The crystal structure of HIV-1 Nef protein bound to the Fyn kinase SH3 domain suggests a role for this complex in altered T cell receptor signaling

BACKGROUND: Human immunodeficiency virus (HIV) Nef protein accelerates virulent progression of acquired immunodeficiency syndrome (AIDS) by its interaction with specific cellular proteins involved in signal transduction and host cell activation. Nef has been shown to bind specifically to a subset of the Src family of kinases. The structures of free Nef and Nef bound to Src homology region 3 (SH3) domain are important for the elucidation of how the affinity and specificity for the Src kinase family SH3 domains are achieved, and also for the development of potential drugs and vaccines against AIDS. RESULTS: We have determined the crystal structures of the conserved core of HIV-1 Nef protein alone and in complex with the wild-type SH3 domain of the p59fyn protein tyrosine kinase (Fyn), at 3.0 A resolution. Comparison of the bound and unbound Nef structures revealed that a proline-rich motif (Pro-x-x-Pro), which is implicated in SH3 binding, is partially disordered in the absence of the binding partner; this motif only fully adopts a left-handed polyproline type II helix conformation upon complex formation with the Fyn SH3 domain. In addition, the structures show how an arginine residue (Arg77) of Nef interacts with Asp 100 of the so-called RT loop within the Fyn SH3 domain, and triggers a hydrogen-bond rearrangement which allows the loop to adapt to complement the Nef surface. The Arg96 residue of the Fyn SH3 domain is specifically accommodated in the same hydrophobic pocket of Nef as the isoleucine residue of a previously described Fyn SH3 (Arg96-->lle) mutant that binds to Nef with higher affinity than the wild type. CONCLUSIONS: The three-dimensional structures support evidence that the Nef-Fyn complex forms in vivo and may have a crucial role in the T cell perturbating action of Nef by altering T cell receptor signaling. The structures of bound and unbound Nef reveal that the multivalency of SH3 binding may be achieved by a ligand induced flexibility in the RT loop. The structures suggest possible targets for the design of inhibitors which specifically block Nef-SH3 interactions.     

Nuclear cardiology, Part III: Scintigraphic evaluation of cardiac perfusion

OBJECTIVE: After reading Part III of this series of nuclear cardiology articles, the technologist should be able to: (a) compare and contrast radiopharmaceuticals used for myocardial perfusion imaging; (b) describe imaging protocols used for detecting coronary artery disease; and (c) describe imaging patterns seen following reconstruction of myocardial images.     

Is compulsive internet use related to sensitivity to reward and punishment,and impulsivity?

Aim of the present study was to examine whether the personality correlates sensitivity to reward and to punishment, and impulsivity predict compulsive internet use (CIU). Furthermore, the predictive value of these personality correlates was compared to the predictive value of factors relating to psychosocial wellbeing. The results showed that particularly rash spontaneous impulsivity predicts CIU and that this personality factor is more important than psychosocial wellbeing factors. Sensitivity to reward, which is supposed to play a role in craving processes associated with substance abuse and eating disorders, could not be related to CIU. The data suggest that internet users who are characterized by an impulsive personality feature, are less able to control their use of the internet, which makes them more vulnerable to develop CIU.     

Pulsed corona for breaking up air bubbles in water

The injection of gas into water, through a hollow needle, gives a stream of bubbles. These bubbles can be effectively broken up by application of HV pulses. Our experimental work shows that this mechanism works in demineralized water and even in conducting tap water if the pulses have a short rise time. Bubble diameters are as low as 50 μm. Our pulse source uses either a HV tetrode or a spark gap as switching element. Extremely small bubbles are produced at a high rate and a low power input. Applications for this method can be found in chemical process technology     

Information management in an intensive care unit supported by AIDA

In an internal intensive care unit (ICU), management data is needed for various reasons. For instance, the allocation of resources needs timely information, so management can anticipate changes quickly. Using AIDA, a fourth-generation software package, a simple but adequate system has been created, with which day-to-day operational data is managed, and management information is generated.     

Selective capacity of glass-wool filtration for the separation of human spermatozoa with condensed chromatin: a possible therapeutic modality for male-factor cases?

PURPOSE: The aim of this study was to evaluate chromatin condensation of human spermatozoa following swim-up compared to glass-wool separation. Semen aliquots from men attending an andrological outpatient clinic were processed by means of a swim-up procedure and glass-wool filtration. Chromatin condensation was recorded using aniline blue staining and results were reported according to color intensity of stained sperm heads. Morphometric measurements of sperm heads were performed on stained sperm samples. RESULTS: Glass-wool filtration resulted (i) in a significantly higher total motile sperm count (P < 0.0005) compared to swim-up and (ii) in a significantly higher percentage of normal chromatin-condensed spermatozoa compared to the ejaculate (P < 0.01). CONCLUSION: In contrast, comparing swim-up to the ejaculate, the percentage of matured nuclei (unstained spermatozoa) retrieved following swim-up was significantly lower (P < 0.005). Glass-wool filtration separates human spermatozoa according to motility and size of the sperm head. The size of the sperm head closely correlated with the chromatin condensation quality.     

