Occurrence of somnolence and respiratory depression induced by pregabalin and mirogabalin use and the influence of opioid treatment using the Japanese adverse drug event report database.

Background: Gabapentinoid anticonvulsants are standard treatment for neuropathic pain and are often combined with opioids for treating cancer. It is assumed that this combination may heighten somnolence and respiratory depression due to the inhibitory effects of opioids on the central nervous system. Although pregabalin, a gabapentinoid, is known to increase somnolence frequency during opioid therapy, whether mirogabalin exerts similar effects on somnolence frequency under opioid therapy remains unknown. This study examined the signals of somnolence and respiratory depression in response to pregabalin and mirogabalin use by utilizing data from the Japanese Adverse Drug Event Report database and assessed their interaction with strong opioid analgesics. Methods: Information was obtained from the JADER database from April 2004 to August 2023 via the Pharmaceuticals and Medical Devices Agency website. The study focused on neuropathic pain medications, specifically "pregabalin" and "mirogabalin besilate." Adverse events were defined using preferred terms (PTs) from the Medical Dictionary for Regulatory Activities version 26.1. The PTs considered were "Somnolence (10041349)" and "Respiratory depression (10038678)." To investigate the effect of the combination of strong opioid analgesics with pregabalin and mirogabalin on the occurrence of somnolence, a multivariable logistic regression analysis was conducted. Results: Signals for somnolence were detected with the use of both drugs (pregabalin: information component (IC) [95% confidence intervals (CIs)]: 2.89 [2.70 to 3.08]; mirogabalin: IC [95% CIs] 2.50 [1.85 to 3.16]). When evaluating respiratory depression, a typical and serious adverse event of opioid analgesic use, a signal was detected with pregabalin use but not with mirogabalin use (pregabalin: (IC [95% CIs] 1.28 [0.83 to 1.73]; mirogabalin: IC [95% CIs] -0.15 [-2.20 to 1.89]). Multivariable analysis indicated that the use of strong opioid analgesics increased the occurrence of somnolence when combined with pregabalin but not when combined with mirogabalin (p = 0.004). Conclusion: While the safety of concomitant administation of mirogabalin with opioids remains controversial, caution should be exercised when using pregabalin, especially in combination with opioids for neuropathic pain, compared to that for mirogabalin.

Activation of the p21(CIP1/WAF1) promoter by bone morphogenetic protein-2 in mouse B lineage cells.

BMPs exert a negative growth effect on various types of cells. We have previously reported that BMP-2 inhibited the growth of HS-72 mouse hybridoma cells by inducing p21(CIP1/WAF1) expression. In the present study, we demonstrated that BMP-2 activated the mouse p21(CIP1/WAF1) promoter in HS-72 cells, and that a 29-base pair (b) region of the promoter (-1928/-1900 relative to the TATA box), conserved between mice and humans, was responsive to BMP-2 as well as expression of Smad1, Smad4, and constitutively active mutants of BMP type I receptors. Furthermore, an oligonucleotide containing the 29-b region was found to be associated with Smad4 and phosphorylated Smad1 in the nuclear extract of BMP-2-stimulated HS-72 cells. These results suggested that BMP-2 might activate p21(CIP1/WAF1) transcription by inducing a binding of Smad4 and Smad1 to the 29-b region in HS-72 cells.

Correlation between Bcl-X expression and B-cell hybridoma apoptosis induced by activin A.

In this study, we examined the role of bcl-XL and bcl-XS in apoptotic cell death of HS-72 cells induced by activin A. Immunoblot analysis revealed that a band of Bcl-XL was detected in HS-72 cells cultured with or without activin A. Although untreated HS-72 cells did not express Bcl-XS, the expression of Bcl-XS was significantly increased when cultured with activin A. We also investigated the expression of Bcl-XS and Bcl-XL in HS-72 cells cultured with activin A in the presence of protein kinase C inhibitor 1-(5-isoquinotinesulfonyl)-2-methylpiperazine dihydrochloride (H7), which suppressed apoptosis in HS-72 cells induced by activin A. Exposure to H7 apparently increased the level of Bcl-XL in HS-72 cells cultured with or without activin A. In contrast, no detectable band of Bcl-XS was found in HS-72 cells cultured with activin A and H7. These findings indicate that Bcl-XL upregulation and Bcl-XS downregulation induced by H7 might correlate with the suppression of activin A-induced apoptosis in B-lineage cells.

