoo's Calling, a week before the trade paperback release of her first adult novel, The Casual Vacancy. Could the Sunday Times of London journalist who broke the story of the pseudonymous "Robert Galbraith" have been played like a fiddle to maximize sales for two titles instead of one?
Republished from the Winnipeg Free Press print edition July 18, 2013 ??65534
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Chop the full-flavoured and delicate fish into large, bite-sized pieces and refrigerate (keeping the full-flavoured fish separate from the delicate).
Put the rinsed fish heads and bones in a large soup pot, pour the water over, bring to a boil, and simmer twenty minutes.
Strain, reserving the liquid.
Heat the oil in the same pot and fry the onions, leeks, and fennel until soft, about 10 minutes.
Add the tomatoes, garlic, bouquet garni, fennel seed, orange peel, and saffron.
Season with salt and pepper.
Pour over the fish stock, and simmer half an hour.
Remove the bouquet garni and the orange peel.
Bring the liquid to a rolling boil.
Add the full-flavoured fish and cook 5 minutes.
Add the delicate fish and the splash of Pernod and continue cooking until all the fish is tender, about 5 minutes longer.
Ladle into warm soup plates, sprinkle irreverently with parsley if you like, and top with a rouille-smeared croûte.
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Nation's largest gun show promoter criticizes proposed firearms legislation
Laura Seitz, Deseret News
SANDY — The owner of the largest gun show promoter in the country said he wouldn't support tougher background checks at his exhibitions.
Bob Templeton, owner and CEO of Kaysville-based Crossroads of the West, was preparing Friday for this weekend's showcase at the South Towne Exposition Center.
Templeton said his Jan. 10 visit with Vice President Joe Biden did little to change his stance on gun control. Biden had invited him to Washington, D.C., to discuss issues surrounding gun violence in the U.S.
"It was a cordial meeting," Templeton said. "Vice President Biden is an engaging guy. But generally there was no agreement on how to move forward."
Templeton said he blames the bulk of gun violence on the lack of enforcement of existing laws and a failed mental health system rather than illicit purchases at shows such as Crossroads of the West.
He pointed out that more than 72,000 Americans failed background checks in 2010, but only 44 were prosecuted for providing false information.
"The vice president told me, 'We don't have the resources or personnel to prosecute everyone who lies on a background check form,'" Templeton said. "(I asked), 'How then can we be expected to support new legislation that adds to that burden?' His response was, 'Well, we'll address that when we get to it. Next question.'"
Templeton, who is also the founder and president of the National Association of Arms Shows, said tightening the grip on gun show sales would only impact law-abiding gun owners. He said his customers and business partners are being demonized, that existing laws are sufficient to solve the problem, and that American gun violence has more to do with mental health than firearms.
"There are over 100 million responsible gun owners in America. We are the scapegoats for the violent acts of one unstable man who committed unspeakable tragedies," he said, referring to the recent school shooting in Newtown, Conn.
Salt Lake City Police Chief Chris Burbank also recently visited the White House, joining other police chiefs from throughout the country to discuss gun control with President Barack Obama.
Burbank said he doesn't believe instituting more stringent background checks and banning specific weapons violates the Constitution.
"There are limitations to constitutional rights," he said. "We need to … realize that the Second Amendment doesn't guarantee you access to any firearm you want and the ability to carry it anywhere you want. Reasonableness says we're going to limit access."
The issue is divisive even among vendors at Templeton's gun show.
Russ Garrison regularly sells first-aid equipment at gun shows, including Crossroads of the West, and said most of his customers are avid gun owners. However, he supports a ban on assault weapons and high-capacity magazines.
"I could see a ban happening and I don't have any problem with that," Garrison said. "I'm an (Automatic Rifle) owner and I use 30 round magazines, but I could have enough fun with 10 rounds."
But Garrison is less optimistic that closing the so-called gun show loophole will reduce violent crime. He said too many gun-control advocates are trying to paint America's gun-owning population as violence-loving and paranoid.
