On-Site Pilot-Scale Microalgae Cultivation Using Industrial Wastewater for Bioenergy Production: A Case Study towards Circular Bioeconomy.

This case study assesses the valorization of industrial wastewater streams for bioenergy generation in an industrial munition facility. On-site pilot-scale demonstrations were performed to investigate the feasibility of algal growth in the target wastewater on a larger outdoor scale. An exploratory field study followed by an optimized one were carried out using two 1000 L open raceway ponds deployed within a greenhouse at an industrial munition facility. An online system allowed for constant monitoring of operational parameters such as temperature, pH, light intensity, and dissolved oxygen within the ponds. The original algal seed evolved into an open-air resilient consortium of green microalgae and cyanobacteria that were identified and characterized successfully. Weekly measurements of the level of nutrients in pond liquors were performed along with the determination of the algal biomass to quantitatively evaluate growth yields. After harvesting algae from the ponds, the biomass was concentrated and evaluated for oil content and biochemical methane potential (BMP) to provide an estimate of the algae-based energy production. Additionally, the correlation among biomass, culturing conditions, oil content, and BMP was evaluated. The higher average areal biomass productivity achieved during the summer months was 23.9 ± 0.9 g/m2d, with a BMP of 350 scc/gVS. An oil content of 22 wt.% was observed during operation under low nitrogen loads. Furthermore, a technoeconomic analysis and life cycle assessment demonstrated the viability of the proposed wastewater valorization scenario and aided in optimizing process performance towards further scale-up.

Tuberculosis specific responses following therapy for TB: Impact of HIV co-infection.

Characterizing perturbations in the immune response to tuberculosis in HIV can develop insights into the pathogenesis of coinfection. HIV+ TB+ and TB monoinfected (TB+) subjects recruited from clinics in Bamako prior to initiation of TB treatment were evaluated at time-points following initiation of therapy. Flow cytometry assessed CD4+/CD8+ T cell subsets and activation markers CD38/HLA-DR. Antigen specific responses to TB proteins were assessed by intracellular cytokine detection and proliferation. HIV+ TB+ subjects had significantly higher markers of immune activation in the CD4+ and CD8+ T cells compared to TB+ subjects. HIV+ TB+ had lower numbers of TB-specific CD4+ T cells at baseline. Plasma IFNγ levels were similar between HIV+ TB+ and TB+ subjects. No differences were observed in in-vitro proliferative capacity to TB antigens between HIV+ TB+ and TB+ subjects. Subjects with HIV+ TB+ coinfection demonstrate in vivo expansion of TB-specific CD4+ T cells. Immunodeficiency associated with CD4+ T cell depletion may be less significant compared to immunosuppression associated with HIV viremia or untreated TB infection.

Facile functional analysis of insect odorant receptors expressed in the fruit fly: validation with receptors from taxonomically distant and closely related species.

With the advent of genomic sequences and next-generation sequencing technologies (RNA-Seq), multiple repertoires of olfactory proteins in various insect species are being unraveled. However, functional analyses are lagging behind due in part to the lack of simple and reliable methods for heterologous expression of odorant receptors (ORs). While the Xenopus oocyte recording system fulfills some of this lacuna, this system is devoid of other olfactory proteins, thus testing only the "naked" ORs. Recently, a moth OR was expressed in the majority of neurons in the antennae of the fruit fly using Orco-GAL4 to drive expression of the moth OR. Electroantennogram (EAG) was used to de-orphanize the moth OR, but generic application of this approach was brought to question. Here, we describe that this system works with ORs not only from taxonomically distant insect species (moth), but also closely related species (mosquito), even when the fruit fly has highly sensitive innate ORs for the odorant being tested. We demonstrate that Orco-GAL4 flies expressing the silkworm pheromone receptor, BmorOR1, showed significantly higher responses to the sex pheromone bombykol than the control lines used to drive expression. Additionally, we show that flies expressing an OR from the Southern house mosquito, CquiOR2, gave significantly stronger responses to the cognate odorants indole and 2-methylphenol than the "background noise" recorder from control lines. In summary, we validate the use of Orco-GAL4 driven UAS-OR lines along with EAG analysis as a simple alternative for de-orphanization and functional studies of insect ORs in an intact olfactory system.

