[A case of abscess-associated hepatocellular carcinoma with massive lymphoid cell infiltration].

A 76-year-old Japanese man was admitted to our hospital with chills and fever. Computed tomography revealed a 10-cm cystic tumor with peripheral ring enhancement in the left lobe of the liver and several small low-density areas with early peripheral enhancement in both lobes. The large liver mass was diagnosed as a pyogenic abscess and treated with antibiotics. However, elevation of the tumor marker, PIVKA-II, raised the possibility of hepatocellular carcinoma. A fine-needle aspiration biopsy was performed, and malignant hepatic cells were identified. The patient underwent left hepatectomy. Histological analyses of the resected surgical specimen confirmed necrotic liver abscess and residual hepatocellular carcinoma with massive lymphoid cell infiltration. Immunohistochemical analyses revealed that the lymphoid cells were positive for CD3 and CD8. The PIVKA-II level returned to normal after surgery and the hepatic lesions disappeared within 10 months. These findings suggest that the liver abscess stimulated cancer immunity, resulting in the proliferation of cytotoxic T lymphocytes and, subsequently, tumor regression.

A Circumference-Measurement Method Using a Model of a Leg and a 3D Camera.

The circumference of a limb is an important parameter in the follow-up of an edema. Recently, several methods of measuring the circumference on a limb using 3D cameras have been proposed. However, the 3D cameras used are expensive and difficult to implement in general medical facilities. In this study, we propose a circumference-measurement method using a Structure Sensor. First, the leg is photographed and unnecessary background objects are removed from the obtained point cloud. Next, a cross-sectional view is obtained by slicing the point cloud at the specified leg height. Finally, the circumference measurement at a specified leg height is performed by calculating the circumference using the acquired cross-sectional view. Using this method, the leg circumferences of two healthy subjects were measured at two points. For comparison, circumferences were also measured with a measuring tape. The difference between the values estimated using our method and the measured values was generally less than 0.5 cm.

Pharmacological characterization of a novel potent, selective, and orally active phosphodiesterase 10A inhibitor, PDM-042 [(E)-4-(2-(2-(5,8-dimethyl-[1,2,4]triazolo[1,5-a]pyrazin-2-yl)vinyl)-6-(pyrrolidin-1-yl)pyrimidin-4-yl)morpholine] in rats: potential for the treatment of schizophrenia.

Recently, we identified a novel phosphodiesterase 10A (PDE10A) inhibitor, PDM-042 ((E)-4-(2-(2-(5,8-dimethyl-[1,2,4]triazolo[1,5-a]pyrazin-2-yl)vinyl)-6-(pyrrolidin-1-yl)pyrimidin-4-yl)morpholine). PDM-042 showed potent inhibitory activities for human and rat PDE10A with IC 50 values of less than 1 nmol/L and more than 1000-fold selectivity against other phosphodiesterases. Tritiated PDM-042, [3H]PDM-042, had high affinity for membranes prepared from rat striatum with a Kd value of 8.5 nmol/L. The specific binding of [3H]PDM-042 was displaced in a concentration-dependent manner by PDM-042 and another structurally unrelated PDE10A inhibitor, MP-10. In rat studies, PDM-042 showed excellent brain penetration (striatum/plasma ratio = 6.3), occupancy rate (86.6% at a dose of 3 mg/kg), and good oral bioavailability (33%). These data indicate that PDM-042 is a potent, selective, orally active, and brain-penetrable PDE10A inhibitor. In behavioral studies using rat models relevant to schizophrenia, PDM-042 significantly antagonized MK-801-induced hyperlocomotion (0.1-0.3 mg/kg) without affecting spontaneous locomotor activity and attenuated the conditioned avoidance response (CAR) (0.3-1 mg/kg). In tests for adverse effects, PDM-042 had a minimal effect on catalepsy, even at a much higher dose (10 mg/kg) than the minimal effective dose (0.3 mg/kg) in the CAR. Furthermore, PDM-042 had no effect on prolactin release or glucose elevation up to 3 mg/kg, while risperidone increased prolactin release and olanzapine enhanced glucose levels at doses near their efficacious ones in the CAR. Our results suggest that PDM-042 is a good pharmacological tool that can be used to investigate the role of PDE10A and may have therapeutic potential for the treatment of schizophrenia.

1(OH) vitamin D3 supplementation improves the sensitivity of the immune-response during Peg-IFN/RBV therapy in chronic hepatitis C patients-case controlled trial.

