Enhancement of human basophil histamine release by interleukin 5.

Substantial evidence indicates a close relationship between eosinophils and basophils. We examined whether interleukin 5 (IL-5), known to be eosinophilopoietic and capable of selectively regulating eosinophil functions, has an affect on basophil functions. IL-5 enhanced basophil histamine release evoked by anti-IgE, formyl-methionyl-leucyl-phenylalanine, or ionophore A23187 at picomolar concentrations. Direct action of IL-5 on human basophils was confirmed using highly purified basophil populations. These observations reveal the novel fact that IL-5 is able to modulate basophil functions as well as eosinophil functions, and suggest that normal human basophils possess functional receptors for IL-5.

Human neuroblastoma cells express alpha and beta platelet-derived growth factor receptors coupling with neurotrophic and chemotactic signaling.

Both platelet-derived growth factor (PDGF) A- and B-chains are expressed in mammalian neurons, but their precise roles still remain to be clarified. In the present studies, we examined the expression of two PDGF receptor genes in human tumor cell lines derived from neural crest. The expression of alpha and/or beta PDGF receptors was detected in a wide variety of neural crest-derived human tumor cell lines such as neuroblastoma, primitive neuroectodermal tumor, and Ewing's sarcoma by RNA blot analysis, and confirmed by immunoblot analysis. We have also demonstrated that PDGF receptors on the human neuroblastoma cell lines were biologically functional. Accordingly, chemotactic and mitogenic activities were induced by either PDGF-AA or PDGF-BB in serum-free medium. PDGF isoforms as well as nerve growth factor induced morphological changes showing neuronal cell maturation. Moreover, PDGF coordinately increased the levels of the transcript of the midsize neurofilament gene. The neuroblastoma cell lines also expressed the transcripts of PDGF A- and B-chains. These findings suggest that PDGF isoforms are involved not only in the promotion of the neuroblastoma cell growth, but also in neuronal cell migration, growth, and differentiation in human brain development.

Cholecystokinin-B/gastrin receptor: a novel molecular probe for human small cell lung cancer.

The brain-gut hormones, gastrin and cholecystokinin, have a trophic effect on the gastrointestinal mucosa in vivo and promote the growth of several neoplastic cell lines. In this study, cholecystokinin-B/gastrin receptor has been demonstrated to provide a novel molecular marker for the diagnosis of small cell lung cancer by using biopsy specimens. Physiological expression of the receptor mRNA is detectable in particular areas of the human brain, stomach, and pancreas but not in the lung. The receptor mRNA was detected selectively in all small cell lung cancer (10 cases) with a RT-PCR assay. By contrast, it was detectable in only 1 of 13 squamous cell carcinomas or 21 adenocarcinomas of the lung. Thus, the cholecystokinin-B/gastrin receptor could be an attractive therapeutic target for small cell lung cancer.

Enhancement of tissue-type plasminogen activator (t-PA) activity by purified t-PA receptor expressed in human endothelial cells.

We demonstrated previously that tissue-type plasminogen activator (t-PA) bound to its specific receptor (t-PAR) on human umbilical vein endothelial cells (HUVEC) in suspension and that t-PAR of mol wt. 20 kDa interacted only with t-PA to form 90 kDa complex (Fukao, H., Hagiya, Y., Nonaka, T., Okada, K., and Matsuo, O. (1992) Biochem. Biophys. Res. Commun. 187, 956-962). In the present study, 20 kDa t-PAR was purified from HUVEC and the function of the t-PAR was investigated by analyzing its effect on plasminogen activation by t-PA. About 2.2 microg t-PAR protein was purified from cell lysate of 1.0 X 10(9) HUVEC as a single band on sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) by gel filtration with TSK-3000SW and reversed phase separation with high performance liquid chromatography (HPLC). 125I-t-PA but not 125I-plasminogen specifically bound to the purified t-PAR in ligand blot assay. Plasminogen activation by t-PA in the presence of purified t-PAR in solution was increased. Furthermore, t-PA bound to immobilized t-PAR efficiently expressed its plasminogen activation activity. Kinetic analysis revealed that t-PA in the presence of soluble t-PAR and t-PA bound to immobilized t-PAR exhibited 34- and 90-fold increase in plasminogen activation, respectively. The t-PAR did not interact with anti-annexin II antibody. These findings indicate that the 20 kDa t-PAR is a novel molecule which immobilizes t-PA and enhances its proteolytic activity on the cell surface of endothelial cells.

