An early modern human presence in Sumatra 73,000-63,000 years ago.

Genetic evidence for anatomically modern humans (AMH) out of Africa before 75 thousand years ago (ka) and in island southeast Asia (ISEA) before 60 ka (93-61 ka) predates accepted archaeological records of occupation in the region. Claims that AMH arrived in ISEA before 60 ka (ref. 4) have been supported only by equivocal or non-skeletal evidence. AMH evidence from this period is rare and lacks robust chronologies owing to a lack of direct dating applications, poor preservation and/or excavation strategies and questionable taxonomic identifications. Lida Ajer is a Sumatran Pleistocene cave with a rich rainforest fauna associated with fossil human teeth. The importance of the site is unclear owing to unsupported taxonomic identification of these fossils and uncertainties regarding the age of the deposit, therefore it is rarely considered in models of human dispersal. Here we reinvestigate Lida Ajer to identify the teeth confidently and establish a robust chronology using an integrated dating approach. Using enamel-dentine junction morphology, enamel thickness and comparative morphology, we show that the teeth are unequivocally AMH. Luminescence and uranium-series techniques applied to bone-bearing sediments and speleothems, and coupled uranium-series and electron spin resonance dating of mammalian teeth, place modern humans in Sumatra between 73 and 63 ka. This age is consistent with biostratigraphic estimations, palaeoclimate and sea-level reconstructions, and genetic evidence for a pre-60 ka arrival of AMH into ISEA. Lida Ajer represents, to our knowledge, the earliest evidence of rainforest occupation by AMH, and underscores the importance of reassessing the timing and environmental context of the dispersal of modern humans out of Africa.

Benthic primary producers are key to sustain the Wadden Sea food web: stable carbon isotope analysis at landscape scale.

Coastal food webs can be supported by local benthic or pelagic primary producers and by the import of organic matter. Distinguishing between these energy sources is essential for our understanding of ecosystem functioning. However, the relative contribution of these components to the food web at the landscape scale is often unclear, as many studies lack good taxonomic and spatial resolution across large areas. Here, using stable carbon isotopes, we report on the primary carbon sources for consumers and their spatial variability across one of the world's largest intertidal ecosystems (Dutch Wadden Sea; 1460 km2 intertidal surface area), at an exceptionally high taxonomic (178 species) and spatial resolution (9,165 samples from 839 locations). The absence of overlap in δ13 C values between consumers and terrestrial organic matter suggests that benthic and pelagic producers dominate carbon input into this food web. In combination with the consistent enrichment of benthic primary producers (δ13 C -16.3‰) relative to pelagic primary producers (δ13 C -18.8) across the landscape, this allowed the use of a two-food-source isotope-mixing model. This spatially resolved modelling revealed that benthic primary producers (microphytobenthos) are the most important energy source for the majority of consumers at higher trophic levels (worms, molluscs, crustaceans, fish, and birds), and thus to the whole food web. In addition, we found large spatial heterogeneity in the δ13 C values of benthic primary producers (δ13 C -19.2 to -11.5‰) and primary consumers (δ13 C -25.5 to -9.9‰), emphasizing the need for spatially explicit sampling of benthic and pelagic primary producers in coastal ecosystems. Our findings have important implications for our understanding of the functioning of ecological networks and for the management of coastal ecosystems.

Detection of Epstein-Barr virus DNA by polymerase chain reaction in blood and tissue biopsies from patients with Sjogren's syndrome.

Polymerase chain reaction has been used to detect increased levels of EBV DNA in salivary gland (SG) biopsies and PBL from patients with Sjogren's syndrome (SS). These results suggest that EBV, which has a normal site of latency in a small number of SG epithelial cells, may be reactivated in SS patients and provide a target for immune attack. The great sensitivity of polymerase chain reaction (PCR) and the ability to analyze very small tissue biopsies (37) make this technique well suited for clinical diagnosis. Specific methods to prevent artefactual contamination of tissue biopsy DNA with viral DNA of other samples (i.e., lyophilization of samples before DNA extraction) and the use of an internal positive control (i.e., inclusion of primers for a single copy human gene) during PCR amplification are presented. Since EBV reactivation occurs with markedly increased frequency in patients with lymphoproliferative and immunodeficiency diseases, as well as transplant recipients receiving cyclosporin A (10), rapid methods of viral detection such as PCR may allow better monitoring of medications and early detection of EBV-related lymphomas that may arise in these patients.

