Deoxyribonucleic Acid Measurements in Transitional Cell Carcinomas: Comparison of Flow and Image Cytometry Techniques

Purpose

We compared the results of deoxyribonucleic acid (DNA) ploidy determinations performed by flow cytometry and image cytometry in transitional cell carcinomas of the bladder.

Materials and Methods

In 81 cases of transitional cell carcinoma of the bladder DNA indexes were measured by flow cytometry of bladder washings and tumor tissue samples, and by image cytometry of imprints from tumor tissue samples.

Results

There was good correlation between bladder washings and tissue samples analyzed by flow cytometry in 68 cases but aneuploidy was missed with bladder washings in 13 (16 percent). There was also good correlation between flow and image cytometry in 75 cases (92.59 percent) regarding the detection of aneuploidy. There was agreement between detection of aneuploidy and DNA index in 45 cases (55.5 percent), while both methods detected an aneuploid population in 30 (37 percent) but there was disagreement regarding DNA index. Aneuploid populations were missed by flow cytometry in 6 cases (7.4 percent). Furthermore, in 10 cases peridiploid peaks were found on the image cytometry histograms, which were not visible on flow cytometry. However, it was not possible to assess accurately if these were true peridiploid populations.

Conclusions

There is good overall correlation between DNA content measured by flow and image cytometry but image cytometry has the advantage of visual discrimination, permitting preferential selection and analysis of tumor cells. However, certain problems remain with image cytometry, particularly in the case of peridiploid peaks, which cannot be classified accurately as showing true peridiploid or right shifted diploid populations.     

Therapeutic Potential of Secreted Molecules Derived from Human Amniotic Fluid Mesenchymal Stem/Stroma Cells in a Mice Model of Colitis

Inflammatory bowel diseases (IBDs) are the result of pathological immune responses due to environmental factors or microbial antigens into a genetically predisposed individual. Mainly due to their trophic properties, a mounting interest is focused on the use of human mesenchymal stem/stromal cells (hMSCs) to treat IBD disease in animal models. The aim of the study is to test whether the secreted molecules, derived from a specific population of second trimester amniotic fluid mesenchymal stem/stromal cells, the spindle-shaped MSCs (SS-AF-MSCs), could be utilized as a novel therapeutic, cell free approach for IBD therapy. Induction of colitis was achieved by oral administration of dextran sulphate sodium (DSS) (3 % w/v in tap water), for 5 days, to 8-week-old NOD/SCID mice. The progression of colitis was assessed on a daily basis through recording the body weight, stool consistency and bleeding. Conditioned media (CM) derived from SS-AF-MSCs were collected, concentrated and then delivered intraperitoneally into DSS treated mice. To evaluate and determine the inflammatory cytokine levels, histopathological approach was applied. Administration of CM derived from SS-AF-MSCs cells reduced the severity of colitis in mice. More importantly, TGFb1 protein levels were increased in the mice received CM, while TNFa and MMP2 protein levels were decreased, respectively. Accordingly, IL-10 was significantly increased in mice received CM, whereas TNFa and IL-1b were decreased at mRNA level. Our results demonstrated that CM derived from SS-AF-MSCs cells is able to ameliorate DSS-induced colitis in immunodeficient colitis mouse model, and thus, it has a potential for use in IBD therapy.     

Placental isoform glutathione S‐transferase and P‐glycoprotein expression in advanced nonsmall cell lung cancer

BACKGROUND.

Increased expression of the glutathione S‐transferase placental isoform (GST‐pi) and of P‐glycoprotein (P‐gp) in tissues from patients with nonsmall cell lung cancer (NSCLC) has been associated with poor antineoplastic drug sensitivity, response to treatment, and survival. However, the diagnosis of advanced NSCLC often is based on cytology. The objectives of the current study were to examine GST‐pi and P‐gp expression in cytologic specimens from patients with unresectable NSCLC and to determine the association of that expression with response to chemotherapy and survival.

METHODS.

