The long-term expansion and recession of human populations.

Over the last 12,000 y, human populations have expanded and transformed critical earth systems. Yet, a key unresolved question in the environmental and social sciences remains: Why did human populations grow and, sometimes, decline in the first place? Our research builds on 20 y of archaeological research studying the deep time dynamics of human populations to propose an explanation for the long-term growth and stability of human populations. Innovations in the productive capacity of populations fuels exponential-like growth over thousands of years; however, innovations saturate over time and, often, may leave populations vulnerable to large recessions in their well-being and population density. Empirically, we find a trade-off between changes in land use that increase the production and consumption of carbohydrates, driving repeated waves of population growth over thousands of years, and the susceptibility of populations to large recessions due to a lag in the impact of humans on resources. These results shed light on the long-term drivers of human population growth and decline.

The diffusion of maize to the southwestern United States and its impact.

Our understanding of the initial period of agriculture in the southwestern United States has been transformed by recent discoveries that establish the presence of maize there by 2100 cal. B.C. (calibrated calendrical years before the Christian era) and document the processes by which it was integrated into local foraging economies. Here we review archaeological, paleoecological, linguistic, and genetic data to evaluate the hypothesis that Proto-Uto-Aztecan (PUA) farmers migrating from a homeland in Mesoamerica introduced maize agriculture to the region. We conclude that this hypothesis is untenable and that the available data indicate instead a Great Basin homeland for the PUA, the breakup of this speech community into northern and southern divisions approximately 6900 cal. B.C. and the dispersal of maize agriculture from Mesoamerica to the US Southwest via group-to-group diffusion across a Southern Uto-Aztecan linguistic continuum.

Should I stay or should I go? The emergence of partitioned land use among human foragers.

Taking inspiration from the archaeology of the Texas Coastal Plain (TCP), we develop an ecological theory of population distribution among mobile hunter-gatherers. This theory proposes that, due to the heterogeneity of resources in space and time, foragers create networks of habitats that they access through residential cycling and shared knowledge. The degree of cycling that individuals exhibit in creating networks of habitats, encoded through social relationships, depends on the relative scarcity of resources and fluctuations in those resources. Using a dynamic model of hunter-gatherer population distribution, we illustrate that increases in population density, coupled with shocks to a biophysical or social system, creates a selective environment that favors habitat partitioning and investments in social mechanisms that control the residential cycling of foragers on a landscape. Our work adds a layer of realism to Ideal Distribution Models by adding a time allocation decision process in a variable environment and illustrates a general variance reduction, safe-operating space tradeoff among mobile human foragers that drives social change.

Guest aggregation within poly(L-lactic acid)/pluronic P104 thin films.

Rhodamine 6G (R6G) doped thin films composed of poly(L-lactic acid) (PLLA) and Pluronic P104 were spin cast onto glass microscope slides and characterized by ultraviolet-visible, steady-state, and time-resolved fluorescence spectroscopy. The results show that R6G aggregation within the film increases as the R6G concentration and P104 loading increases. These results suggest an approach for studying drug distributions (monomers, aggregates) within biodegradable polymer formulations.

Differential orientation and conformation of surface-bound keratinocyte growth factor on (hydroxyethyl)methacrylate, (hydroxyethyl)methacrylate/methyl methacrylate, and (hydroxyethyl)methacrylate/methacrylic acid hydrogel copolymers.

