Real-world safety profile of eculizumab in patients with paroxysmal nocturnal hemoglobinuria, atypical hemolytic uremic syndrome, or generalized myasthenia gravis: an integrated analysis of post-marketing surveillance in Japan.

Eculizumab is a C5 inhibitor approved for the treatment of paroxysmal nocturnal hemoglobinuria (PNH), atypical hemolytic uremic syndrome (aHUS), and anti-acetylcholine receptor antibody-positive generalized myasthenia gravis (AChR + gMG) in Japan. We report integrated safety data from post-marketing surveillance in these three indications, focusing on commonly occurring adverse events (AEs) and infection-related AEs. Of 1219 patients registered, 1055 (PNH: 780; aHUS: 192; AChR + gMG: 83) had available safety data. Total eculizumab exposure was 3977.361 patient-years. AEs were reported in 74.03% of patients. AEs with an incidence of  ≥ 1.0 per 100 patient-years included hemolysis, headache, nasopharyngitis, renal impairment, anemia, pneumonia, upper respiratory tract inflammation, influenza, condition aggravated, and infection. The incidence of infection-related AEs was 21.30 per 100 patient-years, the most frequent types (≥ 1.0 per 100 patient-years) being nasopharyngitis, pneumonia, influenza, and infection. Meningococcal infections were reported in four patients (0.10 per 100 patient-years). Two patients died from meningococcal sepsis, with a mortality rate of 0.05 per 100 patient-years. This is the largest safety dataset on eculizumab in Japan derived from more than 10 years of clinical experience. No new safety signals were observed and the safety profile of eculizumab was consistent with that in previous clinical trials and international real-world safety analyses.

Cytototoxic constituents from the bark of Chisocheton cumingianus (Meliaceae).

A new lanostane-type triterpenoid, 3ß-hydroxy-25-ethyl-lanost-9(11),24(24')-diene (1), along with 3ß-hydroxy-lanost-7-ene (2) and ß-sitosterol-3-O-acetate (3) was isolated from the stem bark of C. cumingianus. The chemical structure of the new compound was elucidated on the basis of spectroscopic data. All of the compounds were evaluated for their cytotoxic effects against P-388 murine leukemia cells. Compounds 1-3 showed cytotoxicity against P-388 murine leukemia cells with IC50 values of 28.8 ± 0.10, 4.29 ± 0.03, and 100.18 ± 0.16 µg/ml, respectively.

Frontier studies on highly selective bio-regulators useful for environmentally benign agricultural production.

Fungal metabolites active for insects were obtained from fermentation products using okara media. The mechanisms of action of these compounds against insects were clarified using voltage clamp electrophysiology. The branching factor inducing hyphal branching in arbuscular mycorrhizal (AM) fungi was isolated from the root exudates of Lotus japonicus and identified as 5-deoxystrigol. Strigolactones were originally identified as seed germination stimulants of parasitic weeds; therefore, synthetic strigolactones were developed to exhibit the inducing activity of hyphal branching in AM fungi and diminish the stimulating activity of seed germination of parasitic weeds. Signaling molecules, acylhomoserine lactones (AHLs), in quorum sensing were identified in the fungal strain Mortierella alpina A-178, and the true producer of AHLs was clarified as symbiotic bacteria in the fungus. Since acyl-(S)-adenosylmethionine analogs may be good candidates for competitive inhibitors of AHL synthases, intermediate mimics in the biosynthesis of AHLs have been synthesized.

An acyl-SAM analog as an affinity ligand for identifying quorum sensing signal synthases.

N-Acylhomoserine lactones (AHLs) are quorum sensing signals produced by Gram-negative bacteria. We here report the affinity purification of AHL synthases using beads conjugated with an enzyme inhibitor, which was designed based on the catalytic intermediate acyl-SAM.

Confirming stereochemical structures of strigolactones produced by rice and tobacco.

