The apoptotic and proliferation rate of tumour budding cells in colorectal cancer outlines a heterogeneous population of cells with various impacts on clinical outcome

AIMS In colorectal cancer (CRC), tumour buds represent an aggressive cell type at the invasive front with apparently low proliferation. The aim of this study was to determine proliferation and apoptotic rates of buds in comparison to tumour centre, front and mucosa. METHODS AND RESULTS Whole tissue sections from 188 CRC patients underwent immunohistochemistry for Ki67. Ten high-power fields (HPFs) were evaluated in mucosa, tumour centre, tumour front and tumour buds (total = 40 HPFs/case). Caspase-3 and M30 immunohistochemistry were performed on a multipunch tissue microarray from the same cohort. Ki67, caspase-3 and M30 immunoreactivity were correlated with outcome. The average percentage of cells showing Ki67 positivity was 5.2% in mucosa, and was not significantly different between the centre and front of the tumour (38.2% and 34.9%; P < 0.0001); 0.3% of buds showed Ki67 positivity (P < 0.0001). Caspase-3 expression was similar in mucosa, tumour centre and tumour front, but lower in tumour buds (<0.1%; P < 0.0001). M30 staining in buds was decreased (0.01%; P < 0.0001) in comparison to other areas. Ki67 positivity in buds was detrimental to survival in univariate (P = 0.0352) and multivariate (P = 0.0355) analysis. Caspase-3-positive tumours showed better outcome than negative tumours (P = 0.0262); but tumours with caspase-3-positive buds showed a worse outcome than those with caspase-3-negative buds (P = 0.0235). CONCLUSIONS Ki67, caspase-3 and M30 staining is absent in most tumour buds, suggesting decreased proliferation and apoptosis. However, the fact that Ki67 and caspase-3 immunoreactivity was associated with unfavourable prognosis points to a heterogeneous population of tumour buds.

Eomesodermin, a target gene of Pou4f2, is required for retinal ganglion cell and optic nerve development in the mouse.

The mechanisms regulating retinal ganglion cell (RGC) development are crucial for retinogenesis and for the establishment of normal vision. However, these mechanisms are only vaguely understood. RGCs are the first neuronal lineage to segregate from pluripotent progenitors in the developing retina. As output neurons, RGCs display developmental features very distinct from those of the other retinal cell types. To better understand RGC development, we have previously constructed a gene regulatory network featuring a hierarchical cascade of transcription factors that ultimately controls the expression of downstream effector genes. This has revealed the existence of a Pou domain transcription factor, Pou4f2, that occupies a key node in the RGC gene regulatory network and that is essential for RGC differentiation. However, little is known about the genes that connect upstream regulatory genes, such as Pou4f2 with downstream effector genes responsible for RGC differentiation. The purpose of this study was to characterize the retinal function of eomesodermin (Eomes), a T-box transcription factor with previously unsuspected roles in retinogenesis. We show that Eomes is expressed in developing RGCs and is a mediator of Pou4f2 function. Pou4f2 directly regulates Eomes expression through a cis-regulatory element within a conserved retinal enhancer. Deleting Eomes in the developing retina causes defects reminiscent of those in Pou4f2(-/-) retinas. Moreover, myelin ensheathment in the optic nerves of Eomes(-/-) embryos is severely impaired, suggesting that Eomes regulates this process. We conclude that Eomes is a crucial regulator positioned immediately downstream of Pou4f2 and is required for RGC differentiation and optic nerve development.

A pure population of lung alveolar epithelial type II cells derived from human embryonic stem cells.

