Re-Evaluation of the Cross-Reactions of the Antibody against the Causative Agent for Paracoccidioidomycosis Ceti; Paracoccidioides ceti and the Related Fungal Species.

Paracoccidioidomycosis ceti (PCM-C) is a chronic granulomatous keloidal dermatitis in cetaceans that has been reported worldwide and is caused by Paracoccidioides ceti. Serological cross-reactions among highly pathogenic fungal infections and related diseases have been reported. However, the true cross-reaction of antibodies against P. ceti has remained unknown due to the use of positive control sera from infected dolphins. This study aimed to re-evaluate antibodies from mechanically dislodged fungal cells in the infected tissue of a PCM-C case and demonstrate the actual cross-reaction. The results revealed a limited cross-reaction between PCM-C and paracoccidioidomycosis, while the antibodies did not react with other pathogens such as Coccidioides posadasii, Histoplasama capsulatum, and Arthrographis kalrae. Thus, the method for evaluation of the antibody against PCM-C is reliable, and there is potential for epidemiological study.

Seroprevalences Against Paracoccidioides cetii: A Causative Agent for Paracoccidiomycosis Ceti (PCM-C) and Coccidioides posadasii; for Coccidioidomycosis (CCM) in Dall's Porpoise (Phocoenoides dalli) and Harbor Porpoise (Phocoena phocoena) Stranded at Hokkaido, Japan.

Paracoccidiodomycosis ceti (PCM-C) is a zoonotic mycosis characterized by chronic granulomatous cutaneous lesions in cetaceans. It is distributed worldwide and is caused by an unculturable fungus; Paracoccidioides cetii. On the other hand, coccidioidomycosis (CCM), caused by Coccidioides spp., is also a zoonotic and highly pathogenic fungal infection endemic in both American continents. Even though the Far East is not an endemic area of CCM, an autochthonous case has been reported in China. Although the seroprevalence against P. cetii in captive dolphins was 61.0%, there is no information on wild dolphins living in cold waters. The present study aimed to investigate the seroprevalence against P. cetii and C. posadasii in 15 Dall's porpoises (Phocoenoides dalli) and 11 harbor porpoises (Phocoena phocoena) stranded in Hokkaido, Japan. The seroprevalence against P. cetii in the above dolphins was 26.9% (7/26), which was recorded only in Dall's porpoises (7/15), and that against C. posadasii was 15.4% (4/26), three in Dall's porpoises and one in harbor porpoise. The present study demonstrated positive seroprevalence against P. cetii and C. posadasii in wild cetaceans living in the subarctic areas of the Far East as the first records, and would issue the warning those who live in the area were exposed to the causative agent of CCM from seawater.

A 1-bp deletion in Mc1r in a Norway rat (Rattus norvegicus) from Sado Island, Japan gives rise to a yellowish color variant: an insight into mammalian MC1R variants.

The melanocortin-1 receptor gene (MC1R) controls production of the pigments eumelanin and pheomelanin. Changes in MC1R lead to variation in coat color in mammals, which can range from entirely black (melanism) to yellowish. In this study, we report a case of a wild-caught Norway rat (Rattus norvegicus) from Sado Island, Japan with a yellowish coat color. Upon sequencing the whole coding region of the Mc1r gene (954 bp), we found a 1-bp deletion at site 337 (c.337del), indicative of a frameshift mutation, which was characterized as a severe loss-of-function or null mutation. A spectrophotometer was used to measure coat color, revealing that the rat had a distinctly lighter coat, based on lightness score, than mice with homozygous similar loss-of-function mutations. This implies that loss-of-function mutations can yield different phenotypes in murine rodents. The loss-of-function-mutant rat exhibited a contrasting coat pattern consisting of darker and lighter colors along its dorsal and ventral sides, respectively. Similar patterns have been observed in homozygous MC1R-deficient mutants in other mammals, implying that the countershading pattern can still be expressed despite the absence of MC1R in the melanocyte.

Discovery of a sugar-based nanoparticle universally existing in boiling herbal water extracts and their immunostimulant effect.