Major strigolactones (SLs) produced by rice (Oryza sativa L. cv. Nipponbare) and tobacco (Nicotiana tabacum L. cv. Michinoku No. 1) were purified and their stereochemical structures were determined by comparing with optically pure synthetic standards for their NMR and CD data and retention times and mass fragmentations in ESI-LC/MS and GC-MS. SLs purified from root exudates of rice plants were orobanchol, orobanchyl acetate, and ent-2'-epi-5-deoxystrigol. In addition to these SLs, 7-oxoorobanchyl acetate and the putative three methoxy-5-deoxystrigol isomers were detected by LC-MS/MS. The production of 7-oxoorobanchyl acetate seemed to occur in the early growth stage, as it was detected only in the root exudates collected during the first week of incubation. The root exudates of tobacco contained at least 11 SLs, including solanacol, solanacyl acetate, orobanchol, ent-2'-epi-orobanchol, orobanchyl acetate, ent-2'-epi-orobanchyl acetate, 5-deoxystrigol, ent-2'-epi-5-deoxystrigol, and three isomers of putative didehydro-orobanchol whose structures remain to be clarified. Furthermore, two sorgolactone isomers but not sorgolactone were detected as minor SLs by LC-MS/MS analysis. It is intriguing to note that rice plants produced only orobanchol-type SLs, derived from ent-2'-epi-5-deoxystrigol, but both orobanchol-type and strigol-type SLs, derived from 5-deoxystrigol were detected in tobacco plants.

[A case of advanced colon cancer successfully treated with cetuximab diagnosed as EGFR-positive by liver biopsy].

Cetuximab, a monoclonal antibody used to target the epidermal growth factor receptor(EGFR), was approved for refractory and metastatic colon cancer expressing EGFR, and the EGFR expression must be confirmed. But the EGFR expression may become false negative by immunohistochemistry. When we used a past operation specimen for a search in particular, a correct evaluation may be difficult for prolonged formalin fixation. We report a case successfully treated by cetuximab, who was diagnosed as EGFR-negative by the past operation specimen, but as EGFR-positive by the liver biopsy specimen. A 51- year old woman with multiple organ metastases, who had experienced failure with prior oxaliplatin, irinotecan(CPT-11), 5- FU and bevacizumab regimens, was administered cetuximab plus CPT-11 because it was EGFR-positive by liver biopsy, and the tumor was obviously reduced. It is useful to obtain another specimen such as by liver biopsy, when the EGFR expression is negative by a past operation specimen.

Three austin family compounds from Penicillium brasilianum exhibit selective blocking action on cockroach nicotinic acetylcholine receptors.

Austin (AT) and its derivatives (dehydroaustin (DAT) and acetoxydehydroaustin (ADAT)) produced by Penicillium brasilianum MG-11 exhibit toxicity to insects, yet their targets are unknown. Here, we used whole-cell patch-clamp electrophysiology to investigate the action of AT family compounds on cockroach acetylcholine (ACh), γ-aminobutyric acid (GABA) and l-glutamate receptors expressed in the American cockroach (Periplaneta americana) neuron. U-tube application of AT or its derivatives did not induce any current amplitudes, suggesting that they did not act as agonist of these three receptors. In the second step of experiments, they were bath-applied for 1min before co-application with the corresponding ligand. We found that AT and its derivatives had no effect on GABA and l-glutamate-induced currents, whereas they significantly reduced ACh- and epibatidine-induced currents, showing that these compounds acted as selective antagonists of nicotinic acetylcholine receptors (nAChRs) expressed in the cockroach neuron. Of the compounds, DAT showed the highest blocking potency for nAChRs, differentially attenuating the peak and slowly desensitizing current amplitude of ACh-induced responses with pIC(50) (=-logIC(50) (M)) values of 6.11 and 5.91, respectively. DAT reduced the maximum normalized response to ACh without a significant shift in EC(50), suggesting that the blocking action is not competitive with ACh.

[Combination therapy with rituximab and cladribine for patients with follicular lymphoma].