Alveolar epithelial type II (ATII) cells are small, cuboidal cells that constitute approximately 60% of the pulmonary alveolar epithelium. These cells are crucial for repair of the injured alveolus by differentiating into alveolar epithelial type I cells. ATII cells derived from human ES (hES) cells are a promising source of cells that could be used therapeutically to treat distal lung diseases. We have developed a reliable transfection and culture procedure, which facilitates, via genetic selection, the differentiation of hES cells into an essentially pure (>99%) population of ATII cells (hES-ATII). Purity, as well as biological features and morphological characteristics of normal ATII cells, was demonstrated for the hES-ATII cells, including lamellar body formation, expression of surfactant proteins A, B, and C, alpha-1-antitrypsin, and the cystic fibrosis transmembrane conductance receptor, as well as the synthesis and secretion of complement proteins C3 and C5. Collectively, these data document the successful generation of a pure population of ATII cells derived from hES cells, providing a practical source of ATII cells to explore in disease models their potential in the regeneration and repair of the injured alveolus and in the therapeutic treatment of genetic diseases affecting the lung.

The impact of cancer on the population of Salvador-Bahia, Brazil

The impact of cancer on the population of Salvador-Bahia, Brazil was studied using mortality data available from the Brazilian National Bureau of Vital Statistics. Average annual site, age, and gender specific and adjusted cancer mortality rates were determined for the years 1977-83 and contrasted with United States cancer mortality rates for the year of 1977. The accuracy of the cancer mortality rates generated by this research was determined by comparing the underlying causes of death as coded on death certificates to pathology reports and to hospital diagnosis of a sample of 966 deaths occurring in Salvador during the year of 1983. To further explore the information available on the death certificate, a population based decedent case control study was used to determine the relationship between type of occupation (proxy for exposure) and mortality by cancer sites known to be occupationally related.^ The rates in Salvador for cancer of the stomach, oral cavity, and biliary passages are, on average, two fold higher than the U.S. rates.^ The death certificate was found to be accurate for 65 percent of the 485 cancer deaths studied. Thirty five histologically confirmed cancer deaths were found in a random sample of 481 deaths from other causes. This means that, approximately 700 more deaths may be lost among the remainder 10,073 death certificates stating a cause other than cancer.^ In addition, despite the known limitations of decedent case-control studies, cancers of the oral cavity OR = 2.44, CI = 1.17-5.09, stomach OR = 2.31, CI = 1.18-4.52, liver OR = 4.06, CI = 1.27-12.99, bladder OR = 6.77, CI = 1.5-30.67, and lymphoma OR = 2.55, CI = 1.04-6.25 had elevated point estimates, for different age strata indicating an association between these cancers and occupations that led to exposure to petroleum and its derivates. ^

Behavioral and psychosocial factors associated with sexual risk in a population of crack cocaine users

The purpose of this dissertation was to examine the relationship between key psychosocial and behavioral components of the Transtheoretical Model and the Theory of Reasoned Action for sexual risk reduction in a population of crack cocaine smokers and sex workers, not in drug treatment. ^ The first study examined the results of an analysis of the association between two principal constructs in the Transtheoretical Model, the processes of change and the stages of change for condom use, in a high risk population. In the analysis of variance for all respondents, the overall F-test revealed that people in different stages have different levels of experiential process use, F(3,317) = 17.79, p = 0.0001 and different levels of behavioral process use, F(3,317) = 28.59, p = .0001. For the experiential processes, there was a significant difference between the precontemplation/contemplation stage, and both the action, and maintenance, stages.^ The second study explored the relationship between the Theory of Reasoned Action “beliefs” and the stages-of-change in the same population. In the analysis of variance for all participants, the results indicate that people in different stages did value the positive beliefs differently, F(3,502) = 15.38, p = .0001 but did not value the negative beliefs differently, F(3,502) = 2.08, p = .10. ^ The third study explored differences in stage-of-change by gender, partner type drug use, and HIV status. Three discriminant functions emerged, with a combined χ2(12) = 139.57, p = <.0001. The loading matrix of correlations between predictors and discriminant functions demonstrate that the strongest predictor for distinguishing between the precontemplation/contemplation stage and the preparation, action, and maintenance stages (first function) is partner type (.962). The loadings on the second discriminant function suggest that once partner type has been accounted for, ever having HIV/AIDS (.935) was the best predictor for distinguishing between the first three stages and the maintenance stage. ^ These studies demonstrate that behavioral change theories can contribute important insight to researchers and program planners attempting to alter HIV risk behavior in high-risk populations. ^