The development of hydrogels for protein delivery requires protein-hydrogel interactions that cause minimal disruption of the protein's biological activity. Biological activity can be influenced by factors such as orientational accessibility for receptor binding and conformational changes, and these factors can be influenced by the hydrogel surface chemistry. (Hydroxyethyl)methacrylate (HEMA) hydrogels are of interest as drug delivery vehicles for keratinocyte growth factor (KGF) which is known to promote re-epithelialization in wound healing. The authors report here the surface characterization of three different HEMA hydrogel copolymers and their effects on the orientation and conformation of surface-bound KGF. In this work, they characterize two copolymers in addition to HEMA alone and report how protein orientation and conformation is affected. The first copolymer incorporates methyl methacrylate (MMA), which is known to promote the adsorption of protein to its surface due to its hydrophobicity. The second copolymer incorporates methacrylic acid (MAA), which is known to promote the diffusion of protein into its surface due to its hydrophilicity. They find that KGF at the surface of the HEMA/MMA copolymer appears to be more orientationally accessible and conformationally active than KGF at the surface of the HEMA/MAA copolymer. They also report that KGF at the surface of the HEMA/MAA copolymer becomes conformationally unfolded, likely due to hydrogen bonding. KGF at the surface of these copolymers can be differentiated by Fourier-transform infrared-attenuated total reflectance spectroscopy and time-of-flight secondary ion mass spectrometry in conjunction with principal component analysis. The differences in KGF orientation and conformation between these copolymers may result in different biological responses in future cell-based experiments.

Tailored delivery of active keratinocyte growth factor from biodegradable polymer formulations.

We report the results of a high throughput screening campaign that is aimed to develop a biodegradable polymer-based formulation to deliver active keratinocyte growth factor (KGF) and provide a means to tune the KGF delivery rate. A statistical design strategy was used to prepare and screen a series of polymer blends that were composed of poly(lactic acid) (PLA), poly(glycolic acid) (PGA), and the surfactant sodium bis(ethylhexyl)sulfosuccinate (Aerosol-OT, AOT). Chloroform was the solvent. Our high throughput screening method used a two-tiered assessment strategy. At Level 1, we identified "lead" KFG-loaded formulations that exhibited KGF emission spectra that were the most similar to the native KGF spectrum recorded in buffer. At Level 2, we used steady-state emission and a homogeneous polarization immunoassay strategy to determine the concentration of total and active KGF, respectively, liberated from the lead formulations during biodegradation. After preparing and screening 2500 formulations, we identified several viable, lead formulations. An analysis of the data showed that the combination of PLA, PGA, and AOT were important to yield a high fraction of active KGF upon release from the formulation; no combination of any two together produced an effect as good as the ternary formulation. The optimum formulations that yielded the highest fraction of active KGF upon release had the following general features: PLA/PGA (w/w) near unity, AOT loading of 100-200 mM, water/AOT mole ratio of 10-20, and a pH between 6 and 8. PLA alone cast from chloroform delivered KGF, but that KGF did not bind to anti-KGF antibodies (i.e., it was inactive). We can tune the KGF release kinetics by more than two orders of magnitude while maintaining the KGF activity upon liberation from the formulation by adjusting the PLA molecular weight.

Interleukin-1 facilitates airway epithelial migration in response to injury.

OBJECTIVE: To test the hypothesis that interleukin (IL)-1 plays a permissive role in respiratory epithelial cell migration and proliferation. STUDY DESIGN: Primary cultures of porcine respiratory epithelial cells or tracheal organ explants were cultured in the presence or absence of function-blocking antibodies to IL-1. Areas of epithelial cell outgrowth were determined in control and antibody-treated organ explants daily for 4 days. At intervals, cultured cells were collected for cell counting and viability determination. Time course and dose-response curves were constructed for control and antibody-treated groups. RESULTS: Interleukin-1 secretion into culture supernatants increased sharply from days 3 to 7. Outgrowths from tracheal explants were reduced by greater than 60% by single antibody treatment, and by over 90% by treatment with antibodies to both IL-1alpha and -1beta by day 4 of culture. Function-blocking antibodies to IL-1 significantly reduced cell number by day 7 of culture. CONCLUSIONS: Interleukin-1 is produced by respiratory epithelial cells in culture during log phase growth and plays a permissive role in cell migration and proliferation.

Possible impedance of luminal reepithelialization by tracheal cartilage metalloproteinases.

