Measuring the response of human head and neck squamous cell carcinoma to irradiation in a microfluidic model allowing customized therapy

Radiotherapy is the standard treatment for head and neck squamous cell carcinoma (HNSCC), however, radioresistance remains a major clinical problem despite significant improvements in treatment protocols. Therapeutic outcome could potentially be improved if a patient's tumour response to irradiation could be predicted ex vivo before clinical application. The present study employed a bespoke microfluidic device to maintain HNSCC tissue whilst subjecting it to external beam irradiation and measured the responses using a panel of cell death and proliferation markers. HNSCC biopsies from five newly-presenting patients [2 lymph node (LN); 3 primary tumour (PT)] were divided into parallel microfluidic devices and replicates of each tumour were subjected to single-dose irradiation (0, 5, 10, 15 and 20 Gy). Lactate dehydrogenase (LDH) release was measured and tissue sections were stained for cytokeratin (CK), cleaved-CK18 (cCK18), phosphorylated-H2AX (λH2AX) and Ki.67 by immunohistochemistry. In addition, fragmented DNA was detected using terminal deoxynucleotidyl transferase dUTP nick end labelling (TUNEL). Compared with non.irradiated controls, higher irradiation doses resulted in elevated CK18-labelling index in two lymph nodes [15 Gy; 34.8% on LN1 and 31.7% on LN2 (p=0.006)] and a single laryngeal primary tumour (20 Gy; 31.5%; p=0.014). Significantly higher levels of DNA fragmentation were also detected in both lymph node samples and one primary tumour but at varying doses of irradiation, i.e., LN1 (20 Gy; 27.6%; p=0.047), LN2 (15 Gy; 15.3%; p=0.038) and PT3 (10 Gy; 35.2%; p=0.01). The λH2AX expression was raised but not significantly in the majority of samples. The percentage of Ki.67 positive nuclei reduced dose-dependently following irradiation. In contrast no significant difference in LDH release was observed between irradiated groups and controls. There is clear interand intra-patient variability in response to irradiation when measuring a variety of parameters, which offers the potential for the approach to provide clinically valuable information

Effect of the source charge on charged-beam interferometry

We investigate quantal perturbations of the interferometric correlations of charged bosons by the Coulomb field of an instantaneous, charged source. The source charge increases the apparent source size by weakening the correlation at non-zero relative momenta. The effect is strongest for pairs with a small total momentum and is stronger for kaons than for pions of the same momenta. The experimental data currently available are well described by this effect without invoking Pratt's exploding source model. A simple expression is proposed to account for the effect.Comment: 9 pages TEX, 3 Postscript figures available at http://www.krl.caltech.edu/preprints/MAP.htm

The NuMI Neutrino Beam and Potential for an Off-Axis Experiment

The Neutrinos at the Main Injector (NuMI) facility at Fermilab is under construction and due to begin operations in late 2004. NuMI will deliver an intense $\nu_{\mu}$ beam of variable energy 2-20 GeV directed into the Earth at 58 mrad. Several aspects of the design are reviewed, and potential limitations to the ultimate neutrino flux are described. In addition, potential measurements of neutrino mixing properties are described.Comment: talk given at NuFact '02, Imperial College London, proceedings to appear in J. Phys. G, revised to add a referenc

Neutrons from multiplicity-selected Au-Au collisions at 150, 250, 400, and 650 AMeV

We measured neutron triple-differential cross sections from multiplicity-selected Au-Au collisions at 150, 250, 400, and 650 \AMeV. The reaction plane for each collision was estimated from the summed transverse velocity vector of the charged fragments emitted in the collision. We examined the azimuthal distribution of the triple-differential cross sections as a function of the polar angle and the neutron rapidity. We extracted the average in--plane transverse momentum $\langle P_x\rangle$ and the normalized observable $\langle P_x/P_\perp\rangle$, where $P_\perp$ is the neutron transverse momentum, as a function of the neutron center-of-mass rapidity, and we examined the dependence of these observables on beam energy. These collective flow observables for neutrons, which are consistent with those of protons plus bound nucleons from the Plastic Ball Group, agree with the Boltzmann--Uehling--Uhlenbeck (BUU) calculations with a momentum--dependent interaction. Also, we calculated the polar-angle-integrated maximum azimuthal anisotropy ratio R from the value of $\langle P_x/P_\perp\rangle$.Comment: 20 LaTeX pages. 11 figures to be faxed on request, send email to sender's addres

