Genome-wide association study identifies a potent locus associated with human opioid sensitivity.

Opioids, such as morphine and fentanyl, are widely used as effective analgesics for the treatment of acute and chronic pain. In addition, the opioid system has a key role in the rewarding effects of morphine, ethanol, cocaine and various other drugs. Although opioid sensitivity is well known to vary widely among individual subjects, several candidate genetic polymorphisms reported so far are not sufficient for fully understanding the wide range of interindividual differences in human opioid sensitivity. By conducting a multistage genome-wide association study (GWAS) in healthy subjects, we found that genetic polymorphisms within a linkage disequilibrium block that spans 2q33.3-2q34 were strongly associated with the requirements for postoperative opioid analgesics after painful cosmetic surgery. The C allele of the best candidate single-nucleotide polymorphism (SNP), rs2952768, was associated with more analgesic requirements, and consistent results were obtained in patients who underwent abdominal surgery. In addition, carriers of the C allele in this SNP exhibited less vulnerability to severe drug dependence in patients with methamphetamine dependence, alcohol dependence, and eating disorders and a lower 'Reward Dependence' score on a personality questionnaire in healthy subjects. Furthermore, the C/C genotype of this SNP was significantly associated with the elevated expression of a neighboring gene, CREB1. These results show that SNPs in this locus are the most potent genetic factors associated with human opioid sensitivity known to date, affecting both the efficacy of opioid analgesics and liability to severe substance dependence. Our findings provide valuable information for the personalized treatment of pain and drug dependence.

Relationship between gagging severity and its management in dentistry.

The purpose of this study was to investigate how grading according to our new gagging reflex index correlated with patient background and subsequent management. After obtaining institutional approval and informed consent, 110 patients with a gagging problem were enrolled. The patients completed the State-Trait Anxiety Inventory (STAI), the Dental Anxiety Scale (DAS), and a health questionnaire at initial consultation. On the second visit, an intra-oral examination was carried out and the severity of gag reflex determined according to our new, 5-level Classification of Gagging Problem (CGP) index: normal gagging but not desensitised (G1 = score 1); mild gagging (G2 = score 2); moderate gagging (G3 = score 3); severe gagging (G4 = score 4); and very severe gagging (G5 = score 5). No difference was found in grade based on age or STAI or DAS scores. The CGP score in male patients was significantly higher than that in female. The management classification method and degree of desensitisation were investigated retrospectively in each patient at 3 months and 1 year after initial consultation. The higher the CGP grade, the more often intravenous sedation or general anaesthesia was required due to difficultly in desensitisation. The present results suggest that determining whether it is possible to examine the molar area without inducing the gag reflex offers the key to deciding the treatment strategy.

Emerging ST121/agr4 community-associated methicillin-resistant Staphylococcus aureus (MRSA) with strong adhesin and cytolytic activities: trigger for MRSA pneumonia and fatal aspiration pneumonia in an influenza-infected elderly.

The pathogenesis of community-associated methicillin-resistant Staphylococcus aureus (CA-MRSA) pneumonia in influenza-infected elderly individuals has not yet been elucidated in detail. In the present study, a 92-year-old man infected with influenza developed CA-MRSA pneumonia. His CA-MRSA was an emerging type, originated in ST121/agr4 S. aureus, with diversities of Panton-Valentine leucocidin (PVL)(-)/spat5110/SCCmecV(+) versus PVL(+)/spat159((etc.))/SCCmec (-), but with common virulence potentials of strong adhesin and cytolytic activities. Resistance to erythromycin/clindamycin (inducible-type) and gentamicin was detected. Pneumonia improved with the administration of levofloxacin, but with the subsequent development of fatal aspiration pneumonia. Hence, characteristic CA-MRSA with strong adhesin and cytolytic activities triggered influenza-related sequential complications.

Induction of an eosinophil chemotactic factor production from T lymphocytes by a B cell lymphoma line.

