Evolution of lung breathing from a lungless primitive vertebrate.

Air breathing was critical to the terrestrial radiation and evolution of tetrapods and arose in fish. The vertebrate lung originated from a progenitor structure present in primitive boney fish. The origin of the neural substrates, which are sensitive to metabolically produced CO2 and which rhythmically activate respiratory muscles to match lung ventilation to metabolic demand, is enigmatic. We have found that a distinct periodic centrally generated rhythm, described as "cough" and occurring in lamprey in vivo and in vitro, is modulated by central sensitivity to CO2. This suggests that elements critical for the evolution of breathing in tetrapods, were present in the most basal vertebrate ancestors prior to the evolution of the lung. We propose that the evolution of breathing in all vertebrates occurred through exaptations derived from these critical basal elements.

Isoflurane causes concentration-dependent inhibition of medullary raphé 5-HT neurons in situ.

BACKGROUND: Anesthetics have a profound influence on a myriad of autonomic processes. Mechanisms of general anesthesia, and how these mechanisms give rise to the multifaceted state of anesthesia, are largely unknown. The ascending and descending serotonin (5-HT) networks are key modulators of autonomic pathways, and are critically involved in homeostatic reflexes across the motor, somatosensory, limbic and autonomic systems. These 5-HT networks are thought to contribute to anesthetic effects, but how anesthetics affect 5-HT neuron function remains a pertinent question. We hypothesized that the volatile anesthetic isoflurane inhibits action potential discharge of medullary raphé 5-HT neurons. METHODS: We conducted extracellular recordings on individual neurons in the medullary raphé region of the unanesthetized in situ perfused brainstem preparation to determine how exposure to isoflurane affects 5-HT neurons. We examined changes in 5-HT neuron baseline firing in response to treatment with either 1, 1.5, or 2% isoflurane. We measured isoflurane concentrations by gas chromatography-mass spectrometry (GC-MS) analysis. RESULTS: Exposure to isoflurane inhibited action potential discharge in raphé 5-HT neurons. We document a concentration-dependent inhibition over a range of concentrations approximating isoflurane MAC (minimum alveolar concentration required for surgical anesthesia). Delivered concentrations of isoflurane were confirmed using GC-MS analysis. CONCLUSIONS: These findings illustrate that halogenated anesthetics greatly affect 5-HT neuron firing and suggest 5-HT neuron contributions to mechanisms of general anesthesia.

Cesium accumulation by aquatic organisms at different trophic levels following an experimental release into a small reservoir.

The rates of accumulation and subsequent loss of stable cesium (¹³³Cs) by organisms at different trophic levels within plankton-based and periphyton-based food chains were measured following the addition of ¹³³Cs into a small reservoir near Aiken, South Carolina, USA. An uptake parameter u (L kg⁻¹ d⁻¹ dry mass) and a loss rate parameter k (d⁻¹) were estimated for each organism using time-series measurements of ¹³³Cs concentrations in water and biota, and these parameters were used to estimate maximum concentrations, times to maximum concentrations, and concentration ratios (C(r)). The maximum ¹³³Cs concentrations for plankton, periphyton, the insect larva Chaoborus punctipennis, which feeds on plankton, and the snail Helisoma trivolvis, which feeds on periphyton, occurred within the first 14 days following the addition, whereas the maximum concentrations for the fish species Lepomis macrochirus and Micropterus salmoides occurred after 170 days. The C(r) based on dry mass for plankton and C. punctipennis were 1220 L kg⁻¹ and 5570 L kg⁻¹, respectively, and were less than the C(r) of 8630 L kg⁻¹ for periphyton and 47,700 L kg⁻¹ for H. trivolvis. Although the C(r) differed between plankton-based and periphyton-based food chains, they displayed similar levels of biomagnification. Biomagnification was also indicated for fish where the C(r) for the mostly nonpiscivorous L. macrochirus of 22,600 L kg⁻¹ was three times less than that for mostly piscivorous M. salmoides of 71,500 L kg⁻¹. Although the C(r) for M. salmoides was greater than those for periphyton and H. trivolvis, the maximum ¹³³Cs concentrations for periphyton and H. trivolvis were greater than that for M. salmoides.

Lung respiratory rhythm and pattern generation in the bullfrog: role of neurokinin-1 and mu-opioid receptors.

