Increased glucose production in mice overexpressing human fructose-1,6-bisphosphatase in the liver

Increased endogenous glucose production (EGP) predominantly from the liver is a characteristic feature of type 2 diabetes, which positively correlates with fasting hyperglycemia. Gluconeogenesis is the biochemical pathway shown to significantly contribute to increased EGP in diabetes. Fructose-1,6-bisphosphatase (FBPase) is a regulated enzyme in gluconeogenesis that is increased in animal models of obesity and insulin resistance. However, whether a specific increase in liver FBPase can result in increased EGP has not been shown. The objective of this study was to determine the role of upregulated liver FBPase in glucose homeostasis. To achieve this goal, we generated human liver FBPase transgenic mice under the control of the transthyretin promoter, using insulator sequences to flank the transgene and protect it from site-of-integration effects. This resulted in a liver-specific model, as transgene expression was not detected in other tissues. Mice were studied under the following conditions: 1) at two ages (24 wk and 1 yr old), 2) after a 60% high-fat diet, and 3) when bred to homozygosity. Hemizygous transgenic mice had an approximately threefold increase in total liver FBPase mRNA with concomitant increases in FBPase protein and enzyme activity levels. After high-fat feeding, hemizygous transgenics were glucose intolerant compared with negative littermates (P < 0.02). Furthermore, when bred to homozygosity, chow-fed transgenic mice showed a 5.5-fold increase in liver FBPase levels and were glucose intolerant compared with negative littermates, with a significantly higher rate of EGP (P < 0.006). This is the first study to show that FBPase regulates EGP and whole body glucose homeostasis in a liver-specific transgenic model. Our homozygous transgenic model may be useful for testing human FBPase inhibitor compounds with the potential to treat patients with type 2 diabetes.     

Inelastic behavior in repeated shearing of bovine white matter

Understanding the brain's response to multiple loadings requires knowledge of how straining changes the mechanical response of brain tissue. We studied the inelastic behavior of bovine white matter and found that when this tissue is stretched beyond a critical strain threshold, its reloading stiffness drops. An upper bound for this strain threshold was characterized, and was found to be strain rate dependent at low strain rates and strain rate independent at higher strain rates. Results suggest that permanent changes to tissue mechanics can occur at strains below those believed to cause physiological disruption or rupture of axons. Such behavior is characteristic of disentanglement in fibrous-networked solids, in which strain-induced mechanical changes may result from fiber realignment rather than fiber breakage.     

Tuning the Au(I)-mediated inhibition of cathepsin B through ligand substitutions

It has been over 80 years since the antiarthritic properties of gold(I) complexes were first recognized. However, a detailed understanding of their mechanism of action has been slow to develop. One likely biological target of gold(I) is the cathepsin family of lysosomal cysteine proteases, enzymes involved in the inflammation and joint destruction that are hallmarks of rheumatoid arthritis (RA). We have previously shown that analogs of auranofin, a clinically available antiarthritic drug, inhibit cathepsin B. In this study, the extent to which the steric and electronic properties of the phosphine ligand can be modified to obtain enhanced potency against cathepsin B is investigated.     

Japan’s Hidden Youths: Mainstreaming the Emotionally Distressed in Japan

One of the most talked-about social issues in Japan in recent years has been the problem of the nation's purportedly one million "hidden" youths, known as hikikomori (literally, "the withdrawn"). Most observers agree that the category of hikikomori encompasses a wide range of problems and provocations. The fact that these various dilemmas lead to the shared outcome of shutting oneself away at home is the point of departure here. The article explores the spheres of mental health care, education and family, focusing on the reluctance to highlight underlying psychological dimensions of hikikomori and the desire on the part of schools and families to "mainstream" Japanese children, accommodating as many as possible within standardized public education. Hikikomori can perhaps be seen as a manifestation of Japanese democracy, in which the good society is imagined as cohesive, protective and secure, rather than one in which the individual can freely exercise the right to be different. Schools, families and the sphere of mental health care have focused on producing social inclusion but have discouraged citizens from being labeled as "different" -- even when such a distinction might help them. The dearth of facilities and discourse for caring for the mentally ill or learning disabled is, in many respects, the darker side of Japan's successes. Those who cannot adjust are cared for through the institutions of families, companies and various other spheres that offer spaces to rest and to temporarily "drop out"; however, the expectation is that rest will eventually lead to a re-entry into mainstream society. Often the psychological problem or disability that led to the problem goes unnamed and untreated (hikikomori, psychiatry, special education, youth, family, Japan).     