BACKGROUND: Rapid reepithelialization of respiratory epithelium after injury to the large conducting airway (eg, trachea and bronchus) is poor. Our laboratory has developed an in vitro model of the trachea that allows us to examine reepithelialization in a complex culture system. We previously described how the presence of cartilage inhibited respiratory epithelial cell (REC) migration/proliferation. In the present study, we examined the effect of cartilage-conditioned medium (CCM) on REC proliferation. We hypothesized that a potential cause of delayed reepithelialization of the large conducting airway after injury could be excessive or aberrant secretion of matrix metalloproteinases (MMPs) by cartilage. DESIGN: We assessed cartilage-derived MMP production and effects on REC proliferation by adding CCM to primary cultures of porcine RECs on type I collagen and determining the cell number and viability. Cartilage-conditioned medium-derived MMP activity was determined by means of gelatin zymography in pooled samples from different times during in vitro cartilage culture. RESULTS: We detected MMP-2 and a small amount of MMP-9 in CCM. Enzyme activity was abolished by EDTA, confirming MMP identity. Cartilage-conditioned medium inhibited REC attachment and proliferation. Addition of the MMP inhibitor GM6001 to cartilage cultures yielded CCM that did not inhibit REC growth, indicating a role for cartilage-derived MMPs in modulating REC proliferation. CONCLUSION: Cartilage production and activity of MMP after injury to the large conducting airway may be a factor in the failure of luminal reepithelialization, resulting in aberrant repair.

Keratinocyte growth factor and autocrine repair in airway epithelium.

BACKGROUND: Delayed or nonreepithelialization of the large conducting airway (ie, trachea and bronchus) is a clinically recognized but poorly understood result of airway trauma. This delay results in granulation tissue formation and scarring, which impairs mucocilliary transport and can critically compromise gas exchange. Keratinocyte growth factor (KGF) is a known epithelial cell mitogen that is derived from mesenchymal cells. We previously observed its expression in injured tracheal explants, and in the present study we investigated its origin. DESIGN: Freshly isolated porcine tracheal epithelial cells were cytospun onto glass slides for immunohistochemical identification and localization of KGF and for in situ hybridization localization of its messenger RNA. Polymerase chain reaction analysis for KGF was also performed. RESULTS: Freshly isolated respiratory epithelial cells were identified as being of epithelial origin and uncontaminated by fibroblasts, as evidenced by stains that were positive for AE3 and negative for vimentin. Immunohistochemical analysis and in situ hybridization revealed a subset of cells that were positive for both the protein and the message for KGF. CONCLUSION: This subset of KGF-expressing respiratory epithelial cells may participate in a hitherto undescribed autocrine loop for stimulating KGF production in response to injury.

Conditional prerequisites for microchannel cytologic analysis on wet mount (fluid-based) biopsies.

BACKGROUND: Advanced capabilities in genomic sequencing developed in the research sector will soon enter the clinical arena. Issues such as the proportioning of patient specimen material for traditional bright-field microscopic evaluation or dedication for molecular analysis will intensify, particularly in situations of small core biopsies. Microfluidics appears aptly suited as a platform capable of allowing traditional cytologic diagnostics and downstream molecular analysis from the same specimen. However, clarification is needed to determine that forces which act on cells in a fluidic environment do not drastically alter their cytologic features. METHODS: Cells were processed for flow-through in a microfluidic channel and evaluated qualitatively and quantitatively for alterations due to fluid-shear stress or anoikis. RESULTS: Processing caused separation of cells from cohesive clusters to smaller groups and individual cells, leading to greater variation in parameters associated with the nucleus in nontumor cells but no significant change in tumor cells. These differences were most readily apparent by quantitative measures, and to a lesser extent, qualitative evaluation. Time-dependent processing played a larger role in cytologic alteration than fluid-shear stress for nontumor cells. CONCLUSIONS: Passage of cells through a microfluidic channel is a feasible approach that can be integrated into future platforms intent on integrating cytologic assessment of cells with recovery of the same cells for downstream assays.

Determining the protein drug release characteristics and cell adhesion to a PLLA or PLGA biodegradable polymer membrane.

