Limulus amebocyte lysate assay for detection of endotoxin in patients with sepsis syndrome. AMCC Sepsis Project Working Group

Clinical predictions alone are insufficiently accurate to identify patients with specific types of bloodstream infection; laboratory assays might improve such predictions. Therefore, we performed a prospective cohort study of 356 episodes of sepsis syndrome and did Limulus amebocyte lysate (LAL) assays for endotoxin. The main outcome measures were bacteremia and infection due to gram-negative organisms; other types of infection were secondary outcomes. Assays were defined as positive if the result was > or = 0.4 enzyme-linked immunosorbent assay units per milliliter. There were positive assays in 119 (33%) of 356 episodes. Assay positivity correlated with the presence of fungal bloodstream infection (P < .003) but correlated negatively with the presence of gram-negative organisms in the bloodstream (P = .04). A trend toward higher rates of mortality in the LAL assay-positive episodes was no longer present after adjusting for severity. Thus, results of LAL assay did not correlate with the presence of bacteremia due to gram-negative organisms or with mortality after adjusting for severity but did correlate with the presence of fungal bloodstream infection.     

Mice lacking dopamine D4 receptors are supersensitive to ethanol, cocaine, and methamphetamine

The human dopamine D4 receptor (D4R) has received considerable attention because of its high affinity for the atypical antipsychotic clozapine and the unusually polymorphic nature of its gene. To clarify the in vivo role of the D4R, we produced and analyzed mutant mice (D4R-/-) lacking this protein. Although less active in open field tests, D4R-/- mice outperformed wild-type mice on the rotarod and displayed locomotor supersensitivity to ethanol, cocaine, and methamphetamine. Biochemical analyses revealed that dopamine synthesis and its conversion to DOPAC were elevated in the dorsal striatum from D4R-/- mice. Based on these findings, we propose that the D4R modulates normal, coordinated and drug-stimulated motor behaviors as well as the activity of nigrostriatal dopamine neurons.     

Identification of a gene cluster of biosynthetic genes of rubradirin substructures in S. achromogenes var. rubradiris NRRL3061

Rubradirin, an ansamycin antibiotic has been purified from Streptomyces achromogenes var. rubradiris NRRL3061. It consists of four distinct structural moieties, rubransarol, 3-amino-4-hydroxy-coumarin, dihydroxydipicolinic acid, and 2,6-dideoxynitrosugar (DNS). Polymerase chain reaction (PCR) primers were designed based on consensus sequences of dTDP-D-glucose 4,6-dehydratase, one of enzymes involved in the biosynthesis of 2,6-dideoxysugar. A PCR product was obtained from S. achromogenes var. rubradiris. Hybridization of the PCR product to a cosmid library constructed from S. achromogenes genomic DNA has led to the identification of three unlinked regions of DNA. One of three kinds of cosmid clones contains homologues of dTDP-D-glucose 4,6-dehydratase, 3-amino-5-hydroxybenzoic acid (AHBA) synthase, and eryA genes. The size of the gene homologous to eryA is 30 kb, and the AHBA synthase gene homologue resides between the eryA homologous genes. A gene cluster of rubransarol and 2,6-dideoxynitrosugar is around 50 kb. Sequencing of the PCR product from the AHBA synthase gene homologue isolated from S. achromogenes revealed 85% amino acid sequence homology (73/86) with the AHBA synthase from a rifamycin-producer. dTDP-D-glucose 4,6-dehydratase gene homologue was subcloned from one of the isolated cosmid clones and sequenced. It showed 65% homology (43/66) with dTDP-D-glucose 4,6-dehydratase from a streptomycin-producer.     

Caspase-3-generated fragment of gelsolin: effector of morphological change in apoptosis

The caspase-3 (CPP32, apopain, YAMA) family of cysteinyl proteases has been implicated as key mediators of apoptosis in mammalian cells. Gelsolin was identified as a substrate for caspase-3 by screening the translation products of small complementary DNA pools for sensitivity to cleavage by caspase-3. Gelsolin was cleaved in vivo in a caspase-dependent manner in cells stimulated by Fas. Caspase-cleaved gelsolin severed actin filaments in vitro in a Ca2+-independent manner. Expression of the gelsolin cleavage product in multiple cell types caused the cells to round up, detach from the plate, and undergo nuclear fragmentation. Neutrophils isolated from mice lacking gelsolin had delayed onset of both blebbing and DNA fragmentation, following apoptosis induction, compared with wild-type neutrophils. Thus, cleaved gelsolin may be one physiological effector of morphologic change during apoptosis.     