"Nobody likes to be labeled. There are so many good people involved with guns who are big into hunting and camping and fishing," Garrison said. "And I'm going to support them. I'm still a believer it's not the gun that kills the person; it's the person who kills the person."
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RBI Keeps Rates on Hold: What Experts Say
Sep 30, 2014
Little Chance of Rate Cut in FY15: Axis Bank
Sep 29, 2014
RBI May Cut Rates Early Next Year: Rangarajan
Thursday December 18, 2014
The latest policy statement has clearly given indication if the price behaviour is along the trajectory envisage...I expect probably changes will come in the early part of next year. It could be any time.
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Tuesday December 16, 2014
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Friday December 5, 2014
At the monetary policy review earlier this week, RBI Governor Raghuram Rajan chose to hold rates, saying it is premature to go in for a cut, but painted an optimistic picture, saying inflation is coming under control. However, he hinted of a cut in e...
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Debt Markets Start Pricing in Early 2015 Rate Cut; Rally Seen to Continue
Tuesday December 2, 2014
Traders said they expect the debt and swap market rally to continue in the near-term, though the pace of the rally may slow until there is more clarity on the timing of the rate move.
What Raghuram Rajan Said at RBI Policy Meet Today
Tuesday December 2, 2014
Governor Raghuram Rajan signaled a softening of stance on interest rates, which could pave the way for a rate cut early next year.
Sensex, Nifty Choppy After RBI Keeps Rates Steady
Tuesday December 2, 2014
At 12:12 pm, Sensex was down 133 points to 28,426 while Nifty fell 40 points to 8,515. Selling pressure was seen in rate sensitive banking, real estate and auto stocks.
Sensex Recovers After Dipping Briefly; Early Rate Cut Hopes Boost
Tuesday December 2, 2014
Traders said a rate cut in early 2015 was more or less certain now unless global commodity prices moved adversely.
RBI Opens Door For Rate Cut Early Next Year
Tuesday December 2, 2014
Forty-one of 45 economists polled by Reuters had forecast that the RBI would keep the repo rate at 8.00 per cent, while four expected a reduction of 25 basis points.
Raghuram Rajan Holds Repo Rate Despite Government Pressure
Tuesday December 2, 2014
Most economists had expected Dr Rajan to hold rates despite pressure from Finance Minister Arun Jaitley, who had pushed for a cut in interest rates in a bid to lift demand and drive growth. The Indian economy is struggling to emerge from its slowest ...
Despite Government Pressure, RBI Unlikely to Cut Rates: 10 Facts
Tuesday December 2, 2014
The Reserve Bank of India is expected to keep interest rates unchanged on Tuesday, staying focused on containing inflation.
Rates Have Come Down But Banks Haven't Passed On: Raghuram Rajan
Tuesday December 2, 2014
Reserve Bank Governor Raghuram Rajan on Tuesday held repo rate at 8 per cent as expected. The central bank has not cut rates since May 2013 despite retail inflation easing to a record low of 5.5 per cent in October.
RBI Announcement at 11 am Unlikely to Cut Interest Rates
Tuesday December 2, 2014
Forty-one of 45 economists saw the RBI keeping the repo rate at 8 per cent at its policy review, while four expected a reduction of 25 basis points (0.25 per cent).
Why Raghuram Rajan May Not Oblige Arun Jaitley With a Rate Cut
Tuesday December 2, 2014
Reserve Bank Governor Raghuram Rajan will be under the burden of huge expectations when he delivers his bi-monthly monetary policy on Tuesday. Indian stock markets have run up to record highs on expectations of a rate cut.
RBI Chief Rajan's Tryst With Tuesdays: Preferred Day for Policy Reviews
Sunday November 30, 2014
It may be sheer coincidence that all monetary policy reviews by Reserve Bank of India (RBI) Governor Raghuram Govind Rajan this fiscal year have taken place on Tuesdays and this would be the case for the upcoming review as well.
RBI May Keep Policy Rate Unchanged on December 2
Sunday November 30, 2014
The Reserve Bank of India (RBI) may keep policy rate unchanged in its upcoming monetary policy review on Tuesday even as the Finance Minister and industry have clamoured for the rate cut to prop the economy after GDP growth slipped to 5.3 per cent in...