A patient tumour-on-a-chip system for personalised investigation of radiotherapy based treatment regimens

Development of personalised cancer models to predict response to radiation would benefit patient care; particularly in malignancies where treatment resistance is prevalent. Herein, a robust, easy to use, tumour-on-a-chip platform which maintains precision cut head and neck cancer for the purpose of ex vivo irradiation is described. The device utilises sintered discs to separate the biopsy and medium, mimicking in vivo microvascular flow and diffusion, maintaining tissue viability for 68 h. Integrity of tissues is demonstrated by the low levels of lactate dehydrogenase release and retained histology, accompanied by assessment of cell viability by trypan blue exclusion and flow cytometry; fluid dynamic modelling validates culture conditions. An irradiation jig is described for reproducible delivery of clinically-relevant doses (5 × 2 Gy) to newly-presenting primary tumours (n = 12); the addition of concurrent cisplatin is also investigated (n = 8) with response analysed by immunohistochemistry. Fractionated irradiation reduced proliferation (BrdU, p = 0.0064), increased DNA damage (ƴH2AX, p = 0.0043) and caspase-dependent apoptosis (caspase-cleaved cytokeratin-18) compared to control; caspase-dependent apoptosis was further increased by concurrent cisplatin compared to control (p = 0.0063). This is a proof of principle study showing the response of cancer tissue to irradiation ex vivo in a bespoke system. The novel platform described has the potential to personalise treatment for patients in a cost-effective manner with applicability to any solid tumour

Very Long Baseline Neutrino Oscillation Experiment for Precise Measurements of Mixing Parameters and CP Violating Effects

We analyze the prospects of a feasible, Brookhaven National Laboratory based, very long baseline (BVLB) neutrino oscillation experiment consisting of a conventional horn produced low energy wide band beam and a detector of 500 kT fiducial mass with modest requirements on event recognition and resolution. Such an experiment is intended primarily to determine CP violating effects in the neutrino sector for 3-generation mixing. We analyze the sensitivity of such an experiment. We conclude that this experiment will allow determination of the CP phase $\delta_{CP}$ and the currently unknown mixing parameter $\theta_{13}$, if $\sin ^2 2 \theta_{13} \geq 0.01$, a value $\sim 15$ times lower than the present experimental upper limit. In addition to $\theta_{13}$ and $\delta_{CP}$, the experiment has great potential for precise measurements of most other parameters in the neutrino mixing matrix including $\Delta m^2_{32}$, $\sin^2 2\theta_{23}$, $\Delta m^2_{21}\times \sin 2 \theta_{12}$, and the mass ordering of neutrinos through the observation of the matter effect in the $\nu_\mu \to \nu_e$ appearance channel.Comment: 12 pages, 10 figure