Nonindigenous Plant Advantage in Native and Exotic Australian Grasses under Experimental Drought, Warming, and Atmospheric CO2 Enrichment.

A general prediction of ecological theory is that climate change will favor invasive nonindigenous plant species (NIPS) over native species. However, the relative fitness advantage enjoyed by NIPS is often affected by resource limitation and potentially by extreme climatic events such as drought. Genetic constraints may also limit the ability of NIPS to adapt to changing climatic conditions. In this study, we investigated evidence for potential NIPS advantage under climate change in two sympatric perennial stipoid grasses from southeast Australia, the NIPS Nassella neesiana and the native Austrostipa bigeniculata. We compared the growth and reproduction of both species under current and year 2050 drought, temperature and CO2 regimes in a multifactor outdoor climate simulation experiment, hypothesizing that NIPS advantage would be higher under more favorable growing conditions. We also compared the quantitative variation and heritability of growth traits in populations of both species collected along a 200 km climatic transect. In contrast to our hypothesis we found that the NIPS N. neesiana was less responsive than A. bigeniculata to winter warming but maintained higher reproductive output during spring drought. However, overall tussock expansion was far more rapid in N. neesiana, and so it maintained an overall fitness advantage over A. bigeniculata in all climate regimes. N. neesiana also exhibited similar or lower quantitative variation and growth trait heritability than A. bigeniculata within populations but greater variability among populations, probably reflecting a complex past introduction history. We found some evidence that additional spring warmth increases the impact of drought on reproduction but not that elevated atmospheric CO2 ameliorates drought severity. Overall, we conclude that NIPS advantage under climate change may be limited by a lack of responsiveness to key climatic drivers, reduced genetic variability in range-edge populations, and complex drought-CO2 interactions.

Molecular strain typing of Mycobacterium tuberculosis complex in Bamako, Mali.

OBJECTIVE: To identify strains of Mycobacterium tuberculosis complex (MTC) circulating in Bamako and to examine the relationship between the strains and their drug susceptibility profiles. METHODS: Between 2006 and 2010, we conducted a cross-sectional study using spoligotyping to identify strains of MTC recovered from 126 tuberculosis (TB) patients under treatment in Bamako, Mali. RESULT: Three members of the MTC were isolated: M. tuberculosis (71.4%), M. africanum (27.8%) and M. bovis (0.8%). Of these, three strains were found to be the most prevalent: M. tuberculosis T1 (MTB T1; 38.9%), M. africanum F2 (MAF2; 26.2%) and M. tuberculosis Latin American and Mediterranean 10 (MTB LAM 10; 10.3%). MAF2 and MTB LAM 10 strains have a lower risk of multidrug resistance (MDR) than MTB T1 (respectively OR 0.1, 95%CI 0.03-0.4 and OR 0.1, 95%CI 0.01-0.8). Age ≥ 32 years (OR 1.4, 95%CI 0.4-3.9), negative human immunodeficiency virus status (OR 0.4, 95%CI 0.1-2.5) and male sex (OR 4, 95%CI 0.9-16.5) were not associated with MDR. The prevalence of MDR among treatment and retreatment failure patients was respectively 25% and 81.8% compared to new patients (2.9%). CONCLUSION: This study indicates a low level of primary drug resistance in Bamako, affirms the importance of using correct drug regimens, and suggests that the MTB T1 strain may be associated with the development of resistance.

