Structure,
Journal Year:
2025,
Volume and Issue:
unknown
Published: Feb. 1, 2025
Highlights•A
software
suite
for
automated
analysis
of
lipid
membranes
in
electron
micrographs•Includes
segmentation,
shape
identification,
and
membrane
properties•Applied
to
datasets
with
different
lipids
protein-induced
changes•Features
an
intuitive
GUI
batch
micrograph
analysisSummaryImaging
structures
associated
protein
complexes
using
cryoelectron
microscopy
(cryo-EM)
is
a
common
visualization
structure
determination
technique.
The
quantitative
the
structures,
however,
not
routine
time
consuming
particular
when
large
amounts
data
are
involved.
Here,
we
introduce
image-processing
cryo-vesicle
image
analyzer
(CryoVIA)
that
parametrizes
from
cryo-EM
images.
This
toolkit
combines
identification
methods
automatically
perform
large-scale
local
global
properties
such
as
bilayer
thickness,
size,
curvature
including
classifications.
We
included
analyses
exemplary
compositions
changes
through
endosomal
sorting
required
transport
III
(ESCRT-III)
remodeling
protein.
opens
new
possibilities
systematically
study
structural
their
modifications
images.Graphical
abstract
bioRxiv (Cold Spring Harbor Laboratory),
Journal Year:
2025,
Volume and Issue:
unknown
Published: Jan. 19, 2025
Abstract
The
big
potassium
(BK)
channels
remain
open
with
a
small
limiting
probability
of
P
o
∼
10
-7
at
minimal
Ca
2+
and
negative
voltages
<
−100
mV.
molecular
origin
functional
significance
such
“intrinsic
opening”
are
not
understood.
Here
we
combine
atomistic
simulations
electrophysiological
experiments
to
show
that
the
intrinsic
opening
BK
is
an
inherent
property
vapor
barrier,
generated
by
hydrophobic
dewetting
inner
pore
in
deactivated
state.
barrier
only
gives
rise
finite
free
energy
8
kcal/mol,
cannot
completely
shut
down
K
+
flow
even
when
voltage
sensor
domains
fully
deactivated.
This
results
leaking
currents
can
be
measured
as
indication
opening.
shallow
slope
primarily
from
electric
field
effects
on
permeating
ion
through
barrier.
We
further
demonstrate
perturbed
mutations
truncation
cytosolic
domains,
leading
predicable
changes
measurements.
Therefore,
channels,
possibly
other
opens
up
opportunity
experimentally
study
gating.
Our
suggest
fundamental
basis
for
allosteric
mechanism
activation
both
.
Journal for ImmunoTherapy of Cancer,
Journal Year:
2025,
Volume and Issue:
13(1), P. e010005 - e010005
Published: Jan. 1, 2025
Background
A
number
of
immunotherapeutic
approaches
have
been
developed
and
are
entering
the
clinic.
Bispecific
antibodies
(BsAbs)
one
these
modalities
induce
robust
efficacy
by
endogenous
T
cells
in
several
hematological
malignancies.
However,
most
treated
patients
experience
only
a
temporary
benefit.
Currently
available
BsAbs
provide
anti-CD3
antibody-mediated
T-cell
stimulation,
but
not
costimulation
or
cytokine
signaling
essential
for
full
activation.
Here,
we
hypothesized
that
simultaneous
input
more
comprehensive
signals
would
elicit
durable
effector
functions.
Methods
We
genetically
engineered
leukemia
cell
line
K562
to
express
BsAbs,
costimulatory
ligands,
cytokines,
blocking
against
immune
checkpoint
molecules
on
surface,
from
which
obtained
plasma
membrane
fractions
mechanical
homogenization
subsequent
isolation
steps.
Plasma
membranes
were
reconstituted
poly
(lactic-co-glycolic
acid)
surface
generate
membrane-coated
nanoparticles
(NPs).
Alternatively,
nano-sized
vesicles
(MVs)
generated
ultrasonic
dispersion
isolated
membranes.
The
antitumor
function
NPs
MVs
loaded
with
various
immunomodulatory
factors
was
evaluated
vitro
vivo.
Results
Both
induced
BsAb-mediated
antigen-specific
cytotoxic
activity
non-specific
cells,
inducing
slightly
better
response
Importantly,
activation
elicited
presence
target
tumor
providing
safety
advantage
clinical
use.
expressing
(CD80/4-1BBL)
cytokines
(interleukin
(IL)-7/IL-15)
further
enhanced
therapeutic
In
addition,
inflammatory
IL-12
IL-18
objective
responses
solid
models
partly
converting
immunosuppressive
macrophages
proinflammatory
phenotypes
infiltration
into
tumor.
Finally,
showed
also
activate
natural
killer
(NK)
loading
multiple
ligands.
4-1BBL,
IL-15,
IL-21
NK-cell
an
manner.
Conclusions
efficiently
vivo
simultaneously
delivering
immunostimulatory
cells.
This
platform
enables
delivery
desired
combinations
NK
Structure,
Journal Year:
2025,
Volume and Issue:
unknown
Published: Feb. 1, 2025
Highlights•A
software
suite
for
automated
analysis
of
lipid
membranes
in
electron
micrographs•Includes
segmentation,
shape
identification,
and
membrane
properties•Applied
to
datasets
with
different
lipids
protein-induced
changes•Features
an
intuitive
GUI
batch
micrograph
analysisSummaryImaging
structures
associated
protein
complexes
using
cryoelectron
microscopy
(cryo-EM)
is
a
common
visualization
structure
determination
technique.
The
quantitative
the
structures,
however,
not
routine
time
consuming
particular
when
large
amounts
data
are
involved.
Here,
we
introduce
image-processing
cryo-vesicle
image
analyzer
(CryoVIA)
that
parametrizes
from
cryo-EM
images.
This
toolkit
combines
identification
methods
automatically
perform
large-scale
local
global
properties
such
as
bilayer
thickness,
size,
curvature
including
classifications.
We
included
analyses
exemplary
compositions
changes
through
endosomal
sorting
required
transport
III
(ESCRT-III)
remodeling
protein.
opens
new
possibilities
systematically
study
structural
their
modifications
images.Graphical
abstract
