Myostatin is a TGF-ss family member that normally acts to limit skeletal muscle mass. Follistatin is a myostatin-binding protein that can inhibit myostatin activity in vitro and promote muscle growth in vivo. Mice homozygous for a mutation in the Fst gene have been shown to die immediately after birth but have a reduced amount of muscle tissue, consistent with a role for follistatin in regulating myogenesis. Here, we show that Fst mutant mice exhibit haploinsufficiency, with muscles of Fst heterozygotes having significantly reduced size, a shift toward more oxidative fiber types, an impairment of muscle remodeling in response to cardiotoxin-induced injury, and a reduction in tetanic force production yet a maintenance of specific force. We show that the effect of heterozygous loss of Fst is at least partially retained in a Mstn-null background, implying that follistatin normally acts to inhibit other TGF-ss family members in addition to myostatin to regulate muscle size. Finally, we present genetic evidence suggesting that activin A may be one of the ligands that is regulated by follistatin and that functions with myostatin to limit muscle mass. These findings potentially have important implications with respect to the development of therapeutics targeting this signaling pathway to preserve muscle mass and prevent muscle atrophy in a variety of inherited and acquired forms of muscle degeneration.

A 56-yr-old man with lung adenocarcinoma presented with subsegmental pulmonary thrombosis. Platelet count on presentation was 531x10(9)/L. The patient was anticoagulated with subcutaneous low molecular weight heparin (LMWH). Next day, oral anticoagulation was initiated with 5 mg of warfarin once daily with LMWH and LMWH was discontinued at third hospital day. On the third day of oral anticoagulation therapy, he complained of left leg swelling and prolonged painful penile erection of 24 hr-duration. His platelet count reached a nadir 164x10(9)/L at that time, and the patient had a deficiency of protein C and S, with an activity level of 16% and 20% of normal value. Warfarin was stopped and he underwent penile aspiration. The next day, left leg edema and penile erection was disappeared, but penile and glans penis necrosis was started. This case illustrates that processes underlying heparin-induced thrombocytopenia (HIT) may also underlie warfarin-induced skin necrosis.

The authors report a case of acute kidney injury (AKI) resulting from menstruation-related disseminated intravascular coagulation (DIC) in an adenomyosis patient. A 40-yr-old woman who had received gonadotropin for ovulation induction therapy presented with anuria and an elevated serum creatinine level. Her medical history showed primary infertility with diffuse adenomyosis. On admission, her pregnancy test was negative and her menstrual cycle had started 1 day previously. Laboratory data were consistent with DIC, and it was believed to be related to myometrial injury resulting from heavy intramyometrial menstrual flow. Gonadotropin is considered to play an important role in the development of fulminant DIC. This rare case suggests that physicians should be aware that gonadotropin may provoke fulminant DIC in women with adenomyosis.

The purpose of this study was to analyze the effects of smoking on corpus callosum volume. In addition, the relationships between smoking duration, smoking frequency, and corpus callosum volume were analyzed. Magnetic resonance brain images were acquired for 58 normal Korean men (30 smokers (age 32.82+/-14.12 years) and 28 non-smokers (age 35.49+/-13.11 years)). The corpus callosum volume was measured using Brain Voyager 2000 S/W and was normalized by intracranical volume, which was calculated using cerebral sizes. The corpus callosum volume for smokers was significantly smaller than that for non-smokers. Also, there was a negative correlation between corpus callosum volume and smoking duration. The change of white matter volume (e.g., corpus callosum) might be a primary factor for characterizing the effects of smoking.

We investigated the thermal effects on heart rate, hemodynamics, and response of vitelline arteries of stage-18 chicken embryos. Heart rate was monitored by a high-speed imaging method, while hemodynamic quantities were evaluated using a particle image velocimetry (PIV) technique. Experiments were carried out at seven different temperatures (36-42 degrees C with 1 degrees C interval) after 1h of incubation to stabilize the heart rate. The heart rate increased in a linear manner (r=0.992). Due to the increased cardiac output (or heart rate), the hemodynamic quantities such as mean velocity (U(mean)), velocity fluctuation (U(fluc)), and peak velocity (U(peak)) also increased with respect to the Womersley number (Omega) in the manner r=0.599, 0.693, and 0.725, respectively. This indicates that the mechanical force exerting on the vessel walls increases. However, the active response (or regulation) of the vitelline arteries was not observed in this study.

