Production of Recombinant Chemokines and Validation of Refolding.

The diverse roles of chemokines in normal immune function and many human diseases have motivated numerous investigations into the structure and function of this family of proteins. Recombinant chemokines are often used to study how chemokines coordinate the trafficking of immune cells in various biological contexts. A reliable source of biologically active protein is vital for any in vitro or in vivo functional analysis. In this chapter, we describe a general method for the production of recombinant chemokines and robust techniques for efficient refolding that ensure consistently high biological activity. Considerations for initiating development of protocols consistent with Current Good Manufacturing Practices (cGMPs) to produce biologically active chemokines suitable for use in clinical trials are also discussed.

Incidence of Gastric Cancer in Marrakech and Casablanca, Morocco.

Gastric cancer is the fifth most common cancer globally with over 70% of new cases occurring in developing countries. In Morocco, oncologists in Marrakech suspected higher frequency of gastric cancer compared to Casablanca, a city 150 kilometers away. This study calculated age-specific, sex-specific, and total incidence rates of gastric cancer in Marrakech and was compared to the Casablanca population-based cancer registry. Using medical records from Center Hospital University Mohammad VI and reports from 4 main private pathology laboratories in Marrakech, we identified 774 patients for the period 2008-2012. Comparison of rates showed higher age-specific incidence in Marrakech in nearly all age groups for both genders. A higher total incidence in Marrakech than in Casablanca was found with rates of 5.50 and 3.23 per 100,000, respectively. Incidence was significantly higher among males in Marrakech than males in Casablanca (7.19 and 3.91 per 100,000, resp.) and females in Marrakech compared to females in Casablanca (3.87 and 2.58 per 100,000, resp.). Future studies should address possible underestimation of gastric cancer in Marrakech, estimate incidence in other regions of Morocco, and investigate possible risk factors to explain the difference in rates.

Measuring the effect of improvement in methodological techniques on data collection in the Gharbiah population-based cancer registry in Egypt: Implications for other Low- and Middle-Income Countries.

The purpose of this study was to describe and quantify procedures and methods that maximized the efficiency of the Gharbiah Cancer Registry (GPCR), the only population-based cancer registry in Egypt. The procedures and measures included a locally-developed software program to translate names from Arabic to English, a new national ID number for demographic and occupational information, and linkage of cancer cases to new electronic mortality records of the Ministry of Health. Data was compiled from the 34,058 cases from the registry for the years 1999-2007. Cases and registry variables about demographic and clinical information were reviewed by year to assess trends associated with each new method or procedure during the study period. The introduction of the name translation software in conjunction with other demographic variables increased the identification of detected duplicates from 23.4% to 78.1%. Use of the national ID increased the proportion of cases with occupation information from 27% to 89%. Records with complete mortality information increased from 18% to 43%. Proportion of cases that came from death certificate only, decreased from 9.8% to 4.7%. Overall, the study revealed that introducing and utilizing local and culture-specific methodological changes, software, and electronic non-cancer databases had a significant impact on data quality and completeness. This study may have translational implications for improving the quality of cancer registries in LMICs considering the emerging advances in electronic databases and utilization of health software and computerization of data.

Amyloid precursor-like protein 2 (APLP2) affects the actin cytoskeleton and increases pancreatic cancer growth and metastasis.

Amyloid precursor-like protein 2 (APLP2) is aberrantly expressed in pancreatic cancer. Here we showed that APLP2 is increased in pancreatic cancer metastases, particularly in metastatic lesions found in the diaphragm and intestine. Examination of matched human primary tumor-liver metastasis pairs showed that 38.1% of the patients had positive APLP2 expression in both the primary tumor and the corresponding liver metastasis. Stable knock-down of APLP2 expression (with inducible shRNA) in pancreatic cancer cells reduced the ability of these cells to migrate and invade. Loss of APLP2 decreased cortical actin and increased intracellular actin filaments in pancreatic cancer cells. Down-regulation of APLP2 decreased the weight and metastasis of orthotopically transplanted pancreatic tumors in nude mice.

Measuring the Effect of Improved Medical Facilities and Focused Training on Data Quality and Completeness: An Example from the Gharbiah Population-Based Cancer Registry, Egypt.

Cancer rates are increasing in low- and middle- income countries. There are a limited number of populationbased cancer registries in Africa and the Egyptian population-based registry in Gharbiah is one of those registries. This registry has followed the standard international registration process and methods since 1999 and has been included in Cancer Incidence in Five Continents volumes IX and X. This article illustrates the reflection of improving medical care in the geographic region of the registry and focused training on enhancing the registry data. The registry area has seen advancement in medical care and cancer diagnostic facilities during the study period. The focused training included 8 different international training sessions over 8 different years for the registrars, administrators, and directors as well as continuing on-the-job training for other registry personnel. These improvements resulted in an overall 40% increase in nonmicroscopic diagnosis of hepatocellular carcinoma, as well as 20%, 10%, and 10% increases in microscopic diagnosis of pancreatic, brain, and lung cancers, respectively, over 9 years. An overall increase of 5% to 10% in subsite diagnosis was also seen for lung, colon, brain, bladder, and breast cancers for the same 9 years. An increase of 3% in grading was seen for solid tumors while 11% was seen for lymphoma. This study showed that low- and middle- income countries can observe higher data quality for cancer registries with improvement in medical care and focused training.

Impact of beta 2-microglobulin on tapasin expression and covalent association.

Cellular immunity is dependent on major histocompatibility complex (MHC) class I molecules enabling cytotoxic T cell recognition of malignant and infected cells. Loading of antigenic peptides onto MHC class I is assisted by a peptide-loading protein complex including tapasin. We found that tapasin expression is enhanced by beta 2-microglobulin via both transcriptional and post-transcriptional mechanisms. In addition, using conditions which preserve the tapasin-ERp57 disulfide-bonded conjugate, we demonstrated that beta 2-microglobulin increases tapasin-containing protein complexes, and reduces the level of MHC class I/ERp57 complexes lacking tapasin. Overall, our results provide a new perspective on the regulation of tapasin expression and association.