Bax targets mitochondria by distinct mechanisms before or during apoptotic cell death: a requirement for VDAC2 or Bak for efficient Bax apoptotic function.

In non-apoptotic cells, Bak constitutively resides in the mitochondrial outer membrane. In contrast, Bax is in a dynamic equilibrium between the cytosol and mitochondria, and is commonly predominant in the cytosol. In response to an apoptotic stimulus, Bax and Bak change conformation, leading to Bax accumulation at mitochondria and Bak/Bax oligomerization to form a pore in the mitochondrial outer membrane that is responsible for cell death. Using blue native-PAGE to investigate how Bax oligomerizes in the mitochondrial outer membrane, we observed that, like Bak, a proportion of Bax that constitutively resides at mitochondria associates with voltage-dependent anion channel (VDAC)2 prior to an apoptotic stimulus. During apoptosis, Bax dissociates from VDAC2 and homo-oligomerizes to form high molecular weight oligomers. In cells that lack VDAC2, constitutive mitochondrial localization of Bax and Bak was impaired, suggesting that VDAC2 has a role in Bax and Bak import to, or stability at, the mitochondrial outer membrane. However, following an apoptotic stimulus, Bak and Bax retained the ability to accumulate at VDAC2-deficient mitochondria and to mediate cell death. Silencing of Bak in VDAC2-deficient cells indicated that Bax required either VDAC2 or Bak in order to translocate to and oligomerize at the mitochondrial outer membrane to efficiently mediate apoptosis. In contrast, efficient Bak homo-oligomerization at the mitochondrial outer membrane and its pro-apoptotic function required neither VDAC2 nor Bax. Even a C-terminal mutant of Bax (S184L) that localizes to mitochondria did not constitutively target mitochondria deficient in VDAC2, but was recruited to mitochondria following an apoptotic stimulus dependent on Bak or upon over-expression of Bcl-xL. Together, our data suggest that Bax localizes to the mitochondrial outer membrane via alternate mechanisms, either constitutively via an interaction with VDAC2 or after activation via interaction with Bcl-2 family proteins.

Magnetism in single metalloorganic complexes formed by atom manipulation.

The magnetic properties of molecular structures can be tailored by chemical synthesis or bottom-up assembly at the atomic scale. We used scanning tunneling microscopy to study charge and spin transfer in individual complexes of transition metals with the charge acceptor, tetracyanoethylene (TCNE). The complexes were formed on a thin insulator, Cu2N on Cu(100), by manipulation of individual atoms and molecules. The Cu2N layer decouples the complexes from Cu electron density, enabling direct imaging of the TCNE molecular orbitals as well as spin-flip inelastic electron tunneling spectroscopy. Results were obtained at low temperature down to 1 K and in magnetic fields up to 7 T in order to resolve splitting of spin states in the complexes. We also performed spin-polarized density functional theory calculations to compare with the experimental data. Our results indicate that charge transfer to TCNE leads to a change in spin magnitude, Kondo resonance, and magnetic anisotropy for the metal atoms.

Experimental demonstration of scanned spin-precession microscopy.

We present a new tool for imaging spin properties. We show that a spatially averaged spin signal, measured as a function of a scanned magnetic probe's position, contains information about the local spin properties. In this first demonstration we map the injected spin density in GaAs by measuring spin photoluminescence with a resolution of 1.2 µm. The ultimate limit of the technique is set by the gradient of the probe's field, allowing for a resolution beyond the optical diffraction limit. Such probes can also be integrated with other detection methods. This generality allows the technique to be extended to buried interfaces and optically inactive materials.

Photonic band structures of periodic arrays of pores in a metallic host: tight-binding beyond the quasistatic approximation.

We have calculated the photonic band structures of metallic inverse opals and of periodic linear chains of spherical pores in a metallic host, below a plasma frequency ωp. In both cases, we use a tight-binding approximation, assuming a Drude dielectric function for the metallic component, but without making the quasistatic approximation. The tight-binding modes are linear combinations of the single-cavity transverse magnetic (TM) modes. For the inverse-opal structures, the lowest modes are analogous to those constructed from the three degenerate atomic p-states in fcc crystals. For the linear chains, in the limit of small spheres compared to a wavelength, the results bear some qualitative resemblance to the dispersion relation for metal spheres in an insulating host, as calculated by Brongersma et al. [Phys. Rev. B 62, R16356 (2000)]. Because the electromagnetic fields of these modes decay exponentially in the metal, there are no radiative losses, in contrast to the case of arrays of metallic spheres in air. We suggest that this tight-binding approach to photonic band structures of such metallic inverse materials may be a useful approach for studying photonic crystals containing metallic components, even beyond the quasistatic approximation.

