Asymptomatic neonatal herpes simplex virus infection in mice leads to long-term cognitive impairment.

Neonatal herpes simplex virus (nHSV) is a devastating infection impacting approximately 14,000 newborns globally each year. Infection is associated with high neurologic morbidity and mortality, making early intervention and treatment critical. Clinical outcomes of symptomatic nHSV infections are well-studied, but little is known about the frequency of, or outcomes following, sub-clinical or asymptomatic nHSV. Given the ubiquitous nature of HSV infection and frequency of asymptomatic shedding in adults, subclinical infections are underreported, yet could contribute to long-term neurological damage. To assess potential neurological morbidity associated with subclinical nHSV infection, we developed a low-dose (100 PFU) HSV infection protocol in neonatal C57BL/6 mice. At this dose, HSV DNA was detected in the brain by PCR but was not associated with acute clinical symptoms. However, months after initial inoculation with 100 PFU of HSV, we observed impaired mouse performance on a range of cognitive and memory performance tasks. Memory impairment was induced by infection with either HSV-1 or HSV-2 wild-type viruses, but not by a viral mutant lacking the autophagy-modulating Beclin-binding domain of the neurovirulence gene γ34.5. Retroviral expression of wild type γ34.5 gene led to behavioral pathology in mice, suggesting that γ34.5 expression may be sufficient to cause cognitive impairment. Maternal immunization and HSV-specific antibody treatment prevented offspring from developing neurological sequelae following nHSV-1 infection. Altogether, these results support the idea that subclinical neonatal infections may lead to cognitive decline in adulthood, with possible profound implications for research on human neurodegenerative disorders such as Alzheimer's Disease.

Genetic insight on ice sheet history.

Octopus DNA reveals timing of the most recent collapse of the West Antarctic Ice Sheet.

SEA-LEVEL RISE. Sea-level rise due to polar ice-sheet mass loss during past warm periods.

Interdisciplinary studies of geologic archives have ushered in a new era of deciphering magnitudes, rates, and sources of sea-level rise from polar ice-sheet loss during past warm periods. Accounting for glacial isostatic processes helps to reconcile spatial variability in peak sea level during marine isotope stages 5e and 11, when the global mean reached 6 to 9 meters and 6 to 13 meters higher than present, respectively. Dynamic topography introduces large uncertainties on longer time scales, precluding robust sea-level estimates for intervals such as the Pliocene. Present climate is warming to a level associated with significant polar ice-sheet loss in the past. Here, we outline advances and challenges involved in constraining ice-sheet sensitivity to climate change with use of paleo-sea level records.

Ice volume and sea level during the last interglacial.

During the last interglacial period, ~125,000 years ago, sea level was at least several meters higher than at present, with substantial variability observed for peak sea level at geographically diverse sites. Speculation that the West Antarctic ice sheet collapsed during the last interglacial period has drawn particular interest to understanding climate and ice-sheet dynamics during this time interval. We provide an internally consistent database of coral U-Th ages to assess last interglacial sea-level observations in the context of isostatic modeling and stratigraphic evidence. These data indicate that global (eustatic) sea level peaked 5.5 to 9 meters above present sea level, requiring smaller ice sheets in both Greenland and Antarctica relative to today and indicating strong sea-level sensitivity to small changes in radiative forcing.

Enhancement of meniscal repair in the avascular zone using mesenchymal stem cells in a porcine model.

We hypothesised that meniscal tears treated with mesenchymal stem cells (MSCs) together with a conventional suturing technique would show improved healing compared with those treated by a conventional suturing technique alone. In a controlled laboratory study 28 adult pigs (56 knees) underwent meniscal procedures after the creation of a radial incision to represent a tear. Group 1 (n = 9) had a radial meniscal tear which was left untreated. In group 2 (n = 19) the incision was repaired with sutures and fibrin glue and in group 3, the experimental group (n = 28), treatment was by MSCs, suturing and fibrin glue. At eight weeks, macroscopic examination of group 1 showed no healing in any specimens. In group 2 no healing was found in 12 specimens and incomplete healing in seven. The experimental group 3 had 21 specimens with complete healing, five with incomplete healing and two with no healing. Between the experimental group and each of the control groups this difference was significant (p < 0.001). The histological and macroscopic findings showed that the repair of meniscal tears in the avascular zone was significantly improved with MSCs, but that the mechanical properties of the healed menisci remained reduced.