Production of an eosinophil chemotactic factor (ECF) from human mononuclear leukocytes (MNL) was induced by coculture with an irradiated B cell lymphoma line, BALL-1. BALL-1 induced ECF production from OKT4-positive T lymphocytes without evident IL-2 production. Treatment of MNL with anti-IL-2 antibody failed to suppress the BALL-1-induced ECF production, whereas the treatment strongly inhibited IL-2-induced ECF production. Control supernatants of BALL-1 cells alone did not induce ECF production. BALL-1 fixed with periodate-lysine-paraformaldehyde, but not acetone or ethanol, induced evident ECF production. The isoelectric point of BALL-1-induced ECF (m.w. 10-30 kD) was around pI 7, whereas that of the IL-2-induced ECF was around pI 5. A combination of monoclonal antibodies against IL-3, IL-5, and GM-CSF suppressed the activity of the IL-2-induced ECF but not that of the BALL-1-induced ECF. BALL-1-induced ECF suppressed a respiratory burst from an eosinophilic cell line (YY-1) induced by N-formyl-L-methionyl-L-leucyl-L-phenylalanine, whereas the IL-2-induced ECF did not, suggesting that the biological function of these two ECF is different, at least in the effect on respiratory burst of eosinophils. From the present results we propose that one reason for infiltration of eosinophils into the stroma of tumors is that some tumor cells can stimulate OKT4-positive T lymphocytes to produce an ECF, and that eosinophils attracted by this ECF exhibit biological functions which are different from those of eosinophils attracted by other ECF.

Production of fibroblast proliferative cytokines from T lymphocytes stimulated by a B cell lymphoma line and their functional heterogeneity.

Human mononuclear leukocytes (MNL) produced several factors with fibroblast proliferation activity (FPA) for HFL-1, a human lung fibroblast cell line, when MNL were cocultured with irradiated BALL-1, a B cell lymphoma line (BCLL), but not with other BCLL. The cellular source of BALL-1-induced FPA seemed to be CD4-positive T lymphocytes. On isoelectric electrophoresis, major activity of BALL-1-induced FPA was detected in the fractions around pH 4-5, and minor activity was present in the fractions around pH 6-7. Major BALL-1-induced FPA consisted of at least 4 different fibroblast proliferation factors (FPFs) according to their molecular weight; 320-600 kDa (P-I), 50-110 kDa (P-II), 22-38 kDa (P-III) and 4.6-11 kDa (P-IV). P-I had affinity to heparin though the rest had little or no affinity. FPA of P-I was suppressed by an antibody against acidic FGF, and FPA of P-III was suppressed by an antibody against IL-6. On the other hand, FPA of P-II and P-IV was suppressed by none of the antibodies against cytokines with FPA, such as FGF, IL-4, IL-6, IFN-gamma, TGF-beta and TNF-alpha. It was thus suggested that P-I was acidic FGF, that P-III was IL-6, and that P-II and P-IV were different cytokines from those described above. Furthermore, it was found that P-II and P-IV failed to exhibit proliferation activity for human umbilical vein endothelial cells (HUVEC).(ABSTRACT TRUNCATED AT 250 WORDS)

Monocyte chemoattractant protein-1 in idiopathic pulmonary fibrosis and other interstitial lung diseases.

Macrophages play a crucial role in the pathogenesis of idiopathic pulmonary fibrosis (IPF). To examine the mechanisms for increased monocyte/macrophage recruitment in IPF and nonIPF interstitial lung diseases (nonIPF) the localization of monocyte chemoattractant protein-1 (MCP-1) was investigated in 14 cases of IPF, seven cases of nonIPF, and seven normal control lungs (CTRL) by immunohistochemistry using a specific anti-MCP-1 monoclonal antibody, F9. By double immunohistochemical staining using F9 and one of the cell type specific antibodies significant differences in the staining pattern of MCP-1 were observed between IPF and nonIPF. In IPF MCP-1 was observed in cuboidal and flattened metaplastic epithelial cells, alveolar macrophages, and vascular endothelial cells. In contrast, no epithelial cells were stained for MCP-1 in nonIPF cases, although alveolar macrophages and vascular endothelial cells were labeled. Northern hybridization analysis of selected cases showed marked expression of MCP-1 messenger RNA (mRNA) in IPF and nonIPF compared with CTRL. These findings suggest that the MCP-1 production in IPF and nonIPF plays an important role in the recruitment of monocyte/macrophages. Monocyte chemoattractant protein-1 production by epithelial cells in IPF may be caused by the metaplastic nature of the epithelial cells and may be one of the key factors inducing the irreversible progression of IPF.