Location of the lung respiratory rhythm generator (RRG) in the bullfrog brainstem was investigated by examining neurokinin-1 and mu-opioid receptor (NK1R, muOR) colocalization by immunohistochemistry and characterizing the role of these receptors in lung rhythm and episodic pattern generation. NK1R and muOR occurred in brainstems from all developmental stages. In juvenile bullfrogs a distinct area of colocalization was coincident with high-intensity fluorescent labeling of muOR; high-intensity labeling of muOR was not distinctly and consistently localized in tadpole brainstems. NK1R labeling intensity did not change with development. Similarity in colocalization is consistent with similarity in responses to substance P (SP, NK1R agonist) and DAMGO (muOR agonist) when bath applied to bullfrog brainstems of different developmental stages. In early stage tadpoles and juvenile bullfrogs, SP increased and DAMGO decreased lung burst frequency. In juvenile bullfrogs, SP increased lung burst frequency, episode frequency, but decreased number of lung bursts per episode and lung burst duration. In contrast, DAMGO decreased lung burst frequency and burst cycle frequency, episode frequency, and number of lung bursts per episode but increased all other lung burst parameters. Based on these results, we hypothesize that NK1R and muOR colocalization together with a metamorphosis-related increase in muOR intensity marks the location of the lung RRG but not necessarily the lung episodic pattern generator.

Responses of benthic fish exposed to contaminants in outdoor microcosms--examining the ecological relevance of previous laboratory toxicity tests.

Previous laboratory studies indicate that coal combustion wastes (a mixture composed of fly ash and other lower volume wastes such as bottom ash; hereafter collectively referred to as ash) adversely affect the health of benthic fish (Erimyzon sucetta; lake chubsucker), but fish in these studies were provided with ample uncontaminated food resources. Because aquatic disposal of ash can also adversely affect food resources for benthic fish, we hypothesized that changes in resources might exacerbate the effects of ash on fish observed in laboratory studies. We exposed juvenile E. sucetta in outdoor microcosms to water, sediment, and benthic resources from an ash-contaminated site or a reference site for 45 days and compared our findings to previous laboratory studies. Benthic invertebrate biomass was nearly three times greater in controls compared to ash microcosms. Total organic content of control sediment (41%) was also greater than in ash sediments (17%), suggesting that additional benthic resources may have also been limited in ash microcosms. Benthic invertebrates isolated from the ash microcosms had trace element concentrations (As, Cd, Co, Cr, Cs, Se, Sr, and V) up to 18 times higher than in weathered ash used in laboratory studies. The concentrations of trace elements accumulated by fish reflected the high dietary concentrations encountered in the ash microcosms and were associated with reduced growth (final mass = 0.07 g) and survival (25%) compared to controls (0.37 g and 67%, respectively). Accumulation of trace elements, as well as reductions in growth and survival, were more pronounced than in previous laboratory studies, suggesting that resource conditions may be important in mediating ash toxicity. Taken together, our studies suggest that ash discharge into aquatic systems is a more serious threat to the health of benthic fish than previously predicted based upon laboratory toxicity tests.

Do semiclassical zero temperature black holes exist?

The semiclassical Einstein equations are solved to first order in epsilon = Planck's over 2pi/M2 for the case of a Reissner-Nordström black hole perturbed by the vacuum stress energy of quantized free fields. Massless and massive fields of spin 0, 1/2, and 1 are considered. We show that in all physically realistic cases, macroscopic zero temperature black hole solutions do not exist. Any static zero temperature semiclassical black hole solutions must then be microscopic and isolated in the space of solutions; they do not join smoothly onto the classical extreme Reissner-Nordström solution as epsilon-->0.

The respiratory central pattern generator of Lymnaea: a model, measured and malleable.

Great progress has been made, and continues to be made in our understanding of the neuronal mechanisms underlying respiration in a wide variety of model systems. The central pattern generator (CPG) controlling aerial respiration in the pond-snail Lymnaea is a particularly well-studied model. Using in vitro and semi-intact preparations, the neural circuitry that controls aerial respiration has been characterized as consisting of three identified interneurons. Furthermore, insight has been gained into the behavioural, cellular and synaptic mechanisms by which this circuit controls respiratory rhythmogenesis. It has also been demonstrated that aerial respiratory behaviour can be modified both by experience and by environmental factors. Studies have shown that, in a behavioural hierarchy, respiration is subservient to the whole-body withdrawal response that respiratory behaviour can be modified through operant conditioning, and that respiratory behaviour is altered by hypoxia. Through research on the Lymnaea respiratory CPG we are coming to a better understanding of the construction and malleability of a CPG network. The malleability of this CPG is of particular interest. No longer can neuronal networks underlying respiratory behaviour be considered hard-wired; they have inherent plasticity.

Use of isolated digestive-gland cells in the study of biochemical and physiological processes in gastropod molluscs.

We describe a method for preparation and maintenance of isolated digestive-gland cells in the abalone, Haliotis kamtschatkana. Viability of the isolated cells was confirmed by the fact that 18 h after preparation the cells exhibited less than 5% staining with trypan blue and actively synthesized glycogen following the addition of glucose substrate. Use of the method in a 15-month study of metabolic activity of the digestive gland of H. kamtschatkana showed significant differences in oxygen consumption of isolated-cell preparations correlated with seasonal differences in somatic and gametogenetic growth, and with relative size of the digestive gland.