Expression of the cellular FLICE-inhibitory protein (c-FLIP) protects Hodgkin's lymphoma cells from autonomous Fas-mediated death.

Hodgkin's lymphoma (HL) is characterized by the presence of malignant so-called Hodgkin's/Reed-Sternberg (HRS) cells, which display resistance to certain apoptotic stimuli, including a lack of sensitivity to Fas-mediated cell death. However, the mechanisms responsible for their resistance to apoptosis inducers have not been elucidated. Here we confirm that both HL-derived cell lines and the HRS cells of primary HL tissues express Fas ligand (FasL) along with the inhibitory c-FLIP protein. Down-regulation of cellular FLICE (FADD-like IL-1beta-converting enzyme)-inhibitory protein (c-FLIP) through the use of specific small inhibitory RNAs (siRNAs) leads to reduced viability of the L428 and L591 HL-derived cell lines. To determine whether endogenous FasL was responsible for the reduction in cell viability observed after down-regulation of c-FLIP, L428 and L591 cells were treated with c-FLIP-specific siRNAs with and without siRNAs directed to FasL. Treatment of these cells with both c-FLIP- and FasL-specific siRNAs in combination restored cell viability to near control levels. Our results provide a mechanism whereby HRS cells are protected from autonomous FasL-mediated cell death while preserving their ability to evade immunosurveillance. Targeting c-FLIP could provide a novel approach to the treatment of HL.

NAADP receptors are present and functional in the heart.

Alongside the well-studied inositol 1,4,5 trisphosphate and ryanodine receptors, evidence is gathering that a new intracellular release mechanism, gated by the pyridine nucleotide nicotinic acid adenine dinucleotide phosphate (NAADP), is present in numerous organisms, ranging from plant to mammalian cells (reviewed in [1]). Most cells have been shown to express at least two Ca(2+)-release mechanisms controlled by different messengers, and this can lead to redundancy, convergence, or divergence of responses. One exception appears to be muscle and heart contractile tissues. Here, it is thought that the dominant intracellular channel is the ryanodine receptor, while IP(3) receptors are poorly expressed and their role appears to be negligible. We now report that NAADP receptors are functional and abundant in cardiac microsomes. NAADP binds specifically and with high affinity (130 pM and 4 nM) to two sites on cardiac microsomes and releases Ca(2+) with an apparent EC(50) of 323 +/- 14 nM. Furthermore, binding experiments show that this receptor displays both positive and negative cooperativity, a peculiarity unique among intracellular Ca(2+) channels. Therefore, we show that the heart possesses multiple mechanisms to increase the complexity of Ca(2+) signaling and that NAADP may be integral in the functioning of this organ.

An accurate, convective energy equation based automated meshing technique for analysis of blood vessels and tissues.

An automated three-element meshing method for generating finite element based models for the accurate thermal analysis of blood vessels imbedded in tissue has been developed and evaluated. The meshing method places eight noded hexahedral elements inside the vessels where advective flows exist, and four noded tetrahedral elements in the surrounding tissue. The higher order hexahedrals are used where advective flow fields occur, since high accuracy is required and effective upwinding algorithms exist. Tetrahedral elements are placed in the remaining tissue region, since they are computationally more efficient and existing automatic tetrahedral mesh generators can be used. Five noded pyramid elements connect the hexahedrals and tetrahedrals. A convective energy equation (CEE) based finite element algorithm solves for the temperature distributions in the flowing blood, while a finite element formulation of a generalized conduction equation is used in the surrounding tissue. Use of the CEE allows accurate solutions to be obtained without the necessity of assuming ad hoc values for heat transfer coefficients. Comparisons of the predictions of the three-element model to analytical solutions show that the three-element model accurately simulates temperature fields. Energy balance checks show that the three-element model has small, acceptable errors. In summary, this method provides an accurate, automatic finite element gridding procedure for thermal analysis of irregularly shaped tissue regions that contain important blood vessels. At present, the models so generated are relatively large (in order to obtain accurate results) and are, thus, best used for providing accurate reference values for checking other approximate formulations to complicated, conjugated blood heat transfer problems.