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On June 15, in the same moment the Bruins won something great, they lost something great. Game 7 of the Stanley Cup finals was Mark Recchi's last career game, so whether the Bruins entered the 2011-12 season as Stanley Cup champions or simply Eastern Conference champs, they would be doing so without one of their alternate captains from a historic season.
With Recchi now retired, someone else will join captain Zdeno Chara and longest-tenured Bruin Patrice Bergeron as bears to bare a letter other than the spoked 'B' on their jerseys.
There are three obvious candidates who come to mind: a two-time Cup champion and resident bruiser (Shawn Thornton), the longest-tenured Bruin not named Tim Thomas or Marc Savard to not have a letter on his sweater (Andrew Ference) and the young star who brings the two things fans love in goals and fights (Milan Lucic).
All three are deserving of the distinction, with Thornton and Ference perhaps more deserving based on their time in the league. Here's a look at the argument for each one.
Thornton is the everyman in a room full of down-to-earth guys. He's great with the media, as he answered a question about pressure when down in the conference semifinals vs. the Canadiens by saying, "pressure is five kids and no job." He is the ultimate believer in the "never get too high, never get too low" mindset, and it's a lesson his teammates have listened to.
To best illustrate how wide-ranging Thornton's impact is on the Bruins, consider that Recchi spent the night before Game 7 having a glass of wine with No. 22, Recchi told ESPN Boston this summer. Thornton also sat just a couple spots down from rookie Tyler Seguin in the B's room, and it was Thornton's motivational talks that Seguin said kept him upbeat during his healthy scratches, despite the fact that the two were essentially competing for the final roster spot. And who did Brad Marchand play with for the first three months of the season?
Thornton's impact when put in the lineup in Game 3 of the Cup finals was enormous, and it was definitely felt throughout the lineup. Yet it's just that that might hurt his case when it comes to getting a letter. If the B's find themselves in a similar situation next year regarding the decision to sit Seguin or Thornton, it sure would look weird if they made their alternate captain a healthy scratch, wouldn't it?
At the end of the day, Thornton is going to be a major leader on this Bruins team no matter what. An 'A' sure would let the rest of the league know, though.
If you were on the Bruins to the result of the team's work turn into the selection of Zach Hamill (eighth overall in 2007), you have seen a lot, and you've seen them come a long way.
Andrew Ference, like Bergeron, Chara, Tim Thomas, and David Krejci to an extent, is one such Bruin. Acquired in February of 2007, Ference was with the team when it was terrible, and he's been there as the organization has turned it around.
Ference may stand out given his political activism and his outspoken nature, but he did not cause a ruckus in the dressing room over the course of the championship season. Yes, that is including him saying Daniel Paille's hit on Raymond Sawada was dirty and had to be taken out of the game. To be completely honest, I didn't hear him say it because I was with Steve Buckley and Bergeron, the latter of whom said the exact same thing. That was an issue that only some in the media chose to make a big deal of, but it didn't impact the B's.
In fact, the only extra stuff he's brought have been T-shirts in support of Adam McQuaid (his daughter coined "Darth Quaider," causing him to customize a T-shirt online. He also sported a Kenny Powers shirt, given the similarities in mullet between McQuaid and Danny McBride's HBO character) as well as the disgusting late-'80s Bruins jacket he purchased on eBay for $35, the latter of which became a big part of the Cup run and now resides with Recchi as a retirement gift.
While he may not always be on the ice given his history of nagging injuries (he played 60 games or less in three straight seasons before playing 70 last year), the selling point when it comes to Ference's candidacy that, as much as it may surprise some given the comments regarding Paille, is that he always has his teammates' backs, and he proved it with his fists rather than his words. Even if Ference couldn't possibly have a great angle on the play, if he didn't like the way his teammate went down, somebody was going to pay for it. It's a trait instilled in some other Bruins (Thornton and Gregory Campbell immediately come to mind), but it was the most obvious with Ference last season.
