The binding of substrates and inhibitors to the metal center of myoinositol monophosphatase

The synthetic substrate anthraniloyl-beta-glycerol-P binds to myoinositol monophosphatase with a Kd = 5 microM at pH 7.5. The anthraniloyl chromophore, excited at 330 nm, sensitizes the long lived luminescence of bound Tb(III) at 490, 545, 585 and 620 nm. Assuming a mechanism of radiationless energy transfer, the actual distance of separation between the donor-acceptor pair was calculated to be R = 10 A. Tb(III) binds to the monophosphatase with a Kd = 2 microM, whereas Ca-(II) displaces the lanthanide at concentrations above 0.1 mM. The binding studies support the notion that Tb(III), Ca(II) and Mg(II) interact with a common binding site on the protein. Phosphate ion, a strong competitive inhibitor, perturbs the luminescence of bound Tb(III), whereas the substrate beta-glycero-P has no effect on the luminescence yield and long-lived emission of bound Tb(III). It is suggested that the phosphate group of the substrate is not in direct contact with the metal ion coordinated to several amino acid residues of the enzyme.     

Abnormal behavioral responses to fenfluramine in patients with affective and personality disorders. Correlation with increased serotonergic responsivity

Serotonergic responsivity was assessed in 20 psychiatric patients by the prolactin response to a fenfluramine challenge test. During the fenfluramine challenge 6 of 20 patients (30%) spontaneously reported psychopathologic reactions that included: increased anxiety/agitation, psychotic symptoms, illusions, mood elevation, and anergia. The time of peak behavioral symptoms (2.5 +/- 0.8 hrs) corresponded closely to the time of peak increase in prolactin levels (3.0 +/- 1.1 hr). Abnormal behavioral responders had statistically significant greater increases in prolactin 1 to 4 hr after fenfluramine when compared to normal responders. Patients who developed an abnormal psychopathologic response to fenfluramine were characterized by higher levels of anxiety and agitation at the time of admission to the hospital but otherwise were not distinguishable on the basis of severity of other psychiatric symptoms. This study suggests that increased serotonergic transmission may trigger anxiety, psychosis, and mood elevation in specific vulnerable individuals, whereas other patients with similar psychiatric illnesses are not affected.     

Accuracy of using computed tomography to identify pedicle screw placement in cadaveric human lumbar spine

STUDY DESIGN: Utility of using computed tomography to predict pedicle screw misplacement. OBJECTIVE: This study defines the sensitivity and specificity of predicting pedicle screw placement by experienced clinicians using a CT scan image. SUMMARY OF BACKGROUND DATA: In clinical and research settings, the method most commonly used to evaluate pedicle screws placement has been computed tomography. However, no current literature describes the accuracy of this method of evaluating screw placement. METHOD: Cobalt-chrome and titanium alloy pedicle screws of identical size were placed in six cadaveric human lumbar spine. Wide laminectomy was performed to allow complete visualization of the pedicles. Three consecutive lumbar levels were instrumented in each spine, giving 36 pedicle screw placements to identify. The instrumented spines were imaged, and four orthopaedic spine surgeons and a musculoskeletal radiologist were asked to read the images to identify the accuracy of screw placement within the pedicles. RESULTS: The sensitivity rate of identifying a misplaced screw was 67 +/- 6% for cobalt-chrome screws compared with 86 +/- 5% for titanium screws (P < 0.005). The specificity rates of radiographic diagnosis of misplaced pedicle screws were 66 +/- 10% for cobalt-chrome screws and 88 +/- 8% for titanium screws (P < 0.005). Similarly, a statistically significant difference was found in the sensitivity rates of identifying screws placed correctly in the pedicle: 70 +/- 10% for cobalt-chrome screws versus 89 +/- 8% for titanium screws (P < 0.005). Overall accuracy rates were 68 +/- 7% for cobalt chrome screws versus 87 +/- 3% for titanium screws (P < 0.002). CONCLUSION: Reliance on the computed tomography scan data alone in determining accuracy of pedicle screws can lead to inaccuracies in both clinical and research conditions.     

A case-mix classification system for medical rehabilitation

Dissatisfaction with Medicare's current system of paying for rehabilitation care has led to proposals for a rehabilitation prospective payment system, but first a classification system for rehabilitation patients must be created. Data for 36,980 patients admitted to and discharged from 125 rehabilitation facilities between January 1, 1990, and April 19, 1991, were provided by the Uniform Data System for Medical Rehabilitation. Classification rules were formed using clinical judgment and a recursive partitioning algorithm. The Functional Independence Measure version of the Function Related Groups (FIM-FRGs) uses four predictor variables: diagnosis leading to disability, admission scores for motor and cognitive functional status subscales as measured by the Functional Independence Measure, and patient age. The system contains 53 FRGs and explains 31.3% of the variance in the natural logarithm length of stay for patients in a validation sample. The FIM-FRG classification system is conceptually simple and stable when tested on a validation sample. The classification system contains a manageable number of groups, and may represent a solution to the problem of classifying medical rehabilitation patients for payment, facility planning, and research on the outcomes, quality, and cost of rehabilitation.     