Biodegradable polymers are of interest for developing controlled protein drug delivery platforms. In this study, two poly (alpha-hydroxy) esters were formulated with Aerosol-OT, a surfactant stabilizer, to encapsulate the protein keratinocyte growth factor (KGF) for controlled release KGF is involved in a number of crucial biologic processes, most notably epithelial growth and repair. The concentration of KGF that caused a biological response in vitro was determined (optimally 10 ng/mL) and compared with the release of KGF from the two biodegradable polymer membrane formulations. Each polymer formulation released biologically relevant levels, 10 ng/mL of active KGF, although with different times release kinetics. The membrane composed of PLGA/AOT/KGF exhibited a faster release rate of KGF into solution after 120 h of degradation time than the release rate of the PLLA/AOT/KGF matrices. Cell seeding assays showed that both polymer matrices, when formulated with AOT, sustained cell growth. Time of Flight Secondary Ion Mass Spectrometry (ToF-SIMS) was used to characterize the distribution of AOT and KGF through the polymer membrane. (c) 2010 Wiley Periodicals, Inc. J Biomed Mater Res, 2010.

Tubulin glutamylation regulates ciliary motility by altering inner dynein arm activity.

How microtubule-associated motor proteins are regulated is not well understood. A potential mechanism for spatial regulation of motor proteins is provided by posttranslational modifications of tubulin subunits that form patterns on microtubules. Glutamylation is a conserved tubulin modification [1] that is enriched in axonemes. The enzymes responsible for this posttranslational modification, glutamic acid ligases (E-ligases), belong to a family of proteins with a tubulin tyrosine ligase (TTL) homology domain (TTL-like or TTLL proteins) [2]. We show that in cilia of Tetrahymena, TTLL6 E-ligases generate glutamylation mainly on the B-tubule of outer doublet microtubules, the site of force production by ciliary dynein. Deletion of two TTLL6 paralogs caused severe deficiency in ciliary motility associated with abnormal waveform and reduced beat frequency. In isolated axonemes with a normal dynein arm composition, TTLL6 deficiency did not affect the rate of ATP-induced doublet microtubule sliding. Unexpectedly, the same TTLL6 deficiency increased the velocity of microtubule sliding in axonemes that also lack outer dynein arms, in which forces are generated by inner dynein arms. We conclude that tubulin glutamylation on the B-tubule inhibits the net force imposed on sliding doublet microtubules by inner dynein arms.

Phase separation at the surface of poly(ethylene oxide)-containing biodegradable poly(L-lactic acid) blends.

The surface chemistry and in-depth distribution of the composition of a poly(ethylene oxide) (PEO)-containing biodegradable poly(L-lactic acid) (PLLA) blend matrix system have been investigated using X-ray photoelectron spectroscopy (XPS). This study reports detailed quantitative compositional information using a novel numerical method for determining depth profiles. The PEO system studied is an amphiphilic Pluronic P104 surfactant, PEO-b-poly(propylene oxide) (PPO)-b-PEO. The extent of phase separation is analyzed by determining the surface enrichment of the PEO component via measurement of chemical composition at the polymer-air interface. For this blend system, the combination of the PPO component in the Pluronic surfactants drives the formation of a surface excess of Pluronic in the blends with PLLA. The surface excess profile shows a rapid increase in Pluronic surface composition versus bulk Pluronic mass fractions of 1-5%, but the profile levels off above bulk Pluronic mass fractions of 5%.

Targeted gene disruption of dynein heavy chain 7 of Tetrahymena thermophila results in altered ciliary waveform and reduced swim speed.

Tetrahymena thermophila swims by the coordinated beating of hundreds of cilia that cover its body. It has been proposed that the outer arm dyneins of the ciliary axoneme control beat frequency, whereas the inner arm dyneins control waveform. To test the role of one of these inner arms, dynein heavy chain 7 protein (Dyh7p), a knockout mutant was generated by targeted biolistic transformation of the vegetative macronucleus. Disruption of DYH7, the gene which encodes Dyh7p, was confirmed by PCR examination of both genomic and cDNA templates. Both intact and detergent extracted, reactivated cell model preparations of these mutants, which we call DYH7neo3, displayed swim speeds that were almost half that of wild-type cells. Although the DYH7neo3 mutants were slower than wild type, they were able to modulate their swim speed and show ciliary reversal in response to depolarizing stimuli. High-speed video microscopy of intact, free-swimming DYH7neo3 mutants revealed an irregular pattern of ciliary beat and waveform. The mutant cilia appeared to be engaging in less coordinated, swiveling movements in which the typical shape, periodicity and coordination seen in wild-type cilia were absent or disturbed. We propose that the axonemal inner arm dynein heavy chain 7 proteins contribute to the formation of normal ciliary waveform, which in turn governs the forward swimming velocity of these cells.

