Mutant and chimeric recobinant plasminogen activatorsproduction in eukaryotic cellsand preliminary characterization

Mutant urokinase-type plasminogen activator (u-PA) genes and hybrid genes between tissue-type plasminogen activator (t-PA) and u-PA have been designed to direct the synthesis of new plasminogen activators and to investigate the structure-function relationship in these molecules. The following classes of constructs were made starting from cDNA encoding human t-PA or u-PA: 1) u-PA mutants in which the Arg156 and Lys158 were substituted with threonine, thus preventing cleavage by thrombin and plasmin; 2) hybrid molecules in which the NH2-terminal regions of t-PA (amino acid residues 1-67, 1-262, or 1-313) were fused with the COOH-terminal region of u-PA (amino acids 136-411, 139-411, or 195-411, respectively); and 3) a hybrid molecule in which the second kringle of t-PA (amino acids 173-262) was inserted between amino acids 130 and 139 of u-PA. In all cases but one, the recombinant proteins, produced by transfected eukaryotic cells, were efficiently secreted in the culture medium. The translation products have been tested for their ability to activate plasminogen after in situ binding to an insolubilized monoclonal antibody directed against urokinase. All recombinant enzymes were shown to be active, except those in which Lys158 of u-PA was substituted with threonine. Recombination of structural regions derived from t-PA, such as the finger, the kringle 2, or most of the A-chain sequences, with the protease part or the complete u-PA molecule did not impair the catalytic activity of the hybrid polypeptides. This observation supports the hypothesis that structural domains in t-PA and u-PA fold independently from one to another

The Burden Patients with Myasthenia Gravis Experience in Terms of Breathing, Fatigue, Sleep, Mental Health, Discomfort and Usual Activities in Comparison to the General Population

Introduction: Myasthenia gravis (MG) is a rare neuromuscular disorder marked by a variable combination of weakness of eye, bulbar, respiratory, axial, and limb muscles. This study compared the experience of people with MG regarding breathing, fatigue, sleep, pain/discomfort, mental health, and usual activities with the general population. Methods: The MyRealWorld-MG digital, multinational study enrolled patients with MG and collected demographics, PROMIS-Dyspnea, PROMIS-Sleep Disturbance, FACIT-Fatigue, EQ-5D-5L, Health Utilities Index (HUI-3), Hospital Anxiety and Depression Scale (HADS), MG-Activities of Daily Living (MG-ADL), and MG-Quality-of-Life (MG-QoL-15r). Comparisons with the general population were based on PROMIS population norms, published literature, or on data from a digital, multinational, observational study which enrolled a representative sample of the general population (POPUP). Results: In MyRealWorld-MG (N = 2074), patients experienced higher intensity, frequency, and duration of PROMIS shortness of breath than a US population (p &lt; 0.0001). Patients with MG had higher PROMIS-Sleep Disturbance scores than POPUP (53.7 vs 50.0, p &lt; 0.0001), and 54.9% of patients had clinically severe FACIT-Fatigue scores vs 6.8% in POPUP (p &lt; 0.0001). Among patients with MG, 69.6% and 18.5% had moderate-to-severe HADS-Anxiety and HADS-Depression compared to 20.3% and 6.9% in POPUP (p &lt; 0.001). Statistically significant and strong associations were found between fatigue, sleep, dyspnea, usual activities, and emotions. All outcomes worsened with more severe disease. Conclusion: A considerable burden was observed in this comparison of breathing, sleep, fatigue, mental health, and usual activities between patients with MG and the general population, using data from two international studies and published population norms. Even mildly affected patients had significantly worse outcomes than the general population.</p

People Diagnosed with Myasthenia Gravis have Lower health-related quality of life and Need More Medical and Caregiver Help in Comparison to the General Population:Analysis of Two Observational Studies

Introduction: Myasthenia gravis (MG) is a neuromuscular disease causing extreme muscular fatigue, triggering problems with vision, swallowing, speech, mobility, dexterity, and breathing. This analysis intended to estimate the health-related quality-of-life impact, the medical burden, and the need for caregiver help of people diagnosed with MG. Methods: MyRealWorld-MG (MRW) is an observational study among adults diagnosed with MG in 9 countries. The General Population Norms (POPUP) observational study enrolled representative members of the general population in 8 countries. In both digital studies, respondents entered personal characteristics and provided data on medical conditions, EQ-5D-5L, HUI3, MG-Activities of Daily Living (MG-ADL), sick leave, caregiver help, and medical care utilization. Results: In MRW (n = 1859), 58.4% of respondents had moderate-to-severe MG. Average utility values were lower in MRW versus POPUP (0.739 vs. 0.843 for EQ-5D-5L; 0.493 vs. 0.746 for HUI3), and declined with more severe disease (0.872, 0.707, 0.511 EQ-5D-5L utilities and 0.695, 0.443, 0.168 HUI3 utilities for mild, moderate, and severe MG, respectively). Taking sick leave in the past month was 2.6 times more frequent among people diagnosed with MG compared to the general population (34.4% vs. 13.2%) and four times more people diagnosed with MG reported needing help from a caregiver (34.8% vs. 8.3%). Use of medical care was twice as likely in MRW in comparison with POPUP (51.9% vs. 24.6%). Conclusion: This direct comparison of people diagnosed with MG and the general population using two large international studies revealed significant negative impact of MG. Results were consistent across all outcomes, in all countries.</p

