Effects of nine weeks of unloading on neuromuscular activities in adult rats

Effects of 9-week hindlimb suspension and 8-week recovery on locomotor performance and electromyogram (EMG) activities of soleus (Sol), plantaris (Pl), lateral gastrocnemius (LG), and tibialis anterior (TA), were studied in adult rats. Hyperextension of knee and ankle joints, noted after nine weeks of suspension, did not recover during 8-week ambulation. Growth of Sol was fully inhibited by suspension and did not recover completely within 8 weeks of ambulation. The EMG levels in Sol, Pl, and LG 1 day after suspension were 52-76% less than the pre-suspension level (resting on the floor). These activities were recovered to or near the pre-suspension level around 1 week, but decreased again to 10-29% of controls from 7 to 9 weeks. The integrated EMG of TA was elevated during the first week of suspension but then gradually declined to control levels within four weeks. At the end of suspension, the Sol and Pl, not the LG, EMGs remained reduced and the TA EMG remained hyperactive. Co-activation of dorsi- and plantar- flexors occurred often during quadripedal walking following suspension. Such a phenomenon was not observed in the control rats. These phenomena were recovered within 1 week. It is suggested that the unloading-related alterations of neuromuscular activities and/or locomotion, but not the hyperextension of knee and ankle joints, in rats are reversible.     

Human brain activity during illusory visual jitter as revealed by functional magnetic resonance imaging

One central problem in vision is how to compensate for retinal slip. A novel illusion (visual jitter) suggests the compensation mechanism is based solely on retinal motion. Adaptation to visual noise attenuates the motion signals used by the compensation stage, producing illusory jitter due to the undercompensation of retinal slip. Here, we investigated the neural substrate of retinal slip compensation during this illusion using high-field fMRI and retinotopic mapping in flattened cortical format. When jitter perception occurred, MR signal decreased in lower stages of the visual system but increased prominently in area MT+. In conclusion, visual areas as early as V1 are responsible for the adaptation stage, and MT+ is involved in the compensation stage. The present finding suggests the pathway from V1 to MT+ has an important role in stabilizing the visual world.     

Early Onset of Lipofuscin Accumulation in Dystrophin-Deficient Skeletal Muscles of DMD Patients and mdx Mice

Lipofuscin, the so-called ageing pigment, is formed by the oxidative degradation of cellular macromolecules by oxygen-derived free radicals and redox-active metal ions. Usually it accumulates in post-mitotic, long-lived cells such as neurons and cardiac muscle cells. In contrast, it is rarely seen in either normal or diseased skeletal muscle fibres. In this paper, we report that lipofuscin accumulates at an early age in both human and murine dystrophic muscles. Autofluorescent lipofuscin granules were localized, using confocal laser scanning microscopy and electron microscopy, in dystrophin-deficient skeletal muscles of X chromosome-linked young Duchenne muscular dystrophy (DMD) patients and of mdx mice at various ages after birth. Age-matched normal controls were studied similarly. Autofluorescent lipofuscin granules were observed in dystrophic biceps brachii muscles of 2-7-year-old DMD patients where degeneration and regeneration of myofibres are active, but they were rarely seen in age-matched normal controls. In normal mice, lipofuscin first appears in diaphragm muscles nearly 20 weeks after birth but in mdx muscles it occurs much earlier, 4 weeks after birth, when the primary degeneration of dystrophin-deficient myofibres is at a peak. Lipofuscin accumulation increases with age in both mdx and normal controls and is always higher in dystrophic muscles than in age-matched normal controls. At the electron microscopical level, it was confirmed that the localisation of autofluorescent granules observed by light microscopy in dystrophin-deficient skeletal muscles coincided with lipofuscin granules in myofibres and myosatellite cells, and in macrophages accumulating around myofibres and in interstitial connective tissue. Our results agree with previous biochemical and histochemical data implying increased oxidative damages in DMD and mdx muscles. They indicate that dystrophin-deficient myofibres are either more susceptible to oxidative stress, or are subjected to higher intra- or extracellular oxidative stress than normal controls, or both.     