The new work of the nonprofit board.

Too often, the board of a nonprofit organization is little more than a collection of high-powered people engaged in low-level activities. But that can change, the authors say, if trustees are willing to discover and take on the new work of the board. When they perform the new work, a board's members can significantly advance the institution's mission and long-term welfare. Doing the new work requires a board to engage in new practices. First, the board must go beyond rubber-stamping management's proposals and find out what issues really matter to the institution. It can do that by making the CEO paint the big picture of the organization's strategic concerns, by understanding key stakeholders, by consulting experts, and by deciding what needs to be measured in order to judge the institution's performance. Second, a board doing the new work must take action: the board must not only set policy but also work with management to implement it. Third, the board must go beyond strictly functional organization: the new work requires flexibility and encourages ad hoc arrangements. Finally, board meetings--where boards underperform most visibly--should be driven by goals, not by processes. The authors give many examples of boards that have successfully embraced the new work. The stakes are high: if boards demonstrate that they can change effectively, the professional staff at the institutions they serve just may follow suit.

Depurination of plant ribosomes by pokeweed antiviral protein.

Mammalian ribosomes have been shown to be enzymatically modified by ribosomal inactivating protein (RIPs) via specific depurination of rRNA. Here we report that ribosomes isolated from wheat germ contain intact and undepurinated rRNA and are depurinated by pokeweed antiviral protein (PAP). Pokeweed ribosomes isolated under the same conditions are depurinated. Total RNA isolated from pokeweed in the presence of strong denaturants was found to pbe partially depurinated. We conclude that wheat germ ribosomes are resistant to the endogenous RIP, tritin, but are sensitive to PAP and that pokeweed ribosomes can be depurinated by the N-glycosidase activity of endogenous PAP during isolation.

Ostwald ripening of clays and metamorphic minerals.

Analyses of particle size distributions indicate that clay minerals and other diagenetic and metamorphic minerals commonly undergo recrystallization by Ostwald ripening. The shapes of their particle size distributions can yield the rate law for this process. One consequence of Ostwald ripening is that a record of the recrystallization process is preserved in the various particle sizes. Therefore, one can determine the detailed geologic history of clays and other recrystallized minerals by separating, from a single sample, the various particle sizes for independent chemical, structural, and isotopic analyses.

Etoposide compared with the combination of vincristine, doxorubicin, and cyclophosphamide in the treatment of small cell lung cancer.

One hundred and three patients with small cell lung carcinoma were stratified according to stage of disease (47 limited disease, 56 extensive disease) and then randomised to receive etoposide 300 mg/m2 alone for two days or a combination (VAC) of vincristine 1 mg/m2, doxorubicin (Adriamycin) 50 mg/m2, and cyclophosphamide 1000 mg/m2. The drugs were given at three week intervals. Patients were assessed after three cycles of treatment and continued with the same regimen if in complete remission and with the alternative regimen if in partial remission; they were withdrawn if the disease had progressed. Twenty four patients (23%) achieved complete remission and this occurred more often when patients were receiving VAC (19 of 82) than etoposide (5 of 75). There was no difference, however, in overall survival between those initially treated with etoposide and those having combination chemotherapy, whether for limited disease (both 8 months) or extensive disease (7 and 5.5 months). Toxicity was less with etoposide. Survival was disappointing, especially with limited disease, even in patients who showed a complete response to treatment.

Stimulus factors influencing the identification of voiced stop consonants by normal-hearing and hearing-impaired adults.

The effects of mild-to-moderate hearing impairment on the perceptual importance of three acoustic correlates of stop consonant place of articulation were examined. Normal-hearing and hearing-impaired adults identified a stimulus set comprising all possible combinations of the levels of three factors: formant transition type (three levels), spectral tilt type (three levels), and abruptness of frequency change (two levels). The levels of these factors correspond to those appropriate for /b/, /d/, and /g/ in the /ae/ environment. Normal-hearing subjects responded primarily in accord with the place of articulation specified by the formant transitions. Hearing-impaired subjects showed less-than-normal reliance on formant transitions and greater-than-normal reliance on spectral tilt and abruptness of frequency change. These results suggest that hearing impairment affects the perceptual importance of cues to stop consonant identity, increasing the importance of information provided by both temporal characteristics and gross spectral shape and decreasing the importance of information provided by the formant transitions.

Cost management techniques. Part II.

Reducing the cost of purchased products and services is of continuing importance to hospital purchasing and materials managers. In the first of three articles, the authors addressed the life cycle costing approach applied to purchasing as a means of evaluating total costs over the life of a product rather than only evaluating quoted prices. Life cycle costing is most appropriate for longer lived products, those with maintenance, repair, and disposal considerations. This article continues the discussion of cost management techniques focusing on ways to identify and manage total costs of purchasing and acquisition. The third article will present integrated cost and value management techniques.