Partial redirection of transgenic human growth hormone secretion from rat salivary glands

Regulated secretory pathway proteins, when delivered as transgenes to salivary glands, are secreted predominantly into saliva. This is not useful for those proteins whose therapeutic function is required systemically, for example, human growth hormone (hGH). One strategy to improve the efficiency of hGH secretion into the bloodstream involves manipulation of existing sorting signals. The C terminus of hGH is highly conserved and contains a domain similar to the regulated pathway sorting domain of pro-opiomelanocortin (POMC). We hypothesized that, similar to POMC, mutation of this domain would divert hGH secretion from the regulated to the constitutive pathway, which in salivary glands leads to the bloodstream. Several mutations were made in the C terminus of the hGH cDNA and tested in vitro. One biologically active mutant containing E174A and E186A substitutions, and with an included C-terminal extension, was studied in greater detail. Compared with wild-type hGH, we found that this mutant hGH accumulated in the Golgi/trans-Golgi network and showed increased basal secretion in AtT20 cells, a model endocrine cell line. Importantly, in vivo, the mutant hGH displayed a relative increase in the proportion of constitutive pathway secretion seen from rat salivary glands, with a significantly lower saliva-versus-serum secretion ratio (p=0.03). Although this mutant is unlikely to be therapeutically beneficial, these results suggest that the final destination of a transgenic secretory protein may be controlled by reengineering its sorting determinants.     

Pre-referral practices: Under office of civil rights scrutiny

Generally, pre-referral interventions have been emphasized as a means of keeping students in the regular classroom unless such attempts fail and formal evaluation is warranted for a possible special education placement. The Office of Civil Rights, examining a number of pre-referral practices, has identified violations of guaranteed rights and has called for substantial changes. Recommendations are provided on how to preserve pre-referral practices and yet operate within legal mandates.     

Influence of pain and quality of life on desire for hastened death in patients with advanced cancer

OBJECTIVE: to assess the relationship between quality of life, pain and desire for hastened death in advanced cancer patients. METHODS: 120 Greek patients with advanced cancer were interviewed with the European Organization for Research and Treatment of Cancer Quality of Life Questionnaire C30 version 3.0 (EORTC QLQ-C30), the Greek Brief Pain Inventory (G-BPI), the Greek Hospital Anxiety and Depression Scale (G-HADS) and the Greek Schedule of Attitudes toward Hastened Death (G-SAHD). RESULTS: statistically significant associations were found between total G-SAHD scores and scores for the worst level of pain in the previous 24 hours (G-BP13) (r = 0.279, P = 0.002), and between total G-SAHD scores and scores for the level of pain relief obtained in the last 24 hours (G-BP18) (r = -0.326, P = 0.0005). The strongest correlations were found between G-SADH and emotional functioning (r = 0.569, P<0.0001) and global quality of life (r = -0.331, P<0.0001) from EORTC QLQ-C30. In multivariate analyses, emotional functioning, social functioning, financial impact, and the interference of pain in general activity and mood were significant predictors of G-SAHD (all P<0.0001). CONCLUSION: quality of life and pain appeared to have a statistically significant relationship with desire for hastened death. Adequate palliative care should alleviate pain and the desire for hastened death, improving quality of life.     

The Application of Science- and Risk-Based Concepts to Drug Substance Stability Strategies

The International Conference on Harmonization (ICH) has provided practical guidance on the amount and type of drug substance stability data needed to support marketing applications (International Conference on Harmonization 2001, 2002, 2003a, b). Additional guidance has been issued by the World Health Organization (WHO 2009). Recent scientific advances and practices have resulted in improved scientific understanding of the chemical and physical attributes that contribute directly or indirectly to drug substance stability. Combining this improved understanding with the science- and risk-based approaches detailed in ICH Q8, Q9, and Q10 allows for alternative and more scientifically driven approaches to meet the scientific and regulatory objectives for drug substance stability (International Conference on Harmonization 2005, 2008, 2009). In this paper, proposals are presented to more fully leverage enhanced product knowledge to design improved stability strategies. The chemical and physical attributes that potentially impact drug substance stability are discussed, and strategies that leverage accelerated stability studies are presented.     

The impact of olive oil consumption pattern on the risk of acute coronary syndromes: The CARDIO2000 case-control study

BACKGROUND: According to epidemiological and metabolic studies monounsaturated fatty acids (MUFAs) seem to exert a protection against coronary heart disease (CHD) risk. The aim of the present study was to evaluate the association between the pattern of edible oils and fats consumption and the prevalence of a first, nonfatal event of an acute coronary syndrome (ACS) in a Greek sample. METHODS: Seven hundred males and 148 females patients with first event of an ACS and 1078 population-based controls, age and sex matched, were randomly selected. Detailed information regarding their medical records, alcohol intake, physical activity and smoking habits was recorded. Nutritional habits were evaluated with a semi-quantitative food-frequency questionnaire and use of oils in daily cooking or preparation of food was also recorded. Multiple logistic regression analysis estimated the odds ratio (OR) of having ACS by types of oil used, after taking into account the effect of several confounders. RESULTS: Exclusive use of olive oil was associated with 47% (95% confidence interval (CI) 0.4-0.71) lower likelihood of having ACS, compared to nonuse, after adjusting for BMI, smoking, physical activity level, educational status, the presence of family history of CHD, as well as hypertension, hypercholesterolemia and diabetes. Consumption of olive oil in combination with other oils or fats was not significantly associated with lower odds of ACS compared to no olive oil consumption (p=0.14). CONCLUSIONS: Exclusive use of olive oil during food preparation seems to offer significant protection against CHD, irrespective of various clinical, lifestyle and other characteristics of the participants.     

Expression of ets-1 is not affected by N-ras or H-ras during oral oncogenesis

PURPOSE: To determine whether ras-activated cascades lead to activation of ets-1 expression in sequential histological stages of oral oncogenesis in an experimental animal model. METHODS: Thirty-seven Syrian golden hamsters were divided into three experimental groups (A, B, C) and one control group. The hamsters' buccal pouches in experimental groups were treated with 0.5% 9, 10-dimethyl-1, 2-benzanthracene (DMBA) for 14 weeks and were excised at 10, 14, 19 weeks, respectively. The biopsies were classified pathologically (normal mucosa, hyperkeratosis, hyperplasia, dysplasia, early invasion, well and moderately differentiated carcinoma) and studied immunohistochemically. The two-tailed Student's t test was performed for each animal group and for each histological category. RESULTS: The ets-1 expression increased in early stages of oral tumor formation and invasion. The expression of N-ras gradually decreased during oral oncogenesis, as previously observed with H-ras. CONCLUSIONS: Neither N-ras nor H-ras affects ets-1 expression in contrast to other types of cancer in which N-ras and ets-1 are implicated in the same signalling pathway. Therefore, the existing pathway implicating these proteins might be somehow altered in oral cancer. It seems that ets-1 is a good prognostic marker for invasiveness and progression of oral cancer.