OBJECTIVE: 1,25(OH)2 vitamin D3 can affect immune cells. However, the mechanism responsible for the favorable effects of 1(OH) vitamin D3, which becomes 1,25(OH)2 vitamin D3 in the liver, is not clear. The aim of this study is to analyze the immunological response of 1(OH) vitamin D3 supplementation in CH-C patients. DESIGN: Forty-two CH-C patients were treated with 1(OH) vitamin D3/Peg-IFNα/RBV. Forty-two case-matched controls were treated with Peg-IFNα/RBV. The expression of Interferon-stimulated genes (ISGs)-mRNA in the liver biopsy samples and JFH-1 replicating Huh-7 cells were quantified by real-time PCR. Ten kinds of cytokines in the plasma were quantified during treatment by using a suspension beads array. A trans-well co-culture system with peripheral blood mononuclear cells (PBMCs) and Huh-7 cells was used to analyze the effect of 1(OH) vitamin D3. The activities of the Th1 response were compared between subjects treated with 1(OH) vitamin D3/Peg-IFN/RBV and those treated with Peg-IFN/RBV therapy alone. RESULTS: 1(OH) vitamin D3/Peg-IFN/RBV treatment could induce rapid viral reduction, especially in IL28B T/T polymorphism. Several kinds of cytokines including IP-10 were significantly decreased after 4 weeks of 1(OH) vitamin D3 treatment (p<0.05). Th1 responses in the subjects treated with 1(OH) vitamin D3/Peg-IFN/RBV were significantly higher than those treated with Peg-IFN/RBV at 12 weeks after Peg-IFN/RBV therapy (p<0.05). The expression of ISGs in the patient's liver biopsy samples was significantly lower than in those treated without 1(OH) vitamin D3 (p<0.05). CONCLUSION: 1(OH) vitamin D3 could improve the sensitivity of Peg-IFN/RBV therapy on HCV-infected hepatocytes by reducing the IP-10 production from PBMCs and ISGs expression in the liver.

Gnarled-trunk evolutionary model of influenza A virus hemagglutinin.

Human influenza A viruses undergo antigenic changes with gradual accumulation of amino acid substitutions on the hemagglutinin (HA) molecule. A strong antigenic mismatch between vaccine and epidemic strains often requires the replacement of influenza vaccines worldwide. To establish a practical model enabling us to predict the future direction of the influenza virus evolution, relative distances of amino acid sequences among past epidemic strains were analyzed by multidimensional scaling (MDS). We found that human influenza viruses have evolved along a gnarled evolutionary pathway with an approximately constant curvature in the MDS-constructed 3D space. The gnarled pathway indicated that evolution on the trunk favored multiple substitutions at the same amino acid positions on HA. The constant curvature was reasonably explained by assuming that the rate of amino acid substitutions varied from one position to another according to a gamma distribution. Furthermore, we utilized the estimated parameters of the gamma distribution to predict the amino acid substitutions on HA in subsequent years. Retrospective prediction tests for 12 years from 1997 to 2009 showed that 70% of actual amino acid substitutions were correctly predicted, and that 45% of predicted amino acid substitutions have been actually observed. Although it remains unsolved how to predict the exact timing of antigenic changes, the present results suggest that our model may have the potential to recognize emerging epidemic strains.

Knowledge of Vibrio vulnificus infection among Japanese patients with liver diseases: a prospective multicenter study.

BACKGROUND: Vibrio vulnificus (V. vulnificus) is a seafood-borne infectious pathogen that can be lethal to humans. The infection has been correlated with pre-existing liver disease, particularly liver cirrhosis. Awareness of V. vulnificus infection among Japanese citizens is low, despite the increasing number of patients with hepatocellular carcinoma (HCC). The present study was conducted to assess the level of knowledge of patients with liver disease regarding V. vulnificus infection. MATERIAL/METHODS: Questionnaires were sent to patients with chronic liver disease who had been treated by liver specialists at 14 medical institutes. RESULTS: Of 1,336 patients, 304 (22.8%) had liver cirrhosis, and 732 (54.8%) had comorbidities of this disease. Only 14.5% (194/1,336) of patients had knowledge of V. vulnificus infection. Of 304 patients with liver cirrhosis, 17.4% (53/304) of the patients had knowledge of V. vulnificus infection. Of 60 patients with liver cirrhosis and diabetes mellitus, 11 (18.3%) patients had knowledge of V. vulnificus infections. Even when the patients with high risk factors such as liver cirrhosis and diabetes mellitus had knowledge of V. vulnificus infections, most ate raw seafood without regard to season. CONCLUSIONS: Patients with chronic liver diseases and their physicians need to be better educated about V. vulnificus infection and its prevention.

Pegylated interferon plus ribavirin for genotype Ib chronic hepatitis C in Japan.