Treatment strategies for follicular lymphoma have not been established. We report the outcome after combination therapy with rituximab and cladribine (RC) for 8 patients with follicular lymphoma treated between January 2005 and December 2006 in our hospitals. Median patient age was 57 (range 42 approximately 73) years. There were 4 males and 4 females. Only 1 patient had refractory disease, while the others had untreated disease. On the follicular lymphoma international prognostic index, 4 patients were in the low-risk group, 3 in the intermediate-risk group and 1 in the high-risk group. The median follow-up period was 36 (range 22 approximately 90) weeks. The RC protocol consisted of intravenous rituximab at a dose of 375 mg/m (2) on day 1 and cladribine at a dose of 0.1 mg /kg per day for 2-hours on day 1 through 5. The median number of RC courses was 5 (range 3 approximately 8). The median interval between the 2 courses was 7 (range 3 approximately 26) weeks. The overall response rate was 87.5%. Grade 3 neutropenia was observed in 50% patients, although G-CSF was not needed. There was no apparent thrombocytopenia or anemia. Herpes zoster was observed after treatment in 1 patient. RC is considered highly effective and well tolerated.

Prediction of gastroesophageal reflux in otitis media with effusion in adults.

CONCLUSIONS: The measurement of pepsinogen I (PGI) in middle-ear effusions (MEEs) and a questionnaire on the frequency symptoms of gastroesophageal reflux (GER) disease are tools that can be used to screen for the existence of GER. OBJECTIVE: To seek methods that would be beneficial as a screen for the presence of GER among adult patients with OME. MATERIALS AND METHODS: Fifty-eight adult outpatients with OME were asked to answer a questionnaire of the frequency scale for symptoms of GER disease. Samples of MEEs were obtained from each subject and were measured for concentrations of PGI and PGII. Some patients were followed up after being treated with a proton pump inhibitor. RESULTS: The percentage of patients with high PGI concentrations in their MEEs was higher in those with GER-related symptoms than in those without GER-related symptoms. Moreover, OME was present bilaterally in a higher percentage of patients with GER-related symptoms. There were patients in whom PGI levels decreased after receiving treatment for GER.

Comparison of the quantity of cochlear melanin in young and old C57BL/6 mice.

OBJECTIVE: To elucidate the functional relationship between cochlear melanin and aging. DESIGN: Melanin has been described in the cochlear labyrinth and has been suggested to protect the cochlea from various types of trauma. The quantity of melanin has been shown to change with aging in several organs; however, to our knowledge, aging changes in the cochlea have not been documented. Therefore, we chemically quantified cochlear eumelanin and pheomelanin contents and compared these in young and old C57BL/6 mice using high-performance liquid chromatography. Because melanin deposits in the cochlea present most extensively in the stria vascularis, we morphologically examined the stria using transmission electron microscopy. SUBJECTS: Cochleae from an inbred strain of C57BL/6 male and female mice; 6 at the age of 10 weeks and 5 at the age of 100 weeks were studied. RESULTS: The quantities of cochlear eumelanin and pheomelanin were 421 and 480 ng per cochlea in young mice, and 2060 and 765 ng per cochlea in old mice, respectively. Under transmission electron microscopy, the number of pigmented granules seemed to be greater in older mice compared with younger mice, especially in marginal cells. CONCLUSION: To our knowledge, our findings are the first quantitative evidence to show an age-related overexpression of cochlear melanin and an alteration in the proportion of eumelanin and pheomelanin with aging, suggesting a possible otoprotective function of eumelanin against age-related cochlear deterioration.

Otitis media in adults as a symptom of gastroesophageal reflux.

OBJECTIVE: To investigate the clinical relationship between gastroesophageal reflux (GER) and otitis media with effusion (OME) in adults. STUDY DESIGN AND SETTING: Sixty patients with OME with unknown causes were asked to answer a new questionnaire specific for the diagnosis of GER disease; pepsinogen (PG) levels in their middle-ear effusions (MEEs) were measured. RESULTS: The percentage of patients with high PG concentrations in their MEEs was significantly higher in those with questionnaire-positive GER than in those with questionnaire-negative GER. OME was present bilaterally in a significantly higher percentage of patients with questionnaire-positive GER. The PG levels decreased in some of the patients after receiving proton pump inhibitors, which also decreased the symptoms of GER. CONCLUSION: The presence of PG in MEEs supports the existence of GER; treatment for GER should be considered in patients with ear complaints, especially in those who have GER-related symptoms.