The role of activin type I receptors in activin A-induced growth arrest and apoptosis in mouse B-cell hybridoma cells.

Activins transduce their signals by binding to activin type I receptors and activin type II receptors, both of which contain a serine/threonine kinase domain. In this study, we established stable transfectants expressing two types of activin receptors, ActRI and ActRIB, to clarify the role of these receptors in activin signalling for growth inhibition in HS-72 mouse B-cell hybridoma cells. Over-expression of ActRI suppressed activin A-induced cell-cycle arrest in the G1 phase caused by inhibition of retinoblastoma protein phosphorylation through induction of p21CIP1/WAF1, a cyclin-dependent kinase inhibitor, and subsequent apoptosis. In contrast, HS-72 clones that over-expressed ActRIB significantly facilitated activin A-induced apoptosis. These results indicate that ActRI and ActRIB are distinct from each other and that the ActRI/ActRIB expression ratio could regulate cell-cycle arrest in the G1 phase and subsequent apoptosis in HS-72 cells induced by activin A.

Activin A induction of cell-cycle arrest involves modulation of cyclin D2 and p21CIP1/WAF1 in plasmacytic cells.

Activins, members of the transforming growth factor-beta family, have been implicated in the regulation of growth and differentiation of various types of cells. We have recently found that activin A induces apoptotic cell death of plasmacytic cells including B cell hybridoma cells and myeloma cells. In the present study, we demonstrated that activin A caused cell-cycle arrest in the G1 phase before appearance of apoptotic cells in mouse B cell hybridoma cells. Phosphorylation of retinoblastoma protein (Rb) and in vitro Rb kinase activity of cyclin-dependent kinase (CDK)4 was inhibited in activin A-treated cells. Analysis of expression of genes regulating Rb phosphorylation revealed that activin A suppressed cyclin D2, the sole D-type cyclin gene expressed in the hybridoma cells, and activated p21CIP1/WAF1 but had no effect on expression of cyclin-dependent kinases (CDK2, CDK4, CDK6) and other CDK inhibitors (p27KIP1, p16INK4a, p15INK4b). Modulation of cyclin D2 and p21CIP1/WAF1 expression resulted in a decrease in level of cyclin D2-CDK4 complex and an increase in level of CDK4 complexed with p21CIP1/WAF1. Moreover, overexpression of cyclin D2 partially abrogated inhibition of Rb phosphorylation and G1 arrest in the hybridoma cells.

Activin A-induced apoptosis is suppressed by BCL-2.

Activin A, a member of TGF beta superfamily has various activities including induction of apoptosis in mammalian cells. However, it remains unknown how activin A induces cell death. To clarify this, we investigated the expression of BCL-2 and BAX, and the effect of BCL-2 overexpression on activin A-induced apoptosis in B cell hybridoma cell lines. The activin A-sensitive cell lines expressed BAX but not BCL-2 and that activin A did not increase BAX levels. Overexpression of human BCL-2 suppressed activin A-induced apoptosis in these cells. Thus, activin A has been shown to induce apoptosis by a BCL-2-inhibitable mechanism without activating BAX.

Induction of apoptosis in B lineage cells by activin A derived from macrophages.

A factor produced by P388D1 cell line murine macrophages showed a profound suppressive effect on the in vitro proliferation of B lineage cells. It was purified to homogeneity from conditioned media of P388D1 cells stimulated with phorbol 12-myristate 13-acetate for 48 h by a three-step procedure. The purified factor gave a single band of protein with a molecular mass of 16 kD on SDS-polyacrylamide gel electrophoresis. We show here that exposure of B lineage cells to this factor results in the induction of a cytotoxic effect and a significant increase in the proportion of fragmented DNA. DNA fragmentation was detected in B lineage cells after 3 h culture with the factor in the quantitative colorimetric determination. The mechanism of cell death was characterized by a ladder-like electrophoretic pattern of degraded chromosomal DNA, indicating that the factor induces apoptosis. The NH2-terminal amino acid sequence of this factor was identical with that of activin A over the 26 amino acid residues identified. We sought to determine whether apoptosis could be modulated by two kinds of inhibitor of protein kinases, H7 and HA1004, in concentrations that are below their toxicity limits. Apoptosis induced by the factor was suppressed by H7 but was relatively unaffected by HA1004. These findings suggest that the signals by protein kinases may regulate apoptotic B cell death by the factor activin A, derived from macrophages.