Patients with unresectable, cytologically diagnosed NSCLC were eligible for the study. Diagnosis was made by fiberoptic bronchoscopy, and staging was done according to international standards. All patients received sequential chemoradiotherapy and were re‐evaluated for treatment response. GST‐pi and P‐gp expression levels were evaluated by immunocytochemistry and immunohistochemistry of bronchial brushing/washing and bronchial tissue biopsy, respectively. Survival was defined as the time between diagnosis and death or last follow‐up at 24 months.

RESULTS.

Thirty‐nine patients were included in the study. There were 35 men and 4 women, and the mean patient age (±standard deviation was 61.4 years (±9.1 years). There were 4 patients with stage IIIA NSCLC, 32 patients with stage IIIB NSCLC, and 3 patients with stage IV NSCLC. Cytologic evaluation of GST‐pi and P‐gp expression paralleled expression determined in pathology specimens. GST‐pi and P‐gp expression levels were associated inversely with response to chemotherapy and survival.

CONCLUSIONS.

Cytologic evaluation of GST‐pi and P‐gp expression may predictor the response to treatment and the survival of patients with advanced NSCLC. Cancer (Cancer Cytopathol) 2008. © 2008 American Cancer Society.     

Inverted papilloma of the renal pelvis

The authors describe a case of inverted papilloma. It is relatively rare and generally appears as a benign tumour.     

Prophylactic effect of brinzolamide–brimonidine fixed combination on intraocular pressure spikes after intravitreal anti-VEGF injections

International Ophthalmology - To evaluate the effect of topical prophylaxis with brinzolamide–brimonidine fixed combination on short-term intraocular pressure (IOP) elevation after...     

Influence of environmental factors in the development of inflammatory bowel diseases

Idiopathic inflammatory bowel diseases(IBD), Crohn's disease(CD) and ulcerative colitis(UC), are multifactorial diseases that are manifested after disruption of a genetic predisposed individual and its intestinal microflora through an environmental stimulus. Urbanization and industrialization are associated with IBD. Epidemiological data, clinical observations and family/immigrants studies indicate the significance of environmental influence in the development of IBD. Some environmental factors have a different effect on the subtypes of IBD. Smoking and appendectomy is negatively associated with UC, but they are aggravating factors for CD. A westernized high fat diet, full of refined carbohydrates is strongly associated with the development of IBD, contrary to a high in fruit, vegetables and polyunsaturated fatty acid-3 diet that is protective against these diseases. High intake of nonsteroidal antiinflammatory drug and oral contraceptive pills as well as the inadequacy of vitamin D leads to an increased risk for IBD and a more malignant course of disease. Moreover, other factors such as air pollution, psychological factors, sleep disturbances and exercise influence the development and the course of IBD. Epigenetic mechanism like DNA methylation, histone modification and altered expression of miR NAS could explain the connection between genes and environmental factors in triggering the development of IBD.     

Risk factors for gastroesophageal reflux disease and analysis of genetic contributors

Gastroesophageal reflux disease (GERD) is a common gastrointestinal disorder with an increasing prevalence. GERD develops when the reflux of stomach contents causes troublesome typical and atypical symptoms and/or complications. Several risk factors of GERD have been identified and evaluated over the years, including a considerable amount of genetic factors. Multiple mechanisms are involved in the pathogenesis of GERD including: (1) motor abnormalities, such as impaired lower esophageal sphincter (LES) resting tone, transient LES relaxations, impaired esophageal acid clearance and delayed gastric emptying; and (2) anatomical factors, such as hiatal hernia and obesity. Genetic contribution seems to play a major role in GERD and GERD- related disorders development such Barrett’s esophagus and esophageal adenocarcinoma. Twin and family studies have revealed an about 31% heritability of the disease. Numerous single-nucleotide polymorphisms in various genes like FOXF1, MHC, CCND1, anti-inflammatory cytokine and DNA repair genes have been strongly associated with increased GERD risk. GERD, Barrett’s esophagus and esophageal adenocarcinoma share several genetic loci. Despite GERD polygenic basis, specific genetic loci such as rs10419226 on chromosome 19, rs2687201 on chromosome 3, rs10852151 on chromosome 15 and rs520525 on the paired related homeobox 1 gene have been mentioned as potential risk factors. Further investigation on the risk genes may elucidate their exact function and role and demonstrate new therapeutic approaches to this increasingly common disease.