Structure and function in rhodopsin: rhodopsin mutants with a neutral amino acid at E134 have a partially activated conformation in the dark state

The Glu-134-Arg-135 residues in rhodopsin, located near the cytoplasmic end of the C helix, are involved in G protein binding, or activation, or both. Furthermore, the charge-neutralizing mutation Glu-134 to Gln-134 produces hyperactivity in the activated state and produces constitutive activity in opsin. The Glu/Asp-Arg charge pair is highly conserved in equivalent positions in other G protein-coupled receptors. To investigate the structural consequences of charge-neutralizing mutations at Glu-134 and Arg-135 in rhodopsin, single spin-labeled side chains were introduced at sites in the cytoplasmic domains of helices C (140), E (227), F (250), or G (316) to serve as "molecular sensors" of the local helix bundle conformation. In each of the spin-labeled rhodopsins, a Gln substitution was introduced at either Glu-134 or Arg-135, and the electron paramagnetic resonance spectrum of the spin label was used to monitor the structural response of the helix bundle. The results indicate that a Gln substitution at Glu-134 induces a photoactivated conformation around helices C and G even in the dark state, an observation of potential relevance to the hyperactivity and constitutive activity of the mutant. In contrast, little change is induced in helix F, which has been shown to undergo a dominant motion upon photoactivation. This result implies that the multiple helix motions accompanying photoactivation are not strongly coupled and can be induced to take place independently. Gln substitution at Arg-135 produces only minor structural changes in the dark- or light-activated conformation, suggesting that this residue is not a determinant of structure in the regions investigated, although it may be functionally important.     

Assessment of a radioactive waste disposal site at Enewetak Atoll

The 43 nuclear tests conducted at Enewetak Atoll by the United States between 1948 and 1958 produced close-in fallout that contaminated the islands and lagoon of the atoll with radioactive fission and activation products, and unfissioned nuclear fuel. In 1972, the U.S. government announced that it would conduct a cleanup and restoration operation to return the atoll to the Enewetak people. The radiological cleanup began in 1977 and lasted to 1980 and focused on reducing the concentration of the transuranium elements (238,239,240Pu and 241Am = TRU) in soils on some of the islands that might eventually be used for residence or for subsistence agricultural. The cleanup plan called for relocating soil and some other contaminated debris to Runit Island on the eastern perimeter of the Atoll. Some of the contaminated soil was mixed with cement and the mixture placed below the water level in the Cactus Crater that was formed by a nuclear explosion in 1958. The remainder of the contaminated material was mixed with concrete and placed above ground over the crater in the shape of a dome. A concrete cap was constructed over the dome of soil. Concern has been expressed by the people of Enewetak and by others over the possible aquatic impacts from the radionuclides entombed in the crater. A National Academy of Sciences committee examined the dome and concluded that the containment structure and its contents present no credible health hazard to the people of Enewetak, either now or in the future. The committee suggested that "at least part of the radioactivity contained in the structure is available for transport to the groundwater and subsequently to the lagoon and it is important to determine whether this pathway may be a significant one." Therefore, a surveillance program was started in 1980, in conjunction with other research efforts, to study the radionuclides in samples of fish, groundwater, and lagoon seawater. Our data and conclusions support the findings suggested by the National Academy committee over a decade ago in that any assumption of rapid remobilization of all or any of the dome's transuranics or other radionuclides is an extreme one. Any fear that this structure contains amounts of activity whose release would cause damage to the environment that will result in greater effect on human health is unfounded.     

In vivo bone strain patterns in the zygomatic arch of macaques and the significance of these patterns for functional interpretations of craniofacial form

It has been proposed that the mammalian facial skeleton is optimized for countering or dissipating masticatory stress. As optimized load-bearing structures by definition exhibit maximum strength with a minimum amount of material, this hypothesis predicts that during chewing and biting there should be relatively high and near uniform amounts of strain throughout the facial skeleton. If levels of strain in certain areas of the facial skeleton are relatively low during these behaviors, this indicates that the amount of bone mass in these areas could be significantly reduced without resulting in the danger of structural failure due to repeated masticatory loads. Furthermore, and by definition, this indicates that these areas are not optimized for countering masticatory stress, and instead their overall morphology and concentration of bone mass has most likely been selected or influenced mainly by factors unrelated to the dissipation or countering of chewing and biting forces. An analysis of in vivo bone strain along the lateral aspect of the zygomatic arch of macaques indicates the clear absence of a high and near uniform strain environment throughout its extent. Instead, there is a steep strain gradient along the zygomatic arch, with the highest strains along its anterior portion, intermediate strains along its middle portion, and the lowest strains along its posterior portion. These data, in combination with earlier published data (Hylander et al., 1991), indicate that levels of functional strains during chewing and biting are highly variable from one region of the face to the next, and therefore it is unlikely that all facial bones are especially designed so as to minimize bone tissue and maximize strength for countering masticatory loads. Thus, the functional significance of the morphology of certain facial bones need not necessarily bear any important or special relationship to routine and habitual cyclical mechanical loads associated with chewing or biting. Furthermore, the presence of these steep strain gradients within the facial skeleton suggests that the amount of bone mass in the low-strain areas may be largely determined by factors unrelated to processes frequently referred to as "functional adaptation," or conversely, that the "optimal strain environment" of bone varies enormously throughout the facial skeleton (cf., Rubin et al., 1994). Based solely on anatomical considerations, it is likely that the zygomatic arch is bent in both the parasagittal and transverse planes and twisted about its long axis. Due to constraints on rosette position, the strain data are incapable of determining if one or more of these loading conditions predominate. Instead, the strain data simply provide limited support for the possible presence of all of these loading regimes. Finally, as the masseter muscle is concentrated along the anterior portion of the zygomatic arch and as the arch has fixed ends, the largest shearing forces and the largest bending and twisting moments are located along its anterior portion. This in turn explains why the largest strains are found along the anterior portion of the zygomatic arch.     