Proliferation of embryonic cardiomyocytes in zebrafish requires the sodium channel scn5Lab.

In mice, homozygous deletion of the cardiac sodium channel Scn5a results in defects in cardiac morphology and embryonic death before robust sodium current can be detected. In zebrafish, morpholino knockdown of cardiac sodium channel orthologs scn5Laa and scn5Lab perturbs specification of precardiac mesoderm and inhibits growth of the embryonic heart. It is not known which developmental processes are perturbed by sodium channel knockdown and whether reduced cell number is from impaired migration of cardiac progenitors into the heart, impaired myocyte proliferation, or both. We found that embryos deficient in scn5Lab displayed defects in primary cardiogenesis specific to loss of nkx2.5, but not nkx2.7. We generated kaede reporter fish and demonstrated that embryos treated with anti-scn5Lab morpholino showed normal secondary differentiation of cardiomyocytes at the arterial pole between 30 and 48 h post-fertilization. However, while proliferating myocytes were readily detected at 48 hpf in wild type embryos, there were no BrdU-positive cardiomyocytes in embryos subjected to anti-scn5Lab treatment. Proliferating myocytes were present in embryos injected with anti-tnnt2 morpholino to phenocopy the silent heart mutation, and absent in embryos injected with anti-tnnt2 and anti-scn5Lab morpholinos, indicating cardiac contraction is not required for the loss of proliferation. These data demonstrate that the role of scn5Lab in later heart growth does not involve contribution of the secondary heart field, but rather proliferation of cardiomyocytes, and appears unrelated to the role of the channel in cardiac electrogenesis.

Walkie talkies cause more electromagnetic interference to medical equipment than mobile phones.

It is generally realised that mobile phones can interfere with medical electrical equipment and many hospitals have policies which aim to minimise the risk to their patients from this cause. Walkie talkies are also used in hospitals, but very little information is available concerning their ability to interfere with hospital equipment. Two walkie talkies and three mobile phones have been used to study and compare interference in 29 items of patient-connected equipment in a large hospital. Test results show that with some equipment there will be signal distortion and/or false alarms, and a Power off/on Reset may be necessary. In rare cases, when equipment is subjected to extreme signal strengths, it is possible to observe equipment failure requiring the replacement of failed components. For these tests the walkie talkies were set at 4W of output power and they caused significantly more interference than the mobile phones. The observed effects have been classified according to the maximum distance at which they were observed and according to the criticality of the possible outcome for the patient. It is concluded that, except for emergency services, the use of walkie talkies should be restricted in hospital buildings.

Surface-enhanced plasmon splitting in a liquid-crystal-coated gold nanoparticle.

We show that, when a gold nanoparticle is coated by a thin layer of nematic liquid crystal, the nanoparticle surface has a strong effect on the director orientation, but, surprisingly, this deformation can enhance the surface plasmon splitting. We consider three plausible liquid crystal director configurations in zero electric field: boojum pair (north-south pole configuration), baseball (tetrahedral), and homogeneous. From the discrete dipole approximation, we find that the surface plasmon splitting is largest for the boojum pair, and this result is in good agreement with experiment.

Synchronization in disordered Josephson junction arrays: small-world connections and the Kuramoto model.

We study synchronization in disordered arrays of Josephson junctions. In the first half of the paper, we consider the relation between the coupled resistively and capacitively shunted junction (RCSJ) equations for such arrays and effective phase models of the Winfree type. We describe a multiple-time-scale analysis of the RCSJ equations for a ladder array of junctions with non-negligible capacitance in which we arrive at a second order phase model that captures well the synchronization physics of the RCSJ equations for that geometry. In the second half of the paper, motivated by recent work on small-world networks, we study the effect on synchronization of random, long-range connections between pairs of junctions. We consider the effects of such shortcuts on ladder arrays, finding that the shortcuts make it easier for the array of junctions in the nonzero voltage state to synchronize. In two-dimensional (2D) arrays we find that the additional shortcut junctions are only marginally effective at inducing synchronization of the active junctions. The differences in the effects of shortcut junctions in 1D and 2D can be partly understood in terms of an effective phase model.