Freezing denaturation of ovalbumin at acid pH.

The effects of rapid freezing and thawing at acid pH on the physiochemical properties of ovalbumin were examined. At low pH (around 2), UV difference spectra showed microenvironmental changes around the aromatic amino acid residues; elution curves by gel permeation chromatography showed decreasing numbers of monomers after neutralization. These changes depended on the incubation temperature (between -196 and -10 degrees C) and the protein concentration (0.5-10 mg/ml), and a low concentration of ovalbumin incubated at around -40 degrees C suffered the most damage to its conformation. With freezing and then incubation at -40 degrees C, three of the four sulfhydryl groups in the ovalbumin molecule reacted with 2,2'-dithiodipyridine. The CD spectra showed these changes in the secondary structure, but they were smaller than those when guanidine hydrochloride was used for denaturation. Supercooling at -15 degrees C or freezing at -196 degrees C had little or no effect on the conformation of the ovalbumin molecule. Thus, irreversible conformational changes of ovalbumin were caused under the critical freezing condition at an acid pH. These changes arose from partial denaturation and resembled those with thermal denaturation of ovalbumin at neutral pH.

Conformational changes in ovalbumin at acid pH.

Several physicochemical parameters of ovalbumin were examined at acid pH. The intrinsic viscosity and far UV-CD spectrum at pH 2 did not differ from those at pH 7. But the near UV-CD spectrum, difference absorption spectrum around 250-320 nm, and fluorescence spectrum showed micro-environmental changes around the aromatic amino acid residues in acid solution. The reactivity of one of the four sulfhydryl groups with 2,2'-dithiodipyridine increased at pH below 5. The rate of denaturation by urea and that of surface tension decay were high in the low pH range. We concluded that at low pH (around 2), ovalbumin molecules kept their native globular conformation, but that their chain flexibility increased and they were very susceptible to denaturation. This state might be equivalent to the molten-globule state observed with some globular proteins in acidic region.

Role of an intrachain disulfide bond in the conformation and stability of ovalbumin.

Ovalbumin, which contains one intrachain disulfide bond and four cysteine sulfhydryls, was reduced with dithiothreitol under non-denaturing conditions, and its conformation and stability were compared with those of the disulfide-bonded form. The CD spectrum in the far-UV region revealed that the overall conformation of the reduced form is similar to that of the disulfide-bonded one. Likewise, the inaccessibility to trypsin and the non-reactivity of the four cysteine sulfhydryls, exhibited by the native disulfide-bonded ovalbumin, were still retained in the disulfide-reduced form. Thus, the reduced ovalbumin appeared to substantially take the native-like conformation. However, the near-UV CD spectrum slightly differed between the native and disulfide-reduced forms. Protein alkylation with a fluorescent dye and subsequent sequence analysis showed that the two sulfhydryls (Cys73 and Cys120) originating from the disulfide bond are highly reactive in the reduced form. Furthermore, upon proteolysis with subtilisin, the N-terminal side of Cys73 was cleaved in the reduced form, but not in the disulfide-bonded one. Upon heat denaturation, the transition temperature of the reduced form was lower, by 6.8 degrees C, than that of the disulfide-bonded one. Thus, we concluded that ovalbumin has a native-like conformation in its disulfide-reduced form, but that the local conformation of the reduced form fluctuates more than that of the disulfide-bonded one. Such local destabilization may be related to the decreased stability against heat denaturation.

Two different types of humoral immune response to Actinobacillus actinomycetemcomitans in high-responder periodontitis patients.

Actinobacillus actinomycetemcomitans is considered to be an aetiological agent in various forms of periodontitis, with serotype b-specific carbohydrate being the immunodominant antigen of A. actinomycetemcomitans Y4 in high-responder patients. Lipopolysaccharide (LPS) of the organism may also be an important antigen. The purpose of the present study was to clarify the importance of LPS as an antigen of A. actinomycetemcomitans. Twenty patients who had high antibody titres to strain Y4 were selected, and the reactivity of their sera with LPS was determined by ELISA and Western blotting. Two groups of patients were observed: group 1 had high IgG titres only to serotype b strain, whereas group 2 had high IgG titres to serotypes a, b and c strains. The results of adsorption tests showed that anti-A. actinomycetemcomitans Y4 antibody in group 1 patients mostly consisted of antibody reactive with the serotype b-specific carbohydrate, whereas the antibody in group 2 patients mostly consisted of antibody reactive with the LPS of all serotypes. These data show that anti-LPS antibody is present and predominant in anti-A. actinomycetemcomitans Y4 antibody from some high-responder patients, and indicate an important role for LPS as an antigen in the humoral immune response to the organism.