Gene Discovery and Functional Genomics in the Pig

Advances in gene mapping and genomics in farm animals have been considerable over the past decade. Medium resolution linkage and physical maps have been reported, and specific chromosomal regions and genes associated with traits of biological and economic interest have been identified. We have reached an exciting stage in gene identification, mapping and quantitative trait locus discovery in pigs, as new molecular information is accumulating rapidly. Significant progress has been made by identifying candidate gene associations and low-resolution regions containing quantitative trail loci (QTL). However, we are still disadvantaged by the lack of tools available to efficiently use much of this new information. For example, current pig maps are neither of high enough resolution nor sufficiently informative at the comparative level for positional candidate gene cloning within QTL regions. As well, studying biological mechanisms underlying economically important traits such as reproduction is limited by the lack of molecular resources. This is especially important, as reproduction is very difficult to genetically improve by classical breeding methods due to the relatively low heritability and high expense in data collection. Thus, an improved understanding of porcine reproductive biology is of crucial economic importance, yet reproductive processes are poorly characterized at the molecular level. Recently, new methodologies have been brought to bear on a better understanding of pig molecular biology for accelerating genetic improvement in pigs. Several groups are developing molecular information in the pig, and the total Genbank sequence entries for porcine expressed genes have recently topped 100,000. Our Midwest EST Consortium has produced cDNA libraries containing the majority of genes expressed in major female reproductive tissues, and we have deposited nearly 15,000 gene sequences into public databases. These sequences represent over 8,900 different genes, based on sequence comparison among these data. Furthermore, we have developed computer software to automatically extract sequence similarity of these pig genes with their human counterparts, as well as the mapping information of these human homologues. Within our data set, we have identified nearly 1,500 pig genes with strong similarity to mapped human genes, and we are in the process of mapping 700 of these genes to improve the human-pig comparative map. This work and the complementary work of others can now be used to more rapidly understand and identify the genes controlling reproduction, so that genetic improvement of reproduction phenotypes can accelerate

Neutrons from multiplicity-selected La-La and Nb-Nb collisions at 400A MeV and La-La collisions at 250A MeV

Triple-differential cross sections for neutrons from high-multiplicity La-La collisions at 250 and 400 MeV per nucleon and Nb-Nb collisions at 400 MeV per nucleon were measured at several polar angles as a function of the azimuthal angle with respect to the reaction plane of the collision. The reaction plane was determined by a transverse-velocity method with the capability of identifying charged-particles with Z=1, Z=2, and Z > 2. The flow of neutrons was extracted from the slope at mid-rapidity of the curve of the average in-plane momentum vs the center-of-mass rapidity. The squeeze-out of the participant neutrons was observed in a direction normal to the reaction plane in the normalized momentum coordinates in the center-of-mass system. Experimental results of the neutron squeeze-out were compared with BUU calculations. The polar-angle dependence of the maximum azimuthal anisotropy ratio $r(\theta)$ was found to be insensitive to the mass of the colliding nuclei and the beam energy. Comparison of the observed polar-angle dependence of the maximum azimuthal anisotropy ratio $r(\theta)$ with BUU calculations for free neutrons revealed that $r(\theta)$ is insensitive also to the incompressibility modulus in the nuclear equation of state.Comment: ReVTeX, 16 pages, 17 figures. To be published in Physical Review

The energy dependence of flow in Ni induced collisions from 400 to 1970A MeV

We study the energy dependence of collective (hydrodynamic-like) nuclear matter flow in 400-1970 A MeV Ni+Au and 1000-1970 A MeV Ni+Cu reactions. The flow increases with energy, reaches a maximum, and then gradually decreases at higher energies. A way of comparing the energy dependence of flow values for different projectile-target mass combinations is introduced, which demonstrates a common scaling behaviour among flow values from different systems.Comment: 12 pages, 3 figures. Submitted to Physical Review Letter

Maximum Azimuthal Anisotropy of Neutrons from Nb-Nb Collisions at 400 AMeV and the Nuclear Equation of State

We measured the first azimuthal distributions of triple--differential cross sections of neutrons emitted in heavy-ion collisions, and compared their maximum azimuthal anisotropy ratios with Boltzmann--Uehling--Uhlenbeck (BUU) calculations with a momentum-dependent interaction. The BUU calculations agree with the triple- and double-differential cross sections for positive rapidity neutrons emitted at polar angles from 7 to 27 degrees; however, the maximum azimuthal anisotropy ratio for these free neutrons is insensitive to the size of the nuclear incompressibility modulus K characterizing the nuclear matter equation of state.Comment: Typeset using ReVTeX, with 3 ps figs., uuencoded and appende