Novel NMDA antagonists: replacement of the pyridinium ring of 6,11-ethanobenzo[b]quinolizinium cations with heteroisoquinolinium cations

Replacement of the pyridinium ring of 6,11-ethanobenzo[b]quinolizinium cations with thiazolium (4a and 4b) and N-methylimidazolium (4c and 4d) resulted in equipotent compounds in the [3H]TCP binding assay. The corresponding N-methyl-1,2,4-triazolium analogs were less potent in this assay. The thiazolium derivative 4b, with a Ki = 2.9 nM, is being evaluated as a possible neuroprotective N-methyl-D-aspartic acid (NMDA) antagonist.     

A method for preparing chelate-cytokine conjugates with retention of protein structure, biological activity, and pharmacokinetic properties

The chelating agent diethylenetriaminepentaacetic acid (DTPA) has been conjugated site-specifically to the N-terminus of recombinant human interleukin-2 (rhIL-2) by reaction with DTPA dianhydride at an initial pH of 6.0, thus demonstrating broader application of the conjugation method previously described for the structurally related cytokine rhG-CSF (Ralph et al., 1995). Purity of the DTPA-rhIL-2 conjugate, isolated by cation-exchange FPLC, and chelation of 111In were revealed by cation-exchange HPLC. Purity of the conjugate as well as chelation of radiometal were also demonstrated by SDS-PAGE and TLC, respectively. The stoichiometric molar ratio of DTPA to protein for the conjugate was approximately 1:1 as determined by TLC and mass spectrometry. Localization of the DTPA moiety was resolved by a peptide mapping procedure. The protein retained > 95% secondary structure (alpha helicity) following the conjugation. Addition of metal induced an approximate 22% loss of secondary structure for the conjugate. The in vitro biological activity of the protein was unaffected by the conjugated DTPA, even with chelated metal. Pharmacokinetic analysis of DTPA-conjugated cytokines, following chelated 111In, showed clearance and pharmacokinetic parameter values comparable to those of the corresponding unmodified cytokine. DTPA-conjugated cytokines may prove useful in cytokine research, and furthermore may represent a novel class of molecules for imaging, diagnosing, and/or treatment of malignancies where the cytokine receptor is overexpressed.     

T-lymphocytes from AIDS patients are unable to synthesize ribonucleotides de novo in response to mitogenic stimulation. Impaired pyrimidine responses are already evident at early stages of HIV-1 infection

Proliferative defects have been reported at the level of DNA synthesis, even in T-lymphocytes from asymptomatic human immunodeficiency virus type-1+ (HIV-1+) patients. Since purine and pyrimidine ribonucleotide availability is crucial for proliferation, we compared the ability of HIV-1- and HIV-1+ T-lymphocytes (> 95% CD4+ and CD8+) to activate de novo biosynthetic and salvage pathways following phytohemagglutinin stimulation using 14C-labeled precursors. The striking abnormality already detectable in asymptomatic patients' cells was the impaired ability of CTP, UDP-Glc, and UTP pools to expand over 72 h (44-70% of control), although ATP and GTP pools and responses were normal. In symptomatic patients, resting T-cells showed markedly reduced pyrimidine pools (53-74% of control) with no change following activation. Relatively normal ATP, GTP, and NAD pools masked the same impaired response of de novo synthesis to activation, with ATP and GTP being reduced by 50% at 48 h. Purine salvage was more active than the control in unstimulated HIV-1+ cells. This impaired de novo synthesis in HIV-1+ T-lymphocytes severely restricts the availability of ribonucleotides for vital growth-related activities such as membrane expansion and strand break repair as well as DNA and RNA synthesis. The data indicate that resting T-lymphocytes from symptomatic patients survive through enhanced salvage, but the stimulation induces metabolic cell death, and provide an explanation for the activation-associated lymphocyte death seen in HIV-1+ T-lymphocytes.     