For a team like the Bruins, having that trait (and being a defenseman doesn't hurt either) means embodying what the Bruins pride themselves on: disciplined hockey and a damned physical way of bringing it. Throw in the length of time he's spent in Boston and the respect he's earned and he's a great candidate.
Lucic turned in a career year last season, scoring 30 goals as a 22-year old and becoming the first Bruin to reach the mark since Phil Kessel in the 2008-09 season. He also finished with seven fighting majors (tied with Ference for fourth on the team, but just half of Thornton's total).
Back when the Bruins had hit their lowest point in late December, some idiot wrote that Nathan Horton's lack of production (and, at the time, physicality) may have been a bad influence on Lucic, but the truth is that the two feed off each other. Lucic is the type of player a player would want to play with. His scoring will help your plus-minus and the edge he plays with makes those around him tougher.
Above all, he certainly looks like a player who is in the process of growing into a leader. He is a player today's young hockey fans look up to, and talking to a kid like Anthony Camara (the Bruins' third-round pick in June's draft) gives you the impression that Lucic is among the guys young power-forwards-in-the-making try to model their games after. Lucic is also a good representative of the Bruins with the media – he knows what to say and how to say it honestly.
The issue with Lucic is that his motor can often cause him to run a little hot. Whether or not it was preceded by a dangerous hit that went uncalled (and it was high), the sight of Lucic punching an unsuspecting Freddy Meyer in the temple was the last thing either club wanted to see on Dec. 23. (For what it's worth, the hit from Meyer was one of the instances in which Ference took matters into his own hands, as he got a roughing minor on the play, while Lucic got a match penalty). Lucic also punched Victor Hedman in the face late in Game 1 of the conference finals at the end of a play, which escalated things unnecessarily in a lost game that only had nine seconds remaining. It's those little temptations that Lucic needs to avoid when he eventually wears an 'A' on his jersey. Assuming he stays in Boston long, it's a safe bet that he'll eventually wear a letter, even if he doesn't get it this year.
WHO SHOULD GET IT: Ference
To use two clichés in one sentence, the Bruins can't completely replace Recchi, but they really can't go wrong with either of these guys. The idea of Lucic losing his cool with a dangerous play every once in a blue moon while wearing an 'A' isn't the most appetizing thought, which makes him the first to be crossed off the list. Thornton will always be a leader of any team on which he plays, and the fact that he's never needed a letter to do it speaks volumes to the character of Oshawa native.
Ultimately, a coin could be flipped between Thornton and Ference. While the distinction would be a great way to follow what will likely be the best season of Thornton's career (10 goals, 10 assists), Ference brings the complete package and epitomizes the Bruins in a way for which he may not always receive credit. He'll get the slight edge here, though don't be surprised if Lucic ends up getting it.
EAST RUTHERFORD, N.J. — WEEI.com's Mike Petraglia and Chris Price explain how the Patriots held off the Jets, 17-16, Sunday at MetLife Stadium to improve to 12-3 and lock up a first-round bye in the AFC playoffs.
Greg and Steve give their final thoughts and predictions for what is about to go down at MetLife Stadium today.
Curran calls the guys to discuss the game today and gets into the effect of having no Edelman. He talks about Amendola's role, the Pats running game and more.
Jackie MacMullen, ESPN Boston, talks with MFB about Rondo being traded to the Dallas Mavericks, and how this will have an effect on the Celtics rebuild process
How have the Celtics looked after a couple weeks? Could they be a playoff team? These questions and more...
Sam and Ben talk with Greg Dickerson about how fun this year's Celtics team is to watch as well Jeff Green trade ideas.
Mut, Rob, and Alex talk with former Red Sox pitcher Jon Lester. Jon talks about his experiences with free agency, what it would have taken to sign him to the Red Sox last season, and his experiences with fans since signing with the Cubs.
Mut, Rob, and Alex talk with newly signed Red Sox pitcher, Wade Miley. They talk about the story that came out about the D-Backs wanting him to switch to a gluten-free diet, and what he expects coming from the N.L. West to the A.L. East.