Caracterização epidemiológica dos casos de esquistossomose no Município de Feira de Santana, Bahia 2003-2006 / Epidemiological characteristics of cases of schistosomiasis in the city Of Feira de Santana, Bahia 2003-2006 / Características epidemiológicas de los casos de esquistosomiasis en el Municipio de Feira de Santana, Bahia 2003-2006

A esquistossomose é uma doença infecciosa que, a despeito de apresentar uma redução nos indicadores de morbimortalidade, ainda é considerada um problema de saúde pública no nordeste do Brasil. Estudos relatam maior distribuição espacial desta doença no país, relacionada à intensificação do processo de urbanização em muitos municípios brasileiros. O objetivo deste estudo foi caracterizar os casos de esquistossomose no município de Feira de Santana, Bahia, auxiliando na adoção de políticas de saúde pautadas em ações de prevenção e controle dessa enfermidade. Foi realizado o levantamento e a análise estatística de dados relativos à morbidade por esquistossomose provenientes das fichas de notificação da Vigilância Epidemiológica da Secretaria de Saúde municipal. Pôde-se verificar que Feira de Santana é uma área de baixa endemicidade para esquistossomose, pois apresentou prevalências inferiores a 1 porcento e que os indivíduos mais acometidos pela infecção são do sexo masculino, com idade entre 10 e 29 anos, da raça/cor da pele parda e preta, de baixa escolaridade. Concluiu-se que o entendimento da dinâmica da esquistossomose contribui para estimular a utilização de condutas preventivas e o melhor planejamento das ações de saúde em um município.

Schistosomiasis is an infectious disease that, despite presenting a reduction in morbidity and mortality indicators, is still considered a public health problem in northeastern Brazil. Studies report a higher spatial distribution of this disease in Brazil, related to the intensification of the urbanization process in many municipalities. The aim of this study was to characterize thecases of schistosomiasis in Feira de Santana, Bahia, assisting in the adoption of health policies framed by prevention and control of this disease. The survey and statistical analysis of data on morbidity from schistosomiasis from reporting the Epidemiological Monitoring of Municipal Health Secretariat. It was verified that Feira de Santana is a low endemic area for schistosomiasis, as it showed prevalence below 1 percent and that the individuals most affected by the infection are male, aged between 10 and 29 years, race / color mixed and black skin, the less educated. It was concluded that understanding the dynamics of schistosomiasis helps to stimulate the use of preventive measures and better planning of health activities in the municipality of Feira de Santana.

La esquistosomiasis es una enfermedad infecciosa que, a pesar de presentar uma reducción en los indicadores de morbilidad y mortalidad, todavía se considera un problema de salud pública en el noreste de Brasil. Los estudios reportan una distribución espacial más alta de esta enfermedad en Brasil, relacionado con la intensificación del proceso de urbanización em muchos municipios. El objetivo de este estudio fue caracterizar los casos de esquistosomiasis em Feira de Santana, Bahia, ayudando en la adopción de políticas de salud enmarcada en la prevención y el control de esta enfermedad. La encuesta y el análisis estadístico de los datos sobre la morbilidad por esquistosomiasis de informar de la Vigilancia Epidemiológica de la Secretaría Municipal de Salud. Se verificó que de Feira de Santana es un área endémica de baja de la esquistosomiasis, ya que revelan una prevalencia inferior al 1por ciento y que las personas más afectadas por la infección son varones, con edades comprendidas entre 10 y 29 años, raza / color mixtos y negro de la piel, los menos educados. Se concluye que la comprensión de la dinámica de la esquistosomiasis ayuda a estimular el uso de medidas preventivas y una mejor planificación de las actividades de salud enel municipio de Feira de Santana.

Application of spatio-temporal scan statistics for the detection of areas with increased risk for American visceral leishmaniasis in the state of Bahia, Brazil.