Inhalation toxicity of 1,6-hexanediamine dihydrochloride in F344/N rats and B6C3F1 mice

1,6-Hexanediamine (HDA) is a high production volume chemical which is used as an intermediate in the synthesis of paints, resins, inks, and textiles and as a corrosion inhibitor in lubricants. Two- and 13-week studies of the toxicity of the dihydrochloride salt of HDA (HDDC) were conducted in male and female Fischer 344/N rats and B6C3F1 mice using whole-body inhalation exposure. Both species were evaluated for histopathologic and reproductive effects, and rats were examined for clinical chemistry and hematologic changes. In the 2-week inhalation studies, animals were exposed to 10-800 mg HDDC/m3, 6 hr per day. All rats, all female mice, and two of five male mice in the high-exposure group died before the end of the study. Surviving mice in this group had a dose-dependent depression in body weight gain. Clinical signs were primarily related to upper respiratory tract irritation and included dyspnea and nasal discharge in both species. Treatment-related histopathologic lesions included inflammation and necrosis of the laryngeal epithelium of both species and the tracheal epithelium of mice, as well as focal inflammation and ulceration of the respiratory and olfactory nasal mucosa. In the 13-week inhalation studies, animals were exposed to HDDC at concentrations of 1.6-160 mg/m3 for 6 hr per day, 5 days per week. In addition to the base study groups, a supplemental group of rats at each exposure level was included to assess the effect of HDDC on reproduction. No treatment-related changes in organ weights or organ-to-body-weight ratios occurred in rats, and no treatment-related clinical signs or gross lesions were seen in either species. Chemical-related microscopic lesions were limited to the upper respiratory tract (larynx and nasal passages) in the two highest exposure groups and were similar in both species. These lesions included minimal to mild focal erosion, ulceration, inflammation, and hyperplasia of the laryngeal epithelium, in addition to degeneration of the olfactory and respiratory nasal epithelium. HDDC caused no significant changes in sperm morphology or vaginal cytology and no significant adverse effects on reproduction in rats or mice. Hematologic and clinical chemistry changes in rats were minor and sporadic and were not accompanied by related histologic findings. HDDC did not increase the frequency of micronucleated erythrocytes in mice.(ABSTRACT TRUNCATED AT 400 WORDS)     

Desensitization of mutant acetylcholine receptors in transgenic mice reduces the amplitude of neuromuscular synaptic currents

While the slow onset of desensitization of nicotinic acetylcholine receptors (AChRs), relative to the rate of acetylcholine removal, excludes this kinetic state from shaping synaptic responses in normal neuromuscular transmission, its role in neuromuscular disorders has not been examined. The slow-channel congenital myasthenic syndrome (SCCMS) is a disorder caused by point mutations in the AChR subunit-encoding genes leading to kinetically abnormal (slow) channels, reduced miniature endplate current amplitudes (MEPCs), and degeneration of the postsynaptic membrane. Because of this complicated picture of kinetic and structural change in the neuromuscular junction, it is difficult to assess the importance of the multiple factors that may be responsible for the reduced endplate current amplitudes, and ultimately the clinical syndrome. In order to address this we have used a transgenic mouse model for the SCCMS that has slow AChR ion channels and reduced endplate responsiveness in the absence of any of the degenerative changes. We found that the reduction in MEPC amplitudes in these mice could not be explained by either reduced AChR number or by reduced AChR channel conductance. Rather, we found that the mutant AChRs in situ manifested an activity-dependent reduction in sensitivity that caused diminished MEPC and endplate current amplitude with nerve stimulation. This observation demonstrates that the basis for the reduction in MEPC amplitudes in the SCCMS may be multifactorial. Moreover, these findings demonstrate that, under conditions that alter their rate of desensitization, the kinetic properties of nicotinic AChRs can control the strength of synaptic responses.     

Unified equation of state based on the lattice fluid theory for phase equilibria of complex mixtures part I. Molecular thermodynamic framework

Consistent calculation of fugacities of fluid mixtures remains as one of the most important subjects in contemporary molecular thermodynamics. In practice, equations of state (EOSs) and g^E-models have been used. However, most EOSs are erroneous for condensed phases at high densities and g^E-models are inapplicable for pressuresensitive systems. Recently to remedy the shortcomings in both approaches, there has been a surge of new g^E-EOS mixing rules. By equating any set of EOS and g^E-models, the limitations in both approaches could be resolved significantly. However, the self-consistency in the underlying concept of those mixing rules remains controversial. During the last several years, the present authors proposed a new lattice-fluid EOS and its simplification relevant to phase equilibrium calculations. Without employing any g^E-EOS mixing rule and with only two parameters for a pure component and one adjustable interaction energy parameter for a binary mixture, results obtained to date demonstrated that the EOSs are quantitatively applicable to a great variety of phase equilibrium properties of mixtures, especially, for complex and/or macromolecular systems. In the present article we summarize the EOSs and extended the applications to liquid-liquid Equilibria. In part I, we discussed briefly the molecular thermodynamic aspects of general derivation of the EOS and a brief discussion of applying the EOSs to pure fluids while the illustrative application to various real mixture systems is discussed in part II.