Engineering a high-affinity allosteric binding site for divalent cations in kainate receptors

Kainate receptors are allosterically regulated by sodium ions. Removal of Na⁺ from the extracellular solution, or replacement of Na⁺ by larger monovalent cations, inhibits kainate receptor activity. Sodium binds at a negatively charged cavity in the extracellular neurotransmitter binding domain that is capped by a small hydrophobic residue. Prior work revealed that introduction of aspartic acid at this site strongly reduces GluK2 sensitivity to monovalent cations of different size. We found that the GluK2 M739D mutant is also insensitive to substitution of Na⁺ by the large organic cations Tris and NMDG. Because these are excluded from the Na⁺ binding site by steric hindrance, we investigated the possibility that divalent cations can substitute for Na⁺. We show that in Na⁺ free solutions with low concentrations of Ca²⁺ and Mg²⁺ the GluK2 M739D mutant is inhibited by EDTA; that divalent ions in the micromolar concentration range act as positive allosteric modulators; and that the chemistry of the mutant cation binding site is typical of Ca²⁺ and Mg²⁺ binding sites found in protein crystal structures. Hence, the apparent insensitivity of the M739D mutant to monovalent cations is due to the adventitious allosteric effects of divalent ions at physiological concentrations and below.     

Antibody-mediated marrow failure after allogeneic bone marrow transplantation

Marrow graft failure observed in association with histocompatibility differences between donor and recipient is often attributed to rejection mediated by host-derived cytolytic T lymphocytes. The data presented in this report indicate that persistent host antibodies specific for donor antigen may also mediate graft failure, either by antibody-dependent cell-mediated cytotoxicity (ADCC), or complement- mediated cytotoxicity. In the case of HLA Class I disparity, where all donor cells express the target antigen, the presence of alpha-donor antibody was associated with complete graft failure and death. In the case of ABO blood group antigen disparity, the presence of alpha-donor antibody resulted in erythroid hypoplasia. The latter cases proved informative insofar as they established that host antibodies could persist for more than 18 months after chemoradiotherapy and impair marrow function.     

A prosthetic contact lens in the treatment of ocular manifestations of albinism

A patient with severe ocular albinism was fitted with a prosthetic soft contact lens which improved both photophobic symptoms and facial cosmesis.     

Normal deposition of brain iron in childhood and adolescence: MR imaging at 1.5 T

Magnetic resonance (MR) images of the brain in 285 patients between the ages of 2 and 25 years were retrospectively studied to determine the appearance of brain iron accumulation. The globus pallidus, red nucleus, substantia nigra, and dentate nucleus were evaluated with long TR/TE (repetition time/echo time) spin-echo sequences and staged. All four regions in most patients were initially hyperintense compared with white matter (stage I) before becoming isointense (stage II) and subsequently hypointense (stage III). The globus pallidus was the first to reach stage III, the red nucleus and substantia nigra were next, and the dentate nucleus was last. In general, decreased signal intensity (stage III) was not seen in these regions in patients less than 10 years old; in most patients it was seen by age 25 years. The dentate nucleus decreased in signal intensity more slowly and inconsistently; only one-third of patients had reached stage III by age 25 years. The temporal sequence of normal iron deposition as detected with MR imaging is helpful not only in the diagnosis of known iron-deposition diseases but also in the detection of iron-related pathologic changes.     

Anxiety in patients undergoing MR imaging

To determine and quantify the major sources of anxiety for patients undergoing magnetic resonance (MR) imaging and to suggest means by which to eliminate or diminish their negative effects, the authors studied anxiety in 46 subjects. Of these, 20 randomly selected subjects who successfully completed the examination participated in exit interviews. Six subjects who terminated the examination before completion also completed exit interviews. Pre-imaging and postimaging questionnaires (state-trait anxiety inventory) were administered to measure anxiety in the 20 other subjects. Anxiety was associated with the constrictive dimensions of the magnet bore, examination duration, coil noise, and temperature within the bore. Preparation at the point of referral was consistently absent, incomplete, or misleading. Patients used identifiable strategies to cope with the examination: blinding, breathing relaxation techniques, visualization of pleasant images, and performance of mental exercises.     

Cerebrospinal fluid rhinorrhea: depiction with MR cisternography in dogs

Canine cerebrospinal fluid rhinorrhea, which occurs frequently in purebred beagles, was demonstrated in two dogs on magnetic resonance images after cisternal introduction of gadolinium-DTPA dimeglumine.     

核、膜蛋白的基因能抑制禽类流感病毒及其重组株的繁殖

对人流感病毒A/Udorn/72(H_3N_2)株与禽类流感病毒A/Mallard/NY/78/(H_2N_2)重组后的重组株分析表明,仅含禽类病毒的核蛋白(NP)或膜蛋白(M)的RNA片段的重组株,在松鼠猴的呼吸道繁殖是受限制的。另外。仅有禽类的RNAl和NS基因的重组株(Clone 12)在松鼠猴的气管内的繁殖也明显受限制,而只具有其中一个基因的Clone 9, Clone 2, 则限制就不明显。由此表明,禽类流感病毒的NP和M基因在宿主范围的繁殖限制中起主要作用,而RNAI和SN基因的结合,同样起着繁殖受限制作用。