Attention-deficit hyperactivity symptomatology after traumatic brain injury: a prospective study

OBJECTIVE: To study prospectively the course of attention-deficit hyperactivity (ADH) symptomatology in children and adolescents after traumatic brain injury (TBI). It was hypothesized that ADH symptomatology would be significantly related to severity of injury. METHOD: Subjects were children (n = 50) aged 6 to 14 years at the time they were hospitalized after TBI. The study used a prospective follow-up design. Assessments of preinjury psychiatric, behavioral, socioeconomic, family functioning, and family psychiatric history status were conducted. Severity of injury was assessed by standard clinical scales, and neuroimaging was analyzed. RESULTS: The main finding of this study was that change in ADH symptomatology in the first 2 years after TBI in children and adolescents was significantly related to severity of injury. Overall ADH symptomatology during the study was significantly related to a measure of family dysfunction when family psychiatric history, socioeconomic status, and severity of injury were controlled. CONCLUSION: The presence of a positive "dose-response" relationship between severity of injury and change in ADH symptoms, present from the 3-month assessment, was consistent with an effect directly related to brain damage.     

Correlates of gonococcal infection and of antimicrobial-resistant Neisseria gonorrhoeae among female sex workers, Republic of the Philippines, 1996-1997

From 1994 to 1997, the proportion of Neisseria gonorrhoeae highly resistant to ciprofloxacin (MIC >/=4 microg/mL) increased substantially among female sex workers (FSWs) in the Philippines. Among 1499 Filipina FSWs, we evaluated factors associated with gonococcal infection and with gonococcal antimicrobial resistance. By multivariate analysis, gonococcal infection was associated with sex with a new client, self-prescribed prophylactic antimicrobial use, work in a brothel, and inconsistent condom use and was negatively associated with registration status and vaginal hygiene practices. Factors associated with ciprofloxacin-resistant gonococci included: marital status, living alone, duration of sex work, and clinic site. Further, gonococci highly resistant to ciprofloxacin were isolated from 10 (11.5%) of 87 FSWs reporting self-prescribed antimicrobial use versus 44 (3.4%) of 1295 reporting no antimicrobial use (P<.001). Self-prescribed prophylactic antimicrobial use and inconsistent condom use could be important factors in the continued emergence of gonococcal antimicrobial resistance in the Philippines.     

Control of cell growth. II. Requirement of thyroid hormones for the in vivo estrogen-dependent growth of rat pituitary tumor cells

A subclone of rat pituitary tumor cells, designated GH3/C14, was isolated from the parent population of GH3 pituitary cells and was estrogen-dependent for growth in vivo. GH3/C14 cells inoculated into host animals formed tumors in intact females, estrogen-treated ovariectomized females, and estrogen-treated males, but not in untreated intact or castrated males, or untreated ovariectomized females. Exogenous treatment with estrogens supported tumor formation in male animals. Radioimmunoassay of the average serum estradiol concentrations that support tumor growth in intact females, estradiol-treated intact females, and estradiol-treated intact males gave values of 41 +/- 4, 1,130 +/- 150, and 730 +/- 140 pg/ml, respectively. Tumor formation by GH3/C14 cells inoculated into male animals was not supported by either exogenous progesterone or hydrocortisone acetate. Further, these two steroid hormones had no significant effect on the estrogen-promoted growth in males or females. Exogenous testosterone treatment promoted tumor formation in males and ovariectomized females, but dihydrotestosterone was completely ineffective. Radioimmunoassay of the serum from tumor-bearing animals and the medium from the cells in culture showed that the cells produced growth hormone in vivo and in vitro but did not produce measurable amounts of luteinizing hormone or follicle-stimulating hormone. The groth of GH3/C14 cells in culture was examined in medium without added estrogen, and with estradiol added to the level of either the normal intact female rat or the estradiol-treated animals. No direct growth stimulation by estrogens could be detected in culture; the data suggested an indirect growth control mechanism.