Truncated N-terminal mutants of SV40 large T antigen as minimal immortalizing agents for CNS cells

Immortalized central nervous system (CNS) cell lines are useful as in vitro models for innumerable purposes such as elucidating biochemical pathways, studies of effects of drugs, and ultimately, such cells may also be useful for neural transplantation. The SV40 large T (LT) oncoprotein, commonly used for immortalization, interacts with several cell cycle regulatory factors, including binding and inactivating p53 and retinoblastoma family cell-cycle regulators. In an attempt to define the minimal requirements of SV40 T antigen for immortalizing cells of CNS origin, we constructed T155c, encoding the N-terminal 155 amino acids of LT. The p53 binding region is known to reside in the C-terminal region of LT. An additional series of mutants was produced to further narrow the molecular targets for immortalization, and plasmid vectors were constructed for each. In a p53 temperature sensitive cell line model, T64-7B, expression of T155c and all constructs having mutations outside of the first 82 amino acids were capable of overriding cell-cycle block at the non-permissive growth temperature. Several cell lines were produced from fetal rat mesencephalic and cerebral cortical cultures using the T155c construct. The E107K construct contained a mutation in the Rb binding region, but was nonetheless capable of overcoming cell cycle block in T64-7B cell and immortalizing primary cultured cells. Cells immortalized with T155c were often highly dependent on the presence of bFGF for growth. Telomerase activity, telomere length, growth rates, and integrity of the p53 gene in cells immortalized with T155c did not change over 100 population doublings in culture, indicating that cells immortalized with T155c were generally stable during long periods of continuous culture.     

Intracerebral xenografts of mouse bone marrow cells in adult rats facilitate restoration of cerebral blood flow and blood-brain barrier

We examined in the present study alterations in cerebral blood flow (CBF) and blood-brain barrier (BBB) permeability following intrastriatal transplantation of mouse bone marrow stromal cells (BMSCs) or saline infusion in adult Sprague-Dawley rats. Laser Doppler revealed that transplanted animals exhibited near normal cerebral blood flow (CBF, 150 perfusion units) at a much earlier period post-transplantation (day 4) compared to animals that received saline infusion (day 12) (p's<0.05). Similarly, Evans Blue assay demonstrated that transplanted animals exhibited near complete BBB reconstitution at day 5 post-transplantation, whereas animals that received saline infusion continued to display a compromised BBB up to 11 days post-transplantation. Transplanted animals displayed a cell dose-dependent CBF and BBB restoration. Enzyme-linked immunosorbent assay (ELISA) of transplanted BMSCs revealed elevated levels of transforming growth factor-beta superfamily of neurotrophic factors. Moreover, despite the absence of immunosuppression in this cross-species transplantation, at least in the acute phase (12 days post-transplantation), surviving xenografts were detected during periods of restored CBF and BBB permeability. These observations suggest that restoration of CBF and BBB permeability accompanies the reported functional outcomes associated with intracerebral transplantation of BMSCs.     

Management of menstrual problems and contraception in adolescents with mental retardation: a medical, legal, and ethical review with new suggested guidelines

The controversial history of the reproductive rights of the mentally retarded has led to the formulation of laws in the past century designed to protect women from forced sterilization. Significantly, however, in their official ethical guidelines, The American College of Obstetricians and Gynecologists states that "sterilization should not be denied to individuals simply because they also may be vulnerable to coercion" (Int J Gynaecol Obstet 1999; 65:317). Recent advances in medical and surgical methods of contraception and control of menstrual abnormalities have led to a re-evaluation of the management of adolescents with special needs. Physicians, the courts, parents, and caretakers need to be aware of the latest medical and surgical options available, the current applicable laws in each state if such exist, and the ethical guidelines to determine what treatment option is in the best interests of the patient. This review examines the history of the sterilization of the mentally retarded, the latest surgical and pharmacologic treatments available, and the current legal environment and proposes an algorithm to facilitate the management of menstrual hygiene and contraception.     