Pulmonary granulomatosis caused by aspirated green tea.

We describe the first reported case (to our knowledge) of pulmonary granulomatosis caused by aspirated green tea. In this case, we found granulomatous alveolitis with lymph follicles, T lymphocytosis with predominantly CD8+ cells in the bronchoalveolar lavage fluids, positive serum precipitin and proliferative response of peripheral blood lymphocytes to the tea infusion, and efficacy of steroid therapy. These results indicate that the pathogenesis of the disease was due to both humoral and cellular immunities to the aspirated green tea.

Atypical peritracheobronchial vasculitis and an effective treatment.

We report an interesting case of vasculitis in which the inflammatory lesion was limited to the peritracheobronchus. This case showed positive antineutrophil cytoplasmic antibodies, diffuse peritracheobronchial swelling, and vasculitis in its histology. Steroid therapy was effective for both roentgenological and serological findings. Although the biopsy specimen shows only inflammation and does not satisfy the WHO criteria of Wegener's granulomatosis (WG), a possible diagnosis of WG should not be disregarded.

[Nitric oxide inhalation as an effective therapy for acute respiratory distress syndrome due to near-drowning: a case report].

A 16-year-old boy with acute respiratory distress syndrome (ARDS) due to near-drowning was admitted to our hospital. ARDS was treated with low-level nitric oxide (NO) inhalation (ranging from 4 ppm to 1 ppm) for 24 days. Oxygenation was improved and pulmonary hypertension was reduced after NO inhalation, but systemic blood pressure, heart rate, and cardiac output were not affected. PaO2 improved from 153 Torr to 354 Torr under identical ventilating conditions (F1O2 1.0), and mean pulmonary arterial pressure fell from 40 mm Hg to 27 mmHg. It has been reported that NO inhalation alleviates ventilation-flow mismatch and pulmonary hypertension. It is unclear, however, whether this therapy improves the prognosis for ARDS. In our patient, NO inhalation was effective in alleviating the oxygenation impairment and pulmonary hypertension associated with ARDS.

Serodiagnosis of Hansen's disease/leprosy by enzyme-linked immunosorbent assay using cord factor (trehalose 6,6'-dimycolate) as an antigen.

IgG and/or IgM antibodies against mycobacterial cord factor (trehalose 6,6'-dimycolate, TDM) in sera of 65 patients of Hansen's disease (21 cases with smear-positive and 44 cases with smear-negative) and 60 healthy individuals were tested by enzyme-linked immunosorbent assay (ELISA) with TDM purified from Mycobacterium tuberculosis H37Rv as an antigen. Of 65 patients with Hansen's disease, 58 cases (89.2%) had positive results (21 samples from 21 patients, 100% with acid-fast bacilli positive in the lesion, and 37 samples from 44 patients, 84.0% with acid-fast bacilli negative Hansen's disease diagnosed clinically). The sensitivity and specificity of anti-cord factor ELISA were higher than those of anti-phenolic glycolipid-I (PGL-I) agglutination test. Among the total, 34 patients were classified clinically into three types of the disease, lepromatous leprosy (LL), borderline lepromatous (BL) and borderline tuberculoid (BT). The antibody titer showed LL > BL > BT, indicating that the elevation of anti-cord factor antibody titers appeared to be parallel with the degree of humoral immune response against M. leprae. By using semisynthetic cord factor consisting of a single subclass of mycolic acid from M. tuberculosis, it was revealed that sera from patients with Hansen's disease were highly reactive against alpha-mycoloyl cord factor (alpha-TDM) and less reactive against methoxy mycoloyl TDM (methoxy TDM), differed from sera of tuberculosis patients, which were highly reactive against both methoxy and alpha-mycoloyl cord factor (alpha-TDM). Most of sera from patients with Hansen's disease were more reactive against TMM than TDM, differed from sera of tuberculosis patients which were highly reactive against TDM. ELISA using TDM as an antigen is simple, reproducible and useful for the rapid serodiagnosis of Hansen's disease, especially for smear-negative cases.