Prediction of functional outcome after myocardial infarction using BMIPP and sestamibi scintigraphy

We determined the predictive value of combined beta-methyl iodophenyl pentadecanoic acid (BMIPP) and sestamibi scintigraphy for the functional outcome after myocardial infarction and compared the value of this approach with dobutamine echocardiography. METHODS: Rest BMIPP, rest sestamibi and low-dose dobutamine echocardiographic studies were obtained in 18 patients 4 to 10 days after infarction (mean 6.7 +/- 2.0 days). Six months later, a rest echocardiographic study was performed to assess functional outcome. RESULTS: Wall motion improved in 27/33 segments (82%) which showed mismatching but not in 19/21 segments (90%) with matched defects (p < 0.001). The accuracy of combined BMIPP and sestamibi SPECT in predicting segmental functional outcome was higher (85%) than that of sestamibi uptake alone (77%). Wall motion improved in 16/20 segments (80%) showing contractile reserve and not in 21/34 segments (63%) with the negative dobutamine test, giving an accuracy of 69% for dobutamine echocardiography. Combination of the two techniques resulted in higher positive (94%) and negative predictive values (94%). CONCLUSION: Mismatching of BMIPP and sestamibi uptake is predictive for long-term functional recovery after acute myocardial infarction. In contrast, segments with matched defects contain only scar tissue. Combined BMIPP and sestamibi scintigraphy offers increased accuracy compared to dobutamine echocardiography.     

New aspects of sperm-zona pellucida binding

Sperm-zona pellucida binding tests have become a widely used diagnostic application for clinicians to obtain guidance in so far as the therapeutic approach of the subfertile couple is concerned. Expanding the oocyte sources is imperative to ensure the consistent use of sperm-zona binding assays. Sources include oocytes derived from post-mortem ovaries, inseminated non-fertilized IVF oocytes and recycled hemizonae. Identification of specific gamete dysfunction is one of the most formidable tasks and fertilization disorders due to defective sperm-zona pellucida interaction are relatively common in the clinical practice, thereby emphasizing the importance of sperm-zona binding tests as diagnostic/predictive tests. Independent publications from Norfolk (USA), Melbourne (Australia), Tygerberg (South Africa) and Israel of highly comparable results confirm that sperm-zona binding tests are good predictors of fertilization. Studies using solubilized human pellucida recently evaluated the influence of solubilized human pellucida on spermatozoa during the capacitational process and subsequent sperm-zona binding. Involvement of G protein and carbohydrate moieties in sperm-zona pellucida binding emphasized the biological and biochemical properties of lectin and have afforded much weight to their employment as membrane probes to evaluate cell surface components. Attention has been focused on the alterations of sperm surface receptors (oligosaccharides) during the differential pathway, epididymal transit and capacitation.     

Statistics of star-shaped molecules III. Stars with branched nuclei

Number and weight average molecular weights and scattering behaviour of star molecules with extended branched nuclei are calculated by application of cascade theory. The nuclei considered arise from random polycondensation of monomers of the A₃ or of the A—B/C type. Nuclei of the first type are characterised by very large molecular polydispersities (M_w/M_n α M_w), while nuclei of the second type have less broad molecular weight distributions (M_w/M_n α M_n). The rays of the stars are assumed to be either monodisperse or to obey the Schulz—Flory “most probable” length distribution. Analytic expressions are given for M_w, M_n, 〈S²〉_z and the particle scattering factor P_z(h) which was averaged over the ensemble. The results are compared with stars of spherical and uniform nuclei whose molecular weights and mean square radii of gyration equal M_wN and 〈S²〉_zN from the other two types of nuclei. In the limit of very large ray lengths the scattering behaviour is determined solely by the number of branches z. At shorter chain length of the rays structure and polydispersity of the nuclei have marked influence. This influence is still easily noticed from the angular dependence of scattered light at chain lengths where no differences in the 〈S²〉_z versus M_w plot are detectable. The mean square radius of gyration depends only weakly on the number of rays and eventually becomes independent of it if z 15. In that limit 〈S²〉_z depends virtually on the length of the rays alone and its distribution. Stars whose rays have a most probable length distribution exhibit 〈S²〉_z values twice as large as stars with monodisperse rays. A procedure is suggested and discussed for the determination of the number and length of the rays if the scattering behaviour of the isolated nucleus and the isolated linear chains is known.     

Treatment of mild hyperhomocysteinemia in vascular disease patients

Mild hyperhomocysteinemia is recognized as a risk factor for premature arteriosclerotic disease. A few vitamins and other substances have been reported to reduce blood homocysteine levels, but normalization of elevated blood homocysteine concentrations with any of these substances has not been reported. Therefore, we screened 421 patients suffering from premature peripheral or cerebral occlusive arterial disease by oral methionine loading tests for the presence of mild hyperhomocysteinemia. Thirty-three percent of patients with peripheral and 20% of patients with cerebral occlusive arterial disease were identified with mild hyperhomocysteinemia (14% of the men, 34% of the premenopausal women, and 26% of the postmenopausal women). Mildly hyperhomocysteinemic patients were administered vitamin B6 250 mg daily. After 6 weeks methionine loading tests were again assessed to evaluate the effect of treatment. Patients with nonnormalized homocysteine concentrations were further treated with vitamin B6 250 mg daily and/or folic acid 5 mg daily and/or betaine 6 g daily, solely or in any combination. Vitamin B6 treatment normalized the afterload homocysteine concentration in 56% of the treated patients (71% of the men, 45% of the premenopausal women, and 88% of the postmenopausal women). Further treatment resulted in a normalization of homocysteine levels in 95% of the remaining cases. Thus, mild hyperhomocysteinemia, which is frequently encountered in patients with premature arteriosclerotic disease, can be reduced to normal in virtually all cases by safe and simple treatment with vitamin B6, folic acid, and betaine, each of which is involved in methionine metabolism.