A new method for histamine release from purified peripheral blood basophils using monoclonal antibody-coated magnetic beads

A new method for evaluating histamine release from purified basophils was developed. Basophil-containing leukocytes were directly purified from a small amount of peripheral blood using monoclonal antibody BA312-coated magnetic beads. The purified basophils still rosetted to magnetic beads maintained a normal response to anti-IgE and to dust mite allergen in comparison with the conventional method using washed leukocytes. This methodology facilitates the purification of basophils, anti-IgE- and allergen-induced histamine release, and subsequent histamine determination within only 3 h. The released histamine was analyzed by an enzyme-linked immunosorbent assay (ELISA) with a characteristic detection profile. Since all steps were performed in 96-well microplates, many clinical samples could be analyzed at the same time, permitting easy applications in routine laboratories.     

The dopamine agonist cabergoline provides neuroprotection by activation of the glutathione system and scavenging free radicals

Free radicals are involved in the pathogenesis and/or progression of Parkinson's disease (PD). Several ergot derivative dopamine (DA) agonists have been reported to scavenge free radicals in vitro and to show a neuroprotective effect in vivo. We investigated the in vitro free radical scavenging and antioxidant activities of cabergoline, a long-acting ergot DA agonist, as well as its ability to activate glutathione (GSH), catalase (Cat) and superoxide dismutase (SOD) activating effects and its in vivo neuroprotective properties against 6-hydroxydopamine (6-OHDA) intracerebroventricularly (i.c.v.) in mice. The striatal DA turnover induced by i.c.v. injection of 6-OHDA was completely normalized by pretreatment with cabergoline. Moreover, cabergoline scavenged free radicals in vitro and significantly reduced lipid peroxidation in vitro and in vivo. Furthermore, daily administration of cabergoline to mice significantly increased striatal GSH levels by activation of RNA expressions of GSH-related enzymes, although striatal Cat and SOD activities did not change. In addition, our present results suggest that repeated administration of cabergoline attenuates both 6-OHDA-induced nigrostriatal DAergic dysfunction and DA neuronal cell death, since cabergoline also had a neuroprotective effect in the immunohistochemical experiment. In conclusion, our findings indicate that the multiple antioxidant mechanisms of cabergoline, such as activation of the GSH system and the direct free radical scavenging activity, may explain the neuroprotective effect of this ergot DA agonist.     

Volatile substances from larval habitats mediate species-specific oviposition in Anopheles mosquitoes

Oviposition site selection has been recognized as critical both for the survival and population dynamics of mosquitoes. Volatile substances released from larval habitats have been implicated as potential olfactory cues mediating oviposition. In our continuing studies of cues involved in oviposition site selection, we collected material from the larval habitats of Anopheles albimanus Wiedemann and Anopheles vestitipennis Dyar & Knab, i.e., cyanobacterial mats and Typha domingensis Pers. litter, respectively. The volatile compounds were extracted by freeze-drying the material and trapping the volatilized material on a -55 degrees C titanium condenser. For oviposition trials conducted with wild-caught females, the tested volatile materials were pipetted onto filters floating on the surface of distilled water in Teflon beakers that were placed within oviposition cages. For both species, volatile materials in low concentrations increased oviposition, assessed as egg density, whereas there was a shift to reduced oviposition at higher concentrations. Volatile effect was strongly habitat/species-specific as shown by reciprocal treatment tests.     

Macrophage function in the Schistosoma mansoni egg-induced pulmonary granuloma. Role of arachidonic acid metabolites in macrophage Ia antigen expression.

The ability of arachidonic acid (AA) metabolites to regulate I-region-associated (Ia) antigen expression on macrophages from schistosome-egg-induced pulmonary granulomas was examined. The prostaglandin (PG) analog 15-S-15-CH3-PGE1 (M-PGE1) and PGF2 alpha were found to modulate the kinetics of Ia expression when administered in vivo. Methyl-PGE1 significantly suppressed Ia antigen expression by hypersensitivity granuloma macrophages, while PGF2 alpha appeared to potentiate the expression. Lymphokine-induced Ia antigen expression by cultured granuloma macrophages was likewise dramatically inhibited by M-PGE1. Further analysis using systemically administered inhibitors of AA metabolism demonstrated that the cyclooxygenase inhibitor indomethacin caused augmentation of Ia expression. In contrast, lipoxygenase inhibitors significantly reduced both Ia expression and granuloma size. The role of AA metabolites in modulating chronic inflammation is discussed.     

Venodilator effects of adenosine triphosphate and sodium nitroprusside; Comparisons during controlled hypotension

Adenosine triphosphate as well as sodium nitroprusside has been used for hypotensive anesthesia. The purpose of this study was to examine the possibility that two hypotensive drugs may exert different effects on venous capacitance during controlled hypotension. In rats anesthetized with ketamine, mean arterial pressure was lowered to 50mmHg by intravenous infusion of adenosine triphosphate or sodium nitroprusside. Venous capacitance was assessed before and during induced hypotension by measuring the mean circulatory filling pressure (MCFP). MCFP was measured after briefly arresting the circulation by inflating an indwelling balloon in the right atrium. MCFP was lower during adenosine triphosphate-induced as well as sodium nitroprusside-induced hypotension as compared with the respective value at control (P < 0.01 for adenosine triphosphate and sodium nitroprusside). However, the decrease in MCFP by adenosine triphosphate (0.8 ± 0.1mmHg) was less (P < 0.01) than that by sodium nitroprusside (2.3 ± 0.3mmHg). These results suggest that at a comparable level of arterial hypotension venodilator effect of adenosine triphosphate was less than that of sodium nitroprusside. Less venodilatation during adenosine triphosphate-induced hypotension may contribute to the maintenance of cardiac output during hypotensive anesthesia.(Hoka S, Takeshita A, Aishima K et al.: Venodilator effects of adenosine triphosphate and sodium nitroprusside; comparisons during controlled hypotension. J Anesth 1: 144–147, 1987)     