The boys are back at it, Mut, Rob, and Alex talk with former Red Sox pitcher and new Chicago Cub Jon Lester, they then talk with new Red Sox pitcher and ongoing consumer of gluten, Wade Miley.
Pierre joins the show to talk about the Bruins-Wild game, Bruins trade rumors, and the rest of the NHL.
Jack Edwards talks with Dale, Michael and Jerry Thornton and drops the bomb on us that based on how the standings look now, the Bruins could miss the playoffs this season.
Pierre joins the guys to talk about the Bruins and all things NHL
Curtis spoke with some nut jobs outside the courtroom.
The top stories of the day as recounted by Kirk Minihane.
Jeff Goodman discussed the Rondo trade.
Christian quizzes Lou and Tim on which songs were used for torture by the CIA, and Tim is upset he couldn't remember Team America: World Police
Adam Schefter, ESPN, joins MFB to discuss Jim Harbaugh rumors, Rex Ryan's future, and why the Patriots may be better served by not wraping up the number one seed in the AFC this week
Christian shares what he believes will be the three keys to this Sunday's game against the New York Jets
We check in with the playmaker, the hall of famer Michael Irvin for a look at the NFL and all things Patriots as they prepare to face the Jets.
Dale and Michael discuss the trade of Rajon Rondo, and whether it ultimately was a good deal for the Celtics.
We break down Rex and the Jets vs the Patriots with CSNNE's Tom E Curran
Kay comes into the show to try out for the textress role on the program. We learn about her favorite sports and fondness for Rajon Rondo.
Mikey, Mut, Lenny and Kerry Byrne are taling Patriots-Jets and the potential end of Rex Ryan's coaching career with the team.
Tonight marks 9 straight years of the Planet Mikey Show. We play some sound from the takeover which got Planet Mikey rolling .
Jen Royle joins the show to talk about The Taste and what its like working with amazing chefs and how her personality is going over with viewers.
What should Danny Ainge do with these Celtics? Is Anthony Davis the second best player in the NBA? Are the Warriors truly contenders?
Ryan Lambert of Puck Daddy joins DJ to discuss the "uncoachable(?)" Taylor Hall and revisit the Tyler Seguin trade. Ryan also shares his issues with how the Bruins are run.
Rodney Harrison joined the show to discuss the Pats and Jets.More from this show
Curtis spoke with some nut jobs outside the courtroom.More from this show
Jeff Goodman discussed the Rondo trade.More from this show
Adam Schefter, ESPN, joins MFB to discuss Jim Harbaugh rumors, Rex Ryan's future, and why the Patriots may be better served by not wraping up the number one seed in the AFC this weekMore from this show
Dale and Michael are live on the air as the Rondo deal goes down.More from this show


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TV Show (1999 - 2014)
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Local listings for: Virginia [Set TV Provider]Sunday, December 21 2014
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Me and my fiancé have enjoyed following Joel. We have searched, and will continue to search for a church in our area, but until than and always, we LOVE listening and following Joel! Thank you Joel, God bless you! AMEN Report abuse
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Carrie Underwood's Maternity Style
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Michelle Dockery Previews 'Downton Abbey' Season 5
Sony, YouTube To Stream 'The Interview'
Santa's Celebrity Naughty List 2014!
Clint Eastwood & Wife Finalize Divorce
Access Hollywood: December 24, 2014
Andy Cohen Tells All In 'The Andy Cohen Diaries'
Megan Fox & Brian Austin Green Give Back
Fergie Shows Hometown Love In 'L.A. Love (La La)' Music Video
Jeanne Benedict's Holiday Recipes
5 Easy No-Bake Recipes For Kids
Quick & Easy Christmas Brunch!
Access Your Beauty: Birchbox Holiday Gift Ideas
Last Minute Holiday Cocktail Party Ideas
Watch the latest Access Hollywood Live
Astrid McGuire's 'Hot Santa' Workout!
Vivica A. Fox Recalls Working With Joan Rivers On 'Celebrity Apprentice'
October 22, 2012 04:34 PM EDT
Emily VanCamp is showing off her sexy side in the new issue of GQ.
