Extensive Vibrational Characterisation and Long-Term Monitoring of Honeybee Dorso-Ventral Abdominal Vibration signals.

A very common honeybee signal is the dorso-ventral abdominal vibration (DVAV) signal, widely accepted as a modulatory signal meaning: "prepare for greater activity". In this study, using ultra-sensitive accelerometer technology embedded in the honeycomb, we visually confirm the one-to-one relationship between a DVAV signal being produced and the resulting accelerometer waveform, allowing the measurement of DVAV signals without relying on any visual inspection. We then demonstrate a novel method for the continuous in-situ non-invasive automated monitoring of this honeybee signal, not previously known to induce any vibration into the honeycomb, and most often inaudible to human hearing. We monitored a total of three hives in the UK and France, showing that the signal is very common, highly repeatable and occurs more frequently at night, exhibiting a distinct decrease in instances and increase in amplitude towards mid-afternoon. We also show an unprecedented increase in the cumulative amplitude of DVAV signals occurring in the hours preceding and following a primary swarm. We conclude that DVAV signals may have additional functions beyond solely being a foraging activation signal, and that the amplitude of the signal might be indicative of the switching of its purpose.

Conditional expression of a Gi-coupled receptor in osteoblasts results in trabecular osteopenia.

G protein-coupled receptors (GPCRs) coupled to activation of Gs, such as the PTH1 receptor (PTH1R), have long been known to regulate skeletal function and homeostasis. However, the role of GPCRs coupled to other G proteins such as Gi is not well established. We used the tet-off system to regulate the expression of an activated Gi-coupled GPCR (Ro1) in osteoblasts in vivo. Skeletal phenotypes were assessed in mice expressing Ro1 from conception, from late stages of embryogenesis, and after weaning. Long bones were assessed histologically and by microcomputed tomography. Expression of Ro1 from conception resulted in neonatal lethality that was associated with reduced bone mineralization. Expression of Ro1 starting at late embryogenesis resulted in a severe trabecular bone deficit at 12 wk of age (>51% reduction in trabecular bone volume fraction in the proximal tibia compared with sex-matched control littermates; n = 11; P < 0.01). Ro1 expression for 8 wk beginning at 4 wk of age resulted in a more than 20% reduction in trabecular bone volume fraction compared with sex-matched control littermates (n = 16; P < 0.01). Bone histomorphometry revealed that Ro1 expression is associated with reduced rates of bone formation and mineral apposition without a significant change in osteoblast or osteoclast surface. Our results indicate that signaling by a Gi-coupled GPCR in osteoblasts leads to osteopenia resulting from a reduction in trabecular bone formation. The severity of the phenotype is related to the timing and duration of Ro1 expression during growth and development. The skeletal phenotype in Ro1 mice bears some similarity to that produced by knockout of Gs-alpha expression in osteoblasts and thus may be due at least in part to Gi-mediated inhibition of adenylyl cyclase.

The effect of leukocyte interleukin injection (Multikine) treatment on the peritumoral and intratumoral subpopulation of mononuclear cells and on tumor epithelia: a possible new approach to augmenting sensitivity to radiation therapy and chemotherapy in oral cancer--a multicenter phase I/II clinical Trial.