Association between cupular deposits and otosclerosis.

OBJECTIVE: To evaluate whether otosclerosis is an underlying mechanism for the production of cupular deposits and to study the association between cupular deposits and dysequilibrium in otosclerosis. DESIGN: Retrospective human temporal bone (TB) study. The incidence of cupular deposits in these 70 TBs was analyzed. Correlations between cupular deposits and vestibular symptoms, endosteal involvement of the otosclerotic focus, stapedial fixation, and clinical history of stapes surgery were evaluated. SETTING: Otolaryngology laboratory in a tertiary academic medical center. PATIENTS: The study material consisted of 35 human TBs with otosclerosis and 35 age-matched controls. MAIN OUTCOME MEASURES: Morphometric evaluations of the incidence of cupular deposits, endosteal involvement of the otosclerotic focus, and stapedial fixation were made by light microscopy. Clinical records were reviewed retrospectively for clinical history of stapes surgery and prevalence of vestibular symptoms. The incidence of cupular deposits was compared between the otosclerotic and control groups. Correlations between cupular deposits and vestibular symptoms, endosteal involvement of the otosclerotic focus, stapedial fixation, and clinical history of stapes surgery were evaluated in the subjects with otosclerosis. RESULTS: The incidence of cupular deposits in TBs with otosclerosis was significantly higher than in those without whereas there was no correlation between the incidence of the deposits and dysequilibrium in cases of otosclerosis. An increase in deposits did not correlate with stapedial fixation, stapes surgery, or endosteal involvement. CONCLUSIONS: Our results suggest otosclerosis as an underlying mechanism for the production of cupular deposits; however, we did not find an association between these deposits and vestibular symptoms.

Vascular evaluation in laryngeal diseases: comparison between contact endoscopy and laser Doppler flowmetry.

OBJECTIVE: To determine the efficacies of 2 methods for evaluating vascular changes in laryngeal diseases. DESIGN: Prospective comparative study. SETTING: University hospital. PATIENTS: Twenty-four adults planning to undergo laryngomicrosurgery for unilateral lesions in their vocal cords. INTERVENTION: Vascular evaluation was performed using contact endoscopy and laser Doppler flowmetry during laryngomicrosurgery. MAIN OUTCOME MEASURES: Morphological and physiological variables of the vascular changes. RESULTS: Contact endoscopy examinations revealed changes in the fine vascular network patterns in the vocal cords. Hypervascularity was present in patients with polyps but not in those with nodules, and the degree of vascularity was variable in patients with malignant neoplasms. Patients with papillomas and patients in the postradiation therapy group showed hypovascularity with losses in the capillary network. Laser Doppler flowmetry revealed high mean blood flows in patients with polyps, dysplasia, and malignant neoplasms, whereas patients with papillomas and those in the postradiation therapy group had low blood flows. The variables measured by laser Doppler flowmetry and endoscopy were strongly correlated. CONCLUSION: Evaluations of morphological and physiological changes of vascularity in laryngeal diseases are important for understanding their pathophysiology, and combining contact endoscopy and laser Doppler flowmetry is useful in such evaluations.

Large vestibular aqueduct syndrome: a human temporal bone study.

