The Bcl-2 protein family and its role in the development of neoplastic disease

Programmed cell death is the physiological process responsible for shaping organs during embryogenesis, maintaining tissue homeostasis and allowing controlled deletion of potentially harmful cells within the adult organism. The genetics of apoptosis are well conserved in all metazoans and although the evolution of humans and worms separated more than 600 million years ago, basic signaling concepts in apoptosis are highly related in both species. More crucial to humans than worms is the fact that abnormalities in cell death control can contribute to the development of cancer. While C.elegans can easily survive with additional somatic cells that should normally be deleted during development humans may suffer pathological consequences, ranging from tumorigenesis to autoimmunity, as a result of mutations in cell death regulatory genes. Despite the high degree of evolutionary conservation in cell death control, apoptosis signaling in mammals is much more complex than in C.elegans. In mammalian cells, programmed cell death can be induced either by ligand-mediated activation of certain members of the tumor necrosis factor receptor family--so-called 'death receptors'--such as Fas (CD95/Apo-1) and TRAIL or it can be induced in a cell autonomous manner in response to certain stress signals by pro-apoptotic members of the Bcl-2 family. In this review, we focus on general concepts of how the Bcl-2 protein family regulates cell death and how deregulation of this 'intrinsic' apoptotic signaling pathway impinges on the pathogenesis of malignant disease, the major cause of death in the aging population.     

In vitro and in vivo anticancer activity of mitozolomide and sparsomycin in human tumor xenografts,murine tumors and human bone marrow

Summary The colony formation in agar of human tumor xenografts, of murine tumors and of human bone marrow was used as a test system to determine the in vitro activity of the two novel cytostatic agents, mitozolomide and sparsomycin. Mitozolomide was additionally studied in vivo in nine human tumor xenografts. The comparison of in vitro/in vivo activity allows an assessment of the relevant in vitro dose based on in vivo pharmacological behavior of a compound. Both compounds showed clear dose/response effects in vitro. A dose of 3 g/ml mitozolomide, given by continuous exposure, was active (colony number of test <30% of the control group) in 12/42 (29%) human tumor xenografts as well as in the four murine tumors, P388, L1210, B16 melanoma and colon carcinoma 38, whereas the two human bone marrows showed no significant suppression of the ability to form colonies in culture. The comparison of in vitro with in vivo activity suggests that the in vitro dose of 3 g/ml corresponds best to the activity observed in animal experiments. The highest activity was observed in small-cell cancer of the lung (4/5), followed by melanomas (2/7) and non-small-cell cancer of the lung (2/9). Furthermore, activity was found in a cancer of the large bowel, stomach, breast and in one sarcoma. In the treatment of nine human tumor xenografts growing subcutaneously in nude mice, mitozolomide effected a complete or partial remission in 6 out of 9 tumors. In comparison to standard drugs mitozolomide is one of the most effective compounds in these tumors. These data indicate that mitozolomide possesses potent broad-spectrum activity in human tumor xenografts. Sparsomycin (0.1 g/ml, continuous exposure) was active in 11/46 (24%) human tumor xenografts and in 4/5 of the murine tumors, whereas the colony-forming capacity of four human bone-marrows showed no inhibition, suggesting that this dose level may be the relevant in vitro dose. However, the high in vitro activity in murine tumors is incompatible with the in vivo activity. In mice the only responsive tumor was leukemia P388, whereas the L1210, B16 melanoma and colon carcinoma 38 were resistant. At the dose level of 0.03 g/ml only 3/30 (10%) of the human tumor xenografts were sensitive. In an earlier clinical phase I study the dose-limiting adverse effect was eye toxicity and not bone-marrow suppression. This example illustrates that comparing in vitro with in vivo activity in the same tumor results in a more reliable estimation of the relevant in vitro dose than does comparing in vitro activity with in vitro effects on human bone marrow.Abbreviations Mitozolomide 8-carbamoyl-3-(2-chloroethyl)imidazo(5,1-d)-1,2,3,5-tetrazin-4(3H)-one - NSC 353451 formerly known as azolastone - sparsomycin NSC 59 729 - DTIC 5-(3,3-dimethyltriazen-1-yl)-imidazole-4-carboxamide - MTIC 5-(3-methyltriazen-1-yl)imidazole-4-carboxamide - DMSO dimethylsulfoxide Dedicated to Professor Dr. D. Schmähl, Heidelberg, on the occasion of his 65th birthday     