Myocardial 18F-FDG-PET. Experiences with the euglycemic hyperinsulinemic clamp technique

Myocardial positron emission tomography (PET) with 18F-Fluordeoxyglucose (FDG) is increasingly used for the detection of viable tissue in the infarcted myocardium. Previous studies show that the variable metabolic conditions determine the regional distribution of this tracer and that the inhomogeneities of uptake often observed even in the normal myocardium may relate to substrate availability. The authors tried to stimulate the myocardial FDG uptake by either the technically easier method of glucose loading or by the euglycemic hyperinsulinemic clamp (EHC) technique. In their hands both methods could be considered as equally practicable but differing in some important details in regard to both the study protocol (tracer dose, optimal scanning time) and the reproducibility of results. The EHC allows a quick stabilization of the metabolic environment and resulted in an earlier and markedly increased FDG uptake. However, the important standardization of the method was performed by a computer-controlled system only for the glucose and insulin infusions. Their experiences show that the EHC provides a useful framework for assessing altered cardiac metabolism and possibly describes changes after therapeutic interventions more precisely than the commonly used glucose-loading technique.     

Pharmacologic atrial natriuretic peptide reduces human leg capillary filtration

Atrial natriuretic peptide (ANP) is produced and secreted by atrial cells. We measured calf capillary filtration rate with prolonged venous-occlusion plethysmography of supine healthy male subjects during pharmacologic infusion of ANP (48 pmol/kg/min for 15 min; n = 6) and during placebo infusion (n = 7). Results during infusions were compared to prior control measurements. ANP infusion increased plasma [ANP] from 30 +/- 4 to 2,568 +/- 595 pmol/l. Systemic hemoconcentration occurred during ANP infusion: mean hematocrit and plasma colloid osmotic pressure increased 4.6 and 11.3%, respectively, relative to preinfusion baseline values (p < 0.05). Mean calf filtration, however, was significantly reduced from 0.15 to 0.08 ml/100 ml/min with ANP. Heart rate increased 20% with ANP infusion, whereas blood pressure was unchanged. Calf conductance (blood flow/arterial pressure) and venous compliance were unaffected by ANP infusion. Placebo infusion had no effect relative to prior baseline control measurements. Although ANP induced systemic capillary filtration, in the calf, filtration was reduced with ANP. Therefore, pharmacologic ANP infusion enhances capillary filtration from the systemic circulation, perhaps at upper body or splanchnic sites or both, while having the opposite effect in the leg.     

Estimation of genetic parameters for growth, feed consumption, and conformation traits for double-muscled Belgian blue bulls performance-tested in Belgium

For 1,442 Belgian Blue bulls performance-tested at the Centre de Sélection de la Race Blanc-Bleue Belge, nine traits were observed: height at withers at 7 mo, height at withers at 13 mo, weight at 7 mo, weight at 13 mo, average feed consumption of concentrates, average daily gain, average feed consumption of concentrates per average daily gain, average feed consumption of concentrates per mean metabolic weight, and price per kilogram of live weight. This price is based on muscle conformation and is therefore used as muscle conformation score. Restricted maximum likelihood with a derivative-free algorithm was used to estimate (co)variance components because there were different models and missing values per trait. Estimates of heritabilities were above .50 except for average feed consumption per average daily gain (.16) and average feed consumption per mean metabolic weight (.33). Estimates of genetic and phenotypic correlations between height at withers and weight traits were positive and moderate to high. Average daily gain showed a negative genetic correlation with weight at 7 mo (-.68) but had positive correlations with height at withers at 13 mo and weight at 13 mo (.22 and .43). Muscle conformation expressed as price per kilogram of live weight was related to low average feed consumption per average daily gain. Average feed consumption showed high correlations with weight at 7 mo and weight at 13 mo. Average feed consumption per average daily gain had a high negative genetic correlation with average daily gain (-.89).     

Microbial resistance: what is to be done? Information from the panel of experts

This document represents the recommendations of a panel of Spanish experts on antibiotic use and resistance. In a Task Force, under the auspices of the Spanish Ministry of Health and Consumer Affairs that took place in 1994 in Madrid, the members were gravely concerned about the national increase in antibiotic resistance. They analysed the development, evolution and spread of antibiotic resistance among community-acquired human bacterial pathogens in Spain, its relation with antibiotic consumption, and they proposed future surveillance strategies for monitoring the patterns of antibiotic use and consumption. Success will require a collective action among the producers (pharmaceutical industry), prescribers (doctors, veterinarians), dispensers (pharmacists), and consumers (patients). Two similar documents have been recently published by the American Society of Microbiology and the World Health Organization showing the global concern about this topic.