A novel assay for analysis of the regulation of the function of human osteoclasts

BACKGROUND: Very little is known of the regulation of the function of human osteoclasts, largely due to the virtual impossibility of obtaining human osteoclasts ex vivo. It has recently become possible to generate human osteoclasts in vitro, by incubation of peripheral blood mononuclear cells (PBMCs) in macrophage colony-stimulating factor (M-CSF) and receptor activator of nuclear factor-κB ligand (RANKL). However, the assays at present available do not distinguish clearly between the distinct effects of agents on differentiation and function. MATERIALS AND METHODS: We developed a novel assay for resorptive function of human osteoclasts that minimizes inter-assay variability by using each culture as its own baseline, and that minimizes the confounding effects of agents on differentiation by assessing resorptive function over a short test period. In this assay, the development of resorptive activity is monitored in sample cultures. When resorption is underway, bone resorption (measured as the release of the C-terminal telopeptide degradation product of type I collagen (CTX-I) into the supernatant) is compared before vs after incubation for 1–24 h in test agent. RESULTS: Using this assay, we found that changes in bone resorption could be detected using substantially fewer cultures per variable. Moreover, we could detect effects of agents on resorption within 1 h of addition, a time sufficiently short that a change in release is likely to reflect an effect on function rather than on differentiation. CONCLUSION: The assay makes it possible to distinguish the effects of agents on osteoclastic function, independent of their effects on differentiation

Методическая система обучения математике студентов фармацевтических факультетов

МАТЕМАТИКАОБУЧЕНИЕ /МЕТОДЫМЕТОДИКА ПРЕПОДАВАНИЯСТУДЕНТЫ МЕДИЦИНСКИХ УЧЕБНЫХ ЗАВЕДЕНИЙФАРМАЦЕВТИЧЕСКИЕ ФАКУЛЬТЕТЫОБРАЗОВАТЕЛЬНЫЙ ПРОЦЕССЛИЧНОСТНО-ОРИЕНТИРОВАННЫЙ ПОДХОДМЕТОДИЧЕСКИЕ СИСТЕМЫПРЕПОДАВАНИЕ МАТЕМАТИК

Cognitive outcome two years after frontal lobe resection for epilepsy – A prospective longitudinal study

AbstractPurposeTo investigate cognitive outcomes after frontal lobe resection (FLR) for epilepsy in a consecutive single centre series.MethodsNeuropsychological examinations were performed prior to and two years (mean test interval 2.5 years) after surgery in 30 consecutive patients who underwent FLR. Cognitive outcome was evaluated with particular consideration to the site of surgery (lateral, premotor/SMA [supplementary motor area], mesial/orbital). Cognitive domains assessed were speed, language, memory, attention, executive functions and intelligence. 25 healthy controls were assessed at corresponding time points (mean test interval 3.0 years). Analyses were made both at group and individual levels.ResultsAt baseline the patients performed below controls in variables depending on speed, executive functions, global and verbal intelligence. Two years after surgery, the analyses at the subgroup level indicated that the lateral resection group had less improvement than the controls in global intelligence, FSIQ (p=.037). However, at the individual level, the majority of the change scores (74–100%) were classified as within the normal range for all but one variable. The exception was the variable “Comprehension” (measuring verbal reasoning ability) with reliable declines in 44% (8/18) of the patients. This pattern of decline was observed in the lateral (4/7 patients) and premotor/SMA (4/7 patients) resection groups. Seizure outcome and side of surgery did not influence these results.ConclusionThe main finding was cognitive stability at group level two years after FLR. A reliable decline in verbal reasoning ability was rather common at an individual level, but only in the lateral and premotor/SMA resection groups. The lateral resection group also had less improvement than the controls in global intelligence

Synthesis, X-ray Analysis, and Biological Evaluation of a New Class of Stereopure Lactam-Based HIV-1 Protease Inhibitors

In an effort to identify a new class of druglike HIV-1 protease inhibitors, four different stereopure beta-hydroxy gamma-lactam-containing inhibitors have been synthesized, biologically evaluated, and cocrystallized. The impact of the tether length of the central spacer (two or three carbons) was also investigated. A compound with a shorter tether and (3R,4S) absolute configuration exhibited high activity with a K-i of 2.1 nM and an EC50 of 0.64 mu M. Further optimization by decoration of the P1' side chain furnished an even more potent HIV-1 protease inhibitor (K-i = 0.8 nM, EC50 = 0.04 mu M). According to X-ray analysis, the new class of inhibitors did not fully succeed in forming two symmetric hydrogen bonds to the catalytic aspartates. The crystal structures of the complexes further explain the difference in potency between the shorter inhibitors (two-carbon spacer) and the longer inhibitors (three-carbon spacer)

Cognitive outcome two years after frontal lobe resection for epilepsy – A prospective longitudinal study

AbstractPurposeTo investigate cognitive outcomes after frontal lobe resection (FLR) for epilepsy in a consecutive single centre series.MethodsNeuropsychological examinations were performed prior to and two years (mean test interval 2.5 years) after surgery in 30 consecutive patients who underwent FLR. Cognitive outcome was evaluated with particular consideration to the site of surgery (lateral, premotor/SMA [supplementary motor area], mesial/orbital). Cognitive domains assessed were speed, language, memory, attention, executive functions and intelligence. 25 healthy controls were assessed at corresponding time points (mean test interval 3.0 years). Analyses were made both at group and individual levels.ResultsAt baseline the patients performed below controls in variables depending on speed, executive functions, global and verbal intelligence. Two years after surgery, the analyses at the subgroup level indicated that the lateral resection group had less improvement than the controls in global intelligence, FSIQ (p=.037). However, at the individual level, the majority of the change scores (74–100%) were classified as within the normal range for all but one variable. The exception was the variable “Comprehension” (measuring verbal reasoning ability) with reliable declines in 44% (8/18) of the patients. This pattern of decline was observed in the lateral (4/7 patients) and premotor/SMA (4/7 patients) resection groups. Seizure outcome and side of surgery did not influence these results.ConclusionThe main finding was cognitive stability at group level two years after FLR. A reliable decline in verbal reasoning ability was rather common at an individual level, but only in the lateral and premotor/SMA resection groups. The lateral resection group also had less improvement than the controls in global intelligence