The stem cell organisation, and the proliferative and gene expression profile of Barrett's epithelium, replicates pyloric-type gastric glands

Objective: Barrett's oesophagus shows appearances described as ‘intestinal metaplasia’, in structures called ‘crypts’ but do not typically display crypt architecture. Here, we investigate their relationship to gastric glands. Methods: Cell proliferation and migration within Barrett's glands was assessed by Ki67 and iododeoxyuridine (IdU) labelling. Expression of mucin core proteins (MUC), trefoil family factor (TFF) peptides and LGR5 mRNA was determined by immunohistochemistry or by in situ hybridisation, and clonality was elucidated using mitochondrial DNA (mtDNA) mutations combined with mucin histochemistry. Results: Proliferation predominantly occurs in the middle of Barrett's glands, diminishing towards the surface and the base: IdU dynamics demonstrate bidirectional migration, similar to gastric glands. Distribution of MUC5AC, TFF1, MUC6 and TFF2 in Barrett's mirrors pyloric glands and is preserved in Barrett's dysplasia. MUC2-positive goblet cells are localised above the neck in Barrett's glands, and TFF3 is concentrated in the same region. LGR5 mRNA is detected in the middle of Barrett's glands suggesting a stem cell niche in this locale, similar to that in the gastric pylorus, and distinct from gastric intestinal metaplasia. Gastric and intestinal cell lineages within Barrett's glands are clonal, indicating derivation from a single stem cell. Conclusions: Barrett's shows the proliferative and stem cell architecture, and pattern of gene expression of pyloric gastric glands, maintained by stem cells showing gastric and intestinal differentiation: neutral drift may suggest that intestinal differentiation advances with time, a concept critical for the understanding of the origin and development of Barrett's oesophagus

Expression of Mcm2, geminin and Ki67 in normal oral mucosa, oral epithelial dysplasias and their corresponding squamous-cell carcinomas

Proteins necessary for the normal regulation of the cell cycle include minichromosome maintenance protein 2 (Mcm2) and geminin. These are overexpressed in several premalignant and malignant tumours. The Mcm2/Ki67 ratio can be used to estimate the population of cells that are in early G1 (licensed to proliferate), and the geminin/Ki67 ratio can determine the relative length of G1. A high ratio indicates a short G1 and a high rate of cell proliferation. Mcm2 and geminin have been scarcely explored in oral epithelial dysplasia (OED) and oral squamous-cell carcinoma (OSCC). The purpose of this study was to identify the expression pattern of Mcm2, Ki67 and geminin in normal oral mucosa (NOM), OED and their subsequent OSCC, to determine if expression could help predict the prognosis of OED. Paraffin sections of 41 OED cases that progressed to carcinoma, 40 OED without malignant progression, 38 OSCC and 15 NOM were immunostained with antibodies against Mcm2, geminin and Ki67. Labelling indices (LIs) increased progressively from NOM, OED and OSCC (Mcm2, Po0.001; geminin, Po0.001 and Ki67, Po0.001). In all the OED cases (n ¼ 81) the levels of expression of Mcm2 (LI, 73.6), geminin (LI, 24.4) and Ki67 (LI, 44.5) were elevated indicating a constant cellcycle re-entry. When the OED groups were compared, Mcm2 protein expression was higher in the OED with malignant progression (P ¼ 0.04), likewise there was a significant increase in the Mcm2/Ki67 and geminin/Ki67 ratios (P ¼ 0.04 and 0.02 respectively). Mcm2 and geminin proteins seem to be novel biomarkers of growth and may be useful prognostic tools for OED

Extensive telomere erosion is consistent with localised clonal expansions in Barrett’s metaplasia

Barrett’s oesophagus is a premalignant metaplastic condition that predisposes patients to the development of oesophageal adenocarcinoma. However, only a minor fraction of Barrett’s oesophagus patients progress to adenocarcinoma and it is thus essential to determine bio-molecular markers that can predict the progression of this condition. Telomere dysfunction is considered to drive clonal evolution in several tumour types and telomere length analysis provides clinically relevant prognostic and predictive information. The aim of this work was to use high-resolution telomere analysis to examine telomere dynamics in Barrett’s oesophagus. Telomere length analysis of XpYp, 17p, 11q and 9p, chromosome arms that contain key cancer related genes that are known to be subjected to copy number changes in Barrett’s metaplasia, revealed similar profiles at each chromosome end, indicating that no one specific telomere is likely to suffer preferential telomere erosion. Analysis of patient matched tissues (233 samples from 32 patients) sampled from normal squamous oesophagus, Z-line, and 2 cm intervals within Barrett’s metaplasia, plus oesophago-gastric junction, gastric body and antrum, revealed extensive telomere erosion in Barrett’s metaplasia to within the length ranges at which telomere fusion is detected in other tumour types. Telomere erosion was not uniform, with distinct zones displaying more extensive erosion and more homogenous telomere length profiles. These data are consistent with an extensive proliferative history of cells within Barrett’s metaplasia and are indicative of localised clonal growth. The extent of telomere erosion highlights the potential of telomere dysfunction to drive genome instability and clonal evolution in Barrett’s metaplasia

