1,242 research outputs found
Rapid kit-based (68)Ga-labelling and PET imaging with THP-Tyr(3)-octreotate:a preliminary comparison with DOTA-Tyr(3)-octreotate
BACKGROUND: Ge/(68)Ga generators provide an inexpensive source of a PET isotope to hospitals without cyclotron facilities. The development of new (68)Ga-based molecular imaging agents and subsequent clinical translation would be greatly facilitated by simplification of radiochemical syntheses. We report the properties of a tris(hydroxypyridinone) conjugate of the SSTR2-targeted peptide, Tyr(3)-octreotate (TATE), and compare the (68)Ga-labelling and biodistribution of [(68)Ga(THP-TATE)] with the clinical radiopharmaceutical [(68)Ga(DOTATATE)]. METHODS: A tris(hydroxypyridinone) with a pendant isothiocyanate group was conjugated to the primary amine terminus of H(2)N-PEG(2)-Lys(iv-Dde)(5)-TATE, and the resulting conjugate was deprotected to provide THP-TATE. THP-TATE was radiolabelled with (68)Ga(3+) from a (68)Ge/(68)Ga generator. In vitro uptake was assessed in SSTR2-positive 427-7 cells and SSTR2-negative 427 (parental) cells. Biodistribution of [(68)Ga(THP-TATE)] was compared with that of [(68)Ga(DOTATATE)] in Balb/c nude mice bearing SSTR2-positive AR42J tumours. PET scans were obtained 1 h post-injection, after which animals were euthanised and tissues/organs harvested and counted. RESULTS: [(68)Ga(THP-TATE)] was radiolabelled and formulated rapidly in <2 min, in ≥95 % radiochemical yield at pH 5–6.5 and specific activities of 60–80 MBq nmol(−1) at ambient temperature. [(68)Ga(THP-TATE)] was rapidly internalised into SSTR2-positive cells, but not SSTR2-negative cells, and receptor binding and internalisation were specific. Animals administered [(68)Ga(THP-TATE)] demonstrated comparable SSTR2-positive tumour activity (11.5 ± 0.6 %ID g(−1)) compared to animals administered [(68)Ga(DOTATATE)] (14.4 ± 0.8 %ID g(−1)). Co-administration of unconjugated Tyr(3)-octreotate effectively blocked tumour accumulation of [(68)Ga(THP-TATE)] (2.7 ± 0.6 %ID g(−1)). Blood clearance of [(68)Ga(THP-TATE)] was rapid and excretion was predominantly renal, although compared to [(68)Ga(DOTATATE)], [(68)Ga(THP-TATE)] exhibited comparatively longer kidney retention. CONCLUSIONS: Radiochemical synthesis of [(68)Ga(THP-TATE)] is significantly faster, proceeds under milder conditions, and requires less manipulation than that of [(68)Ga(DOTATATE)]. A (68)Ga-labelled tris(hydroxypyridinone) conjugate of Tyr(3)-octreotate demonstrates specificity and targeting affinity for SSTR2 receptors, with comparable in vivo targeting affinity to the clinical PET tracer, [(68)Ga(DOTATATE)]. Thus, peptide conjugates based on tris(hydroxypyridinones) are conducive to translation to kit-based preparation of PET tracers, enabling the expansion and adoption of (68)Ga PET in hospitals and imaging centres without the need for costly automated synthesis modules
Simplified three-dimensional tissue clearing and incorporation of colorimetric phenotyping.