Virus entry and innate immune activation.

Human cytomegalovirus (HCMV) exhibits an exceptionally broad cellular tropism as it is capable of infecting most major organ systems and cell types. Definitive proof of an essential role for a cellular molecule that serves as an entry receptor has proven very challenging. It is widely hypothesized that receptor utilization, envelope glycoprotein requirements and entry pathways may all vary according to cell type, which is partially supported by the data. What has clearly emerged in recent years is that virus entry is not going undetected by the host. Robust and rapid induction of innate immune response is intimately associated with entry-related events. Here we review the state of knowledge on HCMV cellular entry mediators confronting the scientific challenges by accruing a definitive data set. We also review the roles of pattern recognition receptors such as Toll-like receptors in activation of specific innate immune response and discuss how entry events are tightly coordinated with innate immune initiation steps.

Down-regulation of epidermal growth factor receptor-dependent signaling by Porphyromonas gingivalis lipopolysaccharide in life-expanded human gingival fibroblasts.

BACKGROUND AND OBJECTIVE: Human gingival fibroblasts exhibit proliferative responses following epidermal growth factor exposure, which are thought to enhance periodontal regeneration in the absence of bacterial products such as lipopolysacharide. However, lipopolysaccharide challenge activates human gingival fibroblasts to release several inflammatory mediators that contribute to the immune response associated with periodontitis and attenuate wound repair. We tested the hypothesis that Porphyromonas gingivalis lipopolysaccharide-activated signaling pathways down-regulate epidermal growth factor receptor-dependent events. MATERIAL AND METHODS: To study lipopolysaccharide/epidermal growth factor interactions in human gingival fibroblasts, we introduced the catalytic subunit of human telomerase into human gingival fibroblasts, thereby generating a more long-lived cellular model. These cells were characterized and evaluated for lipopolysaccharide/epidermal growth factor responsiveness and regulation of epidermal growth factor-dependent pathways. RESULTS: Comparison of human telomerase-transduced gingival fibroblasts with human gingival fibroblasts revealed that both cell lines exhibit a spindle-like morphology and express similar levels of epidermal growth factor receptor, CD14 and Toll-like receptors 2 and 4. Importantly, human telomerase-transduced gingival fibroblasts proliferation rates are increased 5-9 fold over human gingival fibroblasts and exhibit a longer life span in culture. In addition, human telomerase-transduced gingival fibroblasts and human gingival fibroblasts exhibit comparable profiles of mitogen-activated protein kinase kinase (extracellular signal-regulated kinase 1/2) activation upon epidermal growth factor or P. gingivalis lipopolysaccharide administration. Interestingly, treatment with P. gingivalis lipopolysaccharide leads to a down-regulation of epidermal growth factor-dependent extracellular signal-regulated kinase 1/2, p38 and cyclic-AMP response element binding protein phosphorylation in both cell types. CONCLUSION: These studies demonstrate that human telomerase-transduced gingival fibroblasts exhibit an extended life span and recapitulate human gingival fibroblasts biology. Moreover, this system has allowed for the first demonstration of lipopolysaccharide down-regulation of epidermal growth factor activated pathways in human gingival fibroblasts and should facilitate the analysis of signaling events relevant to the pathogenesis and treatment of periodontitis.

Engagement of the cellular receptor for glycoprotein B of human cytomegalovirus activates the interferon-responsive pathway.