Monoclonal antibody BLT-1, specific for the bovine homologue of CD5, reacts with the majority of mature T cells, a subpopulation of B cells and stimulates T cell proliferation

Monoclonal antibody (MAb) BLT-1, an IgG2a with kappa light chains, reacted strongly with 21% of bovine thymocytes, weakly with 15% of thymocytes, with a subpopulation of peripheral blood B cells that also expressed CD20 and with peripheral blood T cells. Practically all of the reactive thymocytes were of a large cell subpopulation. By immunoprecipitation, BLT-1 was shown to recognize a membrane molecule with a molecular mass of 67 kDa. In competitive assays for lymphocyte surface binding, BLT-1 and MAb CC-29 (which had been shown previously to react with bovine CD5) blocked one another, indicating that the epitopes recognized were identical or extensively overlapping. In contrast, another CD-5-reactive MAb, CC-17, did not block BLT-1 reactivity with lymphocytes although the reactivity of CC-17 was blocked by BLT-1, suggesting partial overlap of the epitopes or steric hindrance by BLT-1 but not by CC-17. BLT-1 was able to induce proliferation of bovine lymphocytes in culture alone if monocytes were present or in the absence of monocytes synergized with PMA. The results indicate that BLT-1 recognizes an epitope of the bovine homologue of CD5 and that perturbation of the epitope by MAb binding results in signal transduction to bovine lymphocytes.     

Age as a factor in critical care unit admissions

BACKGROUND: Although the aged occupy a high proportion of critical care beds nationwide, few multicenter studies have been undertaken to specifically determine physician attitudes toward the elderly in a critical care setting. We attempt to determine the importance of patient age as a factor in the admission of acutely ill medical patients to critical care units. METHODS: In response to a hypothetical case scenario, physicians were asked to admit one of two patients to a last available critical care unit bed. An accompanying questionnaire was used to gain a ranking of several admission factors as compared with age, and to gain demographic data regarding the study population. Data were subjected to nonparametric statistical analysis. RESULTS: When age was the only difference between two patients in a hypothetical case scenario, 80.7% of respondents chose the younger patient (age 56 years) for admission, 13.2% chose the older patient (age 82 years), and 6.2% abstained. Following the provision of more detailed medical and social information, however, only 53.5% chose the younger patient, 41.2% chose the older patient, and 5.3% continued to abstain. In a ranking of several admission factors, age was found to be of less importance than severity of presenting illness, previous medical history, and do not resuscitate status, but of more importance than patient motivation, ability to contribute to society, family support, and ability to pay for care. When asked if they supported a definitive age criterion that would restrict all patients over a certain age from access to critical care units, 95.1% responded that they did not. CONCLUSIONS: Age is a factor considered by physicians in the admission of acutely ill medical patients to critical care units. Other medical and social factors, however, can affect the impact of patient age on treatment decisions. Further study and discussion are needed to clarify the appropriate role of age and other factors in critical care unit admissions.     

The effects of unilateral velocity-specific concentric strength training

There is conflicting evidence regarding the efficacy of various resistance training programs for increasing strength in trained and contralateral limbs. The purposes of this investigation were to examine the effects of unilateral velocity-specific concentric isokinetic training of the extensor and flexor muscles of the elbow and knee on: 1) the carry-over effect in strength increases to velocities other than the training velocity in the trained limbs and 2) the cross-training effect at various velocities in the contralateral limbs. Twelve adult men (mean age +/- SD = 24 +/- 6 years) volunteered to train their nondominant extremities three times per week (six sets of 10 maximal repetitions) for 8 weeks at 120 degrees/sec using a Cybex II isokinetic dynamometer. The subjects were tested for increases in peak torque at 60, 120, 180, 240, and 300 degrees/sec. The training resulted in significant (p < 0.05) increases in peak torque on the trained side of the body for elbow extension and flexion as well as knee extension and flexion at all velocities tested. These findings indicated that the velocity-specific training resulted in increases in peak torque at velocities that were both greater and less than the training velocity. In addition, there was a cross-training effect, with significant (p < 0.05) increases in peak torque on the contralateral side of the body for elbow extension (all velocities except 300 degrees/sec) as well as knee extension and flexion (all velocities). These results indicate that unilateral velocity-specific concentric isokinetic training is adequate stimulus for eliciting strength gains at a wide range of velocities in both the trained and contralateral limbs.(ABSTRACT TRUNCATED AT 250 WORDS)