Effect of material deprivation on Epstein-Barr virus infection in Hodgkin's disease in the West Midlands.

We have used Townsend scores from postcode data to compare levels of material deprivation and Epstein-Barr virus (EBV)-positivity for 223 patients diagnosed with Hodgkin's disease (HD) in the period 1981-1997. The presence of EBV in HD tumours was determined using in situ hybridization to target the abundantly expressed EBV early RNAs. EBV was detected in the malignant Hodgkin and Reed-Sternberg cells in 47/223 HD cases (21%). There was found to be a tendency for higher Townsend scores (indicative of higher levels of material deprivation) in EBV-positive HD patients, but this association was not statistically significant. When various subgroups of patients from the study were examined separately the indication of higher Townsend scores in EBV-positive patients was found to be more marked for patients with mixed cellularity disease (P = 0.09) and for females (P = 0.03). The results of this study suggest that differences in the level of material deprivation are important in determining the likelihood of EBV-positive HD in the UK, particularly for certain subgroups of patients. It is not known what specific socioeconomic factors are responsible for these differences, although alterations in the timing or rate of primary EBV infection, or decline in the level of EBV-specific immunity, may be important.

Reduced-order modeling for hyperthermia: an extended balanced-realization-based approach.

Accurate thermal models are needed in hyperthermia cancer treatments for such tasks as actuator and sensor placement design, parameter estimation, and feedback temperature control. The complexity of the human body produces full-order models which are too large for effective execution of these tasks, making use of reduced-order models necessary. However, standard balanced-realization (SBR)-based model reduction techniques require a priori knowledge of the particular placement of actuators and sensors for model reduction. Since placement design is intractable (computationally) on the full-order models, SBR techniques must use ad hoc placements. To alleviate this problem, an extended balanced-realization (EBR)-based model-order reduction approach is presented. The new technique allows model order reduction to be performed over all possible placement designs and does not require ad hoc placement designs. It is shown that models obtained using the EBR method are more robust to intratreatment changes in the placement of the applied power field than those models obtained using the SBR method.

A comparison of reduced-order modelling techniques for application in hyperthermia control and estimation.

Reduced-order modelling techniques can make important contributions in the control and state estimation of large systems. In hyperthermia, reduced-order modelling can provide a useful tool by which a large thermal model can be reduced to the most significant subset of its full-order modes, making real-time control and estimation possible. Two such reduction methods, one based on modal decomposition and the other on balanced realization, are compared in the context of simulated hyperthermia heat transfer problems. The results show that the modal decomposition reduction method has three significant advantages over that of balanced realization. First, modal decomposition reduced models result in less error, when compared to the full-order model, than balanced realization reduced models of similar order in problems with low or moderate advective heat transfer. Second, because the balanced realization based methods require a priori knowledge of the sensor and actuator placements, the reduced-order model is not robust to changes in sensor or actuator locations, a limitation not present in modal decomposition. Third, the modal decomposition transformation is less demanding computationally. On the other hand, in thermal problems dominated by advective heat transfer, numerical instabilities make modal decomposition based reduction problematic. Modal decomposition methods are therefore recommended for reduction of models in which advection is not dominant and research continues into methods to render balanced realization based reduction more suitable for real-time clinical hyperthermia control and estimation.

A generic tissue convective energy balance equation: Part I--theory and derivation.

A new equation for calculating temperatures in living tissues, the tissue convective energy balance equation (TCEBE), is derived using only a few assumptions. The resulting equation is basic, general and applicable to any tissue. The (unsolved) TCEBE is used: (a) to relate both Pennes' BHTE perfusion-related parameter (W) and the effective thermal conductivity equation's perfusion-related parameter (keff) to the true capillary perfusion Pcap, and (b) to show that both W and keff are defined, nonphysiological variables, which are only related to Pcap in a problem-dependent manner. Finally, the derivation of the relationship between W and Pcap provides a complete derivation of Pennes' BHTE, something that has not been previously done.

The simulation of discrete vessel effects in experimental hyperthermia.

