Genetic demonstration of a role for PKA in the late phase of LTP and in hippocampus-based long-term memory

To explore the role of protein kinase A (PKA) in the late phase of long-term potentiation (L-LTP) and memory, we generated transgenic mice that express R(AB), an inhibitory form of the regulatory subunit of PKA, only in the hippocampus and other forebrain regions by using the promoter from the gene encoding Ca2+/ calmodulin protein kinase IIalpha. In these R(AB) transgenic mice, hippocampal PKA activity was reduced, and L-LTP was significantly decreased in area CA1, without affecting basal synaptic transmission or the early phase of LTP. Moreover, the L-LTP deficit was paralleled by behavioral deficits in spatial memory and in long-term but not short-term memory for contextual fear conditioning. These deficits in long-term memory were similar to those produced by protein synthesis inhibition. Thus, PKA plays a critical role in the consolidation of long-term memory.     

Refinement of gamma delta resolvase reveals a strikingly flexible molecule

BACKGROUND: gamma delta resolvase is a 20.5 kDa enzyme that catalyzes a site-specific recombination in the second step of the transposition of the gamma delta transposon and requires no cofactors other than Mg2+ for activity. Dimers of resolvase bind cooperatively to DNA at three inverted repeat sequences of differing geometry but catalyze recombination at only one site. RESULTS: The structure of the catalytic domain of gamma delta resolvase, which provides the protein-protein interactions in the synaptic complex, has been refined to an R-factor of 20% at 2.3 A resolution. The structures of the three independent monomers in the asymmetric unit are similar but not identical. Differences occur in the positions of surface loops and in the overall twist of the central beta-sheet of the molecule. The crystal also gives two independent structures for the dimeric form of the molecule, which also show significant differences in the relative orientations of their subunits. CONCLUSION: Resolvase is an unusually flexible protein. This conformational adaptability may be necessary to allow each of the 12 resolvase subunits in the synaptic complex to play a different but specific role in wrapping DNA, binding sites of differing geometry and catalyzing recombination.     

Control of the vertical dimension in Class II correction using a cervical headgear and lower utility arch in growing patients. Part I

A retrospective cephalometric study was performed comparing three groups of 30 growing patients with Class II, Division 1 malocclusions. Group 1 was treated with a cervical headgear/lower utility arch combination (CHG/LUA), group 2 was treated with a cervical headgear alone (CHG), and the third group was untreated. The average treatment time was 1 year, 6 months. No other appliances were used during this period. Maxillary and mandibular dental and skeletal treatment responses were compared with an analysis of variance (ANOVA) and a Scheffe's test. In addition, a multiple stepwise regression was performed to determine whether pretreatment measures of facial pattern were accurate predictors of mandibular rotational response. Both treatment groups demonstrated significant reduction in maxillary protrusion. The CHG-only group showed significantly greater anterior descent of the palatal plane as compared with the untreated group. The maxillary molars showed significant distal movement in both treatment groups without any extrusion beyond that seen with normal growth. The maxillary incisor demonstrated significant retroclination in the CHG-only group. There was no statistical difference among the groups for variables commonly used for measuring mandibular rotation or protrusion. The change in vertical position of the lower molar was not significantly different among the groups. A CHG as used in this study produced maxillary orthopedic and orthodontic changes without upper molar extrusion beyond that seen with normal eruption and in the absence of an opening rotation of the mandible, even in subjects with dolicocephalic facial patterns. The LUA did not appear to influence lower molar eruption or mandibular rotational response. None of the commonly used predictors of facial pattern, such as the Y-axis, XY-axis, or MP angle, accurately predicted mandibular rotational response. Further study would be necessary to ascertain whether this was a result of their invalidation as predictors, or a result of the treatment strategy employed.     