AIM: To evaluate the efficacy of pegylated interferon alpha-2b (peg-IFN alpha-2b) plus ribavirin (RBV) therapy in Japanese patients with chronic hepatitis C (CHC) genotype Ib and a high viral load. METHODS: One hundred and twenty CHC patients (58.3% male) who received peg-IFN alpha-2b plus RBV therapy for 48 wk were enrolled. Sustained virological response (SVR) and clinical parameters were evaluated. RESULTS: One hundred (83.3%) of 120 patients completed 48 wk of treatment. 53 patients (44.3%) achieved SVR. Early virological response (EVR) and end of treatment response (ETR) rates were 50% and 73.3%, respectively. The clinical parameters (SVR vs non-SVR) associated with SVR, ALT (108.4 IU/L vs 74.5 IU/L, P = 0.063), EVR (76.4% vs 16.4%, P < 0.0001), adherence to peg-IFN (>or= 80% of planned dose) at week 12 (48.1% vs 13.6%, P = 0.00036), adherence to peg-IFN at week 48 (54.7% vs 16.2%, P < 0.0001) and adherence to RBV at week 48 (56.1% vs 32.1%, P = 0.0102) were determined using univariate analysis, and EVR and adherence to peg-IFN at week 48 were determined using multivariate analysis. In the older patient group (> 56 years), SVR in females was significantly lower than that in males (17% vs 50%, P = 0.0262). EVR and adherence to Peg-IFN were demonstrated to be the main factors associated with SVR. CONCLUSION: Peg-IFN alpha-2b plus RBV combination therapy demonstrated good tolerability in Japanese patients with CHC and resulted in a SVR rate of 44.3%. Treatment of elderly female patients is still challenging and maintenance of adherence to peg-IFN alpha-2b is important in improving the SVR rate.

Two new entodiniomorphid Triplumaria ciliates from the intestine of the wild African white rhinoceros.

Two new species of Triplumaria in the order Entodiniomorphida, T. alluvia n. sp. and T. grypoclunis n. sp., are described from the large intestine of the wild African white rhinoceros. T. alluvia has three bud-shaped caudalia, one broad skeletal plate with a wavy left dorsal edge, and an axe-shaped tail flap. T. grypoclunis has three short arched caudalia, two broad skeletal plates, and a pointed and ventrally curved tail flap. These two new species have a C-shaped adoral polybrachykinety, a slender perivestibular polybrachykinety, and paralabial kineties in their retractable adoral ciliary zone. In T. alluvia, the perivestibular polybrachykinety is joined to both ends of the adoral polybrachykinety and paralabial kineties along the ventral side of the adoral polybrachykinety, showing the same arrangement as in Cycloposthium species. In T. grypoclunis, the perivestibular polybrachykinety is joined only to the right end of the adoral polybrachykinety and paralabial kineties along the left ventral side of the adoral polybrachykinety, showing an arrangement analogous to the Tripalmaria species.

High frequency of amantadine-resistant influenza A (H3N2) viruses in the 2005-2006 season and rapid detection of amantadine-resistant influenza A (H3N2) viruses by MAMA-PCR.

Using the newly designed mismatch amplification mutation assay (MAMA) PCR, we demonstrated the high frequency of amantadine-resistant influenza A (H3N2) viruses isolated during the 2005-2006 season by detecting the mutation at amino acid position 31 of the M2 protein (S31N). Further, phylogenetic analyses of the HA1 sequences of the S31N viruses revealed that they comprised a clonal lineage that would result in the common characteristic amino acid changes at positions 193 (Ser to Phe) and 225 (Asp to Asn) of the HA protein. We also demonstrated that the S31N/S193F/D225N viruses had already emerged in Aichi Prefecture by the end of the previous 2004-2005 season.

Toll-like receptor 4 signal transduction inhibitor, M62812, suppresses endothelial cell and leukocyte activation and prevents lethal septic shock in mice.

Sepsis occurs when microbes activate toll-like receptors (TLRs) stimulating widespread inflammation and activating coagulation cascades. TLR4 signal transduction has been recognized as a key pathway for lipopolysaccharide (LPS)-induced activation of various cells and an attractive target for treatment of sepsis. We found a new benzisothiazole derivative, M62812 that inhibits TLR4 signal transduction. This compound suppressed LPS-induced upregulation of inflammatory cytokines, adhesion molecules and procoagulant activity in human vascular endothelial cells and peripheral mononuclear cells. The half maximal inhibitory concentrations in these assays ranged from 1 to 3 microg/ml. Single intravenous administration of M62812 (10-20 mg/kg) protected mice from lethality and reduced inflammatory and coagulatory parameters in a murine d-galactosamine-sensitized endotoxin shock model. M62812 (20 mg/kg) also prevented mice from lethality in a murine cecal ligation and puncture model. These results suggest that inhibition of TLR4 signal transduction can suppress coagulation as well as inflammation during sepsis and may be clinically beneficial in sepsis treatment.