Tissue clearing methods promise to provide exquisite three-dimensional imaging information; however, there is a need for simplified methods for lower resource settings and for non-fluorescence based phenotyping to enable light microscopic imaging modalities. Here we describe the simplified CLARITY method (SCM) for tissue clearing that preserves epitopes of interest. We imaged the resulting tissues using light sheet microscopy to generate rapid 3D reconstructions of entire tissues and organs. In addition, to enable clearing and 3D tissue imaging with light microscopy methods, we developed a colorimetric, non-fluorescent method for specifically labeling cleared tissues based on horseradish peroxidase conversion of diaminobenzidine to a colored insoluble product. The methods we describe here are portable and can be accomplished at low cost, and can allow light microscopic imaging of cleared tissues, thus enabling tissue clearing and imaging in a wide variety of settings
GROWTH MINDSET IN ENGLISH LANGUAGE LEARNING OF COLLEGE STUDENTS
This study investigated the growth mindset of college students in English language learning and the challenges they encounter in an online setting. Using a researcher-made questionnaire about the growth mindset of college students in English language learning and the challenges they encounter in an online setting, data from the 60 respondents in the study coming from the different colleges or departments of the Notre Dame of Midsayap College were analyzed quantitatively. Findings indicated that students have a high growth mindset and that factors such as motivation, criticism, environment, and effort had played a major role in their English language learning. Major challenges of students in an online setting include finding difficulty in accessing online materials due to poor internet connection, experiencing headaches and body pains due to prolonged exposure to gadgets, and getting bored easily when class time is too lengthy. The least challenges encountered by students in an online setting yet also hampered their learning are lacking gadgets such as a cellphone and laptops, feeling disconnected from the instructor, and excessive workloads. Furthermore, there is no significant difference in a growth mindset in terms of sex and college or department. However, it was found out that College of Nursing (CN) got the highest mean when it comes to their growth mindset. Meanwhile, College of Information Technology, and Engineering (CITE) got the lowest mean. Moreover, there is a significant relationship between the growth mindset of English language learning and the challenges they encountered in an online setting. Article visualizations
All-optical multimode fibre photoacoustic endomicroscopy with scalable spatial resolution and field-of-view
An all-optical, forward-viewing, optical-resolution photoacoustic endomicroscopy probe was developed for guiding minimally invasive procedures. The probe comprises a multimode fibre for the delivery of excitation laser via wavefront shaping, and a fibre-optic ultrasound sensor based on a plane-concave microresonator at the tip of a single-mode fibre. High-resolution photoacoustic microscopy images of mouse red blood cells and mouse ear vasculature were acquired, and the high scalability of the probe in terms of field-of-view and spatial resolution was demonstrated. The ultrathin photoacoustic endomicroscopy probe promises to guide minimally invasive surgery by providing both molecular and microstructural information
Targeting integrin αvβ6 with gallium-68 tris (hydroxypyridinone) based PET probes
Expression of the cellular transmembrane receptor αvβ6 integrin is mostly restricted to malignant epithelial cells in a wide variety of carcinomas, including pancreatic and others derived from epithelial tissues. Thus, this protein is considered an attractive target for tumour imaging and therapy. Two different (68)Ga hexadentate tris (3,4-hydroxypyridinone) (THP) chelators were produced in this study and coupled to the αvβ6 integrin–selective peptide cyclo(FRGDLAFp(NMe)K) via NHS chemistry. Radiolabelling experiments confirmed a high radiochemical yield of the two PET probes. In addition, cellular binding studies showed high binding affinities in the nanomolar range. The two integrin αvβ6-peptide-THP synthesized and radiolabeled in this study will facilitate in vivo monitoring of transmembrane receptor αvβ6 integrin by using the advantage of THP chemistry for rapid, efficient and stable gallium chelation
Pangenomic analysis reveals plant NAD+ manipulation as an important virulence activity of bacterial pathogen effectors
Nicotinamide adenine dinucleotide (NAD+) has emerged as a key component in prokaryotic and eukaryotic immune systems. The recent discovery that Toll/interleukin-1 receptor (TIR) proteins function as NAD+ hydrolases (NADase) links NAD+-derived small molecules with immune signaling. We investigated pathogen manipulation of host NAD+ metabolism as a virulence strategy. Using the pangenome of the model bacterial pathogen Pseudomonas syringae, we conducted a structure-based similarity search from 35,000 orthogroups for type III effectors (T3Es) with potential NADase activity. Thirteen T3Es, including five newly identified candidates, were identified that possess domain(s) characteristic of seven NAD+-hydrolyzing enzyme families. Most Pseudomonas syringae strains that depend on the type III secretion system to cause disease, encode at least one NAD+-manipulating T3E, and many have