Fibroblast growth factor receptor signaling affects development and function of dopamine neurons - inhibition results in a schizophrenia-like syndrome in transgenic mice.

Developing and mature midbrain dopamine (DA) neurons express fibroblast growth factor (FGF) receptor-1 (FGFR1). To determine the role of FGFR1 signaling in the development of DA neurons, we generated transgenic mice expressing a dominant negative mutant [FGFR1(TK-)] from the catecholaminergic, neuron-specific tyrosine hydroxylase (TH) gene promoter. In homozygous th(tk-)/th(tk-) mice, significant reductions in the size of TH-immunoreactive neurons were found in the substantia nigra compacta (SNc) and the ventral tegmental area (VTA) at postnatal days 0 and 360. Newborn th(tk-)/th(tk-) mice had a reduced density of DA neurons in both SNc and VTA, and the changes in SNc were maintained into adulthood. The reduced density of DA transporter in the striatum further demonstrated an impaired development of the nigro-striatal DA system. Paradoxically, the th(tk-)/th(tk-) mice had increased levels of DA, homovanilic acid and 3-methoxytyramine in the striatum, indicative of excessive DA transmission. These structural and biochemical changes in DA neurons are similar to those reported in human patients with schizophrenia and, furthermore, these th(tk-)/th(tk-) mice displayed an impaired prepulse inhibition that was reversed by a DA receptor antagonist. Thus, this study establishes a new developmental model for a schizophrenia-like disorder in which the inhibition of FGF signaling leads to alterations in DA neurons and DA-mediated behavior.

Analysis of the initial burst of drug release coupled with polymer surface degradation.

PURPOSE: Local pH effect on the release of a model pH-inert hydrophobic drug coupled with polymer degradation is described at the induction phase of biodegradable polymer erosion for better understanding the nature of initial burst of a drug. METHODS: Using a novel approach with time-of-flight secondary ion mass spectrometry. both surface concentration of Ph3N and degradation kinetics of PLLA are simultaneously and independently determined from a model Ph3N/PLLA (20:80 wt%) blend matrix (t approximately 0.4 microm on 1.0 cm2). In vitro hydrolysis of the model blend matrix is investigated for short-term periods (<24 h) at physiologic pH and temperature and compared to basic pH. RESULTS: The rate of PLLA degradation is accelerated by a factor of approximately 3 when using basic pH in vitro, but the rate of Ph3N accumulation at the surface is accelerated by a factor of approximately 6. CONCLUSIONS: A new quantitative method has been developed to examine the earliest stages of polymer degradation and drug release. It was applied to a model system that could not be examined by traditional in vitro methods. For the model system studied the release of a low molecular weight hydrophobic drug at the induction phase of polymer erosion is related to but not singularly dependent on degradation kinetics.

Disruption of genes encoding predicted inner arm dynein heavy chains causes motility phenotypes in Tetrahymena.