Predictors and consequences of aggressive behavior by community-based dementia patients

The frequency, nature, context, and caregivers' reactions to aggressive behavior in 213 dementia patients residing in the community was studied. Aggression was reported in 57.2% of the patients and in 10.6% of the caregivers. Predictors of patient aggression were greater frequency of behavior and memory problems, premorbid aggression, and a more troubled premorbid social relationship between patient and caregiver. Patient aggression predicted the decision to discontinue home care.     

Breast cancer screening for older women in a primary care practice

OBJECTIVE: To determine rates of breast cancer screening for older women cared for in a primary care practice and to identify associations between patient and physician characteristics and breast cancer screening. STUDY DESIGN: A retrospective cohort study of older women. SETTING: An urban hospital-based academic general medicine practice. This practice uses a computerized medical record and office procedures that facilitate tracking and ordering of mammograms. PARTICIPANTS: A random sample of 130 women aged 65 to 80 who receive primary care at a hospital-based general medicine practice. MEASUREMENTS: Data were collected from the hospital's computerized medical record. We recorded all clinical breast exams and mammograms performed or recommended during the 2-year study period. RESULTS: The median age of the 130 women studied was 71, and 21% of the women were black. Most patients had no serious comorbid illness (69%) and were independent in their activities of daily living (92%). During the 2-year study period, mammography was recommended for 95% of women and completed for 84%, and clinical breast exam was performed on 75%. Patients of male physicians had higher rates of mammography than patients of female physicians (89% vs. 75%, P = .045). Patients of faculty physicians had higher rates of clinical breast exam than patients of house officers or fellows (83% vs. 56%, P = .001). CONCLUSIONS: We report a very high rate of mammography for women cared for at a hospital-based primary care practice. The systems in place to facilitate ordering and tracking of mammograms probably contributed to the unusually high rate of mammography observed.     

Estrogen replacement decreases the set point of parathyroid hormone stimulation by calcium in normal postmenopausal women

Estrogens decrease serum total and ionized calcium (Ca) concentrations in postmenopausal women with or without primary hyperparathyroidism, but cause little or no increase in serum PTH suggesting a modification of the relationship between the two. In order to define this relationship, we studied the effect of conjugated estrogens on total and ionized serum Ca and serum PTH concentrations in five normal postmenopausal women, before and after 3, 11, and 23 weeks of therapy. Dynamic tests of parathyroid gland function, based on 2-h iv infusions of CaCl2 and NaEDTA, were performed at each time. Total and ionized serum Ca and carboxylterminal PTH were measured every 15 min during the infusions, and parathyroid function was evaluated by a nonlinear 4-parameter mathematical model. Estrogen therapy caused decreases in serum total [2.36 +/- 0.04 (SD) mmol/L, baseline vs. 2.19 +/- 0.05 mmol/L, 23 weeks, P less than 0.005) and ionized calcium (1.27 +/- 0.01 mmol/L, baseline vs. 1.21 +/- 0.02 mmol/L, 23 weeks, P less than 0.005]; the decreases were evident at 3 weeks and persisted for the duration of the study. Serum PTH concentrations did not change (8.94 +/- 1.84 pmol/L, baseline vs. 8.98 +/- 2.38 pmol/L, 23 weeks). Three parameters of the parathyroid function, the maximal response to hypocalcemic stimulation, the nonsuppressible fraction of circulating PTH, and the slope of PTH on calcium at the set point were not affected by estrogen treatment. The fourth parameter, the set point of PTH stimulation by serum total calcium (2.16 +/- 0.04 mmol/L, baseline vs. 1.97 +/- 0.07 mmol/L, 23 weeks, P less than 0.0166) or by serum ionized Ca (1.19 +/- 0.04 mmol/L, baseline vs. 1.12 +/- 0.03 mmol/L, 23 weeks, P less than 0.01), was decreased by estrogen treatment. This was evident at the earliest time point studied and persisted thereafter. The decrease in ionized Ca set point only explained 40% of the decrease in total calcium set point, the remaining 60% being related to hemodilution of plasma protein during therapy. We conclude that estrogen replacement can influence parathyroid function in postmenopausal women by resetting the set point of PTH stimulation by ionized Ca. This in turn could contribute to the estrogen-induced changes in their Ca balance.     