The "Revenge" star, who showed off a revealing look for the November issue of the men's magazine, addressed why she loves playing bad girl Emily Thorne on the ABC drama.
"I always feel like I'm playing a character when I'm in a beautiful dress, with my hair coiffed. I like getting down and dirty. It's really fun," she told the mag. "There's nothing better than setting an a**hole's house on fire. It's those moments when I think 'I love my job!'"
Despite her gorgeous gowns for "Revenge" and her saucy look for GQ, the actress said she identifies with being a good old tomboy.
"Definitely. I did ballet, which taught me how to be poised and graceful, but still, to this day I'd rather be fishing than going to some glamorous party. It's so therapeutic," she said.
Copyright 2014 by NBC Universal, Inc. All rights reserved.This material may not be published, broadcast, rewritten or redistributed.


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Sarah Jane Crawford sits in
Today on the show Sarah Jane asked the listeners 'Who do you think has the best bum?' after a recent poll voted Kim Kardashian to have the best bum in 2010. Listen back to the show to find out who the listners voted for. Plus SJ wanted to have you ever been stuck somewhere you didn't want to be?
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Digital Living
Image after image splashes on the wall of the art exhibit - a snapshot of young people laughing and drinking, a picture of an elephant, an exposed belly of a woman barely covering her breasts with one arm. The photos were taken from their computers without their knowledge through a technological glitch.
Over in a corner, visitors can sort through Facebook profile photos from unwitting users through a website that organises them by gender, country and adjectives such as "sly," ''smug" or "easy going." Think online dating site, for people who don't know they are on it.
The works are part of The Public Private, an art exhibit that explores the gray areas of online privacy, surveillance and data collection in the age of Facebook and Google. The pieces shift across the boundaries between what's public and private, all through the lens of technology. But lines are never clear, if there are any at all, and that can be unsettling.
Its curator, Christiane Paul, says she hopes visitors will walk away with questions. The exhibit's goal, she says, isn't to universally declare Facebook bad or social media evil, but to get people thinking. It's only been nine years, after all, since Facebook's birth and seven since Twitter was created, so art that explores social networking as a subject is just emerging.
"I don't think good art provides easy answers," says Paul, adjunct curator at Whitney Museum of American Art and a media studies professor at The New School, where the exhibit runs through April 17.
Indeed, bringing the concept of online privacy into the physical world can feel both uncomfortable and eye-opening. The internet may lull us into a false sense of security. Posting something on Facebook, for instance, can feel more private than shouting it on a busy street, even if the opposite is true given that a Facebook post might reach hundreds or thousands of people, perhaps millions.
"We are living, really, in a situation where we don't know what we want to have as public," says Paolo Cirio, one of the artists featured. "We don't know what is public and what is not. It's a moment of confusion for a lot of people. The next generation already has another idea for privacy."
After the exhibit ends, some of the projects will remain available online or offline - at least in some form.
Take Cirio's "Street Ghosts," for example. It's nothing more than life-sized images of people caught in Google's street-level mapping feature, plastered on walls and fences in the same physical place that their digital counterparts occupy on Google Maps. Cirio calls the online versions "digital ghosts." Some of the images are pasted on the outside walls of the building housing the exhibit.
They serve as a reminder of what we leave behind on the Internet.
"We lose control (of our digital information) completely when we die," he says "But it's left in the archives of Google, Facebook."
Some of the works in the show, Paul acknowledges, exist in a legal and ethical gray area.
For "The Others," Eva and Franco Mattes gained access to private computer files through "old software that lets you anonymously share files from your computer," Eva Mattes explains. "But people who are not familiar with it mistakenly share their whole computer without realizing it."
Looking inside a stranger's computer feels a bit like peeking into his or her house, she says.
A previous project by the couple - not part of this exhibit - made all the content of their computer available to the public. For three years, anyone with Internet access could look through their photos, emails, even their bank account.
The artists say they didn't feel conflicted about grabbing the photos from private computers.
