Comparison of real-time PCR assays for detection, quantification, and differentiation of campylobacter jejuni and campylobacter coli in broiler neck skin samples

We tested the use of multiplex real-time PCR for detection and quantification of Campylobacter jejuni and Campylobacter coli on broiler carcass neck skin samples collected during 2008 from slaughterhouses in Switzerland. Results from an established TaqMan assay based on two different targets (hipO and ceuE for C. jejuni and C. coli, respectively) were corroborated with data from a newly developed assay based on a single-nucleotide polymorphism in the fusA gene, which allows differentiation between C. jejuni and C. coli. Both multiplex real-time PCRs were applied simultaneously for direct detection, differentiation, and quantification of Campylobacter from 351 neck skin samples and compared with culture methods. There was good correlation in detection and enumeration between real-time PCR results and quantitative culture, with real-time PCR being more sensitive. Overall, 251 (71.5%) of the samples were PCR positive for Campylobacter, with 211 (60.1%) in the hipO-ceuE assays, 244 (69.5%) in the fusA assay, and 204 (58.1%) of them being positive in both PCR assays. Thus, the fusA assay was similarly sensitive to the enrichment culture (72.4% positive); however, it is faster and allows for quantification. In addition, real-time PCR allowed for species differentiation; roughly 60% of positive samples contained C. jejuni, less than 10% C. coli, and more than 30% contained both species. Real-time PCR proved to be a suitable method for direct detection, quantification, and differentiation of Campylobacter from carcasses, and could permit time-efficient surveillance of these zoonotic agents.

Plexiform hybrid granular cell tumor/perineurioma: a novel variant of benign peripheral nerve sheath tumor with divergent differentiation

The descriptive term hybrid peripheral nerve sheath tumor refers to any neoplasm of the neurilemmal apparatus composed of more than one pathologically defined tumoral equivalent derived from its constituent cells. Within this uncommon nosological category, participation of granular cell tumor - a neoplasm of modified Schwann cells - has been reported only exceptionally. We describe a hitherto not documented variant composed of an organoid mixture of granular cell tumor and perineurioma with plexiform growth. A solitary subcutaneous nodule of 1.5 cm diameter was excised from the right ring finger of a 19-year-old female with no antecedents of neurofibromatosis or relevant trauma. Histology revealed a monotonous, yet cytologically dimorphic proliferation of classical granular cells intermingled with flattened, inconspicuous perineurial cells. Immunohistochemical double labeling detected expression of S100 protein in the former and of EMA and GLUT-1 in the latter. While the respective staining patterns for S100 protein and EMA or GLUT-1 tended to be mutually exclusive, a minority of cells exhibited transitional granular cell/perineurial immunophenotype. Electron microscopy permitted direct visualization of a plethora of lysosomes in the granular cell moiety, and of pinocytotic vesicles and tight junctions in perineurial cells. Intratumoral axons were not detected. Expanding intraneurally, the lesion showed discrete encapsulation by the local perineurium, and resulted in plexiform growth. The MIB-1 labeling index averaged 1%. We interpret our findings as supporting evidence for the dual cell lineage to have arisen through metaplasia, with the tumor's dynamics probably having been driven by the granular cell component.

BIRC6 (APOLLON) is down-regulated in acute myeloid leukemia and its knockdown attenuates neutrophil differentiation

Background Inhibitors of apoptosis (IAPs) were intensively investigated in the context of cancer where they promote tumor growth and chemoresistence. Overexpression of the IAP BIRC6 is associated with unfavorable clinical features and negatively impacts relapse-free survival in childhood acute myeloid leukemia (AML). Currently, BIRC6 levels in adult primary AML have not been compared to the expression in normal myeloid cells. Thus, we compared for the first time BIRC6 levels in adult primary AML patient samples to normal myeloid cells and studied its regulation and function during neutrophil differentiation. Findings We found significantly lower BIRC6 levels in particular AML subtypes as compared to granulocytes from healthy donors. The lowest BIRC6 expression was found in CD34+ progenitor cells. Moreover, BIRC6 expression significantly increased during neutrophil differentiation of AML cell lines and knocking down BIRC6 in NB4 acute promyelocytic leukemia (APL) cells significantly impaired neutrophil differentiation, but not cell viability. Conclusion Together, we found an association of low BIRC6 levels with an immature myeloid phenotype and describe a function for BIRC6 in neutrophil differentiation of APL cells.