Polycaprolactone scaffold and reduced rhBMP-7 dose for the regeneration of critical-sized defects in sheep tibiae

The transplantation of autologous bone graft as a treatment for large bone defects has the limitation of harvesting co-morbidity and limited availability. This drives the orthopaedic research community to develop bone graft substitutes. Routinely, supra-physiological doses of bone morphogenetic proteins (BMPs) are applied perpetuating concerns over undesired side effects and cost of BMPs. We therefore aimed to design a composite scaffold that allows maintenance of protein bioactivity and enhances growth factor retention at the implantation site. Critical-sized defects in sheep tibiae were treated with the autograft and with two dosages of rhBMP-7, 3.5 mg and 1.75 mg, embedded in a slowly degradable medical grade poly(ε-caprolactone) (PCL) scaffold with β-tricalcium phosphate microparticles (mPCL–TCP). Specimens were characterised by biomechanical testing, microcomputed tomography and histology. Bridging was observed within 3 months for the autograft and both rhBMP-7 treatments. No significant difference was observed between the low and high rhBMP-7 dosages or between any of the rhBMP-7 groups and autograft implantation. Scaffolds alone did not induce comparable levels of bone formation compared to the autograft and rhBMP-7 groups. In summary, the mPCL–TCP scaffold with the lower rhBMP-7 dose led to equivalent results to autograft transplantation or the high BMP dosage. Our data suggest a promising clinical future for BMP application in scaffold-based bone tissue engineering, lowering and optimising the amount of required BMP.     

Differential diagnosis of cystic and alveolar echinococcosis using an immunochromatographic test based on the detection of specific antibodies

Human cystic and alveolar echinococcoses are zoonotic diseases caused by the larval stages of Echinococcus granulosus and Echinococcus multilocularis, respectively. As the diseases are co-endemic in many areas of the world, a simple and rapid test for the differential diagnosis of cystic echinococcosis (CE) and alveolar echinocoocosis (AE) is needed. Here, we describe the development of an immunochromatographic test (ICT) using crude hydatid cyst fluid and a recombinant 18-kDa protein (rEm18) as antigens for the detection of E. granulosus and E. multilocularis antibodies in serum samples. The ICT was evaluated with serum samples from 195 echinococcosis patients from different endemic areas in northwestern China. These included 144 from CE patients, 51 from AE patients, 67 from patients with other parasitic diseases, 13 from patients with serous hepatic cysts, and 60 from healthy individuals. The sensitivity and specificity of the ICT for CE were 91.0 and 96.9 % and for AE were 98.0 and 99.3 % with diagnostic efficiencies of 94.1 and 99.1 %, respectively. No significant differences and high degrees of agreement were found between the ICT and an enzyme-linked immunosorbent assay for both CE and AE. Five serum samples from cysticercosis patients and one serum sample from a healthy control were found positive for CE with the ICT. These findings indicate that this test allows for discrimination between both forms of human echinococcosis. In conclusion, the ICT developed in this study is a promising tool for the simultaneous detection and discrimination of CE and AE. This test will be useful for serodiagnosis of CE and AE in clinical settings and screening programs.