Cells respond to contact with human cytomegalovirus (HCMV) virions by initiating intracellular signaling and gene expression characteristic of the interferon (IFN)-responsive pathway. Herein, we demonstrate that a principal mechanism of HCMV-induced signal transduction is via an interaction of the primary viral ligand, glycoprotein B (gB), with its cellular receptor. Cells incubated with a purified, soluble form of gB resulted in the transcriptional upregulation of IFN-responsive genes OAS and ISG54 (encoding 2'-5' oligoadenylate synthetase and an IFN-stimulated gene product of 54 kDa) to a comparable level as virions or IFN. Gene induction was an immediate and direct response to gB which did not require de novo protein synthesis. Neither the initial virus attachment site, heparan sulfate proteoglycans, nor the IFN-alpha/beta or IFN-gamma receptors are involved in the response. Pleotropic protein phosphorylation was required for cellular gene induction, and the mitogen-activated protein kinases ERK1 and ERK2 were activated in response to the ligand. Together these data indicate that a principal means by which cytomegalovirus induces intracellular signaling and activation of the interferon-responsive pathway is via an interaction of gB with an as yet unidentified, likely novel cellular receptor that interfaces with the IFN signaling pathway.

Predictors of glycemic control in children with type 1 diabetes: the importance of race.

Diabetes is a common cause of kidney failure and blindness among young adults, particularly of African-American descent. Since glycemic control is a predictor of diabetes complications, we evaluated the impact of multiple factors including a special multidisciplinary management program on glycosylated hemoglobin in children with Type 1 diabetes. Data was collected from pediatric diabetes clinics in New Orleans, LA and Baltimore, MD. In New Orleans, hemoglobin A(1c) was higher in African-American patients 12. 5+/-3.3% (n=71) vs. 10.7+/-2.1% (n=80) in Caucasian children, p<0. 0001. Longer duration of diabetes was also associated with higher hemoglobin A(1c) in both races. The effect of race on hemoglobin A(1c) was independent of the influence of sex, insurance status, body mass index (BMI) z-score, and number of clinic visits. Covariate analysis with mean blood glucose levels indicated that higher hemoglobin A(1c) was attributable to higher mean blood glucose levels in African-American children. From the Baltimore data, a multidisciplinary intervention program led to improved total glycosylated hemoglobin for Caucasian patients but not for African-American children. Poorer glycemic control of African-American children is likely to predispose them to a higher likelihood of developing microvascular complications as they mature. Standard hospital-based multidisciplinary programming for diabetes management may have limited effectiveness in improving glycemic control of African-American children with diabetes. Innovative intervention programs are needed for these high-risk patients.

Analysis of cytomegalovirus ligands, receptors, and the entry pathway.

The process of virus entry accomplishes the delivery of the viral genetic information into the cell so that replication can take place. Entry of enveloped viruses into mammalian cells requires that the virus attach to the host cell surface through an interaction between an envelope component and a cellular molecule that serves as a receptor. After attachment, the virus must cross the plasma membrane during a phase of the life cycle termed penetration. For enveloped viruses, a fusion reaction occurs between the viral envelope in either the endosomal membrane or the plasma membrane. In simple viral systems that have one or two envelope glycoproteins, generally, a single, specific interaction between a viral protein and a cellular constituent is necessary and sufficient to result in infection. The herpesviruses, including human cytomegalovirus (HCMV), are structurally much more complex and often exhibit broad, diverse cellular tropism. Thus, all evidence to date points to the involvement of multiple cellular and viral proteins with overlapping or compensatory mechanisms possibly responsible for entry into specialized cell types.

Human papilloma virus E6/E7 genes can expand the lifespan of human corneal fibroblasts.