Mirroring the global increase of registered cases of American visceral leishmaniasis (AVL), this infection has become a growing public health problem in Brazil during the last several years. As the traditional approach to control employed by the governmental health agencies has failed to reduce the incidence and epidemic outbreaks of this illness, we propose a re-evaluation of the national strategy of intervention and monitoring. Our thinking is based on a series of spatio-temporal scan statistics of the west-central region of the state of Bahia covering the 11-year period from 1994 to 2004. By analyzing the situation, spatially and temporally, we show that the disease is a not only a growing focal threat but that it is also appearing in the form of endemic clusters in the cities. The areas where the disease has been found have been classified according to the degree of risk of infection for humans and canines. The overall objective of this study was to identify areas of increased risk of AVL, including its seasonality, and to suggest ways and means to improve the detection of the disease. The findings presented here should not only be of interest for the efforts to control AVL in the study area but also be useful for developing control strategies in other endemic regions of Brazil.

A new spontaneous mutation in the mouse protocadherin 15 gene.

We have characterized a new allele of the protocadherin 15 gene (designatedPcdh15(av-6J)) that arose as a spontaneous, recessive mutation in the C57BL/6J inbred strain at Jackson Laboratory. Analysis revealed an inframe deletion in Pcdh15, which is predicted to result in partial deletion of cadherin domain (domain 9) in Pcdh15. Morphologic study revealed normal to moderately defective cochlear hair cell stereocilia in Pcdh15(av-6J) mutants at postnatal day 2 (P2). Stereocilia abnormalities were consistently present at P5 and P10. Degenerative changes including loss of inner and outer hair cells were seen at P20, with severe sensory cell loss in all cochlear turns occurring by P40. The hair cell phenotype observed in the 6J allele between P0 and P20 is the least severe phenotype yet observed in Pcdh15 alleles. However, young Pcdh15(av-6J) mice are unresponsive to auditory stimulation and show circling behavior indicative of vestibular dysfunction. Since these animals show severe functional deficits but have relatively mild stereocilia defects at a young age they may provide an appropriate model to test for a direct role of Pcdh15 in mechanotransduction.

Strong spatial genetic structure in peripheral but not core populations of Sitka spruce [Picea sitchensis (Bong.) Carr.].

We examined spatial genetic structure within eight populations of Sitka spruce classified as core or peripheral based on ecological niche, and continuous or disjunct based on species distribution. In each population, 200 trees were spatially mapped and genotyped for eight cDNA-based sequence tagged site (STS) codominant markers. Spatial autocorrelation was assessed by estimating p(ij), the average co-ancestry coefficient, between individuals within distance intervals. The distribution of alleles and genotypes within core populations was almost random, with nonsignificant co-ancestry values among trees as close as 50 m in core populations. In contrast, the distribution of alleles and genotypes within peripheral populations revealed an aggregation of similar multilocus genotypes, with co-ancestry values greater than 0.20 among trees up to 50 m apart and significant, positive values between trees up to 500 m. The relatively high density of reproductive adults in core populations may lead to highly overlapping seed shadows that limit development of spatial genetic structure. However, in peripheral populations with a lower density of adults, the distribution of alleles and genotypes was highly structured, likely due to offspring establishment near maternal trees and subsequent biparental inbreeding, as well as more recent population establishment at the leading edge of post-Pleistocene range expansion. Conserving genetic diversity in peripheral populations may require larger reserves for in situ conservation than required in core populations. These data on spatial genetic structure can be used to provide guidance for sampling strategies for both ex situ conservation and research collections.

Optimal testing parameters for blood cultures.

The effects of volume of blood, number of consecutive cultures, and incubation time on pathogen recovery were evaluated for 37,568 blood cultures tested with the automated BACTEC 9240 instrument (Becton Dickinson Diagnostic Instrument Systems) at a tertiary care center over the period of 12 June 1996 through 12 October 1997. When the results for this study were compared with previous data published for manual broth-based blood culture systems and patient samples obtained in the 1970s and 1980s, the following were found: (1) the percentage increase in pathogen recovery per milliliter of blood is less, (2) more consecutive blood culture sets over a 24-h period are required to detect bloodstream pathogens, and (3) a shorter duration of incubation is required to diagnose bloodstream infections. Guidelines developed in the 1970s and 1980s for processing and culturing blood may require revision.