Herbal medicine is mainly prepared from boiling herbal water extracts. Many epoch-making immunosuppressant drugs, such as glycyrrhizic acid (old example) and FTY720 (current example), were developed from herbal secondary metabolites in the boiling water extract by partition with organic solvents. However, few immunostimulants have been discovered by this method. Instead of the usual method, we aimed to find a novel immunostimulant component by two unique methods in the research of herbal medicine: ultracentrifugation and electron microscopy. The immunostimulant was not a secondary metabolite, as expected, but the structure was a nanoparticle formed by a polysaccharide. In addition, we clarified the immune effect of the nanoparticle. Intake of the nanoparticle by phagocytosis resulted in immunostimulant effects by increasing the genes and proteins of inflammatory cytokines in macrophage cells. The immunostimulant effects were inhibited by a phagocytosis inhibitor, cytochalasin D. To the best of our knowledge, this study is the first to describe the discovery of a nanoparticle in boiling herbal water extracts and its immunostimulant properties. This study will provide additional understanding of the efficacy of herbal medicine, in that the immunostimulant nanoparticle universally exists in boiling herbal water extracts. Thus, traditional herbal medicine may be an oldest known nanomedicine. Furthermore, this study suggests that the immunostimulant nanoparticle simply can be obtained from herbal medicine only by ultracentrifugation. We hope that this simple strategy will substantially contribute to drug development, including vaccine adjuvant, in the future.

Individual variation of daily torpor and body mass change during winter in the large Japanese field mouse (Apodemus speciosus).

Daily torpor is a strategy used by some overwintering small endotherms to aid in energy conservation. However, the pattern of torpor varies among individuals within species and populations, even under the same environmental conditions, with significant implications for survival rate and reproductive success. Body mass is one factor that may influence this variation, especially in some small mammals that accumulate fat stores prior to overwintering. However, to our knowledge there has been no previous study examining the detailed relationships between torpor expression and body mass change in small mammals that hoard food as an energy resource during winter. The large Japanese field mouse, Apodemus speciosus, whose winter survival strategy depends on food caches instead of fat stores, displays daily torpor under artificial winter conditions (short-day photoperiod and cold). The present study clarifies the characteristics and patterns of daily torpor and body mass change in this species in the laboratory. Although expression of daily torpor was facilitated progressively as in other species, the observed patterns of torpor expression and body mass change showed considerable individual variation. Moreover, there was no obvious correlation between body mass and daily torpor expression. Therefore, it is suggested that in A. speciosus body mass may not contribute to individual variation of daily torpor during winter. Daily torpor during winter may be adjusted by not only mechanisms common to other small mammals, but also species-specific factors relating to the external or internal reserves of energy in small mammals.

Individual differences in torpor expression in adult mice are related to relative birth mass.

Daily torpor is a physiological adaptation in small mammals and birds, characterised by drastic reductions in metabolism and body temperature. Energy-constraining conditions, such as cold and starvation, are known to cause the expression of daily torpor. However, the reason for high degrees of inter- and intra-individual variation in torpor expression (TE) in similar situations is not clear. As littermates of altricial animals are exposed to an uneven allocation of maternal resources from conception to weaning, we tested whether early nutritional experiences have long-term effects on TE in adults. We used full-sibling littermates of laboratory mice that as adults were starved overnight to induce torpor. We measured body mass from birth until adulthood as an indicator of nutritional status, and calculated the relative body mass (RBM) as an indicator of the difference in nutritional status within a litter. After maturation, we subjected mice to five repeated torpor induction trials involving 24 h of fasting and 5 days of recovery. Half of the female mice displayed great individual variation in TE whereas male mice rarely exhibited daily torpor. In females, RBM at birth influenced TE, irrespective of body mass in adulthood; thus, female mice born with low RBMs displayed high TE in adulthood. In conclusion, we provide evidence that TE in mice differs among littermates, and that this variation is linked closely to heterogeneous nutritional experiences during the fetal period.

[Mediastinal Pancreatic Pseudocyst with Pancreatic Pleural Effusion].