Human corneal fibroblasts were infected with a retroviral delivery vector containing the E6 and E7 genes from human Papilloma virus type 16 in order to produce cell lines that have an expanded lifespan in culture. Morphologically, some of the transfected corneal fibroblast lines appeared to have the normal spindle-shape morphology of diploid fibroblasts, whereas other lines appeared to have a more elongated morphology. All the cell lines were anchorage-dependent. Cells that had a normal morphology grew at a rate similar to normal diploid human corneal fibroblasts and had a population doubling time of 48 h. All E6/E7 expressing cell lines, regardless of morphology, produce types I, III, and V collagen, at levels similar to those observed in the parent corneal diploid fibroblast. These corneal fibroblast lines will be a useful in vitro system to study collagen expression and fibril formation, as well as normal stroma development. These results also demonstrate that the use of E6/E7 genes to expand a cell's lifespan can be a powerful tool because it does not appear to alter either the growth rate of the cell or collagen expression.

Hearing loss and automobile airbag deployments.

In an on-going research program, on the causation of injuries in motor vehicle accidents, at the University of Michigan Transportation Research Institute, crashes with airbags have been, and are continuing to be, investigated. In order to determine the occurrence, if any, of 'hearing problems' associated with airbag deployments, drivers and passengers who had been involved in 'airbag' automobile crashes were interviewed by telephone. From the crashes investigated, 225 attempts were made to contact drivers who were exposed to airbag deployments. From these telephone interviews, contacts were made with 177 car occupants. Only three reported that they had experienced any type of hearing-related problems subsequent to their crash. One other case is reported of a driver who had pre-crash hearing loss. It appears that permanent hearing deficit due to airbag deployment, both in cars with the steering wheel airbag alone, and in those with a passenger side airbag, is an infrequent event (1.7%) from the data of this study.

Cellular receptor for human cytomegalovirus.

The results of the studies reported strongly suggest that the binding of HCMV to a target cell in vitro is mediated by a glycoprotein membrane receptor having a molecular weight of approximately 30 kd. It has been further concluded that this particular molecule is the principal HCMV receptor, since it has been observed in each of the cell types that have been studied. As for the 28 and 60 kd molecules, the former may be a proteolytic degradation product and the latter a dimer of the 30 kd receptor. The precise contribution of the 92 kd that may allow for viral penetration into a host cell molecule to the initial recognition event is unknown at the present time.

[Use of cultured, virus-infected cells to study the biogenesis of polarity of epithelial cells]. / Utilisation de cultures de cellules infectées par des virus pour l'étude de la biogénèse de la polarité des cellules épithéliales.

The major characteristic of the eucaryote cell is the presence of specialized organelles in which macromolecular components responsible for various subcellular functions are segregated. The membranes of these organelles serve not only as divisions between the various cytoplasmic compartments, but also provide scaffolding within which the macromolecular complexes of the organelle assemble and become functionally integrated. It is obvious that because of the degree of complexity resulting from the existence of numerous compartments and membrane systems, the development of a genetic programme in a eucaryote cell requires not only the transcription of specific genes and translation in the cytoplasma of the resultant messenger RNA, but also the activity of mechanisms which ensure that each polypeptide reaches the site of its function, which may be in the cytosol, in a membrane, or in the luminal cavity of an organelle. In the special case of membrane proteins, such mechanisms must result not only in the specific distribution of polypeptides newly synthesized in the various types of cell membrane, but also the arrangement of them required in the lipid bi-layer necessary for their normal function.

Web-based multimedia courseware for emergency cardiac patient management simulations.

This is a multidisciplinary inter-departmental/faculty project between the departments of computer science, electronic, communications and electrical engineering and nursing and paramedic sciences. The objective is to develop a web based multimedia front end to existing simulations of cardiac emergency scenaria. It will be used firstly in the teaching of nurses. The University of Hertfordshire is the only University in Britain using simulations of cardiac emergency scenaria for nurse and paramedic science education and therefore this project will add the multimedia dimension in distributed courses over the web and will assess the improvement in the educational process. The use of network and multimedia technologies, provide interactive learning, immediate feedback to students' responses, individually tailored instructions, objective testing and entertaining delivery. The end product of this project will serve as interactive material to enhance experiential learning for nursing students using the simulations of cardiac emergency scenaria. The emergency treatment simulations have been developed using VisSim and may be compiled as C code. The objective of the project is to provide a web based user friendly multimedia interface in order to demonstrate the way in which patients may be managed in critical situations by applying advanced technological equipment and drug administration. Then the user will be able to better appreciate the concepts involved by running the VisSim simulations. The evaluation group for the proposed software will be the Department of Nursing and Paramedic Sciences About 200 nurses use simulations every year for training purposes as part of their course requirements.