Multi Path FTIR Agriculture Air Pollution Measurement System

This paper details the design and validation of a Multiple Path OP-FTIR system with elevation and radial scanning ability and demonstrates its capabilities to quantify and monitor gaseous ammonia emitted from agricultural facilities. The OP-FTIR system has a 500 m range (1000 m full path length) and allows 360° radial scan and 45° scan in elevation. To study large scale sources, two or more similar systems may be needed. For comparison purposes, we ran two similar but not identical OP-FTIR systems side-by-side in a controlled lab environment and in a series of field environments. We determined that in a controlled environment, the two systems can attain an NH3 agreement of 1- 3% at concentrations above 500 ppb. Due to the short path length (~10 m) in the lab, 500 ppb was the detection limit of the two systems. Path lengths in a field are much longer, allowing a lower detection limit. Average agreement in the field was 1-6%. This difference in agreement from the laboratory is likely due to the non-homogeneous distribution of the pollutant

Ductal-lobar organisation of human breast tissue, its relevance in disease and a research objective: vector mapping of parenchyma in complete breasts (the Astley Cooper project)

A human breast has many lobes, which are highly variable in size and shape, each with one central duct, its peripheral branches and their associated glandular tissues. Realising the potential of new endoductal approaches to breast diagnosis and improving our understanding of breast cancer precursors will require greatly improved knowledge of this ductal-lobar anatomy and the distribution of cancer precursors within it. This architecture is very challenging to study in its entirety: whole-breast lobe mapping has only been achieved for two human breasts. Clearly, much more efficient techniques are required. Streamlined data capture and visualisation of breast parenchymal anatomy from thin and thick sections in a vector format would allow integrated mapping of whole-breast structure with conventional histology and molecular data. The 'Astley Cooper digital breast mapping project' is proposed as a name for this achievable research objective. Success would offer new insights into the development of breast cancer precursor lesions, allow testing of the important 'sick lobe' hypothesis, improve correlation with imaging studies and provide 'ground truth' for mathematical modelling of breast growth

Loss of heterozygosity on chromosomes 11 and 17 are markers of recurrence in TCC of the bladder

Approximately 2/3 of patients diagnosed with superficial transitional cell carcinoma of the urinary bladder (TCC) will recur within 2 years. Loss of chromosome 9 and loss of heterozygosity (LOH) at 9q34 in index TCCs identify a subset of patients at high risk of recurrence. This study explores genetic alterations on chromosomes 4, 8, 11 and 17 as predictors of recurrence. A total of 109 carcinomas were investigated at 26 loci. DNA was extracted from microdissected archival normal/tumour tissue and was analysed for loss of heterozygosity (LOH). Fluorescent PCR was performed and genotyping carried out on a Perkin Elmer ABI377 sequencer. LOH of D11S490 or D17S928 was significantly more frequent in index carcinomas of patients who experienced recurrence compared to those with no recurrence (P = 0.004 and 0.019 respectively). These results suggest that loss of these regions is associated with recurrence of TCC. Further investigation is required to identify genes in these regions, which might be responsible for driving recurrence in TCC of the urinary bladder. © 2001 Cancer Research Campaign http://www.bjcancer.co

Is chromosome 9 loss a marker of disease recurrence in transitional cell carcinoma of the urinary bladder?