OBJECTIVES/HYPOTHESIS: Large vestibular aqueduct syndrome (LVAS) is one of the common causes of hearing loss (HL). All prior studies have reported some anomalies associated with LVAS by imaging techniques. This study was undertaken to determine prevalence of LVAS in our temporal bone (TB) collection and its relationship to other systemic or otologic anomalies. STUDY DESIGN: Retrospective, TB histopathologic study. METHODS: Anteroposterior diameters of internal (IA) and external (EA) apertures were measured in 40 normal TBs (40 cases). TBs were considered as large vestibular aqueduct (LVA) if width of apertures was 95% greater than "normals." Systemic and otologic anomalies and histopathology of ears with LVAS were noted. RESULT: Of 1,608 non-"normal" TBs, 63 had LVA. There was negative correlation between IA and EA in 48 TBs with only enlarged IA. Fifteen TBs with enlarged EA always had enlarged IA and were therefore considered as LVAS. The most common pathologic condition was congenital heart anomaly. The most common syndrome or dysplasia was Mondini's. The most common anomalies of external and middle ears were dehiscent facial nerves, low set auricles, and ossicular deformities. Inner ear anomalies included modiolar deficiencies, hair cell loss, interscalar septum defects, and strial atrophy. There was no record of family history of HL, head injury, or craniofacial, branchial, or thyroidal abnormalities. CONCLUSION: Because HL associated with LVAS may be attributed to other ear anomalies, it is important to investigate other inner ear problems and system diagnoses that may indicate a syndrome in patients with radiologically diagnosed LVAS.

A novel RFP-RET transgenic mouse model with abundant eumelanin in the cochlea.

We report on the cochlea of a novel metallothionein-I (MT)/RFP-RET transgenic mouse model with severe systemic melanosis. Electron microscopy revealed that these transgenic mice possess abundant quantities of melanin in the intermediate cells of the stria vascularis. High performance liquid chromatography analysis indicated that cochleae of these transgenic mice contained about twice as much eumelanin as cochleae of control C57BL/6 mice and that the amount of pheomelanin was approximately equal in these two strains. Auditory brainstem responses at 2, 4, 8, and 16 kHz were not significantly different between transgenic and control mice. This is the first report on a mouse model of overproduction of cochlear eumelanin, and our results suggest that this transgenic mouse is an excellent model for investigating the effects of overexpression of cochlear eumelanin. In addition, we provide evidence that eumelanin overproduction in the cochlea does not affect normal hearing.

Heat treatment enhances healing process of experimental pseudomonas corneal ulcer.

We investigated the effects of hyperthermia on the healing process of experimental Pseudomonas corneal ulceration (PCU). Hartley guinea pigs were used to develop animal models of PCU. As a heat source, disposable chemical pocket warmers were applied. The healing process of PCU was compared between the heat-treated corneas and the control corneas. The severity of infection and the degree of angiogenesis were classified by a clinical scoring system. The animals were euthanized 14 days after infection and the corneas were submitted for histopathological examination. The expression of vascular endothelial growth factor (VEGF) was examined immunohistochemically. Comparative reverse transcription polymerase chain reaction was performed to measure the expression level of VEGF in the cornea. Hyperthermia significantly promoted corneal epithelization and neovascularization in the PCU model. Heat treatment significantly decreased the number of viable Pseudomonas organisms present in PCU. On immunohistochemistry, the heated cornea demonstrated more intense staining for VEGF. Comparative reverse transcription polymerase chain reaction showed upregulation of the expression level of VEGF mRNA in the heat-treated cornea. Hyperthermia accelerated the healing process of PCU with increased corneal neovascularization. Angiogenesis may play an important role in the PCU healing process, which is enhanced by the heat treatment.

Multifocal fibrosclerosis: retroperitoneal fibrosis associated with a suprasellar tumor and pachymeningitis.

A 42 year-old man with lumbago and fever had hydronephrosis due to a mass surrounding the abdominal aorta, associated with pachymeningitis and a suprasellar tumor. He was given a diagnosis of multifocal fibrosclerosis, involving both retroperitoneal and intracranial fibrosis. He responded to treatment with corticosteroid and cyclophosphamide. The intracranial lesions seemed to be rare, but important manifestations of the systemic disease, and were clearly visualized by magnetic resonance imaging. The phenotype of the infiltrated lymphocytes was CD4+, supporting the hypothesis that autoimmunity was involved pathogenetically. We propose a comprehensive category termed "inflammatory fibrotic disease" including both systemic and localized diseases.