A new sulfonamide resistance gene (sul3) in Escherichia coli is widespread in the pig population of Switzerland

A new gene, sul3, which specifies a 263-amino-acid protein similar to a dihydropteroate synthase encoded by the 54-kb conjugative plasmid pVP440 from Escherichia coli was characterized. Expression of the cloned sul3 gene conferred resistance to sulfamethoxazole on E. coli. Two copies of the insertion element IS15Delta/26 flanked the region containing sul3. The sul3 gene was detected in one-third of the sulfonamide-resistant pathogenic E. coli isolates from pigs in Switzerland.

Male Dominance in the New Zealand Longfin Eel Population of a New Zealand River: Probable Causes and Implications for Management

The endemic New Zealand longfin eel Anguilla dieffenbachi (hereafter, longfin eel), is overfished, and in southern South Island, New Zealand, rivers have recently become predominated by males. This study examined length and age at sexual differentiation in male eels in the Aparima River catchment (area, 1,375 km(2); mean flow, 20 m(3.)s(-1)) and the sex ratio and distribution of eels throughout the catchment. Longfin eels differentiated into males mostly at lengths from 300 to 460 mm and ages from 10 to 25+ years. Females were rare: Of 738 eels examined for sexual differentiation, 466 were males and 5 were females, and a few others, not examined, were large enough to be female. These counts suggest a male : female ratio among differentiated longfin eels of 68:1. Of 31 differentiated shortfin eels A. australis, less common in the Aparima River, 26 were females. Male longfin eels were distributed throughout the main stern and tributaries; undifferentiated eels were more prevalent in lower and middle reaches and in the main stem than in upper reaches and tributaries. In other studies, male longfin eels predominated commercial catches in the Aparima and four other southernmost rivers, by 2.4:1 to 13.6:1 males to females. The Aparima River had the most skewed sex ratio. Longfin eel catches from the Aparima River will become more male predominated because few sublegal-size females were present. The length-frequency distributions of eels in the present samples and in the commercial catches were truncated just above minimum legal size (about 460 mm), showing that few females escape the fishery. Historically, females predominated these rivers. The recent change in sex ratio is attributable partly to selective harvest of females, and partly to changes in the structure of the population from fishing, such that differentiation into males has been favored. Longevity, delayed sexual maturity, semel-parity, and endemism with restricted range make the longfin eel particularly vulnerable to overfishing.

Scent-marking behaviour and social dynamics in a wild population of Eurasian lynx Lynx lynx.

Scent-marking is widespread among mammals and has been observed in many felid species. Although the behaviour is well-described, little is known about its function in wild felid populations. We investigated patterns of scent-marking and its role in intra- and intersexual communication among resident and non-resident Eurasian lynx Lynx lynx by observing interactions among wild lynx at natural marking sites by means of infrared camera traps. Marking activity of resident animals showed a peak during the mating season and was lowest during the time when females gave birth and lactated. Both sexes scent-marked, but male lynx visited marking sites much more often than females and marked relatively more often when visiting a site. Most visits to marking sites were by residents but we also observed scent-marking by non-residents. Juveniles were never observed marking. We found no evidence of lynx regularly renewing scent-marks after a certain 'expiry date' but the presence of a strange scent-mark triggered over-marking. Males responded similarly to the presence of another individual's scent-mark, irrespective of whether it was the top- or the underlying scent-mark in a mixture of scent-marks they encountered. Our results suggest that marking sites could serve as 'chemical bulletin boards', where male lynx advertise their presence and gain information on ownership relationships in a given area. Females placed their urine marks on top of the ones left by resident males, but further studies are needed to explain the functions of over-marking in females.