Regulation of the function of eosinophils and basophils.

Both eosinophils and basophils play active pathogenic roles in the inflammation associated with allergic disorders. Both types of cells share a majority of their cell surface structures, and because of these common surface molecules, both cells can be stimulated with a single ligand simultaneously. The growth of both types of cells is controlled by IL-3, GM-CSF, and IL-5. All three growth factors are also capable of priming both eosinophils and basophils for enhanced biological functions, such as increased mediator release and prolonged survival. Both cells express beta 2 integrins, and in contrast to neutrophils, they also express several beta 1 integrins. Ligation of these adhesion molecules also transduces the intracellular signal leading to regulation of the cellular functions. In this review, we briefly describe the effects of various ligands of surface receptors and several pharmacological compounds on the functions of human eosinophils and basophils.

Effect of heat shock on the expression of urokinase-type plasminogen activator receptor in human umbilical vein endothelial cells.

We investigated the effect of heat shock on the fibrinolytic potential of human umbilical vein endothelial cells (HUVECs) in culture. When cultured at 43 degrees C, the mRNA for heat shock protein 70 (HSP70) was dramatically induced within 120 min with a maximal induction of more than 90-fold compared with that in HUVECs cultured at 37 degrees C. The level of urokinase-type plasminogen activator (u-PA) receptor (u-PAR) mRNA increased up to 2.2-fold in response to heat shock, which was associated with the increased u-PA binding and cell-surface u-PA activity determined by adding exogenous u-PA to acid-treated HUVECs. The increased u-PAR mRNA returned to normal level when HUVECs were further incubated at 37 degrees C for 180 min, and this decline was not affected in the presence of actinomycin D. Though the secreted antigens for tissue-type plasminogen activator (t-PA) and type 1 plasminogen activator inhibitor (PAI-1) in the conditioned medium (CM) of HUVECs were simultaneously increased at 43 degrees C during this period, the increase in the levels of t-PA (about 26.6-fold at 120 min) was greater than that of PAI-1 (1.8-fold at 120 min). The fibrinolytic activity of CM obtained from HUVECs at 43 degrees C was significantly enhanced up to 3-fold, indicating that heat shock induced hyperfibrinolytic states in HUVECs. The secretion of u-PA into CM was also enhanced by heat shock. These results suggested that human endothelial cells respond to hyperthermia by inducing HSP70 followed by hyperfibrinolytic states with the enhanced expression of u-PAR as well as that of t-PA and u-PA.

Determination of maximal power output at neuromuscular fatigue threshold.

The purpose of this study was to determine the maximal power output at the neuromuscular fatigue threshold (EMGFT), as estimated from electromyographic (EMG) data from representative leg muscles during cycling. The rate of rise in integrated EMG activity as a function of time (iEMG slope) was calculated at each of four constant-power-output ergometer bouts for 20 subjects. The iEMG slopes were plotted against work rates that were well described as linear functions (0.84 < R2 < 0.99). This iEMG slope vs. work rate relationship was extrapolated to zero slope to give an intercept on the power axis that was interpreted as the highest work rate sustainable without EMG evidence of neuromuscular fatigue (EMGFT). Each individual EMGFT was then expressed in terms of an O2 output (VO2) equivalent on the basis of the individual delta efficiency calculated during a linearly increasing maximal exercise test on the same bicycle ergometer. Results indicated a highly significant correlation (r = 0.92, P < 0.01) between EMGFT VO2 and anaerobic threshold VO2, as determined by conventional gas exchange criteria. The mean EMGFT VO2 (1.84 +/- 0.55 l/min) was, however, significantly greater (P < 0.01) than the anaerobic threshold VO2 (1.72 +/- 0.54 l/min). It was suggested that the EMGFT may provide an attractive alternative to the measurement of the highest work rate that can be sustained without evidence of neuromuscular fatigue.

Effect of muscle mass on V(O(2)) kinetics at the onset of work.