New entodiniomorphid ciliates from the intestine of the wild African white rhinoceros belong to a new family, the Gilchristidae.

Gilchristia artemis n.g., n.sp. and Digilchristia draconis n.g., n.sp. in the order Entodiniomorphida are described from the large intestine of the African white rhinoceros, and a new family Gilchristidae is proposed to contain them. These new species have a C-shaped adoral polybrachykinety, a slender vestibular polybrachykinety, and paralabial kineties along the ventral side of the adoral polybrachykinety in their retractable adoral ciliary zone, showing the same arrangement as in the rumen ciliates in the family Ophryoscolecidae. G. artemis has two skeletal plates and D. draconis one plate. In both species the dorsal skeletal plate is bow-shaped, folded in half longitudinally, twisting in the anterior part, and lying along the dorsal left side of the macronucleus. The second plate of G. artemis is slender and lies along the ventral side of the macronucleus. G. artemis has three ciliary arches and D. draconis has four arches along the dorsal and ventral sides of the body. Their arches are long and non-retractable, closely resembling those of ciliates in the families, Spirodiniidae and Cycloposthiidae, and are not analogous to the single retractable ciliary arch of the rumen ciliates in the family Ophryoscolecidae.

[Analysis of complications and prognosis for different types of stroke patients registered between 1993 and 2000 in Aichi Prefecture].

PURPOSE: This study was designed to analyze complications and prognosis for different types of stroke patients registered in Aichi Prefecture between 1993 and 2000. METHODS: A total of 23,979 out of 27,304 registered patients in the Aichi stroke patient registration program with 4 type of strokes (cerebral thrombosis (CRT), cerebral embolism (CRE), intracerebral hemorrhage (ICH), and subarachnoid hemorrhage (SAH)) were analyzed for complications and prognosis (survival, disability, dementia), with reference to gender, age-groups, and types of stroke. RESULTS: A total of 13,365 (55.7%) male (65.5 +/- 12.2 yr: Mean +/- SD) and 10,614 (44.3%) female (69.7 +/- 13.3 yr) patients were registered. As the type of stroke, CRT comprised the highest percentage, both in males (49.5%) and females (41.1%), followed by ICH (males: 30.4%, females: 29.8%). The percentage of SAH in females (17.3%) was found to be about twice as high as that in males (8.3%). Analyses of complications revealed hypertension to be the greatest risk factor in both sexes (about 50%), followed by history of stroke (males: 20.1%, females: 16.2%). Male patients had a significantly higher overall survival rate (84.7%) than females (81.0%) (P <0.001). SAH was associated with the lowest survival rate in both sexes (about 60%), with statistical significance (P<0.001). Development of disability or/and dementia as sequela of stroke was higher in females (disability: 54.5%, dementia: 21.1%) than in males (44.2%, 15.1%, respectively) (P< 0.001). Logistic regression analyses revealed that the factors most contributing to death were advanced age, a history of stroke, heart disease, and renal insufficiency. For the development of disability and dementia, being femal, of advanced age, with a history of stroke, heart disease, and renal insufficiency were important. Abnormal lipid metabolism appeared to be a protection factor regarding prognosis (survival, disability, dementia). CONCLUSIONS: This study demonstrated that hypertension is the most frequently reported complication for all types of stroke except CRE, and logistic regression analyses revealed that the factor contributing most to prognosis (survival, disability, dementia) was a history of stroke. The results suggested the importance of: i) removing hypertansion as the most significant risk factor, as well as diabetes and heart disease in order to prevent strokes; and ii) preventing re-attack(s) of stroke in order to improve the prognosis.

Synthesis and evaluation of 4-substituted benzylamine derivatives as beta-tryptase inhibitors.

Since beta-tryptase is considered a critical mediator of asthma, potent tryptase inhibitors may be useful as new agents for the treatment of asthma. We investigated 4-substituted benzylamine derivatives and obtained M58539 (15h) as a potent inhibitor of beta-tryptase (IC50 = 5.0 nM) with high selectivity against other serine proteases, low molecular weight, clog P value less than 5, lack of amidino and guanidino groups, and independence of Zn2+ ion.

Ability of Shiga toxin-producing Escherichia coli and Salmonella spp. to survive in a desiccation model system and in dry foods.