A Neanderthal lower molar from Stajnia Cave, Poland.

The primary aim of this study was to conduct a taxonomic assessment of the second of three isolated human teeth found in the Stajnia Cave (north of the Carpathians, Poland) in 2008. The specimen was located near a human tooth (S5000), which was identified by Urbanowski et al. (2010) as a Neanderthal permanent upper molar. Both of these teeth were excavated from the D2 layer, which belongs to the D stratigraphic complex comprising the archaeological assemblage associated with the Micoquian tradition. An Ursus spelaeus bone and Mammuthus primigenius tooth that were also excavated from the D2 layer were dated to >49,000 years BP (by AMS (14)C) and 52.9 ka BP (by U-Th), respectively. The sediment overlying stratigraphic complex D was dated to 45.9 ka BP by the OSL method. The S4300 tooth is a lower first or second permanent molar belonging to an individual other than that who once possessed the S5000 tooth. The S4300 tooth exhibits a combination of traits typical of Neanderthal lower molars, including a mid-trigonid crest, large anterior fovea, taurodontism and subvertical grooves on the interproximal face, indicating that this tooth belonged to a Neanderthal individual. The S4300 tooth from Stajnia Cave is one of the oldest human remains found in Poland.

Interactive Retro-Deformation of Terrain for Reconstructing 3D Fault Displacements.

Planetary topography is the result of complex interactions between geological processes, of which faulting is a prominent component. Surface-rupturing earthquakes cut and move landforms which develop across active faults, producing characteristic surface displacements across the fault. Geometric models of faults and their associated surface displacements are commonly applied to reconstruct these offsets to enable interpretation of the observed topography. However, current 2D techniques are limited in their capability to convey both the three-dimensional kinematics of faulting and the incremental sequence of events required by a given reconstruction. Here we present a real-time system for interactive retro-deformation of faulted topography to enable reconstruction of fault displacement within a high-resolution (sub 1m/pixel) 3D terrain visualization. We employ geometry shaders on the GPU to intersect the surface mesh with fault-segments interactively specified by the user and transform the resulting surface blocks in realtime according to a kinematic model of fault motion. Our method facilitates a human-in-the-loop approach to reconstruction of fault displacements by providing instant visual feedback while exploring the parameter space. Thus, scientists can evaluate the validity of traditional point-to-point reconstructions by visually examining a smooth interpolation of the displacement in 3D. We show the efficacy of our approach by using it to reconstruct segments of the San Andreas fault, California as well as a graben structure in the Noctis Labyrinthus region on Mars.

An immortalized human fibroblast cell line is permissive for human cytomegalovirus infection.

Human foreskin fibroblasts (HFF) were immortalized via retrovirus-mediated gene transfer of the E6 and E7 genes of human papillomavirus type 16. An immortalized fibroblast (IF) cell line which was morphologically akin to the parental cell line was isolated. The IF cell line was evaluated for permissiveness to human cytomegalovirus (HCMV) infection after the IF cell line surpassed the normal passage limitation of diploid fibroblasts. Western immunoblot analysis of representative HCMV-encoded immediate-early (72-kDa), early (gB), and late (gH) gene products demonstrated that the IF cell line produced these proteins analogous to those produced by the parental HFF cells. Similar quantities of infectious virus were produced in the IF and HFF cell lines as determined in one-step growth curve experiments. Compared with the HFF cells, morphologically identical plaques were produced in the IF cell line in approximately 10 to 12 days postinfection. These findings indicate that fibroblast cell lines immortalized with transforming genes of human papillomavirus retain complete permissiveness to HCMV infection and support plaque formation. The IF cell line will be useful for future genetic analysis of HCMV.