The multi-dynein hypothesis [Asai, 1995: Cell Motil Cytoskeleton 32:129-132] states: (1) there are many different dynein HC isoforms; (2) each isoform is encoded by a different gene; (3) different isoforms have different functions. Many studies provide evidence in support of the first two statements [Piperno et al., 1990: J Cell Biol 110:379-389; Kagami and Kamiya, 1992: J Cell Sci 103:653-664; Gibbons, 1995: Cell Motil Cytoskeleton 32:136-144; Porter et al., 1996: Genetics 144:569-585; Xu et al., 1999: J Eukaryot Microbiol 46:606-611] and there is evidence that outer arms and inner arms play different roles in flagellar beating [Brokaw and Kamiya, 1987: Cell Motil. Cytoskeleton 8:68-75]. However, there are few studies rigorously testing in vivo whether inner arm dyneins, especially the 1-headed inner arm dyneins, play unique roles. This study tested the third tenet of the multi-dynein hypothesis by introducing mutations into three inner arm dynein HC genes (DYH8, 9 and 12) that are thought to encode HCs associated with 1-headed inner arm dyneins. Southern blots, Northern blots, and RT-PCR analyses indicate that all three mutants (KO-8, 9, and 12) are complete knockouts. Each mutant swims slower than the wild-type cells. The beat frequency of KO-8 cells is lower than that of the wild-type cells while the beat frequencies of KO-9 and KO-12 are not different from that of wild-type cells. Our results suggest that each inner arm dynein HC is essential for normal cell motility and cannot be replaced functionally by other dynein HCs and that not all of the 1-headed inner arm dyneins play the same role in ciliary motility. Thus, the results of our study support the multi-dynein hypothesis [Asai, 1995: Cell Motil Cytoskeleton 32:129-132].

Inner arm dynein 1 is essential for Ca++-dependent ciliary reversals in Tetrahymena thermophila.

Cilia in many organisms undergo a phenomenon called ciliary reversal during which the cilia reverse the beat direction, and the cell swims backwards. Ciliary reversal is typically caused by a depolarizing stimulus that ultimately leads to a rise in intraciliary Ca++ levels. It is this increase in intraciliary Ca++ that triggers ciliary reversal. However, the mechanism by which an increase in intraciliary Ca++ causes ciliary reversal is not known. We have previously mutated the DYH6 gene of Tetrahymena thermophila by targeted gene knockout and shown that the knockout mutants (KO6 mutants) are missing inner arm dynein 1 (I1). In this study, we show that KO6 mutants do not swim backward in response to depolarizing stimuli. In addition to being unable to swim backwards, KO6 mutants swim forward at approximately one half the velocity of wild-type cells. However, the ciliary beat frequency in KO6 mutants is indistinguishable from that of wild-type cells, suggesting that the slow forward swimming of KO6 mutants is caused by an altered waveform rather than an altered beat frequency. Live KO6 cells are also able to increase and decrease their swim speeds in response to stimuli, suggesting that some aspects of their swim speed regulation mechanisms are intact. Detergent-permeabilized KO6 mutants fail to undergo Ca++-dependent ciliary reversals and do not show Ca++-dependent changes in swim speed after MgATP reactivation, indicating that the axonemal machinery required for these responses is insensitive to Ca++ in KO6 mutants. We conclude that Tetrahymena inner arm dynein 1 is not only an essential part of the Ca++-dependent ciliary reversal mechanism but it also may contribute to Ca++-dependent changes in swim speed and to the formation of normal waveform during forward swimming.

A comparative study of primary and immortalized cell adhesion characteristics to modified polymer surfaces: toward the goal of effective re-epithelialization.

Facilitating tissue regeneration or replacement requires development of synthetic surfaces that promote cell adhesion, migration, and proliferation. Two successful approaches have been to incorporate minimal cell adhesion recognition sequences at the biomaterial surface and to integrate the entire adhesion molecule into a compatible synthetic matrix. While adhesion assays using immortalized cell lines are important in evaluating synthetic materials, cell type and source play a significant role in the ability of such models to mimic real tissues. Models that utilize multiple cell types or primary cells are more representative of native tissues than models that use single cell types or primary cells. In this study we investigated primary respiratory epithelial cell (REC) adhesion to modified fluoropolymers incorporating simple functional groups and minimal peptide recognition sequences, and we evaluated the potential of hybrid biopolymer materials to support adhesion and proliferation. X-ray photoelectron spectroscopy (XPS) was used to verify substrate surface composition. Significant differences were found in the adhesion characteristics of primary REC and in the A549 lung carcinoma cell line. Model systems composed of multiple cell types and/or primary cells necessarily represent increased levels of complexity for an investigation of cellular responses to synthetic surfaces. When evaluating biomaterials, adhesion studies using immortalized cell lines cannot necessarily be extrapolated to normal cell behavior.