Inhibition of damage-regulated autophagy modulator-1 (DRAM-1) impairs neutrophil differentiation of NB4 APL cells

The damage-regulator autophagy modulator 1 (DRAM-1) is a lysosomal protein that positively regulates autophagy in a p53-dependent manner. We aimed at analyzing the role of DRAM-1 in granulocytic differentiation of APL cells. We observed a significant increase of DRAM-1 expression during all-trans retinoic acid (ATRA)-induced neutrophil differentiation of NB4 APL cells but not in ATRA-resistant NB4-R2 cells. Next, knocking down DRAM-1 in NB4 APL cells was sufficient to impair neutrophil differentiation. Given that DRAM-1 is a transcriptional target of p53, we tested if DRAM-1 is regulated by the p53 relative p73. Indeed, inhibiting p73 prevented neutrophil differentiation and DRAM-1 induction of NB4 cells. In conclusion, we show for the first time that p73-regulated DRAM-1 is functionally involved in neutrophil differentiation of APL cells.

The Effects Of Farnesyltransferase Inhibitor Abt-100 On Murine Helper T-Cell Differentiation And Cytokine Secretion

Farnesyltransferase Inhibitors (FTIs) are a class of drugs known to prevent the farnesylation and subsequent membrane attachment of a number of intracellular proteins. In various studies, the administration of FTIs has been found to play a role in the activation and development of T-cells in the immune system. FTIs have also been found to act as immunomodulators in delaying MHC-II mismatched skin allografts in mice. This study focuses on the effect of the FTI, ABT-100, on the differentiation and cytokine secretion of Th1 and Th2 helper T-cells in BALB/C mice to better understand which immune responses are targeted by FTIs. Splenocytes were isolated from BALB/C mice, skewed towards either a Th1 or a Th2 phenotype with the addition of cytokines, and treated with various concentrations of ABT-100. Splenocytes were also isolated and immediately cultured in the presence of ABT-100 to observe differentiation trends of helper T-cells. Cytokine production was measured using intracytoplasmic flow cytometry analysis. I found that ABT-100 treatment does not block Th1 or Th2 cell differentiation. Instead, ABT-100 treatment appears to affect cytokine production from effector T-cells. I found that ABT-100 causes a decrease in IFN-¿ production in mature Th1 cells yet does not affect IL-4 production in mature Th2 cells. This decrease in cytokine production as a result of ABT-100 treatments provides a potential mechanism for how ABT-100 works to delay MHC-II mismatched allograft rejection.

Assessing Commonality and Differentiation for Packaging-Family Planning with Application to Medication Labels

This thesis presents a metric for assessing the commonality and differentiation of packaging-family planning with application to medical labels along with supporting background research and findings. Consumable products such as medications rely on the package or label to represent the contents. Package confusion has been widely recognized as a major problem for both over-the-counter and pharmacy-dispensed medications with potentially lethal consequences. It is critical to identify a medication as a member of a product family and differentiate its contributing elements based on visual features on the package or label to avoid consumer confusion and reduce dispensing errors. Indices that indicate degrees of commonality and differentiation of features in consumer products such as batteries, light bulbs, handles, etc for platforms have been shown to benefit development of engineered product families [6]. It is possible to take a similar approach for visual features in packaging such as typography, shape/form, imagery and color to benefit packaging-family development. This thesis establishes a commonality differentiation index for prominence of visual features on over-the-counter and pharmacy-dispensed medications based on occurrence, size, and location of features. It provides a quantitative measure to assist package designers in evaluating alternatives to satisfy strategic goals and improve safety. The index is demonstrated with several medications that have been identified by the Institute for Safe Medication Practice as commonly confused.