Validity of observed differences in dietary surveys by two self-administered questionnaires over a 5-year period: Concordance with self-reported change

Differences observed by comparing the responses to two surveys taken 5 years apart were compared with self-reported changes in dietary habits in the second survey to examine the construct validity of dietary change. That is, when an observed difference about a certain food was consistent with a self-reported change, these two methods seemed to show a high validity regarding dietary change. Both surveys used the same self-administered food intake frequency questionnaire, and subjective changes in food intake were evaluated at the second survey (self-reported change). Eight hundred fifty-five males and one thousand females aged 30-69 years were analyzed. Since results by both methods showed a higher frequency of increased intake of green-yellow vegetables in general, pale- colored vegetables in general, carrots, and squash, these results were thought to be of high validity. However, cabbage, lettuce, and seaweeds showed inconsistent results regarding higher frequencies of intake. Changes among quintiles of lower frequency seemed to show lower validity because the results obtained by the two methods were often inconsistent for dairy foods and some other foods. Thus, self-reported changes may reflect respondents’ attitudes toward foods, not their actual behavior.     

Multiple channels for occupational health services to small-scale enterprises in Japan

This study was performed to clarify the characteristics of multiplechannels for occupational health services (OHS) to small-scaleenterprises (SSEs) in Japan employing less than 50 workers byreviewing relevant research papers. OHS were provided to SSEsby both government organizations and non-government organizations.Government organizations included the Japanese Industrial Safety& Health Association, regional occupational health centres,prefectural occupational health promotion centres, and otherhealth centres. Non-government organizations were parent companies,occupational health organizations, hospitals, medical associations,trade associations and health insurance societies. The qualityof OHS in terms of provision of health personnel and the natureof their services differed greatly between the organizations.Health care programmes were more popular than management ofthe work environment or working practices in OHS to SSEs. Feworganizations provided comprehensive OHS to SSEs. At present,parent companies and occupational health organizations appearto provide the best OHS available to SSEs in Japan.     

The occurrence of polymorphism of mannose 6-phosphate/insulin-like growth factor 2 receptor gene in laboratory and wild rats

Carcinogen-resistant inbred DRH rat strain was established from closed colony Donryu rats in the presence of 3'-methyl-4-dimethylaminoazobenzene (3'-Me-DAB). Despite using 3'-Me-DAB during the stage of selection, the DRH rats developed normally and did not show any spontaneous tumor at over 1 year of age. In the present study, we examined the polymorphism in mannose 6-phosphate/insulin-like growth factor 2 receptor (M6p/Igf2r) gene and found that the DRH rat showed CCC (Proline)-type polymorphism in exon 48 and the Donryu rat had GCC (Alanine) sequence. Since the DRH rat was developed from the Donryu rat, we examined whether this polymorphism in exon 48 of M6p/Igf2r gene was due to mutation that occurred at the stage of selection in the presence of 3'-Me-DAB, using several other laboratory and wild rats. We detected the presence of polymorphism at the same site of the M6p/Igf2r gene among these rats. It is likely that the polymorphism in exon 48 of the M6p/Igf2r gene is present broadly in rats since ancient times and not due to the mutation during the course of selection unless this site is a hot spot for chemical carcinogens.     

Synthesis and physico‐chemical properties of peptides in soil humic substances

Abstract: Soil humic substances (HS) are heterologous, polydispersive, and multi‐functional organometallic macromolecules ubiquitous in soils and sediments. They are key players in the maintenance of the belowground ecosystems and in the bioavailability of both organic and inorganic contaminants. It is widely assumed that the peptidic substructures of HS are readily degraded and therefore do not contribute significantly to interactions with contaminants such as toxic metals. To investigate the turnover of humified peptides, laboratory soil aging experiments were conducted with ¹³C‐glucose or ¹⁵N‐nitrate for 8.5 months. Evidence for random‐coil peptidic structures in the labeled HS was obtained from 2‐D nuclear magnetic resonance (NMR), pyrolysis gas chromatography‐mass spectrometry (pyro‐GC‐MS), and circular dichroism data. Interaction of metals with the peptidic carbonyls of labeled HS was rationalized from the solid‐state NMR data. Detailed ¹³C and ¹⁵N labeling patterns of amino acid residues in the acid hydrolysates of HS acquired from NMR and GC‐MS revealed two pools of peptides, i.e. one extant (unlabeled) and the other, newly humified with little isotopic scrambling (fully labeled). The persistence of pre‐existing peptidic structures indicates their resistance to degradation while the presence of fully labeled peptidic amino acids suggests wholesale incorporation of newly synthesized peptides into HS. These findings are contrary to the general notion that humified peptides are readily degraded.