OBJECTIVES/HYPOTHESIS: The main objective of this study was to investigate the effect of the administration of a novel immunoadjuvant, leukocyte interleukin injection, as part of an immuno-augmenting treatment regimen on the peritumoral and intratumoral subpopulations of the tumor infiltrating mononuclear cells and on the epithelial and stromal components, when administered to patients with advanced primary oral squamous cell carcinoma classified as T2-3N0-2M0, as compared with disease-matched control patients (not treated with leukocyte interleukin injection). STUDY DESIGN: Multicenter Phase I/II clinical trial. Fifty-four patients from four clinical centers were included in the dose-escalating study (27 in each group [leukocyte interleukin injection-treated and control groups]). Cumulative leukocyte inter-leukin injection doses were 2400, 4800, and 8000 IU (as interleukin-2 equivalent). METHODS: Paraffin-embedded tumor samples obtained at surgical resection of the residual tumor (between days 21 and 28 after treatment initiation) were used. Histological analysis, necrosis evaluation, and American Joint Committee on Cancer grading were performed from H&E-stained sections. Immunohistochemical analysis was performed on three different tumor regions (surface, zone 1; center, zone 2; and tumor-stroma interface, zone 3). Trichrome staining was used to evaluate connective tissue, and morphometric measurements were made using ImagePro analysis software. Cell cycling was determined by the use of Ki-67 marker. RESULTS: Leukocyte interleukin injection treatment induced a shift from stromal infiltrating T cells toward intraepithelial T cells and posted a significant (P <.05) increase in intraepithelial CD3-positive T cells independent of the leukocyte interleukin injection dose, whereas the increase in CD25 (interleukin-2 receptor alpha [IL-2Ralpha])-positive lymphoid cells was significant only at the lowest leukocyte interleukin injection dose (P <.05). Furthermore, both low- and medium-dose leukocyte interleukin injection treatment induced a significant (P <.05) increase in the number of cycling tumor cells, as compared with control values. CONCLUSION: The results could be highly beneficial for patients with oral squamous cell carcinoma. First, leukocyte interleukin injection treatment induces T-cell migration into cancer nests and, second, noncycling cancer cells may enter cell cycling on administration of leukocyte interleukin injection. This latter effect may modulate the susceptibility of cancer cells to radiation therapy and chemotherapy. The findings may indicate a need to re-evaluate the way in which follow-up treatment (with radiation therapy and chemotherapy) of patients with head and neck cancer is currently approached.

Electric fields induced in a spherical volume conductor by temporally varying magnetic field gradients.

A homogeneous spherical volume conductor is used as a model system for the purpose of calculating electric fields induced in the human head by externally applied time-varying magnetic fields. We present results for the case where magnetic field gradient coils, used in magnetic resonance imaging (MRI), form the magnetic field, and we use these data to put limits on the rates of gradient change with time needed to produce nerve stimulation. The electric field is calculated analytically for the case of ideal longitudinal and transverse linear field gradients. We also show results from computer calculations yielding the electric field maps in a sphere when the field gradients are generated by a real MRI gradient coil set. In addition, the effect of shifting the sphere within each gradient coil volume is investigated. Numerical analysis shows similar results when applied to a model human head.

Stochastic effects on single phase fluid flow in porous media.

The flow encoded PEPI technique has been used to measure the fluid velocity distribution and fluid flow of water passing through a phantom comprising randomly distributed 10 mm glass beads. The object of these experiments is to determine the degree of causality between one steady-state flow condition and another. That is to say, knowing the mean fluid velocity and velocity distribution, can one predict what happens at a higher mean fluid velocity? In a second related experiment flow is established at a given mean fluid velocity. The velocity distribution is measured. The flow is then turned off and later re-established. In both kinds of experiment we conclude that the errors in predicting the flow velocity distribution and the errors in re-establishing a given velocity distribution lie well outside the intrinsic thermal noise associated with velocity measurement. It follows, therefore, that the causal approach to prediction of flow velocity distributions in porous media using the Navier-Stokes approach is invalid.

Direct measurement of porous media local hydrodynamical permeability using gas MRI.

The concept of hydraulic permeability is at the core of modeling single phase or multi-phase flow in heterogeneous porous media, as it is the spatial distribution of the permeability that primarily governs the behavior of fluid flow in the medium. To date, the modeling of fluid flow in porous media has been hampered by poor estimates of local permeability. Magnetic Resonance Imaging is well known for its ability to measure non-invasively the local density and flow rate of different fluids saturating porous media [1,2]. In this paper we demonstrate the first non-invasive method for the direct measurement of a single projection of the local permeability tensor of a porous medium using gas-phase MRI. The potential for three-dimensional imaging of the medium permeability is also discussed. The limitations of the method are listed and results are presented in a model porous medium as well as in a real oil reservoir rock.

Measuring spatially resolved gas transport and adsorption in coal using MRI.

The storage and transport of gases in coal is of tremendous importance in the utilisation of coalbeds, and in particular the recovery of methane. There is also increasing interest in the use of coal mines as sites for carbon dioxide sequestration to alleviate the potentially harmful effects of global warming. This paper demonstrates the use of magnetic resonance imaging to investigate the spatiotemporal dynamics of gas transport in coal. The presence of significant structural heterogeneities in the coal was observed. Dynamical effects displayed a broad range of time constants ranging from minutes to days.