Warm Breze from the Starboard Bow: a new Population of neutral Helium in the Heliosphere

We investigate the signals from neutral helium atoms observed in situ from Earth orbit in 2010 by the Interstellar Boundary Explorer (IBEX). The full helium signal observed during the 2010 observation season can be explained as a superposition of pristine neutral interstellar He gas and an additional population of neutral helium that we call the Warm Breeze. The Warm Breeze is approximately 2 times slower and 2.5 times warmer than the primary interstellar He population, and its density in front of the heliosphere is ~7% that of the neutral interstellar helium. The inflow direction of the Warm Breeze differs by ~19° from the inflow direction of interstellar gas. The Warm Breeze seems to be a long-term, perhaps permanent feature of the heliospheric environment. It has not been detected earlier because it is strongly ionized inside the heliosphere. This effect brings it below the threshold of detection via pickup ion and heliospheric backscatter glow observations, as well as by the direct sampling of GAS/Ulysses. We discuss possible sources for the Warm Breeze, including (1) the secondary population of interstellar helium, created via charge exchange and perhaps elastic scattering of neutral interstellar He atoms on interstellar He+ ions in the outer heliosheath, or (2) a gust of interstellar He originating from a hypothetic wave train in the Local Interstellar Cloud. A secondary population is expected from models, but the characteristics of the Warm Breeze do not fully conform to modeling results. If, nevertheless, this is the explanation, IBEX-Lo observations of the Warm Breeze provide key insights into the physical state of plasma in the outer heliosheath. If the second hypothesis is true, the source is likely to be located within a few thousand AU from the Sun, which is the propagation range of possible gusts of interstellar neutral helium with the Warm Breeze characteristics against dissipation via elastic scattering in the Local Cloud. Whatever the nature of the Warm Breeze, its discovery exposes a critical new feature of our heliospheric environment.

Common target genes of palatal and gingival fibroblasts for EMD: the microarray approach.

BACKGROUND AND OBJECTIVE Connective tissue grafts are frequently applied, together with Emdogain(®) , for root coverage. However, it is unknown whether fibroblasts from the gingiva and from the palate respond similarly to Emdogain. The aim of this study was therefore to evaluate the effect of Emdogain(®) on fibroblasts from palatal and gingival connective tissue using a genome-wide microarray approach. MATERIAL AND METHODS Human palatal and gingival fibroblasts were exposed to Emdogain(®) and RNA was subjected to microarray analysis followed by gene ontology screening with Database for Annotation, Visualization and Integrated Discovery functional annotation clustering, Kyoto Encyclopedia of Genes and Genomes pathway analysis and the Search Tool for the Retrieval of Interacting Genes/Proteins functional protein association network. Microarray results were confirmed by quantitative RT-PCR analysis. RESULTS The transcription levels of 106 genes were up-/down-regulated by at least five-fold in both gingival and palatal fibroblasts upon exposure to Emdogain(®) . Gene ontology screening assigned the respective genes into 118 biological processes, six cellular components, eight molecular functions and five pathways. Among the striking patterns observed were the changing expression of ligands targeting the transforming growth factor-beta and gp130 receptor family as well as the transition of mesenchymal epithelial cells. Moreover, Emdogain(®) caused changes in expression of receptors for chemokines, lipids and hormones, and for transcription factors such as SMAD3, peroxisome proliferator-activated receptor gamma and those of the ETS family. CONCLUSION The present data suggest that Emdogain(®) causes substantial alterations in gene expression, with similar patterns observed in palatal and gingival fibroblasts.

Dynamics of a Tularemia Outbreak in a Closely Monitored Free-Roaming Population of Wild House Mice.