Indices for bioavailability and biotransformation potential of contaminants in soils.

Bioavailability is an important consideration in risk assessment of soil contaminants and in the selection of appropriate remediation technologies for polluted sites. The present study examined the bioavailability and biodegradation potential of phenanthrene with respect to a pseudomonad in 15 different soils through separate measurements of mineralization, transformation, and desorption to a polymeric infinite sink (Tenax) after 180-d sterile pre-equilibration with phenanthrene. Fractions strongly resistant to desorption and mineralization at long times were evident in all cases. After correcting for bioconversion (moles mineralized per mole transformed) determined in aqueous particle-free soil extracts, a correlation was found between the biotransformation-resistant fraction and the Tenax desorption-resistant fraction. Indices are proposed to assess bioavailability (BAt) and biotransformation potential (BTPt) of a compound in a soil based on parallel desorption and degradation studies over a selected period t. The BAt is the ratio of moles biotransformed to moles desorbed to an infinite sink, and it reflects the biotransformation rate relative to the maximal desorption rate. Values of BA30 (30-d values) ranged from 0.64 (for dark gray silt loam) to 1.12 (Wurtsmith Air Force Base [AFB] 2B, Oscoda, MI, USA). The BTPt is the ratio between moles biotransformed and moles of contaminant remaining sorbed after maximal desorption. The BTPt provides an indication of the maximum extent of biotransformation that may be expected in a system, assuming desorption is a prerequisite for biodegradation. Values of BTP30 ranged between 0.3 (Wurtsmith AFB 1B) and 13 (Mount Pleasant silt loam, NY, USA). The combination of BAt and BTPt provides insights regarding the relationship between physical availability (desorption) and biological processes (biotransformation kinetics, toxicity, other soil factors) that occur during biodegradation and are suggested to represent the remediation potential of the chemical. The BA30 values less than 0.9 and BTP30 values less than five indicate poor potential for site remediation.

Effects of calcium and magnesium pretreatment on hyperkalaemic cardiac arrest in rats.

BACKGROUND AND OBJECTIVE: Administration of calcium safely and effectively reverses many of the electrophysiological actions of hyperkalaemia, but it has not been studied for pretreatment. Based on cellular studies, magnesium also has been suggested to prevent the effects of potassium on the heart. As their mechanisms of action differ, a combination of these drugs might have a synergistic protective action. Both compounds are inexpensive and can be administered safely in modest doses. We investigated whether magnesium, calcium or their combination could protect against hyperkalaemic cardiac arrest. METHODS: Twenty-four adult rats were anaesthetized with halothane and randomly pretreated with CaCl2 15 mg kg(-1), MgSO4 30 mg kg(-1), CaCl2 7.5 mg kg(-1) + MgSO4 15 mg kg(-1) or physiological saline. Potassium (0.01 mmol kg(-1) h(-1)) was infused. The times to the first dysrhythmia, mean arterial pressure decrease to <40% of baseline and cardiovascular collapse were measured. RESULTS: Serum potassium concentrations increased to similar values in all groups (to 12.0 +/- 0.2 mmol L(-1) at the time of collapse). No differences in survival times were observed between groups. There was a trend for respiratory values to be better in the group receiving magnesium. CONCLUSIONS: Pretreatment with magnesium, calcium or a combination of both did not influence the time to cardiovascular collapse, and is therefore--at least in our model--not of any benefit in preventing hyperkalaemic cardiac arrest.

Sorption hysteresis of benzene in charcoal particles.