Investigation of transitional cell carcinoma of the urinary bladder (TCC) patients classified by recurrence and/or progression has demonstrated that loss of chromosome 9, as detected by FISH analysis of the pericentromeric classical satellite marker at 9q12, occurs early. A total of 105 TCCs from 53 patients were analysed in situ by two independent observers for loss of chromosome 9 using quantitative fluorescence in situ hybridization (FISH). All 53 primary tumours were evaluated for chromosomes 9, 7 and 17. Normal ranges for chromosomal copy number were defined for normal skin epidermis and bladder epithelium. Values for chromosome 9 copy number outwith the range 1.51-2.10 (mean +/- 3 x s.d. of normal values) were significantly abnormal. Twenty-five TCCs were detected with consistent monosomic scores. Of 89 TCCs, in which multiple tumour areas were analysed, 85 tumours (96%) demonstrated the same chromosome 9 copy number in all areas (2-6) analysed; only three tumours demonstrated heterogeneity for this locus. A total of 36% (12 out of 33) of patients with subsequent disease recurrence demonstrated loss of chromosome 9 in their primary and all subsequent TCCs analysed. Only a single patient (n = 20) with non-recurrent TCC showed loss of chromosome 9 (P = 0.0085). Of 53 primary tumours, eight showed significant elevation of chromosome 17. Of these patients, six demonstrated elevation in chromosome 7 copy number. No abnormalities were observed in non-recurrent patients. This study describes rapid quantitation of chromosomal copy number by FISH using a pericentromeric probe for chromosome 9 in TCC of the urinary bladder. Routinely fixed and processed material was evaluated without disaggregation. Strict quality control of FISH demonstrated that this technique was reproducible in a clinical environment and could be used to detect genetic changes relevant to patient outcome. It is proposed that loss of chromosome 9 from primary TCC of the urinary bladder identified patients at high risk of recurrence and possible progression

Igniter-induced hybrids in the 20-l sphere

Dust explosibility is traditionally described by two parameters, namely the maximum explosion pressure, P$_{max}$, and the deflagration index, K$_{St}$, usually determined through testing in a closed, pressure-resistant spherical vessel, either 20 L or 1 m$^{3}$ in volume. These parameters constitute key variables in the design of explosion protection systems, such as venting, suppression or isolation systems. The potential for overdriving dust combustion with pyrotechnical igniters in the 20-l sphere has been recognized, discussed and analyzed for many years, notably in the determination of the minimum explosible and limiting oxygen concentrations, which has led to specific guidelines regarding the ignition source strength in ASTM standards. The current paper presents new experimental evidence that the energy provided by pyrotechnical igniters may, in some instances, physically alter the dust being tested in the 20-l sphere. K$_{St}$ values can be several times greater in the small vessel compared to those measured in the 1-m$^{3}$ chamber. Further visual evidence is provided to show that high energy ignition can produce a turbulent flame region, possibly consisting of a hybrid mixture of flammable gas (or vapor) and dust, which can propagate faster than the corresponding pure dust. The experiments suggest that K$_{St}$ values measured in the 20-l sphere may no longer be representative of a dust deflagration in a real process environment. We recommend additional tests in a 1-m$^{3}$ chamber when a dust exhibits a low flash point, or when it's K$_{St}$ is above 300 bar m/s in the 20-l sphere.The authors gratefully acknowledge the support of Fike Corporation for their permission to publish this work

Correlates of Physical Activity in Black, Hispanic, and White Middle School Girls

Background: A need exists to better understand multilevel influences on physical activity among diverse samples of girls. This study examined correlates of moderate-to-vigorous physical activity (MVPA) among adolescent girls from different racial/ethnic backgrounds. Methods: 1,180 6th grade girls (24.5% black, 15.7% Hispanic, and 59.8% white) completed a supervised self-administered questionnaire that measured hypothesized correlates of PA. MVPA data were collected for 6 days using the ActiGraph accelerometer. Hierarchical regression analysis was used to examine correlates of PA in each racial/ethnic group. Results: Hispanic girls (n=185) engaged in 21.7 minutes of MVPA per day, black girls (n=289) engaged in 19.5 minutes of MVPA per day, and white girls (n=706) engaged in 22.8 minutes of MVPA per day. Perceived transportation barriers (+; P=.010) were significantly and positively related to MVPA for Hispanic girls. For black girls, Body Mass Index (BMI) (-; P\u3c.005) and social support from friends (+; P=.006) were significant correlates of MVPA. For white girls, BMI (-; P\u3c.001), barriers (-; P=.012), social support from friends (+; P=.010), participation in school sports (+; P=.009), and community sports (+; P=.025) were significant correlates of MVPA. Explained variance ranged from 30% to 35%. Conclusions: Correlates of MVPA varied by racial/ethnic groups. Effective interventions in ethnically diverse populations may require culturally tailored strategies