Scaling properties of random walks on small-world networks.

Using both numerical simulations and scaling arguments, we study the behavior of a random walker on a one-dimensional small-world network. For the properties we study, we find that the random walk obeys a characteristic scaling form. These properties include the average number of distinct sites visited by the random walker, the mean-square displacement of the walker, and the distribution of first-return times. The scaling form has three characteristic time regimes. At short times, the walker does not see the small-world shortcuts and effectively probes an ordinary Euclidean network in d dimensions. At intermediate times, the properties of the walker shows scaling behavior characteristic of an infinite small-world network. Finally, at long times, the finite size of the network becomes important, and many of the properties of the walker saturate. We propose general analytical forms for the scaling properties in all three regimes, and show that these analytical forms are consistent with our numerical simulations.

Body composition changes following laparoscopic gastric banding for morbid obesity.

Most reports of outcome following obesity surgery report weight and co-morbidity changes only. We studied body composition changes in 17 adult patients (15 F, 2 M, age 43+/-2 years, range 28-58 years), with morbid obesity (initial BMI 40.4+/-4.9 kg/m(2), range 34.7-48.8) who were managed surgically by laparoscopically inserting an adjustable gastric band. Body composition was studied before and after surgery (mean interval of 909+/-51 days, range 441-1155 days) using anthropometry (abdominal circumference, AC, sum of four skinfold thicknesses, SFSUM), whole-body potassium counting (TBK), in vivo neutron activation analysis total body nitrogen (TBProtein) and whole-body dual-energy ray absorptiometry (total body percent fat TBF%, and total body bone mineral density TBBMD). Weight loss over the study period was 23.4+/-2.5 kg. ( p<0.0003) with an AC reduction of 20.0+/-4.5 cm ( p<0.008). Both SFSUM and TBF% were significantly reduced ( p<0.02 and p<0.0005 respectively). Both TBK and TBProtein after normalization for sex and height, were significantly ( p<0.0054 and p<0.001 respectively) reduced, but the ratio of loss of fat mass to fat-free mass, at 4.4:1 was usual for weight loss, and there was no significant changes in the ratio of potassium to protein. TBBMD, after normalization relative to a young same sex adult, was not significantly changed. In this group of patients, most of the substantial weight loss over a 2- to 3-year period was due to loss of fat mass, with relatively less reduction in the components of fat-free mass. Adjustable laparoscopic gastric banding induces fat loss without significant other deleterious effects on body composition.

Extensive mosaic structure revealed by the complete genome sequence of uropathogenic Escherichia coli.

We present the complete genome sequence of uropathogenic Escherichia coli, strain CFT073. A three-way genome comparison of the CFT073, enterohemorrhagic E. coli EDL933, and laboratory strain MG1655 reveals that, amazingly, only 39.2% of their combined (nonredundant) set of proteins actually are common to all three strains. The pathogen genomes are as different from each other as each pathogen is from the benign strain. The difference in disease potential between O157:H7 and CFT073 is reflected in the absence of genes for type III secretion system or phage- and plasmid-encoded toxins found in some classes of diarrheagenic E. coli. The CFT073 genome is particularly rich in genes that encode potential fimbrial adhesins, autotransporters, iron-sequestration systems, and phase-switch recombinases. Striking differences exist between the large pathogenicity islands of CFT073 and two other well-studied uropathogenic E. coli strains, J96 and 536. Comparisons indicate that extraintestinal pathogenic E. coli arose independently from multiple clonal lineages. The different E. coli pathotypes have maintained a remarkable synteny of common, vertically evolved genes, whereas many islands interrupting this common backbone have been acquired by different horizontal transfer events in each strain.

Characterizing the structure of small-world networks.

We give exact relations for small-world networks (SWN's) which are independent of the "degree distribution," i.e., the distribution of nearest-neighbor connections. For the original SWN model, we illustrate how these exact relations can be used to obtain approximations for the corresponding basic probability distribution. In the limit of large system sizes and small disorder, we use numerical studies to obtain a functional fit for this distribution. Finally, we obtain the scaling properties for the mean-square displacement of a random walker, which are determined by the scaling behavior of the underlying SWN.

Correlates of prepubertal bone mineral density in cystic fibrosis.