Apoptosis and inactivation of the PI3-kinase pathway by tetrocarcin A in breast cancers

A survival kinase, Akt, is a downstream factor in the phosphatidylinositide-3'-kinase-dependent pathway, which mediates many biological responses including glucose uptake, protein synthesis and the regulation of proliferation and apoptosis, which is assumed to contribute to acquisition of malignant properties of human cancers. Here we find that an anti-tumor antibiotic, tetrocarcin A, directly induces apoptosis of human breast cancer cells. The apoptosis is accompanied by the activation of a proteolytic cascade of caspases including caspase-3 and -9, and concomitantly decreases phosphorylation of Akt, PDK1, and PTEN, a tumor suppressor that regulates the activity of Akt through the dephosphorylation of polyphosphoinositides. Tetrocarcin A affected neither expression of Akt, PDK1, or PTEN, nor did it affect the expression of Bcl family members including Bcl-2, Bcl-X(L), and Bax. These results suggest that tetrocarcin A could be a potent chemotherapeutic agent for human breast cancer targeting the phosphatidylinositide-3'-kinase/Akt signaling pathway.     

MKBP, a Novel Member of the Small Heat Shock Protein Family, Binds and Activates the Myotonic Dystrophy Protein Kinase

Muscle cells are frequently subjected to severe conditions caused by heat, oxidative, and mechanical stresses. The small heat shock proteins (sHSPs) such as αB-crystallin and HSP27, which are highly expressed in muscle cells, have been suggested to play roles in maintaining myofibrillar integrity against such stresses. Here, we identified a novel member of the sHSP family that associates specifically with myotonic dystrophy protein kinase (DMPK). This DMPK-binding protein, MKBP, shows a unique nature compared with other known sHSPs: (a) In muscle cytosol, MKBP exists as an oligomeric complex separate from the complex formed by αB-crystallin and HSP27. (b) The expression of MKBP is not induced by heat shock, although it shows the characteristic early response of redistribution to the insoluble fraction like other sHSPs. Immunohistochemical analysis of skeletal muscle cells shows that MKBP localizes to the cross sections of individual myofibrils at the Z-membrane as well as the neuromuscular junction, where DMPK has been suggested to be concentrated. In vitro, MKBP enhances the kinase activity of DMPK and protects it from heat-induced inactivation. These results suggest that MKBP constitutes a novel stress-responsive system independent of other known sHSPs in muscle cells and that DMPK may be involved in this system by being activated by MKBP. Importantly, since the amount of MKBP protein, but not that of other sHSP family member proteins, is selectively upregulated in skeletal muscle from DM patients, an interaction between DMPK and MKBP may be involved in the pathogenesis of DM.     

Word Decoding of Protein Amino Acid Sequences with Availability Analysis: A Linguistic Approach

The amino acid sequences of proteins determine their three-dimensional structures and functions. However, how sequence information is related to structures and functions is still enigmatic. In this study, we show that at least a part of the sequence information can be extracted by treating amino acid sequences of proteins as a collection of English words, based on a working hypothesis that amino acid sequences of proteins are composed of short constituent amino acid sequences (SCSs) or “words”. We first confirmed that the English language highly likely follows Zipf''s law, a special case of power law. We found that the rank-frequency plot of SCSs in proteins exhibits a similar distribution when low-rank tails are excluded. In comparison with natural English and “compressed” English without spaces between words, amino acid sequences of proteins show larger linear ranges and smaller exponents with heavier low-rank tails, demonstrating that the SCS distribution in proteins is largely scale-free. A distribution pattern of SCSs in proteins is similar among species, but species-specific features are also present. Based on the availability scores of SCSs, we found that sequence motifs are enriched in high-availability sites (i.e., “key words”) and vice versa. In fact, the highest availability peak within a given protein sequence often directly corresponds to a sequence motif. The amino acid composition of high-availability sites within motifs is different from that of entire motifs and all protein sequences, suggesting the possible functional importance of specific SCSs and their compositional amino acids within motifs. We anticipate that our availability-based word decoding approach is complementary to sequence alignment approaches in predicting functionally important sites of unknown proteins from their amino acid sequences.     