Genome sequence of an M3 strain of Streptococcus pyogenes reveals a large-scale genomic rearrangement in invasive strains and new insights into phage evolution.

Group Astreptococcus (GAS) is a gram-positive bacterial pathogen that causes various suppurative infections and nonsuppurative sequelae. Since the late 1980s, streptococcal toxic-shock like syndrome (STSS) and severe invasive GAS infections have been reported globally. Here we sequenced the genome of serotype M3 strain SSI-1, isolated from an STSS patient in Japan, and compared it with those of other GAS strains. The SSI-1 genome is composed of 1,884,275 bp, and 1.7 Mb of the sequence is highly conserved relative to strain SF370 (serotype M1) and MGAS8232 (serotype M18), and almost completely conserved relative to strain MGAS315 (serotype M3). However, a large genomic rearrangement has been shown to occur across the replication axis between the homologous rrn-comX1 regions and between two prophage-coding regions across the replication axis. Atotal of 1 Mb of chromosomal DNA is inverted across the replication axis. Interestingly, the recombinations between the prophage regions are within the phage genes, and the genes encoding superantigens and mitogenic factors are interchanged between two prophages. This genomic rearrangement occurs in 65% of clinical isolates (64/94) collected after 1990, whereas it is found in only 25% of clinical isolates (7/28) collected before 1985. These observations indicate that streptococcal phages represent important plasticity regions in the GAS chromosome where recombination between homologous phage genes can occur and result not only in new phage derivatives, but also in large chromosomal rearrangements.

A multiplex polymerase chain reaction-based diagnostic method for bacterial vaginosis.

OBJECTIVE: To develop a polymerase chain reaction (PCR)-based diagnostic method for bacterial vaginosis using bacterial vaginosis-associated anaerobes. METHODS: A multiple PCR assay was developed using primers specific to 16S ribosomal deoxyribonucleic acid (DNA) (Mobiluncus mulieris and Mobiluncus curtisii), nanH (Bacteroides fragilis), and an internal spacer region of ribosomal DNA (Gardnerella vaginalis). The vaginal swabs from pregnant and nonpregnant women were examined by Gram stain-based Nugent scoring system. One hundred seventy-two samples of 853 Gram stain-interpretable samples were randomly selected and subjected to multiplex PCR assay. RESULTS: The sensitivity of the PCR assay ranged from 10 to 10 colony-forming units per vaginal swab. The prevalence of the bacterial vaginosis, intermediate, and normal categories was found by Nugent scoring system to be 21.6% (184/853), 26.0% (222/853), and 52.4% (447/853), respectively. By the multiplex PCR-based diagnostic method, 20.3% (35/172) of the samples were identified as bacterial vaginosis. The diagnostic sensitivity, specificity, positive predictive value, and negative predictive value of multiplex PCR in comparison with Gram stain examination were 78.4% (95% confidence interval [CI] 65.1%, 91.6%), 95.6% (95% CI 92.1%, 99.0%), 82.9% (95% CI 70.4%, 95.4%), and 94.2% (95% CI 90.3%, 98.1%), respectively. CONCLUSION: This multiplex PCR can be used as a diagnostic or screening test for bacterial vaginosis.

Identification of a type III secretion system in uropathogenic Escherichia coli.

To determine virulence-related genes in uropathogenic Escherichia coli (UPEC) showing invasiveness to T-24 bladder cancer cells, genomic subtractive hybridization was performed between a highly invasive and a less invasive strain. Forty-nine DNA fragments were isolated from the invasive strain. One of them showed homology with Salmonella invA gene. By chromosomal walking of the strain, a type III secretion system that has been described in E. coli O157:H7 was identified on the genome of the invasive strains. Three strains out of 100 UPEC isolates had a type III secretion system inserted at 64 min of the chromosome, corresponding to E. coli K-12 MG1655. This finding suggested that the type III secretion system could play a part in uropathogenicity of UPEC.