Charcoal is found in water, soil, and sediment where it may act as a sorbent of organic pollutants. The sorption of organic compounds to natural solids often shows hysteresis. The purpose of this study was to determine the source of pronounced hysteresis that we found in the sorption of a hydrophobic compound (benzene) in water to a maple-wood charcoal prepared by oxygen-limited pyrolysis at 673 K. Gas adsorption (N2, Ar, CO2), 13C NMR, and FTIR show the charcoal to be a microporous solid composed primarily of elemental (aromatic) C and secondarily of carboxyl and phenolic C. Nonlocal density functional theory (N2, Ar) and Monte Carlo (CO2) calculations reveal a porosity of 0.15 cm3/g, specific surface area of 400 m2/g, and appreciable porosity in ultramicropores < 10 A. Benzene sorption-desorption conditions were chosen to eliminate artificial causes of hysteresis (rate-limiting diffusion, degradation, colloids effect). Charcoal sorbed up to its own weight of benzene at approximately 69% of benzene water solubility. Sorption was highly irreversible over most of the range tested (10(-4)-10(3) microg/mL). A dimensionless irreversibility index (/i) (0 < or = /i < or = 1) based on local slopes of adsorption and desorption branches was evaluated at numerous places along the isotherm. /i decreases as C increases, from 0.9-1 at low concentration to approximately 0 (approximately fully reversible) at the highest concentrations. Using sedimentation and volumetric displacement measurements, benzene is observed to cause pronounced swelling (up to > 2-fold) of the charcoal particles. It is proposed that hysteresis is due to pore deformation by the solute, which results in the pathway of sorption being different than the pathway of desorption and which leads to entrapment of some adsorbate as the polyaromatic scaffold collapses during desorption. It is suggested that intra-charcoal mass transport may be influenced by structural rearrangement of the solid, in addition to molecular diffusion.

Concentration-dependent kinetics of pollutant desorption from soils.

Sorption-desorption kinetics play a major role in transport and bioavailability of pollutants in soils. Contaminant concentration is a potentially important factor controlling kinetics. A previous paper dealt with the effect of solute concentration on fractional uptake rates of phenanthrene and pyrene from a finite aqueous source. In this study we determined the effect of initial phenanthrene sorbed concentration (q(0)) on the fractional mass desorption rates from each of six soils to a zero-concentration solution, approximated by including a polymer adsorbent (Tenax) as a third-phase sink. The soils were preequilibrated with phenanthrene for 180 d. Consistent with theory, the fractional desorption rates determined by empirical curve fitting increased with q(0) provided the isotherm was nonlinear. After 500 to 600 d of desorption at the steepest possible concentration gradient, all soils retained a highly resistant fraction, which ranged from 4 to 31% of q(0), except for one soil at a high q(0). The highly resistant fraction decreased with increasing q(0), for nonlinear isotherm cases, but increased with q(0) for linear or nearly linear isotherm cases. Application of a nonlinear diffusion model, the dual-mode diffusion model (DMDM), to the nonresistant fraction gave reasonably good fits. The DMDM attributes the increase with concentration of the apparent diffusivity to a decrease in the proportion of sorbate occupying immobile sites (holes) in soil organic matter. The concentration-dependent term in the expression for the apparent diffusivity correlated with either of two indices that reflect the linearity of the sorption isotherm. Bunker C oil present in one soil acted as a partition domain. The findings of this study are consistent with heterogeneous models of soil organic matter, and indicate that concentration effects should be taken into account whenever desorption rate is important.

Laparoscopic mesh and staple Burch colposuspension.

Our objective was to describe our experience with laparoscopic Burch colposuspension and to relate our results to traditional open Burch procedures for the treatment of genuine stress incontinence. Retrospective case series were compared to historical controls. Forty-six women found to have only genuine stress incontinence by history, examination and clinical urodynamics, underwent a mesh and staple laparoscopic Burch procedure. Follow-up ranged from 3 to 50 months. Thirty-seven women were dry, 6 were improved and 3 showed no improvement. This compared to between 75% and 90% of women cured of stress incontinence by the traditional open Burch procedure. We concluded that the results of a mesh and staple laparoscopic Burch procedure in a carefully selected population of women with genuine stress incontinence appears comparable to that reported in the literature for an open Burch procedure.

Evaluating degradation rates of chlorinated organics in groundwater using analytical models.