several. We experimentally confirmed the type III-dependent secretion of a novel T3E, named HopBY, which shows structural similarity to both TIR and adenosine diphosphate ribose (ADPR) cyclase. Homologs of HopBY were predicted to be type VI effectors in diverse bacterial species, indicating potential recruitment of this activity by microbial proteins secreted during various interspecies interactions. HopBY efficiently hydrolyzes NAD+ and specifically produces 2′cADPR, which can also be produced by TIR immune receptors of plants and by other bacteria. Intriguingly, this effector promoted bacterial virulence, indicating that 2′cADPR may not be the signaling molecule that directly initiates immunity. This study highlights a host-pathogen battleground centered around NAD+ metabolism and provides insight into the NAD+-derived molecules involved in plant immunity
Probing Unexpected Reactivity in Radiometal Chemistry: Indium-111-Mediated Hydrolysis of Hybrid Cyclen-Hydroxypyridinone Ligands
Chelators based on hydroxypyridinones have utility in
incorporating
radioactive metal ions into diagnostic and therapeutic agents used
in nuclear medicine. Over the course of our hydroxypyridinone studies,
we have prepared two novel chelators, consisting of a cyclen (1,4,7,10-tetraazacyclododecane)
ring bearing two pendant hydroxypyridinone groups, appended via methylene
acetamide motifs at either the 1,4-positions (L1) or 1,7-positions (L2) of the cyclen ring. In radiolabeling reactions of L1 or L2 with the γ-emitting radioisotope, [111In]In3+, we have observed radiometal-mediated hydrolysis of a single
amide group of either L1 or L2. The reaction of either [111In]In3+ or [natIn]In3+ with either L1 or L2, in aqueous alkaline solutions at 80 °C, initially results
in formation of [In(L1)]+ or [In(L2)]+, respectively. Over time, each of these species undergoes In3+-mediated hydrolysis of a single amide group to yield species
in which In3+ remains coordinated to the resultant chelator,
which consists of a cyclen ring bearing a single hydroxypyridinone
group and a single carboxylate group. The reactivity toward hydrolysis
is higher for the L1 complex
compared to that for the L2 complex.
Density functional theory calculations corroborate these experimental
findings and importantly indicate that the activation energy required
for the hydrolysis of L1 is significantly
lower than that required for L2. This is the first reported example of a chelator undergoing radiometal-mediated
hydrolysis to form a radiometalated complex. It is possible that metal-mediated
amide bond cleavage is a source of instability in other radiotracers,
particularly those in which radiometal complexation occurs in aqueous,
basic solutions at high temperatures. This study highlights the importance
of appropriate characterization of radiolabeled products
(S)-(−)-Fluorenylethylchloroformate (FLEC) ; preparation using asymmetric transfer hydrogenation and application to the analysis and resolution of amines
Fluorenylethylchoroformate (FLEC) is a valuable chiral derivatisation reagent that is used for the resolution of a wide variety of chiral amines. Herein, we describe an improved preparation of (S)-(−)-FLEC using an efficient asymmetric catalytic transfer hydrogenation as the key step. We also demonstrate the application of FLEC as a chiral Fmoc equivalent for chiral resolution, with facile deprotection, of tetrahydroquinaldines, and its capacity for inducing regioselective outcomes in nitration reactions
MeerKLASS: MeerKAT Large Area Synoptic Survey
We discuss the ground-breaking science that will be possible with a wide area
survey, using the MeerKAT telescope, known as MeerKLASS (MeerKAT Large Area
Synoptic Survey). The current specifications of MeerKAT make it a great fit for
science applications that require large survey speeds but not necessarily high
angular resolutions. In particular, for cosmology, a large survey over for hours will potentially provide the first
ever measurements of the baryon acoustic oscillations using the 21cm intensity
mapping technique, with enough accuracy to impose constraints on the nature of
dark energy. The combination with multi-wavelength data will give unique
additional information, such as exquisite constraints on primordial
non-Gaussianity using the multi-tracer technique, as well as a better handle on
foregrounds and systematics. Such a wide survey with MeerKAT is also a great
match for HI galaxy studies, providing unrivalled statistics in the pre-SKA era
for galaxies resolved in the HI emission line beyond local structures at z >
0.01. It will also produce a large continuum galaxy sample down to a depth of
about 5\,Jy in L-band, which is quite unique over such large areas and
will allow studies of the large-scale structure of the Universe out to high
redshifts, complementing the galaxy HI survey to form a transformational
multi-wavelength approach to study galaxy dynamics and evolution. Finally, the
same survey will supply unique information for a range of other science
applications, including a large statistical investigation of galaxy clusters as
well as produce a rotation measure map across a huge swathe of the sky. The
MeerKLASS survey will be a crucial step on the road to using SKA1-MID for
cosmological applications and other commensal surveys, as described in the top
priority SKA key science projects (abridged).Comment: Larger version of the paper submitted to the Proceedings of Science,
"MeerKAT Science: On the Pathway to the SKA", Stellenbosch, 25-27 May 201
- …