Mice deficient in growth differentiation factor 11 (GDF11) signaling display anterior transformation of axial vertebrae and truncation of caudal vertebrae. However, the in vivo molecular mechanisms by which GDF11 signaling regulates the development of the vertebral column have yet to be determined. We found that Gdf11 and Acvr2b mutants are sensitive to exogenous RA treatment on vertebral specification and caudal vertebral development. We show that diminished expression of Cyp26a1, a retinoic acid inactivating enzyme, and concomitant elevation of retinoic acid activity in the caudal region of Gdf11(-/-) embryos may account for this phenomenon. Reduced expression or function of Cyp26a1 enhanced anterior transformation of axial vertebrae in wild-type and Acvr2b mutants. Furthermore, a pan retinoic acid receptor antagonist (AGN193109) could lessen the anterior transformation phenotype and rescue the tail truncation phenotype of Gdf11(-/-) mice. Taken together, these results suggest that GDF11 signaling regulates development of caudal vertebrae and is involved in specification of axial vertebrae in part by maintaining Cyp26a1 expression, which represses retinoic acid activity in the caudal region of embryos during the somitogenesis stage.

There is increasing evidence for an association between WMH and large-artery atherosclerosis. We evaluated 268 patients with acute ischemic stroke to assess the relationship between intracranial (IC) atherosclerosis and WMH. The patients were classified into three groups according to the location of the stenosis; IC, extracranial (EC), and non-stenosis (NS) group. WMH were rated using the semiquantitative method of Scheltens and coworkers. The IC group had significantly more WMH score in comparison with the other groups after controlling age. The linear regression analysis showed that age was the factor most strongly associated with the total score of WMH; and the location of stenosis was positively related to WMH, especially in deep white matter. Our data show that IC stenosis is associated with WMH, indicating that IC stenosis, rather than EC stenosis, is likely to cause white matter lesions. These findings raise the possibility that occlusion of penetrating arteries, embolism to border-zone areas and a hemodynamic mechanism associated with IC stenosis leads to the formation of white matter lesions.

Nuchal-type fibroma, first described in 1988 by Enzinger and Weiss, is a rare fibrous growth occurring predominantly in the interscapular and paraspinous regions. It is typically located in the subcutaneous tissue of the posterior neck but may occur in extranuchal soft tissue sites such as the upper back, shoulder, and facial regions. In this article, we describe the magnetic resonance imaging findings of a nuchal-type fibroma that involved the buttock in a 45-year-old woman, and we review and discuss the literature on the subject.

Summary *A comprehensive understanding of the sap flow dynamics and xylem hydraulic properties is essential to unravel the functional features of water transport from roots to shoots in vascular plants. To evaluate quantitatively the safety and efficiency of this system, nondestructive methods to assess the interactions between sap ascent kinetics and xylem structure are required. *In this study, synchrotron X-ray microscopy was employed to observe anatomical structures and sap flow dynamics in rice (Oryza sativa) xylem simultaneously. The phase-contrast imaging technique allowed nondestructive observation of the xylem structural characteristics and the air-water interfaces generated by dehydration-rehydration cycles in excised leaves. *This X-ray microimaging method provided a unique tool to characterize the perforated end walls of vessel elements and to evaluate their influence on hydraulic resistance during the refilling of embolized vessels. The real-time monitoring of the axial and radial sap flow under various environmental conditions highlighted the important role of perforation plates. *In summary, we report a new methodology to study the sap flow dynamics and xylem hydraulic properties with mum spatial and ms temporal resolution using X-ray microscopy. The experimental procedure described herein provides a useful handle to understand key sap transport phenomena in xylem.

Objectives: The objective of this study was to examine associations between maternal and paternal use of corporal punishment (CP) for 3-year-old children and intimate partner aggression or violence (IPAV) in a population-based sample. Methods: The study sample (N = 1997) was derived from wave 3 of the Fragile Families and Child Wellbeing Study. Mother and father reports regarding their use of CP and their IPAV victimization were analyzed. IPAV included coercion and nonphysical and physical aggression. Results: Approximately 65% of the children were spanked at least once in the previous month by 1 or both parents. Of couples who reported any family aggression (87%), 54% reported that both CP and IPAV occurred. The most prevalent patterns of co-occurrence involved both parents as aggressors either toward each other (ie, bilateral IPAV) or toward the child. The presence of bilateral IPAV essentially doubled the odds that 1 or both parents would use CP, even after controlling for potential confounders such as parenting stress, depression, and alcohol or other drug use. Of the 5 patterns of co-occurring family aggression assessed, the "single aggressor" model, in which only 1 parent aggressed in the family, received the least amount of empirical support. Conclusions: Despite American Academy of Pediatrics' recommendations against the use of CP, CP use remains common in the United States. CP prevention efforts should carefully consider assumptions made about patterns of co-occurring aggression in families, given that adult victims of IPAV, including even minor, nonphysical aggression between parents, have increased odds of using CP with their children.