Fibroblast proliferation factors in pulmonary granuloma induced by Trichosporon cutaneum in rabbits: presence of a lymphocyte-derived fibroblast proliferation factor and its functional specificity.

The fibroblast proliferation activity (FPA) in pulmonary granulomatous lesions in rabbits which were exposed once (primary response) or twice (secondary response) to Trichosporon cutaneum was examined using R9ab, a rabbit fibroblast cell line cell, and fibroblasts from the lesions of the primary and secondary responses. The FPA in lung extracts and cell-free culture supernatants of bronchoalveolar lavage cells from the secondary response was more evident than that from the primary response. FPA from the primary response were recovered at about 60, 18, and 4.5 kD and those from secondary response at about 60, 26, 18, and 4.5 kD on Sephadex G-75 gel filtration. Among the FPA, the activity with a molecular weight of 26 kD and a pI of 7.0 was derived from lymphocytes, whereas the other activities were derived from macrophages. The macrophage-derived fibroblast proliferation factors (FPF) enhanced proliferation of fibroblasts from the lesions of both primary and secondary responses, while the lymphocyte-derived FPF enhanced only proliferation from the secondary response. It was further found that lymphocyte-derived FPF could chemotactically attract fibroblasts from the secondary but not the primary response, indicating functional specificity of lymphocyte-derived FPF on fibroblasts in the secondary response. The present results suggest that this lymphokine with fibroblast proliferation and chemotactic activity plays an important role in the granuloma formation in the secondary response to T. cutaneum.

A B cell lymphoma line, BALL-1 stimulates T cells to produce a unique eosinophil chemotactic factor.

Human mononuclear leukocytes (MNL), probably OKT4-positive T cells, produced an eosinophil chemotactic factor (ECF) when they were cocultured with irradiated BALL-1, a B cell lymphoma line. Treatment of MNL, with anti-IL-2 antibody failed to suppress BALL-1-induced ECF production. Periodate-lysine-paraformaldehyde-fixed but not acetone- and ethanol-fixed BCLL induced evident ECF production. These results suggested that some cell surface molecules play a role in the induction of ECF production. Isoelectric point of BALL-1-induced ECF was around pH7, whereas that of IL-2-induced ECF was around pH 5. The molecular weight of BALL-1-induced ECF was between 10 and 30 kD. Although a combination of MoAb against IL-3, IL-5, and GM, CSF suppressed the activity of IL-2-induced ECF, it failed to suppress that of BALL-1-induced ECF. Furthermore, BALL-1-induced ECF suppressed fMLP-induced respiratory bursts of eosinophils, while IL-2-induced ECF failed. We propose that at least one reason for eosinophil infiltrate into the stroma of tumors is that the tumor cells stimulate T cells to produce BALL-1-induced ECF, and the eosinophils attracted by the ECF exhibit different functions from those by other ECF.

Electroporation of cell membrane visualized under a pulsed-laser fluorescence microscope.

Controlled permeability can be conferred to cell membranes by exposing cells to a microsecond electric pulse of sufficient intensity (electroporation). By constructing a fluorescence microimaging system with a submicrosecond time resolution we have been able to resolve temporally and spatially the events in a single cell under a microsecond electric pulse. An enormous membrane conductance, corresponding to a loss of 0.01-0.1% of the membrane area, was observed in those membrane regions where the transmembrane potential induced by the electric pulse exceeded a critical value. The conductance decreased to a low level in a submillisecond after the pulse, leaving a moderately electroporated cell.

Interleukin-1 production by polymorphonuclear leukocytes during the course of acute inflammation in rabbits.

Inflammation-induced immune enhancement was observed when an antigen was introduced during an early period of casein-induced inflammation. An immune potentiation factor was detected at the early inflammatory site of mice and rabbits. This immune potentiation factor of rabbits was identified as interleukin-1 (IL-1) in respect of either its biologic activities or its molecular structure. Expression of IL-1 mRNA was observed only in peritoneal exudate cells from the early inflammatory site in parallel with the generation of the immune potentiation activity. The inflammation-induced immune potentiation was assumed to be mediated by newly synthesized IL-1 at the early inflammatory site. By using immunocytochemical staining, it was definitely proved with respect to a single cell level that the polymorphonuclear leukocytes were the major cells in synthesizing IL-1 at the casein-induced inflammation in rabbits.