High resolution crystal structures of the <i>Cerebratulus lacteus</i> mini-Hb in the unligated and carbomonoxy states

The nerve tissue mini-hemoglobin from Cerebratulus lacteus (CerHb) displays an essential globin fold hosting a protein matrix tunnel held to allow traffic of small ligands to and from the heme. CerHb heme pocket hosts the distal TyrB10/GlnE7 pair, normally linked to low rates of O2 dissociation and ultra-high O2 affinity. However, CerHb affinity for O2 is similar to that of mammalian myoglobins, due to a dynamic equilibrium between high and low affinity states driven by the ability of ThrE11 to orient the TyrB10 OH group relative to the heme ligand. We present here the high resolution crystal structures of CerHb in the unligated and carbomonoxy states. Although CO binds to the heme with an orientation different from the O2 ligand, the overall binding schemes for CO and O2 are essentially the same, both ligands being stabilized through a network of hydrogen bonds based on TyrB10, GlnE7, and ThrE11. No dramatic protein structural changes are needed to support binding of the ligands, which can freely reach the heme distal site through the apolar tunnel. A lack of main conformational changes between the heme-unligated and -ligated states grants stability to the folded mini-Hb and is a prerequisite for fast ligand diffusion to/from the heme

Phase Space Reduction for Star-Products: An Explicit Construction for CP^n

We derive a closed formula for a star-product on complex projective space and on the domain $SU(n+1)/S(U(1)\times U(n))$ using a completely elementary construction: Starting from the standard star-product of Wick type on $C^{n+1} \setminus \{ 0 \}$ and performing a quantum analogue of Marsden-Weinstein reduction, we can give an easy algebraic description of this star-product. Moreover, going over to a modified star-product on $C^{n+1} \setminus \{ 0 \}$, obtained by an equivalence transformation, this description can be even further simplified, allowing the explicit computation of a closed formula for the star-product on \CP^n which can easily transferred to the domain $SU(n+1)/S(U(1)\times U(n))$.Comment: LaTeX, 17 page

Tunneling Spectroscopy of Tl2Ba2CuO6

New results from tunneling spectroscopies on near optimally-doped single crystals of Tl_{2}Ba_{2}CuO_{6} (Tl-2201) junctions are presented. The superconductor-insulator-normal metal (SIN) tunnel junctions are obtained using the point-contact technique with a Au tip. The tunneling conductances reproducibly show a sharp cusp-like subgap, prominent quasiparticle peaks with a consistent asymmetry, and weakly decreasing backgrounds. A rigorous analysis of the SIN tunneling data is performed using two different models for the $d_{x^{2}-y^{2}}$ (d-wave) density of states (DOS). Based on these and earlier results, the tunneling DOS of Tl-2201 have exhibited the most reproducible data that are consistent with a d-wave gap symmetry. We show that the dip feature at $2\Delta$ that is clearly seen in SIN tunneling data of Bi_{2}Sr_{2}CaCu_{2}O_{8+\delta} is also present in Tl-2201, but at a weaker level. The gap values for crystals with a bulk T_c = 86 K are in the range of 19-25 meV.Comment: 7 pages, 5 figure

Predominantly Superconducting Origin of Large Energy Gaps in Underdoped Bi2Sr2CaCu2O8-d from Tunneling Spectroscopy

New tunneling data are reported in underdoped Bi2Sr2CaCu2O8-d using superconductor-insulator-superconductor break junctions. Energy gaps, Delta, of 51+2, 54+2 and 57+3 meV are observed for three crystals with Tc=77, 74, and 70 K respectively. These energy gaps are nearly three times larger than for overdoped crystals with similar Tc. Detailed examination of tunneling spectra over a wide doping range from underdoped to overdoped, including the Josephson IcRn product, indicate that these energy gaps are predominantly of superconducting origin.Comment: 10 pages, 4 figures, 1 tabl

Isolated unilateral absence of the right pulmo nary artery.

A 40-year-old man was referred for evaluation of cough of a few weeks duration and a history of recurrent respiratory tract infections for several years. Clinical examination revealed no abnormalities. Routine hematologic and biochemical evaluation were normal. Contrast-enhanced computed tomography of the thorax showed a hypoplastic right lung, hyperinflation of the left lung with cardiomediastinal shift to the right. Absence of the right pulmonary artery was noted and replaced by an extensive collateral network of hypertrophied vessels originating from bronchial, intercostal and mammaria interna arteries, and right arteria subclavia (Fig. A, B). Discrete bronchiectasis with thickened bronchial walls in a hypoplastic right lung was noted (Fig. C). A normal bronchial tree and normal parenchyma in the left lung was seen