The persistence and fate of organic contaminants in the environment largely depends on their rate of degradation. Most studies of degradation rate are performed in the lab where chemical conditions can be controlled precisely. Unfortunately, literature values for lab degradation studies often are orders of magnitude higher than for field-generated studies, calling into question the relevance of lab-generated values for characterizing the persistence of organic contaminants in the environment. Complicating analysis of this ostensible disparity between lab and field degradation values, field-generated values often do not account for effects of adsorption. Modeling with a newly derived analytical solution for first-order degradation coupled with advective losses and adsorption to solve for degradation constants is insensitive to uncertainties in field properties. Application to field data shows that accounting for advection and adsorption greatly affects the value of calculated degradation constants compared to disappearance constants, which do not account for these phenomena. In fact, degradation constants, calculated using these analytical solutions and field data, are in the range reported for lab-generated data. Using these analytical solutions, for the sulfate-reducing field conditions documented for this site, perchloroethene, trichloroethene, 1,1,1-trichloroethane, 1,1-dichloroethane, and chloroethane all degraded with half-lives ranging from 5 to 115 d. Consistent with other studies of sulfate-reducing conditions, cis-1,2-dichloroethene did not chemically degrade at a measurable rate. When nonaqueous phase 1,1-dichloroethane is present, down-gradient concentrations vary in an annual sinusoidal pattern, apparently because of seasonal variation in dilution from groundwater recharge.

Antimicrobial susceptibility of Abiotrophia adiacens and Abiotrophia defectiva.

The susceptibilities of 27 Abiotrophia adiacens (proposed reclassification Granulicatella adiacens comb.nov., Collins & Lawson, 2000) and 12 Abiotrophia defectiva isolates were tested by microdilution in pyridoxal hydrochloride and lysed horse blood supplemented Mueller-Hinton broth. According to NCCLS interpretative criteria for Streptococcus spp. not Streptococcus pneumoniae, the susceptibilities of A. adiacens and A. defectiva were, respectively: penicillin, 55% and 8%; amoxicillin, 81% and 92%; ceftriaxone, 63% and 83%; meropenem, 96% and 100%; and 100% for both species with clindamycin, rifampin, levofloxacin, ofloxacin, quinupristin/dalfopristin, and vancomycin.

Progression of head and neck cancer in an in vitro model.

OBJECTIVE: To identify alterations in angiogenesis and cell cycle regulation as preneoplastic cells progress to cancer in an in vitro model of head and neck tumor progression. METHODS: Immortal human gingival keratinocyte (IHGK) cells (preneoplastic) were derived from normal oral keratinocytes and were immortalized with human papillomavirus 16. Transformation of IHGK cells with a carcinogen (NNK, 4-[methylnitrosamino]-1-[3-pyridyl]-1-butanone) gave rise to IHGKN cells. We determined the growth rates, cell cycle phase, expression of cell cycle regulators, and expression of vascular endothelial growth factor along with the organotypic features of these cells and compared them with characteristics of head and neck cancer cells. RESULTS: IHGK and IHGKN cells grown in raft culture were morphologically similar to severe dysplasia and carcinoma, respectively. The proportion of cells in G(0)/G(1) was similar between IHGK and IHGKN. However, the proportion of IHGK cells was 35% greater in S phase as compared with the IHGKN cells, while a greater percentage (40%) of IHGKN cells were in G(2)/M. The expression of the other cell cycle regulators tested was unchanged. IHGK cells secreted less vascular endothelial growth factor on day 1 when compared with IHGKN (50.6 vs 245.6 pg/mL), along with a lower overall production rate (79% vs 133%). CONCLUSIONS: Transformation of IHGK cells resulted in the activation of vascular endothelial growth factor associated with angiogenesis. Inactivation of the G(1) cell cycle regulation occurred during immortalization and before transformation, and was sustained after carcinogen exposure. These alterations correspond to changes observed in patients with head and neck squamous cell carcinoma. This model can be useful in testing novel therapeutic and preventive strategies.