Method for measuring local hydraulic permeability using magnetic resonance imaging.

The hydraulic permeability of a porous medium characterizes the ease with which a fluid may be driven through it, and is defined via the classical Darcy law. A method for noninvasive, nondestructive measurement of one projection of the local permeability tensor of a porous medium, using gas phase magnetic resonance imaging, is presented. Results are shown for one-dimensional experiments on dry porous rocks. The limitations of the method are explored, and the extension to three-dimensional imaging of the permeability is discussed.

Identification of phosphorylation sites in the G protein-coupled receptor for parathyroid hormone. Receptor phosphorylation is not required for agonist-induced internalization.

In some G protein-coupled receptors (GPCRs), agonist-dependent phosphorylation by specific GPCR kinases (GRKs) is an important mediator of receptor desensitization and endocytosis. Phosphorylation and the subsequent events that it triggers, such as arrestin binding, have been suggested to be regulatory mechanisms for a wide variety of GPCRs. In the present study, we investigated whether agonist-induced phosphorylation of the PTH receptor, a class II GPCR, also regulates receptor internalization. Upon agonist stimulation, the PTH receptor was exclusively phosphorylated on serine residues. Phosphoamino acid analysis of a number of receptor mutants in which individual serine residues had been replaced by threonine identified serine residues in positions 485, 486, and 489 of the cytoplasmic tail as sites of phosphorylation after agonist treatment. When serine residues at positions 483, 485, 486, 489, 495, and 498 were simultaneously replaced by alanine residues, the PTH receptor was no longer phosphorylated either basally or in response to PTH. The substitution of these serine residues by alanine affected neither the number of receptors expressed on the cell surface nor the ability of the receptor to signal via Gs. Overexpression of GRK2, but not GRK3, enhanced PTH-stimulated receptor phosphorylation, and this phosphorylation was abolished by alanine mutagenesis of residues 483, 485, 486, 489, 495, and 498. Thus, phosphorylation of the PTH receptor by the endogenous kinase in HEK-293 cells occurs on the same residues targeted by overexpressed GRK2. Strikingly, the rate and extent of PTH-stimulated internalization of mutated PTH receptors lacking phosphorylation sites were identical to that observed for the wild-type PTH receptor. Moreover, overexpressed GRK2, while enhancing the phosphorylation of the wild-type PTH receptor, had no affect on the rate or extent of receptor internalization in response to PTH. Thus, the agonist-occupied PTH receptor is phosphorylated by a kinase similar or identical to GRK2 in HEK-293 cells, but this phosphorylation is not requisite for efficient receptor endocytosis.

Fluid flow in porous systems.

Nuclear magnetic resonance (NMR) measurements of water velocity flowing through glass bead packs with a bead diameter of 10 mm have been made using the phi echo-planar imaging (PEPI) sequence. These results indicate that for various flow rates the flow variance is proportional to the mean flow velocity in agreement with the Mansfield-Issa equation. The velocity distributions are approximately Gaussian. Investigation of the slopes of the variance vs. velocity curves as a function of slice thickness indicate some coherence effects in the connectivity of the glass bead system. An extension of an earlier intervoxel coupling model is presented, which seems to explain the observed coherence effects.

Fluid transport in glass beads phantoms: spatial velocity measurements and confirmation of the stochastic model.

A stochastic model of fluid flow in porous rocks has been previously developed to explain the measured distribution of local velocity. The theoretical predictions of this model agree well with experimental results obtained from the magnetic resonance imaging-based measurements of the spatial variation of velocity of water permeating through Bentheimer and Clashach sandstones. To further verify previous results, we have performed new velocity measurement experiments using an efficient velocity encoded pi-echo planar imaging sequence on glass bead phantoms that exhibit more regular pore size distribution than rocks. The results show that velocity distributions in glass bead phantoms also exhibit Gaussian profiles and the linear relationship between the velocity variance and the mean velocity (the Mansfield-Issa equation).