Helicobacter suis KB1 derived from pig gastric lymphoid follicles induces the formation of gastric lymphoid follicles in mice through the activation of B cells and CD4 positive cells

"Helicobacter heilmannii" ("H. heilmannii"), which belongs to the genus Helicobacter, is a group of bacterial species that display a long spiral-shaped morphology. Recent studies have demonstrated that "H. heilmannii" type 1 is actually H. suis, which mainly colonizes the stomachs of various animals and humans. However, the influence of H. suis on gastric diseases remains to be fully elucidated. In this report, we revealed the relationship between natural H. suis infection and follicular gastritis in the pig stomachs. From sequence analysis of the 16S rRNA, urease A, and urease B genes, the presence of H. suis was confirmed in pig gastric lymphoid follicles, and this bacterium was named H. suis KB1. In addition, H. suis KB1 was inoculated into C57BL/6J mice, and the following mouse model of the pathogenesis of follicular gastritis by H. suis infection was established: H. suis KB1 colonizes the mouse stomach, and moreover, induces the development of lymphoid follicles and acquired immune responses characterized by the activation of B cells and CD4 positive cells. These results may lead to better understanding of the relationship between H. suis and gastric diseases, especially follicular gastritis; and furthermore, our findings emphasize the zoonotic aspects of animal-human infection by H. suis.     

Hyperuricemia in Pediatric Malignancies Before Treatment

The objective of this study was to clarify the prevalence and characteristics of hyperuricemia in various pediatric malignancies before the initiation of treatment. We conducted a retrospective analysis of 119 children with various newly diagnosed malignancies between April 2000 and March 2010. On the basis of the reference values previously established in our laboratory, hyperuricemia was defined as uric acid (UA) levels above 2 standard deviations (s.d.) over the mean values at each age. Thirty-six patients (30.3%) showed hyperuricemia. Hyperuricemia was more common in male patients (36.8%) than in female patients (21.6%). The prevalence of hyperuricemia was highest in patients with lymphoma followed by those with acute lymphoblastic leukemia (ALL). When the study population was divided into hyperuricemia-negative and -positive populations, blood urea nitrogen, creatinine, and lactate dehydrogenase levels, and white blood cell counts (only in leukemia) were found to be significantly higher in the latter group by a univariate analysis. This study highlights useful information for identifying patients with malignancies at risk for tumor lysis syndrome (TLS) before starting chemotherapy.     

Canine X-linked muscular dystrophy in Japan (CXMDJ).

The purpose of this study was to develop a strain of canine X-linked muscular dystrophy (CXMD), a model of Duchenne muscular dystrophy, in Japan. A female beagle was artificially inseminated with frozen-thawed spermatozoa derived from an affected golden retriever. Subsequently, two carrier female dogs (G1 carriers) and four normal male littermates were produced. Thereafter, the two G1 carriers were mated with beagle sires. As a result, each bitch whelped three times, and out of 54 pups, 17 affected male descendants, and 11 carrier female descendants (G2 carriers) were detected. One G2 carrier was then mated with a beagle sire and 15 pups in two whelpings were produced, including five affected males and four carrier females (G3 carriers). A total of 10 female beagles were artificially inseminated to evaluate the fertility of the frozen-thawed spermatozoa from the two affected dogs. The whelping rates of the two affected dogs were 4/5 and the litter sizes were 5.0 +/- 1.41 and 6.0 +/- 0.82, respectively. These results indicate that a canine X-linked muscular dystrophy colony has been established in Japan. We called them CXMDJ.