GGA1 acts as a spatial switch altering amyloid precursor protein trafficking and processing

The beta-amyloid (Abeta) precursor protein (APP) is cleaved sequentially by beta-site of APP-cleaving enzyme (BACE) and gamma-secretase to release the Abeta peptides that accumulate in plaques in Alzheimer's disease (AD). GGA1, a member of the Golgi-localized gamma-ear-containing ARF-binding (GGA) protein family, interacts with BACE and influences its subcellular distribution. We now report that overexpression of GGA1 in cells increased the APP C-terminal fragment resulting from beta-cleavage but surprisingly reduced Abeta. GGA1 confined APP to the Golgi, in which fluorescence resonance energy transfer analyses suggest that the proteins come into close proximity. GGA1 blunted only APP but not notch intracellular domain release. These results suggest that GGA1 prevented APP beta-cleavage products from becoming substrates for gamma-secretase. Direct binding of GGA1 to BACE was not required for these effects, but the integrity of the GAT (GGA1 and TOM) domain of GGA1 was. GGA1 may act as a specific spatial switch influencing APP trafficking and processing, so that APP-GGA1 interactions may have pathophysiological relevance in AD.     

Dose-response relationship for a positive inotropic effect of insulin on isolated papillary muscle

Summary Beneficial effect of glucose and insulin on the myocardium are still a matter of discussion. The influence of insulin on isometric force of contraction of right ventricular papillary muscles of guinea pigs war studied. The papillary muscles were mounted vertically in a 95% O₂, 5% CO₂ modified Krebs-Hensuleit solution (31.5° C, 5.5 mM glucose) and stimulated 1/s. A positive inotropic effect of insulin was dedectable at a concentration of 5×10^–4 IU/ml insulin, was half maximal (52% above controle force of contraction) at 8×10^–3 IU/ml and maximal at 10^–1 IU/ml. The maximal positive inotropic effect was observed 4.7±0.6 min after addition of insulin. After the maximum there was a decrease to a steady state level of 109.8±8.5% of control (p<0.05) in 14.6±1.3 min. Higher glucose (16.5 mM) only shifted the half maximal positive inotropic effect to 5.5×10^–3 IU/ml insulin (n.s.). Inhibition of glycolysis with hypoxia or jodoacetate (5×10^–5 M) did not prevent the positive inotropic effect as known as 75% of control force was retained. When glucose transport was blocked with phlorizin (5×10^–3 M) or phloritin (5×10^–4 M) no positive inotropic action of insulin was observed. Therefore we conclude that the positive inotropic effect of insulin in isolated papillary muscles is mediated by inhanced glucose transport.     

The von Hippel–Lindau tumor suppressor gene is required for cell cycle exit upon serum withdrawal

The inactivation of the von Hippel–Lindau (VHL) tumor suppressor gene predisposes affected individuals to the human VHL cancer syndrome and is associated with sporadic renal cell carcinomas (RCC) and brain hemangioblastomas. VHL-negative 786–0 RCC cells are tumorigenic in nude mice which is suppressed by the reintroduction of VHL. Remarkably, this occurs without affecting the growth rate and cell cycle profile of these cells in culture. The 786–0 cell line, like many cancer cells, fails to exit the cell cycle upon serum withdrawal. Here, it is shown that reintroduction of the wild-type VHL gene restores the ability of VHL-negative RCC cancer cells to exit the cell cycle and enter G₀/quiescence in low serum. Both VHL-positive and VHL-negative RCC cells exit the cell cycle by contact inhibition. The cyclin-dependent kinase inhibitor, p27, accumulates upon serum withdrawal, only in the presence of VHL, as a result of the stabilization of the protein. We propose that the loss of wild-type VHL gene results in a specific cellular defect in serum-dependent growth control, which may initiate tumor formation. This is corrected by the reintroduction of wild-type VHL, implicating VHL as the first tumor suppressor involved in the regulation of cell cycle exit, which is consistent with its gatekeeper function in the kidney.     

The Arabidopsis repressor of light signaling, COP1, is regulated by nuclear exclusion: mutational analysis by bioluminescence resonance energy transfer

Bioluminescence resonance energy transfer (BRET) between Renilla luciferase and yellow fluorescent protein has been adapted to serve as a real-time reporter on protein-protein interactions in live plant cells by using the Arabidopsis Constitutive photomorphogenesis 1 (COP1) protein as a model system. COP1 is a repressor of light signal transduction that functions as part of a nuclear E3 ubiquitin ligase. COP1 possesses a leucine-rich nuclear-exclusion signal that resides in a domain implicated in COP1 dimerization. BRET was applied in conjunction with site-directed mutagenesis to explore the respective contributions of the nuclear-exclusion and dimerization motifs to the regulation of COP1 activity in vivo. One specific mutant protein, COP1(L105A), showed increased nuclear accumulation but retained the ability to dimerize, as monitored by BRET, whereas other mutations inhibited both nuclear exclusion and COP1 dimerization. Mutant rescue and overexpression experiments indicated that nuclear exclusion of COP1 protein is a rate-limiting step in light signal transduction.     

Effects of head positioning on cephalometric measurements

Objective

The goal of the present work was to evaluate different head positions for their effects on cephalometric analysis.