PURPOSE: To assess the rate of serious (>Grade 2, CTCAE 3.0) dermatitis in our head-and-neck cancer (HNC) patients undergoing simultaneous integrated boost intensity-modulated radiotherapy with concomitant cetuximab (SIB-IMRT-cetuximab). We hypothesized a positive association between the radiation dose to the skin and the degree of dermatitis in patients receiving cetuximab. METHODS AND MATERIALS: Between April 2006 and December 2009, 99 HNC patients underwent SIB-IMRT-cetuximab. In 69/99 (70%), systemic treatment consisted of concomitant cetuximab only, whereas 30 (30%) were switched from concomitant cisplatin to concomitant cetuximab. Treatment-related dermatitis was prospectively monitored. Ninety-nine patients treated with four to seven concomitant cycles of cisplatin only served as an internal control group. The radiation dose delivered to the skin was measured and related to dermal reactions. RESULTS: Grade 3/4 dermatitis developed in 34% of the cetuximab cohort, which was substantially higher than in the control cohort (3%, p < 0.01). No cases of skin necrosis or other fatal events related to cetuximab have occurred so far. A significantly larger mean skin area was found exposed to high radiation doses in patients with severe cetuximab-related dermatitis, compared with those without (p < 0.01). CONCLUSION: Concomitant cetuximab resulted in a approximately 10-fold increase in the rate of severe transient dermatitis compared with the use of concomitant cisplatin. We found a positive association between the incidence of Grade 3/4 dermatitis and the radiation dose delivered to the skin in patients receiving cetuximab.

Modulation of transforming growth factor-beta (TGF-beta) signaling to promote muscle growth holds tremendous promise for the muscular dystrophies and other disorders involving the loss of functional muscle mass. Previous studies have focused on the TGF-beta family member myostatin and demonstrated that inhibition of myostatin leads to muscle growth in normal and dystrophic mice. We describe a unique method of systemic inhibition of activin IIB receptor signaling via adeno-associated virus (AAV)-mediated gene transfer of a soluble form of the extracellular domain of the activin IIB receptor to the liver. Treatment of mdx mice with activin IIB receptor blockade led to increased skeletal muscle mass, increased force production in the extensor digitorum longus (EDL), and reduced serum creatine kinase. No effect on heart mass or function was observed. Our results indicate that activin IIB receptor blockade represents a novel and effective therapeutic strategy for the muscular dystrophies. Muscle Nerve, 2010.

The discovery and development of new, highly potent anti-coronavirus agents and effective approaches for controlling the potential emergence of epidemic coronaviruses still remains an important mission. Here, we identified tylophorine compounds, including naturally occurring and synthetic phenanthroindolizidines and phenanthroquinolizidines, as potent in vitro inhibitors of enteropathogenic coronavirus transmissible gastroenteritis virus (TGEV). The potent compounds showed 50% maximal effective concentration (EC(50)) values ranging from 8 to 1468nM as determined by immunofluorescent assay of the expression of TGEV N and S proteins and by real time-quantitative PCR analysis of viral yields. Furthermore, the potent tylophorine compounds exerted profound anti-TGEV replication activity and thereby blocked the TGEV-induced apoptosis and subsequent cytopathic effect in ST cells. Analysis of the structure-activity relations indicated that the most active tylophorine analogues were compounds with a hydroxyl group at the C14 position of the indolizidine moiety or at the C3 position of the phenanthrene moiety and that the quinolizidine counterparts were more potent than indolizidines. In addition, tylophorine compounds strongly reduced cytopathic effect in Vero 76 cells induced by human severe acute respiratory syndrome coronavirus (SARS CoV), with EC(50) values ranging from less than 5 to 340nM. Moreover, a pharmacokinetic study demonstrated high and comparable oral bioavailabilities of 7-methoxycryptopleurine (52.7%) and the naturally occurring tylophorine (65.7%) in rats. Thus, our results suggest that tylophorine compounds are novel and potent anti-coronavirus agents that may be developed into therapeutic agents for treating TGEV or SARS CoV infection.