The dependence of O(2) uptake (V(O(2))) kinetics on the muscle mass recruited under conditions when fiber and muscle recruitment patterns are similar following the onset of exercise has not been determined. We developed a motorized cycle ergometer that facilitated one-leg (1L) cycling in which the electromyographic (EMG) profile of the active muscles was not discernibly altered from that during two-leg (2L) cycling. Six subjects performed 1L and 2L exercise transitions from unloaded cycling to moderate [VT) exercise. The 1L condition yielded kinetics that was unchanged from the 2L condition [the phase 2 time constants (tau(1), in s) for 0.05; for >VT: 1L = 26.8 +/- 12.0; 2L = 27.8 +/- 16.1, P > 0.05]. The overall V(O(2)) kinetics (mean response time) was not significantly different for the two exercise conditions. However, the gain of the fast component (the amplitude/work rate) during the 1L exercise was significantly higher than that for the 2L exercise for both moderate and heavy work rates. The slow-component responses evident for heavy exercise were temporally and quantitatively unaffected by the 1L condition. These data demonstrate that, when leg muscle recruitment patterns are unchanged as assessed by EMG analysis, on-transient V(O(2)) kinetics for both moderate and heavy exercise are not dependent on the muscle mass recruited.

Optimal pedaling rate estimated from neuromuscular fatigue for cyclists.

This study was designed to examine the optimal pedaling rate for pedaling exercise at a given work intensity for cyclists. Six college-aged cyclists each performed six sessions of heavy pedaling exercise at individually selected work rates based on their aerobic capacity. The optimal pedaling rate was evaluated on the basis of minimal neuromuscular fatigue as evidenced by the integrated electromyogram (iEMG) slope defined by the changes in iEMG as a function of time. The means of the iEMG slope demonstrated a quadratic curve versus pedaling rate. The mean values at 80 rpm (0.53 (SD 0.20) microV.min-1) and 90 rpm (0.67 (SD 0.23) microV.min-1) were significantly smaller than those values at any other pedaling rate. On the other hand, the mean value of oxygen uptake (VO2) expressed as a percent of the subject's maximal VO2 (% VO2max) at each pedaling rate also showed a quadratic curve with minimal values at about 60 or 70 rpm. VO2 at 70 rpm (84.0 (SD 5.0) % VO2max) was significantly smaller than those values at 80 rpm (86.3 (SD 3.5) % VO2max), 90 rpm (87.4 (SD 3.8) % VO2max), and 100 rpm (90.1 (SD 3.8) % VO2max). These data strongly suggest that the optimal pedaling rate estimated from neuromuscular fatigue in working muscles is not coincident with the pedaling rate at which the smallest VO2 was obtained, but with the preferred pedaling rate of the subjects. Our findings also suggest that the reason that cyclists prefer a higher pedaling rate is closely related to the development of neuromuscular fatigue in the working muscles.

Neuromuscular, metabolic, and kinetic adaptations for skilled pedaling performance in cyclists.

PURPOSE: The purpose of this study was to clarify the reason for the difference in the preferred cadence between cyclists and noncyclists. METHODS: Male cyclists and noncyclists were evaluated in terms of pedal force, neuromuscular activity for lower extremities, and oxygen consumption among the cadence manipulation (45, 60, 75, 90, and 105 rpm) during pedaling at 150 and 200 W. Noncyclists having the same levels of aerobic and anaerobic capacity as cyclists were chosen from athletes of different sports to avoid any confounding effect from similar kinetic properties of cyclists for lower extremities (i.e., high speed contraction and high repetitions in prolonged exercise) on both pedaling performance and preferred cadence. RESULTS: The peak pedal force significantly decreased with increasing of cadence in both groups, and the value for noncyclists was significantly higher than that for cyclists at each cadence despite the same power output. The normalized iEMG for vastus lateralis and vastus medialis muscles increased in noncyclists with rising cadence; however, cyclists did not show such a significant increase of the normalized iEMG for the muscles. On the other hand, the normalized iEMG for biceps femoris muscle showed a significant increase in cyclists while there was no increase for noncyclists. Oxygen consumption for cyclists was significantly lower than that for noncyclists at 105 rpm for 150 W work and at 75, 90, and 105 rpm for 200 W work. CONCLUSIONS: We conclude that cyclists have a certain pedaling skill regarding the positive utilization for knee flexors up to the higher cadences, which would contribute to a decrease in peak pedal force and which would alleviate muscle activity for the knee extensors. We speculated that pedaling skills that decrease muscle stress influence the preferred cadence selection, contributing to recruitment of ST muscle fibers with fatigue resistance and high mechanical efficiency despite increased oxygen consumption caused by increased repetitions of leg movements.