A 72-year-old man with chronic alcohol related pancreatitis was admitted for dyspnea and pain at the upper body. Chest X-ray showed right massive pleural effusion. Chest and abdominal contrast enhanced thin slice computed tomography revealed the route from the pancreatic head reaching the right thoracic cavity via the esophagus hiatus and the communication between the cystic lesion and main pancreatic duct. We drained the pleural effusion that showed abnormally high amylase activity. We diagnosed his illness as mediastinal pancreatic pseudocyst with pancreatic pleural effusion. Endoscopic Nasopancreatic Drainage catheter was placed in the main pancreatic duct, and the pleural effusion disappeared.

Magnitude of food overabundance affects expression of daily torpor.

Many small mammal species use torpor as a strategy for reducing energy expenditure in winter. Some rodent hibernators also hoard food to provide reserves of energy, and individuals with large hoards express less torpor than those with smaller reserves. These facts imply that animals can recognize levels of food availability, but where food is very plentiful, it is unclear whether torpor expression is affected by temporal changes in the extent of food overabundance. Moreover, the relationship between daily torpor and excess food availability has not been clearly established. The large Japanese field mouse Apodemus speciosus caches food for use as a winter energy resource and exhibits daily torpor under artificial winter conditions. The present study examined whether individuals exposed to different magnitudes of overabundant food exhibited differences in expression of daily torpor, and secondly whether torpor expression varied in response to changes in the overall quantity of overabundant food. It was observed that while absolute quantities of overabundant food did not appear to affect daily torpor expression, the mice did respond to changes in food availability, even when food remained overabundant. This suggests that the mice respond to fluctuations in food availability, even where these changes do not place any constraint on energy budgets. Thus recognition of changing food availability cannot be a purely physiological response to shortage or plenty, and may contribute to predictions of future energy availability. The expression of torpor was inhibited in response to increasing food availability, and the mice used shallower torpor when food availability increased to superabundance. These findings suggest that daily torpor may be regulated not only physiologically in response to energy constraints but also psychologically, via recognition of food availability.

Huddling facilitates expression of daily torpor in the large Japanese field mouse Apodemus speciosus.

Small endotherms employ multiple adaptations to maintain energy balance in winter, including spontaneous daily torpor and simultaneous huddling. The relationships between these adaptations have been discussed in several previous studies, but it has not been well-established if huddling actually affects the expression of torpor in small endotherms. We examine whether and how huddling affects the expression of torpor in the large Japanese field mouse Apodemus speciosus, which is known to become torpid under artificial winter conditions. The mice were found to adjust expression of torpor in response to the number of cage mates. Torpor frequency and minimum torpid body temperature were both significantly elevated when the number of cage mates was increased, but there was no significant change in torpor bout length. Rewarming rate on arousal was lower when the number of cage mates was increased, suggesting reduction in endogenous rewarming due to exogenous passive rewarming. Food consumption per mouse decreased significantly with increasing number of cage mates. Thus, our study demonstrates that social thermoregulatory behaviors such as huddling can facilitate expression of spontaneous daily torpor in small rodents. These findings suggest that energy constraints, such as ambient temperature and food availability may not be the only modulating factors on the expression of daily torpor.

Predictors for prehypertension in patients with impaired glucose tolerance.

Prehypertension (PHT) is associated with increased risk of cardiovascular disease and progression to hypertension. Insulin resistance and hyperinsulinemia have been reported among patients with hypertension. In addition, impaired glucose tolerance (IGT) is a strong predictor of not only of type 2 diabetes but also of cardiovascular disease. However, little is known about the impact of insulin resistance on recently defined categories of hypertension and IGT. The aim of this study was to examine associations of surrogate makers of insulin resistance with PHT and IGT. In a total of 102 IGT patients with normotension and PHT (age: 58+/-5 years; mean+/-SD), blood pressure measurement, 75 g oral glucose tolerance testing (OGTT), metabolic analysis and echocardiography were performed. Body mass index was higher in the PHT group than in the normotension group (p<0.05). The fasting immnunoreactive insulin (F-IRI) (p<0.0001), homeostasis model assessment (HOMA) index (p<0.0001), 30 min postload glucose (p<0.05), 60 min postload glucose (p<0.05), 120 min postload glucose (p<0.01), 120 min postload insulin (p<0.0001) and left ventricular mass index (LVMI) (p<0.0005) were higher in the PHT group than in the normotension group. Multivariate logistic analysis revealed that the presence of PHT was independently predicted by F-IRI. Our findings indicate that the presence of PHT was associated with hyperinsulinemia and that the F-IRI was an independent predictor of PHT in these Japanese patients with IGT.