Lectin cytochemical analysis of glycoconjugates in photoreceptor cell membranes of Lampetra japonica.

Seven types of ferritinized lectin were used to examine the distribution of glycoconjugates on the outer segment membranes of lamprey photoreceptor cells. Ultrastructural pre-embedding labeling revealed that peanut agglutinin, soybean agglutinin and Ricinus communis agglutinin I were preferentially bound to the proximal, lateral and luminal surfaces of the long cell outer segments, whereas Griffonia simplicifolia agglutinin II and concanavalin A agglutinin were bound to the corresponding surfaces of the short cell outer segments. The results indicate that there is marked difference in the composition of glycoconjugates over the outer segment membranes between long and short photoreceptors.

A new automatic device for measuring the spinnbarkeit of saliva: the Neva Meter.

OBJECTIVES: We have recently developed a new device for measuring the spinnbarkeit of saliva called the Neva Meter. The purpose of this study was to evaluate this device and to measure spinnbarkeit as well as viscosity, another important property, in the resting saliva of 24 healthy adults. METHODS: We used polyvinyl alcohol (PVA) as a standard solution to establish the reproducibility of spinnbarkeit tests. We collected resting saliva from 24 employees of a business office (16 males and 8 females, average age: 37.8) and investigated the relationship between spinnbarkeit and viscosity. RESULTS: The spinnbarkeit of PVA increased along with the concentration of the solution, and the reproducibility of the values was acceptable. Spinnbarkeit of resting saliva showed a positive correlation with viscosity at a shear rate of 76.6 s(-1) (r = 0.55, P < 0.05) and 191.5 s(-1) (r = 0.59, p < 0.05). CONCLUSIONS: The newly developed Neva Meter was suitable for measuring the spinnbarkeit of saliva quickly and easily at the chair-side in the dental clinic. Results obtained using this new device may be important for understanding and evaluating the condition of the oral cavity.

[Ultrastructural studies of the lamina suprachoroidea in the human eye].

The lamina suprachoroidea of the human choroid was examined by electron microscopy using both thin sections and freeze fracture replicas. The lamina suprachoroidea was composed of 5-10 layers of pigmented cells interspersed with multiple layers of flattened fibroblastic cells. Tight junctions, gap junctions and intermediate junctions were seen between the fibroblastic cells. Tight junctions did not reveal zonula occludens but predominantly isolated types were revealed, as well as several pore-like fenestrations in the attenuated cell processes of the fibroblasts. The characteristic morphological appearance of the lamina suprachoroidea might suggest that the aqueous humour may be able to leak into the extraocular tissues through it. However, the numerous cellular layers and also the junctional complexes between the fibroblasts of the lamina suprachoroidea are likely to produce more resistance to uveoscleral drainage. As the multi-layered arrangement of fibroblastic cells and pigmented cells in the lamina suprachoroidea did not show the cellular barrier, it may be possible that the extraocular substances are able to flow into the eye through it.

[Investigation of Cryptomeria japonica pollinosis in Izu-Oshima].

We surveyed by questionnaire 4,673 residents of Izu-Oshima who were 15 or more years of age with regard to Cryptomeria japonica pollinosis. The response rate was 22.3%. In the early spring, nasal symptoms were reported by 8.9% of the respondents, ocular symptoms by 5.7%, and dermal symptoms by 8.1%. On scratch tests of symptomatic subjects, 13.8% were positive for Cryptomeria japonica antigen, and 33.3% had an IgE RAST score of 2 or more. The peak Cryptomeria japonica pollen concentrations between February and April 1990 were 118/cm2 on March 7 at the Northern Clinic and 271/cm2 at the Southern Clinic. A second questionnaire survey (response rate: 53.1%), designed to estimate the number of persons with Cryptomeria japonica pollinosis among all residents, revealed that 4.7% concurrently had three or more nasal symptoms and two ocular symptoms. By combining these results with those of a telephone survey of 100 randomly selected nonrespondents, 5.64% of all inhabitants were estimated to have suspected Cryptomeria japonica pollinosis.