Infectious disease outbreaks can be devastating because of their sudden occurrence, as well as the complexity of monitoring and controlling them. Outbreaks in wildlife are even more challenging to observe and describe, especially when small animals or secretive species are involved. Modeling such infectious disease events is relevant to investigating their dynamics and is critical for decision makers to accomplish outbreak management. Tularemia, caused by the bacterium Francisella tularensis, is a potentially lethal zoonosis. Of the few animal outbreaks that have been reported in the literature, only those affecting zoo animals have been closely monitored. Here, we report the first estimation of the basic reproduction number R0 of an outbreak in wildlife caused by F. tularensis using quantitative modeling based on a susceptible-infected-recovered framework. We applied that model to data collected during an extensive investigation of an outbreak of tularemia caused by F. tularensis subsp. holarctica (also designated as type B) in a closely monitored, free-roaming house mouse (Mus musculus domesticus) population in Switzerland. Based on our model and assumptions, the best estimated basic reproduction number R0 of the current outbreak is 1.33. Our results suggest that tularemia can cause severe outbreaks in small rodents. We also concluded that the outbreak self-exhausted in approximately three months without administrating antibiotics.

Intendedness of pregnancy among active duty women in the U.S. Army

Objective. The objective of this study was to determine the prevalence of unintended pregnancy and the association between social and demographic factors among a population of active duty women in the U.S. Army giving birth to viable infants at a U.S. Army hospital at Fort Hood, Texas. Prevalence of unintended pregnancy in this group was 50.9% (95% CI 44.0 to 57.9) with 36.3% being mistimed (95% CI 29.8 to 33.2) and 14.6% being unwanted (95% CI 10.2 to 20.1). A further 14.2% of the women experienced ambivalence (95% CI 9.8 to 19.6). ^ The study population was a cross-sectional group of active duty pregnant women who represent the target population of all female soldiers that deliver viable infants in the Army. Using a survey based on previous studies, intendedness of pregnancy at conception was retrospectively determined. Unintended births are further characterized as mistimed or unwanted. Demographic and other exposures were described bivariately. Associations were evaluated using measures of relative risk and chi-square analysis. ^ The results of the research indicate that in the study population, race/ethnicity is not associated with unintended pregnancy and non-commissioned officers had a lower rate of unintended pregnancy than other rank groupings. ^

Association between temperament and physical activity in a population of minority women

Physical activity has been, and remains, a significant public health issue. Thus, increasing physical activity has been identified as a top priority according to Healthy People 2010. Various behavioral variables have been associated with participation in physical activity, including the Type A behavior pattern (TABP). This study was a secondary data analysis of the Women On The Move pilot study data and examined the relationship between Type A behavior with physical activity. The study population consisted of fifty-six (56) adult minority women 40 years of age and above. The Thurstone Activity Scale was adapted for use in this study to measure TABP. Physical activity behavior was measured using an accelerometer (Computer Science Application, [CSA]) and a physical activity diary. All study questions were examined using multiple linear regression analysis. In all analyses age, household income, and level of education were entered as covariates. The results found no association with TABP and exercise or physical activity. More research involving a larger, more active study population is recommended in order to more precisely determine the relationship of TABP and physical activity. ^

Self-injury in a residential treatment population of children and adolescents

Objectives. The aim of this study was to describe the relationship between child-abuse and self-injury among children and adolescents living in a residential treatment center in a large urban area. ^ Methods. A retrospective study was conducted through a chart review of the residents who were placed at the center from 2003-2006. A total of 35 cases (with at least one documented incident of self-injury during placement at the residential treatment center) were age/gender matched with 35 controls (without at least one documented incident of self-injury during placement at the residential treatment center). ^ Results. In this study, the case subjects were far more likely to be victims of sexual abuse than were the controls (74.3% vs. 25.7%, respectively). Self-harm was found to be 9.5 times as frequent in the group that was exposed to sexual abuse in the source population (OR = 9.500 with a 95% CI = 2.292, 84.111). The difference was statistically significant (McNemar's test, x2 = 12.190 with 1 df. The two-tailed P value equals 0.0005). ^ Conclusion. These findings suggest that school-age and early-adolescent children who have a history of sexual abuse may engage in a variety of self-harming behaviors. Clinicians should consider a history of sexual abuse when working with self-harming children. ^