Effects of endothelial cells on activity of staphylokinase.

Staphylokinase (SAK), produced by Staphylococcus aureus, induces fibrinolytic activity in circulation without systemic fibrinolytic activation. Since the effect of blood vessels on the activity of SAK has not yet been clarified, plasminogen activator (PA) activity of SAK in the presence or absence of endothelial cells was analyzed. The endothelial cells used in this experiment were of a cloned established cell line (TKM-33). In the expression of PA activity by SAK or streptokinase (SK), the kinetic constants revealed as Vmax/km were increased about 1.5-fold in the presence of endothelial cells. Furthermore, an initial lag phase which was observed during the plasminogen activation by SAK was markedly shortened in the presence of endothelial cells. In the case of SK, an initial lag phase was not observed in the absence or presence of endothelial cells. Although PA activity of SAK was inhibited by alpha 2-antiplasmin (alpha 2-AP), the inhibitory effect of alpha 2-AP in the presence of endothelial cells was weaker than in the absence of endothelial cells. The cyanogen bromide digested fibrinogen fragment-2 (FCB-2) distinctly enhanced the PA activity of SAK in the absence and the presence of endothelial cells. However, alpha 2-AP and FCB-2 did not cause a significant alteration of PA activity of SK even in the absence or presence of endothelial cells. These findings suggest that PA activity of SAK is enhanced by endothelial cells, but inhibited by alpha 2-AP. Moreover, PA activity of SAK is further enhanced by fibrin clot in the presence of endothelial cells.

Studies on a new antibiotic M-92 produced by Micromonospora. II. Isolation and physicochemical properties of M-92 and its components.

A new antibiotic complex, M-92 was isolated from the whole fermentation broth of Micromonospora verruculosa MCRL 0404. The whole broth was mixed with talc and filtered. The filter cake thus obtained was extracted with acidic methanol to give a crude powder of M-92 complex, which was then separated into A (acidic) and N (neutral or weakly acidic) groups. The A group components are soluble in alkaline water (pH 8.5), while the N group components are not. These components were further separated into six major components designated VA-2, BA-4, BA-5, BN-1, BN-2 and BN-3 by silica gel column chromatography. Components with the letter "V" are reddish violet and those with "B" are blue. The IR and UV spectra of these components suggest that their chromophores may be the same or very closely related to one another. The molecular formula of BN-3 was determined to be C29H23NO9 by mass spectrometry. The results of various spectroscopies on BN-3 suggest that M-92 components consisted of chromophores in which juglone (5-hydroxynaphthoquinone) is conjugated with naphthazarin (5,8-dihydroxynaphthoquinone).

Yuta (shaman) and community mental health on Okinawa.

This study reports on the relations between the yuta (shaman) and the community mental health activities on Okinawa, Japan. Focus is on the process of initiation of the yuta and its meanings from the mental health viewpoints, the functions of the yuta in the particular cultural background of the island, and the importance of admitting the existence of the yuta in its relations to the psychiatric treatment in a mental hospital. The discussion is based on the authors' research findings which were obtained mostly through their therapeutic activities and their field studies. The authors' assumption is that each culture has in it a certain social background that is unique in that culture by which a stress or insanity is increased or brought about, but in the same culture there are ways to decrease the stress and to cure the insanity.

[A case of food-dependent-exercise induced anaphylaxis possibly induced by shellfish (Sulculus Supertexta and Turbo Cornutus)].

A 17 years old girl experienced an anaphylactic reaction of urticaria, dyspnea, syncope and hypotension while riding a bicycle 55 minutes after eating shellfish Lapas shellfish which was a-like Sulculus Supertexta (SS). She recovered within several hours after the emergency treatment. Another attack occurred 3 months later while she was running with a dog 30 minutes after eating shellfish (Turbo Cornutus; TC). RAST scores were 4 for Lapas and 2 for TC. RAST inhibition test by ELISA showed a high crose-reaction between keyhole limpet hemocyanin (KLH) and Lapas, and between KLH and TC, while the cross reaction between Lapas and TC was low. Gel chromatography with sephacryl G-200 revealed that both Lapas and TC had several allergens with different molecules which were detected by ELISA. Exercise challenge produced an immediate fall of FEV1 and a significant increase in plasma histamine levels for 45 minutes.