IUD use and the risk of endometrial cancer

OBJECTIVE: Although the intrauterine device (IUD) is one of the most widely used forms of contraception throughout the world, its potential long-term effects on the uterus have not been thoroughly evaluated. This paper reports the long-term results of IUD use on the incidence of endometrial cancer. STUDY DESIGN: The data is part of a nationwide case-control, pilot study that was undertaken in order to evaluate the possible influence of ovulation induction drugs on the risk of endometrial cancer. The study included 128 living women 35-64 years old, with a histologically confirmed diagnosis of endometrial carcinoma. The controls were 255 women from the same dialing areas selected by random digit dialing. A multivariate logistic model, controlling for age, was used to assess the independent effects of factors found to be significantly associated with endometrial cancer on univariate analysis. RESULTS: The following parameters were found to be independently associated with endometrial cancer controlling for age: nulliparity OR=2.7 (95% CI 1.1-6.5, P=0.03); history of infertility OR=1.8 (95% CI 1.0-3.3, P=0.05); BMI>/=27 OR=2.3 (95% CI 1.4-3.8, P=0.001).The use of oral contraceptives and IUD were found to be protective; OR=0.29 and 0.37, respectively, (95% CI 0.14-0.61, P=0.001, 0.19-0.70, and 0.003, respectively). CONCLUSIONS: IUD use may have a protective effect on endometrial cancer risk. The protective effect of IUD may be either, through the intense inflammatory response that leads to other lisosomal and inflammatory actions, which may include cells responsible for early elimination of hyperplastic endometrial epithelial cells or, the more complete shedding of the endometrium associated with IUD use may decrease hyperplasia of the endometrium, a known risk factor for endometrial carcinoma.     

Towards a critical health psychology practice

The field of critical psychology is exerting an influence in the way various sub-disciplines within psychology operate. In this article we use a critical psychology framework to review the field of health psychology. Through the use of values, assumptions and practices we review progress in health psychology and offer recommendations for aligning contemporary practices with current thinking in critical psychology. We discuss typical expectations, critical formulations and critical practice for interventions with individuals, groups and communities along these dimensions.     

Assessing data adequacy for clinical research: reliability and validity of a surgical database

As clinical databases are utilized more frequently for clinical research, it is essential that researchers assess the quality of databased information. While researchers have begun to report strategies to measure accuracy of databased information, knowledge remains limited. The purpose of this study was to assess the reliability and validity of databased information among selected study variables contained within a computerized coronary artery surgery clinical database using the written patient medical record as an external standard. Both reliability (N = 400) and validity (N = 100) samples were randomly selected from a databased sampling frame of 548 Medicare subjects who underwent coronary artery bypass grafting surgery in 1998. Reliability assessed by consistency rates were age (95%), race (94%), gender (99%), congestive heart failure (CHF) (60.5%), angina (91.5%), renal insufficiency (82%), hypertension (91.7%), diabetes mellitus (93.7%), chronic obstructive pulmonary disease (COPD) (75.5%), clinical status (97%), American Society of Anesthesiologists classification (99%), prior peripheral vascular surgery (15.5%), prior CABGS (99%), and duration of mechanical ventilation (87.5%). These percentages reflected a large portion of missing data for CHF, COPD, and prior peripheral vascular surgery. Validity assessed by sensitivity and specificity analyses were all greater than 80%. The majority of computerized databased information among selected study variables was the same information recorded in the written patient medical record. Using the same external standard to assess both reliability and validity was a significant limitation of this study, which resulted in the same measure of data adequacy by utilizing differing statistical methods.     

Hypertension in hypophosphatemic rickets—role of secondary hyperparathyroidism

Hypertension has been anecdotally reported in children with familial hypophosphatemic rickets (XLH). To better identify and characterize the clinical and laboratory features of hypertensive XLH children, we reviewed the medical records of 41 XLH children, all treated with phosphate and vitamin D analogues. Eight children, who were originally normotensive, developed hypertension during the 2nd decade of life. At diagnosis of hypertension all had persistent secondary/tertiary hyperparathyroidism (HPTD), defined as high serum parathyroid hormone (PTH) for 12 months or longer. Seven had nephrocalcinosis (NC). Analysis of data showed that of 11 children with HPTD, 8 developed hypertension compared with 0 among 30 without HPTD (P<0.001). Of 40 children studied, 18 had NC that was significantly associated with both HPTD (P<0.01) and hypertension (P<0.025). At diagnosis of hypertension, serum calcium was elevated in 2. Plasma renin activity was high in 3 of 4 patients in whom it was measured. Doppler ultrasonography or renal scan was normal in the 5 children studied. Early echocardiography showed left ventricular hypertrophy in only 2 of 5 children studied. In 3 patients who underwent parathyroidectomy, hypertension persisted and 1 progressed to renal failure. Serum creatinine remained normal in all others. Successful treatment of hypertension consisted of beta-adrenergic blockers, angiotensin converting enzyme inhibitors, and Ca channel blockers as monotherapy or in combination. We conclude that hypertension in treated XLH children is closely associated with HPTD. Emphasis should therefore be placed on prevention of the development of HPTD as a complication of XLH treatment, and close monitoring for hypertension in those who do develop HPTD.