Synchronization of Ca(2+) oscillations among primate LHRH neurons and nonneuronal cells in vitro

Periodic release of luteinizing hormone-releasing hormone (LHRH) from the hypothalamus is essential for normal reproductive function. Pulsatile LHRH release appears to result from the synchronous activity of LHRH neurons. However, how the activity of these neurons is synchronized to release LHRH peptide in a pulsatile manner is unclear. Because there is little evidence of physical coupling among LHRH neurons in the hypothalamus, we hypothesized that the activity of LHRH neurons might be coordinated by indirect intercellular communication via intermediary (nonneural) cells rather than direct interneural coupling. In this study, we used an in vitro preparation of LHRH neurons derived from the olfactory placode of monkey embryos to assess whether nonneuronal cells, play a role in coordinating LHRH neuronal activity. We found that cultured LHRH neurons and nonneuronal cells both exhibit spontaneous oscillations in the concentration of intracellular Ca(2+) ([Ca(2+)](i)) at similar frequencies. Moreover, [Ca(2+)](i) oscillations in both types of cell were periodically synchronized. Synchronized [Ca(2+)](i) oscillations spread as intercellular Ca(2+) waves across fields of cells that included LHRH neurons and nonneuronal cells, although waves spread at a higher velocity among LHRH neurons. These results suggest that LHRH neurons and nonneuronal cells are functionally integrated and that nonneuronal cells could be involved in synchronizing the activity of the LHRH neurosecretory network.     

Mineral metabolism in male cyclists during high-intensity endurance training

This study examined the effects of intense endurance training on basal plasma and 24-hour urinary calcium (Ca), magnesium (Mg), iron (Fe), zinc (Zn), and copper (Cu) levels in 9 male competitive cyclists. The supervised training program followed a baseline period and included a volume phase (6 weeks, averaging 87% of maximal heart rate [HR(max)]), an interval phase (18 days, 100% of HR(max)), and a 10-day unloading taper. The primary training outcome measure was 20-km time-trial cycling performance. Subjects ate unrestricted diets and maintained their weight. Compared to baseline, performance improved significantly (p < .05), while mineral metabolism was not significantly different after the volume phase. However, after the interval phase, renal Ca excretion increased (p < .05) and plasma Ca fell slightly below the clinical norm. As compared to the interval phase, urinary Ca decreased (p <.05), plasma Ca increased (p < .05), and performance further improved (p < .05) after the taper. Whereas Mg, Fe, Zn, and Cu metabolism remained unchanged throughout the study, greater renal Ca excretion was associated with very high intensity interval training.     

Different Toll-like receptor agonists induce distinct macrophage responses

We previously reported that gram-negative bacterial lipopolysaccharide (LPS) activates cells via Toll-like receptor (TLR) 4, whereas the mycobacterial cell wall glycolipid lipoarabinomannan (LAM) activates cells via TLR2. We also identified a secreted TLR2 agonist activity in short-term culture filtrates of Mycobacterium tuberculosis bacilli, termed soluble tuberculosis factor (STF). Here we show that STF contains mannosylated phosphatidylinositol (PIM) and that purified PIM possesses TLR2 agonist activity. Stimulation of RAW 264.7 macrophages by LPS, LAM, STF, and PIM rapidly activated nuclear factor (NF)-kappaB, activator protein-1 (AP-1), and mitogen-activated protein (MAP) kinases. These TLR agonists induced similar levels of NF-kappaB and AP-1 DNA-binding activity, as well as trans-activation function. Unexpectedly, these TLR agonists induced tumor necrosis factor alpha secretion, whereas only LPS was capable of inducing interleukin-1beta and nitric oxide secretion. Thus, different TLR proteins are still capable of activating distinct cellular responses, in spite of their shared capacities to activate NF-kappaB, AP-1, and MAP kinases.     

Effective limitation of blood culture use in the burn unit

There is little data to support the use of blood culture (BC) testing in the burn patient. Clinical signs (fever, leukocytosis) may not reliably indicate infection; moreover, BC's are expensive, invasive and plagued by false positive results. A policy of critical evaluation of lab utilization was instituted in our burn unit in 1993. By 1997 the use of blood culture testing had decreased by 50%. A retrospective analysis of our change in BC utilization was undertaken to derive practice guidelines for usage of this test. METHODS: 47 patients with BC testing in 1997 were compared to a cohort of 47 patients from 1993, representing a total of 441 BC episodes. RESULTS: Comparison of 1993 and 1997 patients revealed no significant differences in patient characteristics or outcomes. The mean white blood cell count and maximum temperature on the day of culture were identical for both positive and negative BC episodes. However, BC's drawn during a state of shock were twice as likely to be positive. Patients who experienced positive BC's had larger burns, received more antibiotics, had more indwelling catheters, and had longer lengths of ventilator support and hospital stays. CONCLUSIONS: Higher patient acuity or the presence of indwelling catheters increases the likelihood of a positive BC. Substantial limitation of BC's without observed changes in length of stay, ventilator days, or mortality suggests that this test can be safely limited without compromising patient outcomes.