Apoptotic response of eosinophils in chronic eosinophilic pneumonia.

To clarify the pathogenesis of chronic eosinophilic pneumonia (CEP), the apoptosis of eosinophils from bronchoalveolar lavage (BAL-Eos) was compared with that of eosinophils from peripheral blood (PB-Eos) in six cases of CEP. The survival rate of eosinophils and the percentage of apoptotic cells of both types of eosinophils were examined, and the effects of interleukin 5 (IL-5) were evaluated. The role of Fas expression in apoptosis of these eosinophils was also studied. The survival rate of BAL-Eos on the third day of culture was significantly higher than that of PB-Eos (p < 0.01). This was associated with a lower proportion of apoptotic cells in BAL-Eos than in PB-Eos; the percentages of apoptotic cells in PB-Eos and BAL-Eos after 24 h of incubation were 21.7 +/- 3.4% and 10.6 +/- 1.7% respectively. IL-5 suppressed apoptosis and increased the survival rate of both PB-Eos and BAL-Eos. It was found that the apoptotic character of BAL-Eos differed from that of PB-Eos in at least three ways. Firstly, the positive rate of Fas expression on PB-Eos was increased after 24 h of incubation, whereas that on BAL-Eos did not change. Secondly, the expression of Fas on PB-Eos was suppressed by IL-5 (18.5 +/- 4.2% - 8.3 +/- 3.2%, p < 0.05), whereas IL-5 failed to suppress Fas expression on BAL-Eos (3.3 +/- 1.6% - 3.6 +/- 1.0%). Lastly, binding of antibody to Fas antigen induced apoptosis of PB-Eos, but not of BAL-Eos. These data suggested that Fas seemed to be involved in the apoptosis of PB-Eos, whereas BAL-Eos were Fas-resistant in chronic eosinophilic pneumonia. In conclusion, apoptosis of eosinophils might be suppressed by proinflammatory cytokines such as IL-5 leading to their accumulation in the lung. Chronic stimulation of eosinophils in the alveolar space with IL-5 may play a crucial role chronic eosinophilic disorders.

Alterations in cytokeratin expression by the alveolar lining epithelial cells in lung tissues from patients with idiopathic pulmonary fibrosis.

It is generally recognized that epithelial cytokeratins (CKs) are expressed in tissue-specific patterns and reflect differentiation, functional specialization, and pathological alterations of the cells. Differential epithelial cell types can thus be distinguished from each other by their selective expression of particular sets of CKs. To determine the characteristics of metaplastic and hyperplastic changes of alveolar-lining epithelial cells in the lungs of idiopathic pulmonary fibrosis (IPF), the expression of individual CKs was studied immunohistochemically using monospecific anti-CK monoclonal antibodies (anti-CKs 7, 8, 10, 13, 14, 16, 17, 18, 19). Biopsy specimens from 17 patients with IPF and normal lung tissues (NL) from seven patients with lung cancer were studied. In the IPF specimens, several kinds of altered epithelial cells were observed, which showed characteristic changes in CK expression compared with NL, especially CKs 8, 14, and 17. Hyperplastic type II cells expressed increased CKs 7, 8, and 19, but not CK 17; flattened or stratified squamous metaplastic cells expressed increased CKs 17 and 14, co-expressed with CKs 7, 8, and 19; bronchiolar metaplastic cells expressed increased CKs 7, 8, and 19, co-expressed with CKs 14 and 17; cuboidal metaplastic cells expressed increased CKs 7, 8, 17, and 19. The quantification of individual CKs in the tissues by enzyme-linked immunosorbent assay revealed increased expression of CKs 8, 14, and 17 in IPF lung tissues compared with NL. These results were consistent with the immunohistochemical observations. The hyperplastic and bronchiolar metaplastic phenotypes were characterized by their increased expression of simple CKs without CK alteration. The squamous metaplastic phenotype showed CK alterations, with the appearance of CKs 17 and 14. Epithelial cells are thus altered not only in shape, but possibly also in differentiation and function, with potential implications for the pathogenesis of IPF.