Correlation between structure and magnetic spin state of the manganese cluster in the oxygen-evolving complex of photosystem II in the S2 state: determination by X-ray absorption spectroscopy

The structure of the manganese cluster in the S2 state with the g approximately 4 EPR signal (S2-g4 state) generated by 130 K illumination of photosystem II (PSII) membranes prepared from spinach has been investigated by X-ray absorption spectroscopy. The Mn X-ray absorption K-edge spectra of the S2-g4 state not only show a shift of the inflection point to higher energy from the S1 state but also reveal a different edge shape from that of the S2 state with the multiline signal (S2-MLS state). Extended X-ray absorption fine structure (EXAFS) studies of the Mn K-edge show that the structure of the Mn cluster in the S2-g4 state is distinctly different from those in the S2-MLS or S1 states. In the S2-g4 state, the second shell of back-scatters from the Mn absorber is found to contain two Mn-Mn distances of 2.73 and 2.85 A. We interpret this to indicate the presence of two nonequivalent di-mu-oxo-bridged Mn binuclear structures in the Mn cluster of the S2-g4 state. The third shell of the S2-g4 state at about 3.3 A also contains increased heterogeneity. By contrast, very little distance disorder was found to exist in the second shell of the S1 or S2-MLS states. A mechanism is proposed to explain these results in the context of our model for the Mn cluster and the EPR properties of the Mn complex in the S2 state.     

Characterization of the equine glycoprotein hormone alpha-subunit gene reveals divergence in the mechanism of pituitary and placental expression

The equine glycoprotein hormone alpha-subunit gene is expressed in both pituitary and placenta, unlike that of all other nonprimate mammals studied, in which expression is limited to pituitary. Previous studies of the 5'-flanking region of the equine alpha-subunit promoter have revealed unique characteristics as well as similarities with the human alpha-subunit promoter, which demonstrates a similar pattern of tissue-specific expression. We have cloned and sequenced the equine alpha-subunit gene and have used tissue culture systems and transgenic mice to characterize its expression. Unlike the human promoter, the cloned equine alpha-subunit promoter failed to direct trophoblast-specific expression in either tissue culture or transgenic mouse models, suggesting an entirely different mechanism for expression. In contrast, the equine alpha-subunit promoter was able to direct gonadotroph expression in both tissue culture and transgenic mouse models. In alphaT3-1 cells, 550 base pair (bp) was sufficient for expression. This expression involves promoter elements identified in other species as playing a role in gonadotroph expression, but mutation of these elements reveals differences in their relative contributions to promoter activity. In mice, 2800 bp of 5'-flanking sequence allowed specific expression in gonadotrophs but not in thyrotrophs or placenta. The pattern of estrogen regulation observed in transgenic mice matched neither the repression that has been observed with human and bovine alpha-subunit promoters in transgenic mice nor the stimulation in mRNA levels reported in mares, suggesting a unique mechanism that is not recapitulated in the transgenic model. Thus the equine alpha-subunit promoter uses a combination of conserved and unique features of gene regulation to direct its pattern of tissue-specific expression.     

Nurse managed occupational health services without on-site clinical care delivery: a case example

1. In this program management/community network model of occupational health services, the occupational health nurse is responsible for managing program development and implementation, with vendors providing the clinical services. 2. Occupational health nurses' primary areas of responsibility are occupational health, disability case management, ergonomics, and health promotion. 3. Successful management of program outcomes requires the occupational health nurse to continually assess employee/business needs, maintain communication with employees and management, and partner with the environmental, health, and safety team, other functional work groups, and vendors. 4. Effective management of contracts becomes critical to the process beginning with clear service requirements through the delivery of quality services.     

Hallux valgus due to cuneiform-metatarsal instability

We present a biomechanical rationale for the treatment of severe hallux valgus deformity by realignment and arthrodesis of the first cuneiform metatarsal (CM) joint. We think that this severe hallux valgus deformity represents instability at the CM-1 articulation, since normal motion at that joint is very small. A forefoot compression test was used to assess the foot preoperatively. This was found not only to realign the first metatarsal, but also to align the metatarsophalangeal joint. A surgical procedure has evolved to include a transverse screw between the first and second metatarsal bases, as well as a second screw from first cuneiform to first metatarsal base, along with a small-volume bone graft of the CM-1 articulation. This is a modern variant of the Lapidus procedure.