Postchallenge plasma glucose and glycemic spikes are associated with pulse pressure in patients with impaired glucose tolerance and essential hypertension.

Elevated pulse pressure (PP) is associated with an increased risk of cardiovascular events. We examined whether PP is associated with post-challenge hyperglycemia in Japanese patients with essential hypertension and impaired glucose tolerance (IGT). In a total of 70 untreated essential hypertensive patients (age: 57+/-4 years, mean+/-SD; males=35, females=35), 24-h ambulatory blood pressure (ABP) monitoring, 75 g oral glucose tolerance testing (OGTT), metabolic analysis and echocardiography were performed. Patients were categorized into a high PP group (PP>or=60 mmHg, n=33) or a normal PP group (PP<60 mmHg, n=37). In all patients, 24-h systolic ABP, daytime systolic ABP, nighttime systolic ABP, and nighttime heart rate were significantly higher in the high PP group. Additionally, fasting immunoreactive insulin (F-IRI), homeostasis model assessment (HOMA) index, left ventricular mass index (LVMI) were also elevated in the high PP group. Finally, the high PP group exhibited impaired insulin secretion, increased post-challenge glucose concentrations and greater glucose spikes (PGS) during 75 g OGTT. Of the parameters measured, 24-h PP correlated positively with age, triglyceride, uric acid, F-IRI, HOMA index, 1-h postload glucose and insulin, 2-h postload glucose and insulin, PGS60, PGS120, PGSmax, LVMI, and deceleration time but correlated negatively with HDL-cholesterol and E/A ratio. Multiple regression analysis revealed that PP level was independently predicted by age, LVMI, and PGS120. Our results show that age, LVMI, and PGS120 are significantly associated with high PP in Japanese patients with IGT and essential hypertension.

The role of microalbuminuria and insulin resistance as significant risk factors for white matter lesions in Japanese type 2 diabetic patients.

OBJECTIVE: The presence of white matter lesions (WML) is an important prognostic factor for the development of stroke. Microalbuminuria, which is associated with diabetes, has been flagged as a novel predictor for cerebrovascular events. This preliminary study was therefore designed to test the hypothesis that the presence of WML correlates with microalbuminuria and insulin resistance in patients with type 2 diabetic mellitus (DM) not receiving insulin treatment. PATIENTS AND METHODS: Based on brain magnetic resonance imaging (MRI) findings, 90 type 2 diabetic patients were divided into two groups: a WML-positive group (57+/-8 years, mean+/-SD, n=34) and a WML-negative group (57+/-6 years, n=56). The level of blood glucose was assessed by fasting plasma glucose (FPG), fasting immunoreactive insulin (F-IRI), homeostasis model assessment (HOMA) index, and hemoglobin A(1c) (HbA1c). RESULTS: The body mass index was higher in the WML-positive group than in the WML-negative group (p<0.01). Plasma levels of triglycerides were higher while high-density lipoprotein cholesterol (HDL-C) was lower in the WML-positive group than in the WML-negative group (p<0.05 and p<0.0001, respectively). Fasting plasma glucose (p<0.005), insulin concentrations (p<0.0001), HOMA index (p<0.0001), and urinary albumin excretion (p<0.0001) levels were higher in the WML-positive group than in the WML-negative group. Multivariate logistic analysis revealed that WML was independently predicted by the microalbuminuria and insulin resistance (p<0.005, p<0.0005, respectively). CONCLUSION: The results of this preliminary study indicate that the presence of WML was associated with the microalbuminuria and insulin resistance in these Japanese patients with type 2 DM; larger cohort studies are warranted to confirm these findings.