AIM: To examine early factors in bone mineral accretion in cystic fibrosis (CF). METHODS: In 22 prepubertal children with CF and mild lung disease, the relation between total body bone mineral density (BMD) and measures of body composition, biochemistry, lung function, and physical activity was studied. RESULTS: There was a non-significant mild reduction in mean total body BMD. No relation was found between BMD and anthropometric indices, fat free soft tissue, degree of lung disease, degree of fat malabsorption, dietary energy intake, or level of physical activity. Significant impairments in physical growth were apparent in this population and were found to correlate with degree of lung disease. CONCLUSION: A CF specific factor appears unlikely to be associated with the osteopenia commonly found in CF. Careful attention to general aspects of lifestyle and nutrition is recommended to maximise bone mineral accretion in this population.

Exact results and scaling properties of small-world networks

We study the distribution function for minimal paths in small-world networks. We derive its general form, which we verify numerically, and also discuss its scaling properties. Using the general form of this distribution function, we derive exact expressions for the average minimal distance &lmacr; and its variance sigma(2). Finally, we study the limit of large system sizes and obtain some approximate results.

Genetic analysis of Escherichia coli K1 gastrointestinal colonization.

Strains of Escherichia coli expressing the K1 polysaccharide capsule colonize the large intestine of newborn infants, and are the leading cause of Gram-negative septicaemia and meningitis in the neonatal period. We used signature-tagged mutagenesis (STM) to identify genes that E. coli K1 requires to colonize the gastrointestinal (GI) tract. A total of 2140 mTn5 mutants was screened for their capacity to colonize the GI tract of infant rats, and 16 colonization defective mutants were identified. The mutants have transposon insertions in genes affecting the synthesis of cell surface structures, membrane transporters, transcriptional regulators, enzymes in metabolic pathways, and in genes of unknown function, designated dgc (defective in GI colonization). Three dgcs are absent from the whole genome sequence of E. coli K-12, although related sequences are found in other pathogenic strains of E. coli and in Shigella flexneri. Additionally, immunohistochemistry was used to define the nature of the colonization defect in five mutants including all dgc mutants. STM was successfully applied to examine the factors involved in E. coli K1 colonization, and the findings are relevant to the pathogenesis of other enteric infections.

Total body dual X-ray absorptiometry is a good measure of both fat mass and fat-free mass in liver cirrhosis compared to "gold-standard" techniques. Melbourne Liver Group.

Liver cirrhosis is a condition in which overnutrition, edema, and undernutrition can coexist simultaneously, or successively, over a period of time, giving rise to alterations in body composition, as well as systemic and multiorgan manifestations. We undertook a cross-sectional study of body composition in 198 adult patients with liver cirrhosis (140 males, mean age 53.6, range 31-85 years; and 58 females, mean age 58.4, range 36-79 years). The patients had cirrhosis of differing etiology and different stages of severity. They were gathered from seven different hospital clinics in the city of Melbourne, Australia, but all the body composition measurements were performed in one body composition laboratory. A variety of body composition techniques were used to identify which commonly available ones could best assess both fat-free mass and fat mass relative to a criterion "gold-standard" method available in a specialist laboratory. A gold-standard fat-free mass (FFMGS) was defined as the sum of total body protein, measured by in vivo neutron activation analysis (IVNA), plus total body water, measured by D2O dilution, plus bone mineral content, measured by dual X-ray absorptiometry (DXA). A gold-standard fat mass (FATGS) was defined as the difference between body weight and FFMGS. "Usual" fat mass and fat-free mass were defined by different techniques including DXA, anthropometry (ANT), single-frequency bioelectrical impedance (SFBIA), multiple-frequency bioelectrical impedance spectroscopy (MFBIA), and whole body gamma counting (TBK). The FFMGS was overhydrated in both sexes, relative to the usual value of 0.73, but women were significantly overhydrated compared to men. Relative to the gold-standard deuterium oxide dilution method for measuring total body water, SFBIA slightly overestimated TBW, whereas MFBIA slightly underestimated TBW, with both methods having wide limits of agreement for any single estimate. In comparing FFM to FFMGS, only DXA showed a small negative bias, in both males and females, with modest limits of agreement for any single estimate. All other methods showed a large negative bias (ANT, SFBIA, and MFBIA) or a large positive bias (TBK) relative to FFMGS, with wide limits of agreement. In comparing FAT with the FATGS, only DXA showed a small positive bias, in both males and females, with modest limits of agreement for any single estimate. All other methods showed a large positive bias (ANT, SFBIA, and MFBIA) or a large negative bias (TBK) relative to FATGS, with wide limits of agreement. In cirrhosis, DXA is a good and widely available method to assess both fat mass and fat-free mass. However, it cannot give information about the quality of the FFM, particularly its water content. The bedside methods of anthropometry and bioelectrical impedance, both SFBIA and MFBIA, are poor methods of measuring body composition in patients with liver cirrhosis, whereas whole body gamma counting, although not widely available, also significantly differs from the gold-standard method of assessment of fat-free mass and fat mass in liver cirrhosis.