"The first thing I noticed is how little difference there is with the ones you see online all the time," Franco Mattes says. "These are supposedly private photos, but they're no different than public photos. In a sense, I'm doing what all photographers do: I 'take' photos."
So has anyone in the photos contacted them?
"The first time we showed the work, in Sheffield, UK, a person claimed to be portrayed. To be honest it was hard to tell whether or not he really was the guy in the photos," Eva Mattes says in an email interview. "Anyway, the next day he came back to see the exhibition with his whole family, to show them he was famous."
She says the couple felt relieved.
Brooke Singer, a new media studies professor at Purchase College in New York, says the imagery of "The Others" is in itself uninteresting. It looks exactly like photos people post online.
"So the point of the project is not the content per se but to have us consider the ethics and audacity of the act," she says.
The project, she adds, speaks more broadly about the digital age - the way limits to what can and cannot be replicated are constantly built up and torn down "by various interests."
"And this also points toward our false sense of isolation as we are on our 'personal' computers always connected to the network," Singer says.
Another gray-area piece, Cirio's "Face-to-Facebook", has been taken offline, after Facebook's lawyers complained. But visitors can use a computer at the exhibit to look through the piece. Cease-and-desist letters from Facebook are posted on an adjacent wall.
The project, created in 2011, consists of data that hundreds of thousands of Facebook users shared on the site publicly - including their name, profile photo, the country they live in and groups they belonged to.
Cirio and co-creator Alessandro Ludovico put all the information in a database and used face-recognition software to put people in personality categories such as "funny," ''climber" and "smug." Then, the artists set up a dating website using the photos and information. It was called Lovely Faces, and it didn't last long.
Facebook sent its first cease-and-desist letter in February 2011, complaining that the artists used an automated program to "scrape" Facebook user data, in violation of the site's policy. The artists eventually took the site offline.
"Although it was online only five days, a lot of people understood that what they publish on Facebook eventually could be used by someone else without their authorization," Cirio says. "And they lose control of their data."
Though many of the pieces poke at Google and Facebook, camera surveillance is also a central theme. Wafaa Bilal's 3rdi project takes a look at what he's left behind - literally. The Iraqi artist had a camera surgically implanted into the back of his head.
It had to be removed because of infection risk and because his body rejected it. Now, he attaches it through less invasive means, using a wraparound headgear.
"During my journey from Iraq to Saudi Arabia, on to Kuwait and then the U.S., I left many people and places behind," Bilal writes on his website describing the project. "The images I have of this journey are inevitably ephemeral, held as they are in my own memory."
The twist is that he doesn't see the memory while it's captured, leaving it behind.
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Potential solutions vary on employee transportation issue
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Area business owners acknowledge that tapping into Milwaukee's workforce has been a growing issue, but a more responsive and flexible solution may call for public subsidies or private funding to address the issue. One staffing agency provides transportation for its employees. However, a Milwaukee alderman says suburban companies that want to access Milwaukee workers need to rethink their location. The Milwaukee Business Journal collected some comments on the issue:
Kim Korth
President and chief executive officer, Dickten Masch Plastics LLC, Nashotah
The lack of access to reliable transportation is sometimes an issue when it comes to attracting and retaining workers for Dickten Masch Plastics LLC, a 30- to 35-minute drive from Milwaukee, Korth said.
And it's a growing problem as Dickten Masch continues its growth.
"It's expensive to drive here (from Milwaukee) and if someone finds a job five minutes away, it's probably better for them financially to work there instead of here," Korth said.
Korth admits the issue hasn't been at the forefront of her firm's problems, but she acknowledges the need for a larger discussion in the business community because the transit problem isn't going away.
"Perhaps if we could have some form of reliable public transportation, that would be helpful," Korth said. "Maybe if employees had a transportation subsidy or tax credit that would be available for gas or a car, that might help."
Eric Isbister
President of GenMet Corp., Mequon
"Maybe we should have a coalition of businesses get together and talk about how we're going to solve this problem," Isbister said. "I know we're not the only ones with this problem."
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