In order to determine desiccation tolerances of bacterial strains, the survival of 58 diarrheagenic strains (18 salmonellae, 35 Shiga toxin-producing Escherichia coli [STEC], and 5 shigellae) and of 15 nonpathogenic E. coli strains was determined after drying at 35 degrees C for 24 h in paper disks. At an inoculum level of 10(7) CFU/disk, most of the salmonellae (14/18) and the STEC strains (31/35) survived with a population of 10(3) to 10(4) CFU/disk, whereas all of the shigellae (5/5) and the majority of the nonpathogenic E. coli strains (9/15) did not survive (the population was decreased to less than the detection limit of 10(2) CFU/disk). After 22 to 24 months of subsequent storage at 4 degrees C, all of the selected salmonellae (4/4) and most of the selected STEC strains (12/15) survived, keeping the original populations (10(3) to 10(4) CFU/disk). In contrast to the case for storage at 4 degrees C, all of 15 selected strains (5 strains each of Salmonella spp., STEC O157, and STEC O26) died after 35 to 70 days of storage at 25 degrees C and 35 degrees C. The survival rates of all of these 15 strains in paper disks after the 24 h of drying were substantially increased (10 to 79 times) by the presence of sucrose (12% to 36%). All of these 15 desiccated strains in paper disks survived after exposure to 70 degrees C for 5 h. The populations of these 15 strains inoculated in dried foods containing sucrose and/or fat (e.g., chocolate) were 100 times higher than those in the dried paper disks after drying for 24 h at 25 degrees C.

Evaluation of pulsed-field gel electrophoresis analysis performed at selected prefectural institutes of public health for use in PulseNet Japan.

In order to evaluate reliability of pulsed-field gel electrophoresis (PFGE) analysis performed at different prefectural public health institutes (PHIs) for use in the PulseNet Japan surveillance system to detect enterohemorrhagic Escherichia coli O157, we compared the results of PFGE-typing of 14 selected strains of O157 performed at 8 selected PHIs to evaluate the reliability of different experimental protocols used in these PHIs. PFGE was performed for 14 strains for which there were 14 PFGE types in 3 PHIs, and 13 PFGE types in 5 PHIs by using their own protocols and/or those of the National Institute of Infectious Diseases (NIID). PFGE fingerprints from 5 out of the 8 PHIs were successfully genotyped for all of the 14 strains. A PFGE fingerprint from one PHI was successfully genotyped when the NIID pulsing protocol was used, but was not genotyped when the PHI's own protocols were used. PFGE fingerprints from 2 PHIs failed to be genotyped for one each of the strains. The cause of this genotyping failure was considered to be inappropriate PFGE pulsing protocols or inadequate digestion of chromosomal DNA. These results suggest that PFGE protocols should be standardized for the establishment of PulseNet Japan.

Matrix metalloproteinases in the pathogenesis of asthma and COPD: implications for therapy.

While asthma is an inflammatory disorder of the airways involving mediators released from mast cells and eosinophils, inflammation alone is insufficient to explain the chronic nature of the disease. Recent progress in the understanding of disease pathogenesis has revealed that airway remodeling, which is at least in part due to an excess of extracellular matrix (ECM) deposition in the airway wall, plays a significant role in airflow obstruction. Matrix metalloproteinases (MMPs) have been suggested to be the major proteolytic enzymes to induce airway remodeling in asthma and COPD. It has been widely accepted that different inflammatory processes are involved in asthma and COPD with different inflammatory cells, mediators, and responses to treatments. Despite these different processes, airflow obstruction and airway remodeling characterize these two diseases. MMP-2 and -9 have been reported to be involved in the pathogenesis of airway remodeling in both diseases and MMP-12, in addition to these MMPs, in the pathogenesis of COPD. In this review, we discuss the current views on the role of MMPs in the pathogenesis of bronchial asthma and COPD. Anti-MMP therapy could theoretically be useful to prevent airway remodeling in asthma and COPD. However, to date no clinical data are available regarding the efficacy of anti-MMP therapies in the treatment of patients with asthma and COPD.

Synthesis and evaluation of 1-arylsulfonyl-3-piperazinone derivatives as factor Xa inhibitors III. Effect of ring opening of piperazinone moiety on inhibition.

Compounds containing an ethylenediamine structure in place of the piperazine ring of M55113 (1) and M55551 (2) were synthesized to investigate the effects of a piperazine moiety and evaluated for activity as factor Xa (FXa) inhibitors. Most such compounds, however, exhibited lower activity (1/10-1/100) than that of M55113 and M55551 as FXa inhibitors.