Of marsupials and men: "Backdoor" dihydrotestosterone synthesis in male sexual differentiation

Following development of the fetal bipotential gonad into a testis, male genital differentiation requires testicular androgens. Fetal Leydig cells produce testosterone that is converted to dihydrotestosterone in genital skin, resulting in labio-scrotal fusion. An alternative 'backdoor' pathway of dihydrotestosterone synthesis that bypasses testosterone has been described in marsupials, but its relevance to human biology has been uncertain. The classic and backdoor pathways share many enzymes, but a 3α-reductase, AKR1C2, is unique to the backdoor pathway. Human AKR1C2 mutations cause disordered sexual differentiation, lending weight to the idea that both pathways are required for normal human male genital development. These observations indicate that fetal dihydrotestosterone acts both as a hormone and as a paracrine factor, substantially revising the classic paradigm for fetal male sexual development.

Evolutionary forces shaping genomic islands of population differentiation in humans

Background Levels of differentiation among populations depend both on demographic and selective factors: genetic drift and local adaptation increase population differentiation, which is eroded by gene flow and balancing selection. We describe here the genomic distribution and the properties of genomic regions with unusually high and low levels of population differentiation in humans to assess the influence of selective and neutral processes on human genetic structure. Methods Individual SNPs of the Human Genome Diversity Panel (HGDP) showing significantly high or low levels of population differentiation were detected under a hierarchical-island model (HIM). A Hidden Markov Model allowed us to detect genomic regions or islands of high or low population differentiation. Results Under the HIM, only 1.5% of all SNPs are significant at the 1% level, but their genomic spatial distribution is significantly non-random. We find evidence that local adaptation shaped high-differentiation islands, as they are enriched for non-synonymous SNPs and overlap with previously identified candidate regions for positive selection. Moreover there is a negative relationship between the size of islands and recombination rate, which is stronger for islands overlapping with genes. Gene ontology analysis supports the role of diet as a major selective pressure in those highly differentiated islands. Low-differentiation islands are also enriched for non-synonymous SNPs, and contain an overly high proportion of genes belonging to the 'Oncogenesis' biological process. Conclusions Even though selection seems to be acting in shaping islands of high population differentiation, neutral demographic processes might have promoted the appearance of some genomic islands since i) as much as 20% of islands are in non-genic regions ii) these non-genic islands are on average two times shorter than genic islands, suggesting a more rapid erosion by recombination, and iii) most loci are strongly differentiated between Africans and non-Africans, a result consistent with known human demographic history.

Stem Cells From Umbilical Cord Wharton's Jelly From Preterm Birth Have Neuroglial Differentiation Potential

Objective:The aim of the study is to determine the neuroglial differentiation potential of human Wharton's jelly-derived mesenchymal stem cells (WJ-MSCs) from preterm birth when compared to term delivery.Study Design:The WJ-MSCs from umbilical cords of preterm birth and term controls were isolated and induced into neural progenitors. The cells were analyzed for neuroglial markers by flow cytometry, real-time polymerase chain reaction, and immunocytochemistry. Results:Independent of gestational age, a subset of WJ-MSC displayed the neural progenitor cell markers Nestin and Musashi-1 and the mature neural markers microtubule-associated protein 2, glial fibrillary acidic protein, and myelin basic protein. Neuroglial induction of WJ-MSCs from term and preterm birth resulted in the enhanced transcription of Nestin and Musashi-1.Conclusions:Undifferentiated WJ-MSCs from preterm birth express neuroglial markers and can be successfully induced into neural progenitors similar to term controls. Their potential use as cellular graft in neuroregenerative therapy for peripartum brain injury in preterm birth has to be tested.