The ability of two simple thermal models to predict experimentally measured in vivo temperature profiles was compared. These comparisons were done both with and without the inclusion of separate, discrete blood vessels. The two tissue models were: 1) Pennes' Bio-Heat Transfer equation (BHTE), and 2) an effective thermal conductivity equation (ETCE). The experimental temperature data were measured (Moros, 1990; Moros et al., 1993) in the thighs of anesthetized greyhound dogs under hyperthermic conditions generated by scanned focused ultrasound. Blood vessels were added to the thermal models in counter-current pairs transiting the model domain. The blood vessels in both models were assumed to have a constant heat transfer coefficient, and an axially varying mixed mean temperature. The vessel locations were determined a posteriori, via inspection of the experimental temperature data. Least square error fits of the predicted model temperatures to the experimental temperature data were obtained by adjusting both (a) the mass flow rate within and (b) the position of each blood vessel, and (c) the value of either the perfusion parameter (W) in the BHTE or the effective thermal conductivity parameter (Keff) in the ETCE. When small numbers (3-4) of blood vessel pairs were included, both of the models showed significant improvement in their ability to predict the experimental temperatures. Although both models performed well in terms of predicting temperatures near large vessels, the BHTE had a statistically significant better ability to predict the complete set of measured temperatures at all locations.

Experimental evaluation of two simple thermal models using hyperthermia in muscle in vivo.

The predictions from two simple field equation models for calculating temperature distributions in tissue, namely, the Pennes' bioheat transfer equation (BHTE) and an effective thermal conductivity equation (ETCE), were compared to in vivo experimental temperature measurements made under hyperthermic conditions generated by scanned focused ultrasound. The models were kept simple (i.e. homogenous isotropic properties, no separate blood vessels included) in order to concentrate attention on the predictive abilities of these field equations using a minimum number of free parameters. Simulated results were fitted to the experimental data (multiple, linear temperature profiles in the thigh muscles of greyhound dogs) by minimizing a performance index using a golden section searth. This search determined a value for the single free parameter in each model (blood perfusion in the BHTE, and effective thermal conductivity in the ETCE) which minimized the square error difference between the experimental and simulated temperatures. The results showed that (a) the simple BHTE model could qualitatively reproduce the major features of the temperature patterns seen experimentally better than the ETCE model could, and (b) the simple BHTE model produced better quantitative fits to the experimental data than did the simple ETCE model. In addition, blood perfusion predictions from the BHTE model compared well to measurements done with coloured microspheres. Finally, the experimental results showed that individual, large blood vessels appeared to have a major influence in producing asymmetries in the experimental data in 21% of the measured temperature profiles.

Cataract extraction in a fur seal.

Cataracts are common in pinnipeds, but ocular examination and surgery are complicated because of the small pupil, which is difficult to dilate with mydriatics. Vision was evident after a cataract was removed from a fur seal, using a manual technique of irrigation and aspiration. Access to the lens was limited by the constricted pupil, which could not be dilated with tropicamide, phenylephrine, or atropine.

Transfer of secretory proteins from the endoplasmic reticulum to the Golgi apparatus: discrimination between homologous and heterologous transfer in intact heterokaryons.

To examine aspects of the transfer of secretory proteins from the endoplasmic reticulum to the Golgi apparatus in situ, heterokaryons were formed between Hep G2 human hepatoma cells and WI-38 human fibroblasts. The cells were appropriately treated with cycloheximide before fusion, which emptied them of their respective secretory proteins, serum albumin for the Hep G2 cells and procollagen I for the WI-38 cells. After fusion was complete, the cycloheximide was washed out, protein synthesis was resumed, and the rates of reappearance of serum albumin and procollagen I in the two separated Golgi apparatuses within each heterokaryon were followed by immunofluorescence microscopy. Serum albumin was found to always reappear first in the Golgi apparatus contributed by the Hep G2 half of the heterokaryon, and procollagen I in the Golgi apparatus of the WI-38 half. These results suggest that the endoplasmic reticulum-to-Golgi apparatus transfer in situ is not simply a stochastic process but is either spatially restricted or exhibits cell-type specificity or both.

Occult malignancy presenting as acute fulminant hepatic failure.

Only six cases of hepatic metastatic disease presenting as acute fulminant liver failure have been recorded in the literature. A seventh case is reported here where, after presenting in acute liver failure, it was not possible to establish a tissue diagnosis but evidence of massive liver replacement by tumour was provided by ultrasound imaging and radionuclide scintiscanning.