Trefoil factor 2 (Tff2) deficiency in murine digestive tract influences the immune system

Background & Aims: The gastrointestinal trefoil factor family (TFF1, TFF2, TFF3) peptides are considered to play an important role in maintaining the integrity of the mucosa. The physiological role of TFF2 in the protection of the GI tract was investigated in TFF2 deficiency. Methods: TFF2-/- mice were generated and differential expression of various genes was assessed by using a mouse expression microarray, quantitative real time PCR, Northern blots or immunohistochemistry. Results: On an mRNA level we found 128 differentially expressed genes. We observed modulation of a number of crucial genes involved in innate and adaptive immunity in the TFF2-/- mice. Expression of proteasomal subunits genes (LMP2, LMP7 and PSMB5) involved in the MHC class I presentation pathway were modulated indicating the formation of immunoproteasomes improving antigen presentation. Expression of one subunit of a transporter (TAP1) responsible for importing degraded antigens into ER was increased, similarly to the BAG2 gene that modulates chaperone activity in ER helping proper loading on MHC class I molecules. Several mouse defensin (cryptdin) genes coding important intestinal microbicidal proteins were up-regulated as a consequence of TFF2 deficiency. Normally moderate expression of TFF3 was highly increased in stomach

Therapy Insight: Parenteral Estrogen treatment for Prostate Cancer—a new dawn for an old therapy

Oral estrogens were the treatment of choice for carcinoma of the prostate for over four decades, but were abandoned because of an excess of cardiovascular and thromboembolic toxicity. It is now recognized that most of this toxicity is related to the first pass portal circulation, which upregulates the hepatic metabolism of hormones, lipids and coagulation proteins. Most of this toxicity can be avoided by parenteral (intramuscular or transdermal) estrogen administration, which avoids hepatic enzyme induction. It also seems that a short-term but modest increase in cardiovascular morbidity (but not mortality) is compensated for by a long-term cardioprotective benefit, which accrues progressively as vascular remodeling develops over time. Parenteral estrogen therapy has the advantage of giving protection against the effects of andropause (similar to the female menopause), which are induced by conventional androgen suppression and include osteoporotic fracture, hot flashes, asthenia and cognitive dysfunction. In addition, parenteral estrogen therapy is significantly cheaper than contemporary endocrine therapy, with substantive economic implications for health providers

The effect of circulating antigen on the Biodistribution of the engineered Human antibody hCTM01 in a nude mice model

Clinical studies are currently underway to assess the biodistribution and therapeutic potential of the genetically engineered human antibody hCTM01 directed against polymorphic epithelial mucin (PEM) in patients with ovarian carcinoma. The present study was undertaken to assess the effect of circulating PEM antigen on the biodistribution of the anti-PEM antibody in mice bearing MUC-1 transfected adenocarcinoma cell lines. Tumour xenografts were established from three cell lines: 413-BCR, which expressed antigen on the cell surface and also shed antigen into the circulation, E3P23, which expressed the antigen but did not shed into the circulation, and a negative control (410.4 MUCI). Groups of five mice were injected with 1.0 mg/kg antibody, imaged after 72 h and then sacrificed, followed by assay of tissue uptake. The results showed a clear difference in the tumour and liver uptake, with the non-secreting cell line showing almost twice the tumour uptake and approximately 20% of the liver uptake of the secreting cell line

The expression pattern of MUC1 Glycoforms and other Biomarkers of Endometrial receptivity in fertile and infertile women

Changes in the surface epithelium of the endometrium, characterized in part by alterations in cell-surface molecules, sex steroid receptors and the appearance of pinopodes, coincide with the window of endometrial receptivity in the menstrual cycle. This study was performed to evaluate the usefulness of hematoxylin and eosin staining, scanning and transmission microscopy, and MUC1 glycoform, sex steroid receptor, and interleukin receptor (type 1) expression as biomarkers of endometrial receptivity using carefully characterized clinical fertile and infertile groups of women. Using a combination of immunohistochemistry and scanning electron microscopy (SEM) called scanning immunoelectron microscopy (SIM), we confirmed that MUC1 mucin was not associated with the endometrial pinopodes, which have been linked with embryo adhesion. We also showed that failure of embryo implantation was associated with an abnormal endometrial expression of MUC1 mucin, and retention of nuclear progesterone receptor (PR) particularly in epithelial cells. Hematoxylin and eosin staining, transmission electron microscopy (TEM), SEM in isolation and immunohistochemistry for interleukin receptor were not shown to be useful markers. Progesterone-dependent regulation of MUC1 appears to be an important factor in determining endometrial receptivity

Expression of bcl-2 and p53 Oncoproteins in Schistosomiasis-associated transitional and Squamous Cell Carcinoma of Urinary Bladder