A novel and rapid method for synthesizing positive controls and standards for quantitative PCR

We developed and tested a method to produce DNA standards and controls for quantitative PCR by designing and performing partial hybridization of long oligonucleotides before double stranded DNA fragments were synthesized and subsequently amplified by conventional PCR. This approach does not require any natural DNA template. Applications include the production of standards, which cannot be easily produced from DNA extracted from bacteria or plants.     

Ultrafiltration-based techniques for rapid and simultaneous concentration of multiple microbe classes from 100-L tap water samples

This study focused on ultrafiltration as a technique for simultaneously concentrating and recovering viruses, bacteria and parasites in 100-L drinking water samples. A chemical dispersant, sodium polyphosphate, and Tween 80 were used to increase microbial recovery efficiencies. Secondary concentration was performed to reduce sample volumes to 3-5 mL for analysis using tissue culture, microscopy, and real-time PCR and RT-PCR. At seeding levels of 100-1000 (CFU, PFU, oocysts, or particles), a "high-flux" ultrafiltration procedure was found to achieve mean recoveries of 51-94% of simultaneously seeded MS2 bacteriophage, echovirus 1, Salmonella enterica subsp. enterica serovar Typhimurium, Bacillus atrophaeus subsp. globigii endospores, Cryptosporidium parvum oocysts, and 4.5-mum microspheres. When 4-7% of the final sample concentrate volume was assayed using real-time PCR and RT-PCR, overall method sensitivities were <100 C. parvum oocysts, <240 PFU echovirus 1, <100 CFU Salmonella and approximately 160 CFU B. atrophaeus spores in 100-L drinking water samples. The "high-flux" ultrafiltration procedure required approximately 2 h, including time required for backflushing. Secondary concentration procedures required an additional 1-3 h, while nucleic acid extraction and real-time PCR procedures required an additional 2-2.5 h. Thus, this study demonstrated that efficient recovery and sensitive detection of diverse microbes in 100-L drinking water samples could be achieved within 5-8 h using ultrafiltration, rapid secondary processing techniques, and real-time PCR.     

Ageing does not significantly affect performance in a spatial learning task in the domestic cat (Felis silvestris catus)

A spatial learning task was used to examine cognitive function in 36 cats (1.0–15.1 years of age), with a control for motor function. No significant age-related decline in cognitive function was found. Initial selection of 75 cats showed no significant age differences between young (0–3 years), adult (3.1–8 years), senior (8.1–12 years) and geriatric cats (12.1–15.1 years) locating food rewards in a holeboard box. Consequently, the senior and geriatric groups were combined because of a concern that age effects may not be seen in the 8–12-year-old cats. Differences between these three age groups of cats to learn the position of three food rewards in an array of 30 otherwise empty (food-scented) positions were non-significant. Differences between age groups remained non-significant for retrieval of rewards from these three positions even when masked with a tissue paper cover. There was no significant effect of age on speed of reaching each criterion. Although the total number of errors was similar, there was a significantly increased proportion of working memory errors (when the cat returned to a previously baited position) and decreased proportion of reference memory errors (when the cat put a nose or paw in an empty position) with increasing age. There was no effect of age on motor function, with success in a plank-crossing test related only to plank width. These findings support previous work suggesting that the cat differs from other species in the way in which it demonstrates age-related declines in cognitive function. Further studies using different testing methodologies are required to assess other cognitive functions that may decline with age.