Characterization of a panel of insertion mutants in human cytomegalovirus glycoprotein B.

Glycoprotein B (gB; gpUL55) of human cytomegalovirus (HCMV) plays a critical role in virus entry and cell-to-cell spread of infection. To define the structure-function relationships in gB, a panel of linker-insertion mutations was generated throughout the coding region. This strategy yielded a panel of 22 mutants with four amino acid insertions and 3 large truncation mutants. Assessment of the mutant proteins' biosynthetic properties and folding patterns analyzed in context with predicted secondary features revealed novel insights into gB's structure and trafficking properties. All of the insertion mutants were able to assemble into oligomers, suggesting that oligomerization is tolerant of small insertions and/or that multiple regions of the protein may be involved. Computer algorithm predictions of gB's secondary structure indicate that the furin-recognized cleavage site falls within an exposed loop. This loop may be particularly sensitive to structural alterations, since insertions upstream and downstream of the cleavage site rendered the mutant proteins cleavage defective. In addition, a strong correlation existed between terminal folding and cleavage of gB. Interestingly, terminal folding was not correlated with delivery to the cell surface but may influence the rate of transport to the cell surface. Nine mutants, containing insertions in both the extracellular and intracellular portions of gB, retained wild-type structural properties. This panel of characterized gB mutants, the first of this type for an HCMV protein, will be a useful tool in dissecting the role of gB during HCMV infection.

Interference with annexin II has no effect on entry of human cytomegalovirus into fibroblast cells.

Annexin II has been identified as a human cytomegalovirus (HCMV)-binding protein, shown to be a component of purified virions and proposed as a cellular receptor for the virus. In addition, annexin II is capable of associating with the major HCMV envelope glycoprotein, gB (gpUL55). As one approach to examine the role of annexin II in virus entry, a high-titre polyclonal annexin II-specific antibody was produced and its effects in virus entry and cell-to-cell spread assays were tested. This anti-annexin II serum recognized virion and cell surface annexin II and annexin II-derived peptides. Recombinant annexin II, with characteristic calcium- and phospholipid-binding activities, was also examined. Pretreatment of cells, virions or both with polyclonal anti-annexin II serum, affinity-purified anti-annexin II antibodies or recombinant annexin II protein prior to infection was inconsequential to the entry of HCMV into fibroblasts. HCMV was also able to dose-dependently penetrate annexin II-deficient 293 cells. Furthermore, the spread of HCMV from cell to cell was not inhibited in the presence of polyclonal anti-annexin II antibodies or exogenous annexin II protein. These experiments do not support a direct role of annexin II in virus entry or spread. Alternative roles for the gB-annexin II interaction are proposed.

Effect of herpes simplex virus type 1 on cellular pools of oligosaccharide-lipid.

Incorporation of [3H]mannose into cellular pools of mannosylphosphoryl dolichol (Man-P-Dol), oligosaccharide-lipid, and glycoprotein was measured and compared in herpes simplex virus type 1 (HSV-1)-infected cells and -uninfected cells. While mannose incorporation into the monosaccharide-dolichol fraction was similar in infected or uninfected Vero cells, incorporation into the oligosaccharide-lipid fraction was markedly reduced in HSV-1-infected cells (64% of control levels). In contrast, mannose incorporation into glycoprotein was significantly increased in virus-infected cells versus uninfected cells (194% of control levels). The kinetics of incorporation into the various fractions was examined and it was determined that there was minimal increase in mannose incorporation into oligosaccharide-lipid after 8 hr postinfection in virus-infected cells. This corresponded to the time at which nonglycosylated precursors of the HSV-1 glycoproteins were first detected in association with the nuclear fraction. These data suggest that there is an accelerated turnover of oligosaccharide-lipid in virus-infected Vero cells which is most likely due to extensive glycoprotein synthesis.