Differentiation in Male and Female Speech Styles

From the moment of their birth, a person's life is determined by their sex. Ms. Goroshko wants to know why this difference is so striking, why society is so concerned to sustain it, and how it is able to persist even when certain national or behavioural stereotypes are erased between people. She is convinced of the existence of not only social, but biological differences between men and women, and set herself the task, in a manuscript totalling 126 pages, written in Ukrainian and including extensive illustrations, of analysing these distinctions as they are manifested in language. She points out that, even before 1900, certain stylistic differences between the ways that men and women speak had been noted. Since then it has become possible, for instance in the case of Japanese, to point to examples of male and female sub-languages. In general, one can single out the following characteristics. Males tend to write with less fluency, to refer to events in a verb-phrase, to be time-oriented, to involve themselves more in their references to events, to locate events in their personal sphere of activity, and to refer less to others. Therefore, concludes Ms Goroshko, the male is shown to be more active, more ego-involved in what he does, and less concerned about others. Women, in contrast, were more fluent, referred to events in a noun-phrase, were less time-oriented, tended to be less involved in their event-references, locate events within their interactive community and refer more to others. They spent much more time discussing personal and domestic subjects, relationship problems, family, health and reproductive matters, weight, food and clothing, men, and other women. As regards discourse strategies, Ms Goroshko notes the following. Men more often begin a conversation, they make more utterances, these utterances are longer, they make more assertions, speak less carefully, generally determine the topic of conversation, speak more impersonally, use more vulgar expressions, and use fewer diminutives and more imperatives. Women's speech strategies, apart from being the opposite of those enumerated above, also contain more euphemisms, polite forms, apologies, laughter and crying. All of the above leads Ms. Goroshko to conclude that the differences between male and female speech forms are more striking than the similarities. Furthermore she is convinced that the biological divergence between the sexes is what generates the verbal divergence, and that social factors can only intensify or diminish the differentiation in verbal behaviour established by the sex of a person. Bearing all this in mind, Ms Goroshko set out to construct a grammar of male and female styles of speaking within Russian. One of her most important research tools was a certain type of free association test. She took a list comprising twelve stimuli (to love, to have, to speak, to fuck, a man, a woman, a child, the sky, a prayer, green, beautiful) and gave it to a group of participants specially selected, according to a preliminary psychological testing, for the high levels of masculinity or femininity they displayed. Preliminary responses revealed that the female reactions were more diverse than the male ones, there were more sentences and word combinations in the female reactions, men gave more negative responses to the stimulus and sometimes didn't want to react at all, women reacted more to adjectives and men to nouns, and that, surprisingly, women coloured more negatively their reactions to the words man, to love and a child (Ms. Goroshko is inclined to attribute this to the present economic situation in Russia). Another test performed by Ms. Goroshko was the so-called "defective text" developed by A.A. Brudny. All participants were distributed with packets of complete sentences, which had been taken from a text and then mixed at random. The task was to reconstruct the original text. There were three types of test, the first descriptive, the second narrative, and the third logical. Ms. Goroshko created computer programmes to analyse the results. She found that none of the reconstructed texts was coincident with the original, differing both from the original text and amongst themselves and that there were many more disparities in the male than the female texts. In the descriptive and logical texts the differences manifested themselves more clearly in the male texts, and in the narrative texts in the female texts. The widest dispersal of values was observed at the outset, while the female text ending was practically coincident with the original (in contrast to the male ending). The greatest differences in text reconstruction for both males and females were registered in the middle of the texts. Women, Ms. Goroshko claims, were more sensitive to the semantic structure of the texts, since they assembled the narrative text much more accurately than the other two, while the men assembled more accurately the logical text. Texts written by women were assembled more accurately by women and texts by men by men. On the basis of computer analysis, Ms. Goroshko found that female speech was substantially more emotional. It was expressed by various means, hyperbole, metaphor, comparisons, epithets, ways of enumeration, and with the aid of interjections, rhetorical questions, exclamations. The level of literacy was higher for female speech, and there were fewer mistakes in grammar and spelling in female texts. The last stage of Ms Goroshko's research concerned the social stereotypes of beliefs about men and women in Russian society today. A large number of respondents were asked questions such as "What merits must a woman possess?", "What are male vices and virtues?", etc. After statistical manipulation, an image of modern man and woman, as it exists in the minds of modern Russian men and women, emerged. Ms. Goroshko believes that her findings are significant not only within the field of linguistics. She has already successfully worked on anonymous texts and been able to decide on the sex of the author and consequently believes that in the future her research may even be of benefit to forensic science.