OBJECTIVE: The primary objective of this study is to evaluate the hemodynamic and structural characteristics of the omphalo-mesenteric (vitelline) arteries in stage 18 chicken embryos. The measured results were compared with Murray's law to validate the theoretical prediction on the vascular structure. METHODS: \Variation of hemodynamic parameters such as mean velocity (U(mean)), peak velocity (U(peak)) at the systolic phase, velocity fluctuations (U(fluc)) at the pulsatile frequency, and the Womersley number (Omega) were measured with respect to the geometric parameters including the bifurcation cascade level (BCL), vessel diameter (D), and distance (L) from the first bifurcation. They were assessed by using the time-resolved in vivo micro-PIV (particle image velocimetry) technique and the geometric information was obtained from the microscopic vessel images. RESULTS: The effect of "branching of the vessel" on the variation of hemodynamic characteristics is similar to those of the "increase in distance" from the first bifurcation and the "decrease in vessel diameter". The flow quantities (U(mean), U(peak) and U(fluc)) decrease due to the increase in cross-sectional area ratio (gamma=1.209 = ( summation operatorD(daughter)(2))/D(mother)(2)), and the Womersley number also decreases as the bifurcation cascades (Omega<<1). CONCLUSION: The geometric parameters are closely related to the variation of hemodynamic characteristics. Murray's law with non-constant viscosity hypothesis would provide an insight on the two-phase nature of microvascular blood flows.

Poly(L-lactic acid) (PLLA) microfibrous scaffolds with three-dimensional (3D) structures were fabricated using an electrospinning technique with a subsequent mechanical expansion process. To achieve a 3D fibrous structure, the fusion at the contact points of the as-spun PLLA microfibers was avoided using an appropriate binary solvent system of methylene chloride and acetone. The solvent composition was optimized based on the solvent power, volatility, and viscosity (methylene chloride:acetone = 9:1 volume ratio). The final 3D structure of the electrospun scaffolds was obtained after mechanical expansion of the electrospun microfibrous mats. The pore sizes of the scaffolds were controlled by varying the degree of expansion of the nonbonded microfibrous mats, and they were in the range of several microns up to 400 mum. The 3D scaffolds were examined for their morphological properties and their potential use for the proliferation of osteoblasts. Generally recognized electrospun 2D nanofibrous membranes were also tested in order to compare the cell behaviors using different scaffold geometries. The 3D scaffolds demonstrated a high level of osteoblast proliferation (1.8-fold higher than nanofibrous membranes in a week). The osteoblasts actively penetrated the inside of the 3D scaffold and showed a spatial cell distribution, as confirmed by SEM and H&E staining, while a monolayer formed in the case of the 2D nanofibrous membranes with limited cell infiltration. In vivo results further showed that 3D electrospun microfibrous matrices were a favorable substrate for cell infiltration and bone formation after 2 and 4 weeks, using a rabbit calvarial defect model. In this study, the 3D microfibrous PLLA scaffolds fabricated using electrospinning techniques might be an innovative addition to tissue engineering applications. (c) 2010 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 2010.

Lysophosphatidic acid (LPA) is a lipid mediator that mediates several effects that promote cancer progress. The LPA receptor type 2 (LPA2) expression is often elevated in several types of cancers, including colorectal cancer (CRC). In this study, we investigated the role of LPA(2) in the development of intestinal adenomas by comparing Apc(Min/+) mice and Apc(Min/+)/Lpar2(-/-) mice. There were 50% fewer intestinal adenomas in Apc(Min/+)/Lpar2(-/-) mice than Apc(Min/+) mice. Smaller size adenomas (< 1mm) were found at higher frequencies in Apc(Min/+)/Lpar2(-/-) mice compared with Apc(Min/+) mice at the two age groups examined. The expression level of LPA(2) correlated with increased size of intestinal adenomas. Reduced tumor multiplicity and size in Apc(Min/+)/Lpar2(-/-) mice correlated with decreased proliferation of intestinal epithelial cells. Apc(Min/+)/Lpar2(-/-) mice showed an increased level of apoptosis, suggesting that LPA2-mediated signaling stimulates intestinal tumor development and progress by regulating both cell proliferation and survival. In addition, the expression levels of Krüpple-like factor 5 (KLF5), beta-catenin, cyclin D1, c-Myc, and hypoxia-inducible factor-1alpha (HIF-1alpha) were significantly altered in Apc(Min/+)/Lpar2(-/-) mice compared with Apc(Min/+) mice. In vitro studies using HCT116 cells showed that LPA induced cyclin D1, c-Myc, and HIF-1alpha expression, which was attenuated by knockdown of LPA(2). In summary, intestinal tumor initiated by Apc mutations is altered by LPA(2)-mediated signaling, which regulates tumor growth and survival by altering multiple targets.