Materials and methods

Cephalograms were obtained from a skull phantom adjusted to various degrees of inclination (0°, 2°, 4°, 6°, 8°, 10°), lateral tilting (0°, 2°, 4°, 6°, 8°, 10°), and rotation (0°, 3°, 6°, 9°, 12°, 15°, and 21°). All these combinations resulted in a total of 210?digital cephalograms for assessment. On-screen analysis of these images by an orthodontist was completed within a few days.

Results

Lateral tilting and rotation revealed considerably stronger effects on the values measured than inclination. Starting at 2° of tilting or 3° of rotation, numerous parameters yielded deviations of ≥?2° or 2?mm from the baseline values. Increasing degrees of rotation showed more pronounced value changes than increasing degrees of lateral tilting. Skeletal and dental parameters whose landmarks are mainly located in the median plane were less susceptible than parameters whose reference points had to be averaged. Values of parameters located in the median plane were less affected by rotation when combined with higher degrees of initial lateral tilting (>?6°) than with lower ones.

Conclusion

The usefulness of cephalometric data becomes limited once the head is rotated by 3° or laterally tilted by 2°. Values obtained in the median plane are more stable than those obtained in peripheral locations. Various degrees of inclination applied to the skull phantom with its solid structures had no measurable effects. Accurate head positioning is an essential prerequisite for obtaining meaningful results from cephalograms based on reproducibly identifiable landmarks.     

Gesunderhaltung und Entlastung pflegender Angeh?riger von Demenzkranken durch ein ?initiales Case Management“

Background

When facing the well-known demographic development with an increasing number of people suffering from dementia, there is a need of programmes to support nursing relatives and care at home. Many support services have been established in the past few years but they are rarely used by the relatives and the patients. The purpose of the Lighthouse Project Ulm (ULTDEM Study) was to prove the effectiveness of a single advisory approach in order to provide support services after care level classification and to relieve the burden placed on relatives caring for family members suffering from dementia (“initial case management”).

Methods

The ULTDEM Study is a prospective, open, randomized, controlled, interventional study with different parallel outcome measures (burden of caring, quality of life and mood). After the randomization, the interventional group was given comprehensive, individual advice about available treatment possibilities for dementia patients. Control group participants received standard treatment. Inclusion criteria were application of a care level (0 or 1) as well as dementia diagnosis. All participants (patients/relatives) underwent an initial and a 6?month comprehensive assessment.

Results

Our results show that a single advisory approach does not lead to a significant difference in outcome measures in interventional and control groups. Those tendencies described have to be interpreted as clinically not relevant. Although utilization of support services increases, it remains similar in both study groups. A confirmatory interpretation has not been possible due to a lack of adjustment to the findings regarding multiple testing and an insufficient degree of recruitment. Possible causes will be discussed such as premature intervention during the course of the disease, a lack of intervention blinding, recruitment bias and lack of an influence on adherence with regard to the use of support services.

Implications

The study demonstrates that there is a substantial information deficit for persons affected by dementia and their relatives. Innovative ways still have to be developed to ensure that this information actually reaches the target audience.     

A Putative Novel Hepatitis E Virus Genotype 3 Subtype Identified in Rabbit,Germany 2016

Hepatitis E is an emerging viral disease that is the leading cause of viral hepatitis in the world. The vast majority of hepatitis E cases in developed countries are caused by zoonotic genotypes 3 and 4 of hepatitis E virus (HEV) for which pig and wild boar and to lesser extent rabbits are the main reservoir. According to recent reports rabbits are a source of human HEV infection and highlight the risk of zoonotic foodborne transmission. Here we report the molecular analysis of a novel HEV strain identified in a rabbit during a countrywide surveillance of rabbits and hares in Germany, 2016. The analysis of the complete genome reveals characteristics of a putative novel recombinant subtype of the species Orthohepevirus A within the clade of genotype 3 but not closely related to any known subtypes. Importantly, the genome of this strain possesses a nucleotide exchange in the overlapping region of open reading frames ORF2/ORF3 interfering with a broadly applied diagnostic real-time RT-PCR. In conclusion, a new type of HEV strain was identified in a German rabbit with atypical and novel sequence characteristics.     

The genome sequence of Clostridium tetani,the causative agent of tetanus disease

Tetanus disease is one of the most dramatic and globally prevalent diseases of humans and vertebrate animals, and has been reported for over 24 centuries. The manifestation of the disease, spastic paralysis, is caused by the second most poisonous substance known, the tetanus toxin, with a human lethal dose of approximately 1 ng/kg. Fortunately, this disease is successfully controlled through immunization with tetanus toxoid; nevertheless, according to the World Health Organization, an estimated 400,000 cases still occur each year, mainly of neonatal tetanus. The causative agent of tetanus disease is Clostridium tetani, an anaerobic spore-forming bacterium, whose natural habitat is soil, dust, and intestinal tracts of various animals. Here we report the complete genome sequence of toxigenic C. tetani E88, a variant of strain Massachusetts. The genome consists of a 2,799,250-bp chromosome encoding 2,372 ORFs. The tetanus toxin and a collagenase are encoded on a 74,082-bp plasmid, containing 61 ORFs. Additional virulence-related factors could be identified, such as an array of surface-layer and adhesion proteins (35 ORFs), some of them unique to C. tetani. Comparative genomics with the genomes of Clostridium perfringens, the causative agent of gas gangrene, and Clostridium acetobutylicum, a nonpathogenic solvent producer, revealed a remarkable capacity of C. tetani: The organism can rely on an extensive sodium ion bioenergetics. Additional candidate genes involved in the establishment and maintenance of a pathogenic lifestyle of C. tetani are presented.