Objectives: To analyse the expression of bcl-2 and p53 proteins in schistosomiasis-associated bladder cancer compared to adjacent urothelium showing morphological alteration including hyperplasia, metaplasia and dysplasia. Materials and Methods: Twenty-two formalin-fixed and paraffin-embedded samples of histologically confirmed schistosomiasis-associated bladder tumours were assessed for grade, pathological stage (pT category) and the presence of hyperplasia, metaplasia and dysplasia in adjacent mucosa. There were 11 transitional cell carcinomas (TCCs), 10 squamous cell carcinomas (SCCs) and one diffuse infiltrating (signet-ring cell type) adenocarcinoma. Epithelial hyperplasia was observed in seven cases and metaplasia in 18, most of which were squamous except for a single case of glandular metaplasia. Focal dysplastic areas were observed in eight cases. Sections were stained immunohistochemically for the expression of bcl-2 and p53 proteins. Results: Immunoreactivity for bcl-2 was present in seven of 22 cases (four SCCs and three TCCs) of which three cases (two SCCs and one TCC) were strongly positive, but in contrast to previous studies, there was no increase in the poorly differentiated tumours. bcl-2 was absent in metaplastic and dysplastic epithelium (except in a single case of glandular metaplasia) but it was present at low levels in basal cells of morphologically normal or hyperplastic transitional epithelium in 15 of 22 cases. Nine of 11 TCCs and seven of 10 SCCs showed p53 nuclear immunoreactivity. Heterogenous weak to moderate immunoreactivity for p53 was seen in 13 of 18 cases of metaplasia and in seven of eight cases of dysplasia in the mucosa adjacent to tumours. p53 was absent in normal and hyperplastic urothelium. Co-expression of p53 and bcl-2 was only seen in well differentiated areas of three tumours (two SCCs and one TCC). Conclusion: This study detected the expression of bcl-2 in a subset of bladder cancers (32%), whereas most (72%) of the tumours expressed immunoreactive p53. Additionally, p53 was also detected in metaplastic and dysplastic epithelium. Over-expression of both p53 and bcl-2 in the same tumour was only evident in a minority of schistosomiasis-associated cancers (13%). There was no inverse relationship of p53 and bcl-2 immunoreactivity

Immunolocalisation of Cripto and Amphiregulin in Rectal Cancer: correlation with Prognosis

We have examined the expression of two newly identified members of the EGF family, cripto and amphiregulin (AR), in a series of 58 primary rectal carcinomas and adjacent non-involved mucosa by immunohistochemical staining using rabbit polyclonal antibodies. More than 90% (53/58) of rectal carcinomas showed AR immunoreactivity whereas cripto was found in 41 out of 58 (71%) tumours. Cripto immunoreactivity was most frequently seen in tumours arising in the lower third of the rectum (p\u3c0.01) and in flat and excavated lesions (p\u3c0.05). Out of 54 normal rectal mucosae adjacent to carcinoma 20 (37%) showed cripto immunoreactivity and all showed a trend towards a higher recurrence rate. Ten of these 20 (50%) rectal tumours showed less cripto immuno-reactivity than the adjacent normal mucosa and were penetrating through the bowel wall and recurred within 5 years. There was a correlation between increased cripto immunoreactivity in the normal mucosa and lymph node involvement (p=0.01). AR immunoreactivity was present in the majority (52/58, 94%) of the normal mucosae adjacent to tumours. No correlation was found between AR immunostaining, histology and prognosis

Disruption of the developmentally regulated Rev3l Gene causes Embryonic Lethality

The REV3 gene encodes the catalytic subunit of DNA polymerase (pol) ζ, which can replicate past certain types of DNA lesions [1]. Saccharomyces cerevisiaerev3 mutants are viable and have lower rates of spontaneous and DNA-damage-induced mutagenesis[2]. Reduction in the level of Rev31, the presumed catalytic subunit of mammalian pol ζ, decreased damage-induced mutagenesis in human cell lines [3]. To study the function of mammalian Rev31, we inactivated the gene in mice. Two exons containing conserved DNA polymerase motifs were replaced by a cassette encoding G418 resistance and β-galactosidase, under the control of the Rev3l promoter. Surprisingly, disruption of Rev3lcaused mid-gestation embryonic lethality, with the frequency of Rev3l–/– embryos declining markedly between 9.5 and 12.5days post coitum (dpc). Rev3l–/– embryos were smaller than their heterozygous littermates and showed retarded development. Tissues in many areas were disorganised, with significantly reduced cell density. Rev3lexpression, traced by β-galactosidase staining, was first detected during early somitogenesis and gradually expanded to other tissues of mesodermal origin, including extraembryonic membranes. Embryonic death coincided with the period of more widely distributed Rev3l expression. The data demonstrate an essential function for murine Rev31 and suggest that bypass of specific types of DNAlesions by pol ζ is essential for cell viability during embryonic development in mammals