The purpose of this study was to investigate the effects of knee-flexion angles on subjective discomfort ratings, heart rates, and muscle fatigue using median frequency (MDF) in a static-sustaining task. Thirty healthy participants maintained 13 postures including standing, squatting, sitting, and kneeling postures and then MDFs from the erector spinae, biceps femoris, vastus medialis, gastrocnemius, and tibialis anterior muscles, subjective discomfort, and heart rates were collected every 3 min during a sustained 15 min task. Results showed that the discomfort, heart rate and muscle fatigue were significantly influenced by the body postures. In general, standing and sitting postures showed less discomfort as well as lower heart rates, whereas squatting postures (KF120, KF90, KF60) had higher discomfort and heart rates. Three MDF change trends were reported associated with postures in this study. First, there were less changes of MDFs for standing and sitting postures; second, all patterns of MDFs for KF 150 and KF120 decreased, and lastly some MDFs had increasing trends and others showed decreasing trends for KF30, KF30T, and kneeling postures.

Imaging the pharmacodynamics of anti-cancer drugs may allow early assessment of anti-cancer effects. Increases in 3'-deoxy-3'-[(18)F]fluorothymidine ([(18)F]FLT) uptake early after thymidylate synthase inhibition(TS) inhibition, the so-called flare response, is considered to be due largely to an increase in binding sites for type-1 equilibrative nucleoside transporter. We investigated the induction of thymidine kinase 1 (TK1) after 5-fluorouracil (5-FU) treatment as one of mechanisms for [(18)F]FLT flare. Exposure of 9 cancer cell lines to 5-FU for 24 hrs induced a 2.5 approximately 3.5-fold increase in [(18)F]FLT uptake, significantly higher than the 1.5-fold increase observed 2 hr after treatment. The increase of [(18)F]FLT uptake 24 hr after 5-FU exposure accompanied TK1 induction in most cell lines. In representative cell lines (A431 and HT29), 5-FU time-dependently increased [(18)F]FLT uptake, kinase activity and the levels of protein and mRNA for TK1, sequential cyclin E and A induction, and G(1)-S phase transition. Cycloheximide treatment and knockdown of TK1 completely inhibited 5-FU-induced [(18)F]FLT flare. On the other hand, HCT8 cells showed a biphasic [(18)F]FLT flare with lacked TK1 induction in response to the dosage of 5-FU. Cycloheximide did not inhibit 5-FU-induced [(18)F]FLT flare in this cells. In vivo dynamic [(18)F]FLT-PET and ex vivo analysis in HT29 tumor-bearing mice showed significantly increased [(18)F]FLT flux and TK1 activity of tumor tissue 24 hrs after 5-FU administration (P<0.05). Conclusively, 5-FU induced TK1 and TK1-mediated high [(18)F]FLT flare in most of cell lines. [(18)F]FLT-PET may be used to assess pharmacodynamics of TS inhibitor by a mechanism involving TK1 induction.

Abstract Rationale, aims and objectives Our objective was to describe the factors associated with doctor-rated and patient-rated gout severity to explain how doctor assessment involving patient-reported outcomes can improve the clinical management of gout. Methods Patients completed a newly validated gout-specific health-related quality of life instrument, the Gout Impact Scale (GIS) and other questions regarding their gout. Both patients and their doctors gave an overall gout severity assessment. We conducted correlation analyses between each predictor of interest and the two different severity ratings (doctor-rated severity and patient-rated severity). Stepwise multiple regressions were performed to determine the best predictors for doctor-rated and patient-rated severity, respectively. Results Doctor-rated severity more closely correlated with objective clinical and laboratory findings, particularly the presence of tophi, which was not a leading factor in patient-rated severity assessments. Patient-rated severity more closely correlated with the domains of the GIS, which expressed the impact of gout on health-related quality of life. Conclusion Doctors might have a better understanding of their patients' level of disease impact if they incorporate an instrument such as the GIS in their evaluation of gout severity and their decisions regarding aggressiveness of treatment. The increased use of patient-reported outcomes measures has the potential to improve quality of care and patient satisfaction, as well as reduce costs of health care utilization.

Fluorescent silica nanoparticles deposited with highly monodisperse gold nanoparticles (1-2 nm) were synthesized via the W/O method and intensive ultrasound irradiation. A large surface area of gold-doped fluorescent silica nanoparticle serves as a platform to immobilize a specific binding protein for biomolecules interaction in bioimaging applications.