[Measurement of serum and sputum eosinophil cationic protein concentrations in asthma].

Concentrations of eosinophil cationic protein (ECP) were measured in serum from patients with asthma, patients with chronic obstructive pulmonary disease (COPD), and healthy subjects. The relationships between serum ECP concentration and percent of predicted FEV1 (%FEV1), and between serum and sputum ECP concentrations were also examined in patients with asthma. Serum ECP concentration in asthma was significantly higher than those in COPD patients and healthy subjects. There was a significant inverse correlation between serum ECP concentration and %FEV1 when analysis was restricted to data from asthma patients less than 60 years old. ECP concentration in serum and in sputum were significantly correlated. These results suggest that in asthma patients, eosinophils are activated in serum, and that the degree of eosinophil activation in the airway can be estimated by measuring serum ECP to some extent. Because it does not require bronchial biopsy or bronchoalveolar lavage, measurement of serum ECP may be useful as a minimally invasive monitoring of asthma.

[Results of a survey of Japanese cedar pollinosis in Hachijyojima].

We performed a questionnaire survey of Japanese cedar pollinosis among 7,946 residents of the Sakashita region (Mitsune, Ogago) and Sakaue region (Kashidate, Sueyoshi, Nakanogo) of Hachijyojima who were at least 15 years of age. The response rate was 21.3%. The percentage of respondents who reported three or more nasal symptoms concurrentry with two ocular symptoms in early spring (from the end of February to the end of March) was 1.8% in the Sakashita region and 0.3% in the Sakaue region. About 1.5% of Hachijyojima residents were suspected to have Japanese cedar pollinosis. On scratch tests of symptomatic subjects, 9.2% showed positive reactions for Japanese cedar antigen, and 12.1% had an IgE RAST score of 2 or more. The peak Japanese cedar pollen concentration between February 11 and March 31, 1992 was 74/cm2 on March 6 in the Sakashita region and 127/cm2 on the same day in the Sakaue region. This survey confirmed the presence of a low incidence of Japanese cedar pollinosis in Hachijyojima, an isolated island 290 km from Tokyo.

[The prevalence of orthostatic dysregulation complicated with bronchial asthma].

Orthostatic dysregulation (OD) has been reported to be complicated with childhood bronchial asthma. 42 patients (28 females and 14 males) with adult-onset bronchial asthma were selected randomly to investigate the prevalence of OD. OD was diagnosed by both questionnaire for subjective symptoms and tilting test (Schellong test and upright ECG). Our results revealed that 64.3% (both 64.3% in females and males) of the patients were complicated with OD. There was no significant difference in the duration of asthma, FEV0.1, %FEV1.0, serum IgE level, and severity of asthma between patients with OD and without OD. Furthermore, no significant difference in the results of tilting test were observed. In serum level of theophylline we couldn't detect any subjective difference between the two groups, however there was significant difference between positive patients and negative patients in tilting test. In conclusion, OD is frequently complicated with adult-onset asthma and we should be careful of the subjective symptoms concerned with OD.

[Isolation and characterization of oral treponemes].

The oral treponemes were isolated on nine media and classified according to Anaerobe Laboratory Manual (1977). The four media, which were used with the plate-in-bottle method, were Medium 10 (M10), M10 supplemented with 10% rumen fluid, M10 supplemented with 10% rabbit serum and cocarboxylate, and M10 added with rifampicin. The five media with the anaerobic glove box method were New Oral Spirochetes (NOS) medium, NOS medium added with rifampicin, alternated modified Oral Treponemes Isolation (aOTI) medium, and alternated M10 for anaerobic glove box. The isolation rate of the oral treponemes from 59 subgingival plaque samples of various forms of periodontal diseases was 85 percent. Ninety clinical isolates were subcultured and classified into six groups (I-VI) based on the carbohydrate fermentation and metabolic end products. Three saccaharolytic groups were further divided into nine subgroups by the fermentation patterns of sucrose, lactose, maltose and rhamnose. Two saccaharolytic subgroups were identified as Treponema socranskii, and two asaccharolytic groups were identified as "T. denticola" and "T. vincentii". However, the other subgroups and group could not be identified according to the characteristics of the presently recognized species.