Heterogeneity of eosinophils in chronic eosinophilic pneumonia.

It has previously been shown that patients with chronic eosinophilic pneumonia can be divided into 2 groups according to the chemotactic response of their eosinophils to 5 different eosinophil chemotactic factors (ECFs) and laboratory findings. In contrast, eosinophils obtained by bronchoalveolar lavage from both groups responded to all 5 ECFs. The correlation between the two groups and the expression of several antigens (VLA-4, CD69, ICAM-1 and CD11b) on eosinophils. The VLA-4 expression of group 1 eosinophils was higher than that of group 2 eosinophils. More interestingly, eosinophils that migrated towards ECF-PI9 expressed less CD69 than those that migrated towards other STO-2-derived ECF. The heterogeneous response of eosinophils to STO-2-derived ECFs suggests that the population of eosinophils is heterogeneous.

Chemotactic heterogeneity of eosinophils in Kimura's disease.

We evaluated the chemotactic heterogeneity of eosinophils in Kimura's disease. Patients with Kimura's disease were divided into two groups according to their clinical findings: one group had no other symptoms (KD), and another was accompanied with atopic dermatitis (KD + AD). The chemotactic response of eosinophils from two groups to 5 eosinophil chemotactic factors (ECF) derived from STO-2, an established T cell line. Eosinophils from KD were attracted only by ECF-PI5 and PI6 but not by ECF-PI7, PI8 and PI9. On the other hand, eosinophils from KD + AD responded to all 5 ECF. Eosinophils were further fractionated into normodense and hypodense eosinophils, and assessed for their chemotactic response. We thus found that there was little essential difference in their chemotactic responses to STO-2-derived ECF except ECF-PI9, though random migration of hypodense eosinophils was enhanced. The hypothesis that hypodense eosinophils are in the activated form was not always true, especially in the chemotactic response to ECF.

Expression of apoptosis-related antigen on eosinophils in chronic eosinophilic pneumonia.

We examined the expression of apoptosis-related antigens Fas and bcl-2 on eosinophils from peripheral blood (PB) and bronchoalveolar lavage (BAL) in patients with chronic eosinophilic pneumonia (CEP). The expression of those antigens was assessed before and after culture with or without eosinophil chemotactic factors derived from an established T-cell line (STO-2-derived ECFs; ECF-PI5, 6, 7, 8, and 9), granulocyte-macrophage colony stimulating factor, and interleukin 5 (IL-5). We found that the expression of these antigens on eosinophils from PB increased after 24 h culture without any stimulation. In contrast, little or no change was observed even after 24 h culture in eosinophils from BAL. All STO2-derived ECFs and IL-5 suppressed Fas expression on eosinophils from PB. Furthermore, we found that eosinophils which were attracted by ECF-PI9 expressed Fas and bcl-2 more highly than those attracted by other ECFs and IL-5. Such a heterogeneous response of eosinophils to respective ECFs suggests the possibility of a heterogeneous population of eosinophils in patients with CEP.

Trehalose 6,6'-dimycolate (cord factor) of Mycobacterium tuberculosis induces corneal angiogenesis in rats.

Neovascularization or angiogenesis is required for the progression of chronic inflammation. The mechanism of inflammatory neovascularization in tuberculosis remains unknown. Trehalose 6, 6'-dimycolate (TDM) purified from Mycobacterium tuberculosis was injected into rat corneas. TDM challenge provoked a local granulomatous response in association with neovascularization. Neovascularization was seen within a few days after the challenge, with the extent of neovascularization being dose dependent, although granulomatous lesions developed 14 days after the challenge. Cytokines, including tumor necrosis factor alpha (TNF-alpha), interleukin-8 (IL-8), IL-1beta, and vascular endothelial growth factor (VEGF), were found in lesions at the early stage (within a few days after the challenge) and were detectable until day 21. Neovascularization was inhibited substantially by neutralizing antibodies to VEGF and IL-8 but not IL-1beta. Treatment with anti-TNF-alpha antibodies resulted in partial inhibition. TDM possesses pleiotropic activities, and the cytokine network plays an important role in the process of neovascularization.