Criminal investigation of child sexual abuse: a comparison of cases referred to the prosecutor to those not referred.

OBJECTIVE: The present study sought to identify characteristics of child sexual abuse cases which differentiate cases referred for criminal prosecution ("criminal-action") from those not referred ("dropped") by investigators. METHOD: The study sample consisted of 1043 children who completed a forensic interview for sexual abuse that allegedly occurred at the hands of an adult between January 1, 1993 and December 31, 1996 in Bernalillo County of New Mexico. Data was systematically obtained from forensic interview files and offender records at the local prosecutor's office. RESULTS: Differences between criminal-action and dropped cases were found in relation to the children (age, sex and ethnicity), the alleged offenders (age, sex and relationship to child), and the case characteristics (disclosure and injury to the child). CONCLUSIONS: The present study provided insight into the characteristics of a previously ignored population (reported child sexual abuse cases that are not referred for prosecution). Recommendations are made to address the needs of these children and their families.

Comparison of total body chlorine, potassium, and water measurements in children with cystic fibrosis.

BACKGROUND: Symptoms of cystic fibrosis (CF) may limit the utility of total body chlorine (TBCl) and total body potassium (TBK) measurements for assessing body fluid compartments of children. OBJECTIVE: This study assessed relations among independent measurements of TBCl, TBK, and total body water (TBW) in children with CF. DESIGN: We compared cross-sectional measurements of TBCl by in vivo neutron activation analysis, TBK by whole-body counting of (40)K, TBW by D(2)O dilution [TBW(D(2)O)], and TBW from TBCl and TBK [TBW(Cl + K)] in 19 prepubertal children (13 boys) aged 7.6-12.5 y who had mild symptoms of CF. Body-composition measurements were compared with data from previous studies of healthy children. RESULTS: Subjects with CF had deficits in TBCl, TBK, TBW, and body weight compared with control reference data (P < 0.05). The ratios (TBCl + TBK)/TBW and TBCl/TBK were not significantly different from control reference values, and plasma chlorine and potassium concentrations were within control reference ranges. The sum of TBCl and TBK correlated with TBW(D(2)O) (r(2) = 0.79, P < 0.001), and TBW(Cl + K) correlated with TBW(D(2)O) (r(2) = 0.78, P < 0.001). TBW(Cl + K) was similar to TBW(D(2)O) (mean +/- SEM: 19.0 +/- 0.5 compared with 19.4 +/- 0.5 L; NS). CONCLUSIONS: Prepubertal children with mild symptoms of CF can develop deficits in TBCl, TBK, and TBW that reflect chronic energy malnutrition. Mild symptoms of CF do not appear to affect normal relations among TBCl, TBK, and TBW. Measurements of TBCl and TBK may be used to assess body fluid compartments in these patients.

Familial support as perceived by adult victims of childhood sexual abuse.

With the increasing popularity of family therapies in cases of childhood sexual abuse (CSA), many studies have discussed the importance of familial support in the successful recovery of victims. Therefore, a systematic study exploring family support of victims was undertaken to shed light on the level of family support perceived by adult victims of CSA. It was anticipated that victims of intrafamilial CSA would report more family dissatisfaction and perceive less family support than extrafamilial victims. As hypothesized, intrafamilial victims reported significantly more general family dissatisfaction and specifically, perceived significantly less father, parent, and brother support. Additionally, the gender of the victim played a significant role: Male victims reported less father, parent and sister support and less family protectiveness after disclosure when compared to female victims. Based on the results, clinicians should be aware that issues of gender and relatedness may play a significant part in the effectiveness of their CSA clients' family support systems.