Directional effect on post-stroke motor overflow characteristics

Motor overflow (MO) is an involuntary muscle activation associated with strenuous contralateral movement and may become manifested after stroke. The study was undertaken to investigate physiological correlation underlying atypical directional effect of joint movement on post-stroke MO in the affected upper limb. Thirty patients with unilateral post-stroke hemiparesis and fifteen age-matched healthy controls participated in this study. According to motor function assessed with the Fugl-Meyer arm scale, the patients were categorized into two groups of equal number with better (CVA_G; n = 15) or poorer motor functions (CVA_P; n = 15). Surface electromyography (EMG) was used to record irradiated muscle activation from eight muscles of the affected upper limb when the subjects performed maximal isometric contractions in different directions with the unaffected shoulder, elbow and wrist joints. The results showed that only MO amplitude of the CVA_G and the control groups was more sensitive to variations in direction of joint movement in the unaffected arm than the CVA_P group. The CVA_G group exhibited larger amplitudes of MO than the control analog, whereas this tendency was reversed for the CVA_P group. In terms of EMG polar plots, spatial representations of post-stroke MO were insensitive to direction of contralateral movement. The spatial representations of the CVA_G and CVA_P groups were predominated by potent flexion-abduction synergy, contrary to the typical extension adduction synergy seen in the control analog. In conclusion, post-stroke MO amplitude was subject to contralateral movement direction for healthy controls and stroke patients with better motor recovery. However, alterations in MO spatial pattern due to directional effect were not strictly related to the degree of motor deficits of the stroke victims.     

Ubiquitin-independent proteasomal degradation during oncogenic viral infections

Most eukaryotic proteins destined for imminent destruction are first tagged with a chain of ubiquitin molecules and are subsequently dismantled by the proteasome. Ubiquitin-independent degradation of substrates by the proteasome, however, also occurs. The number of documented proteasome-dependent, ubiquitin-independent degradation events remains relatively small but continues to grow. Proteins involved in oncogenesis and tumor suppression make up the majority of the known cases for this type of protein destruction. Provocatively, viruses with confirmed or suspected oncogenic properties are also prominent participants in the pantheon of ubiquitin-independent proteasomal degradation events. In this review, we identify and describe examples of proteasome-dependent, ubiquitin-independent protein degradation that occur during tumor virus infections, speculate why this type of protein destruction may be preferred during oncogenesis, and argue that this uncommon type of protein turnover represents a prime target for antiviral and anticancer therapeutics.     

Effects of Buprenorphine and Estrous Cycle in a Murine Model of Cecal Ligation and Puncture

The effect of opioids on the immunopathology of sepsis models in mice has been controversial. In previous work, we showed that mortality and various inflammatory parameters did not differ between female mice given saline or buprenorphine after cecal ligation and puncture. To investigate further, we hypothesized that buprenorphine would not affect outcomes of sepsis at any stage of estrous. Female mice were allocated into 4 groups (n = 20 per group) according to stage of estrous. Mice then underwent cecal ligation and puncture and received either buprenorphine or saline. In 3-wk survival studies, overall survival did not differ between buprenorphine- and saline-treated mice. When mice were stratified according to stage of estrous, survival did not vary among saline-treated groups but was lower in buprenorphine-treated mice in metestrus compared with proestrus. To investigate inflammation as a potential mechanism for survival, we measured cell counts and cytokine levels in the peripheral blood and peritoneal lavage fluid at 12 and 24 h after cecal ligation and puncture. At 24 h, buprenorphine-treated mice in proestrus had more circulating neutrophils and monocytes than did saline-treated mice in proestrus and more circulating WBC than did mice in any other stage with or without buprenorphine. Our current results suggest that the effects of buprenorphine on a 50% survival model of sepsis in BALB/c female mice are minimal overall but that the stage of estrous has various effects in this model. Investigators should consider the effects of buprenorphine and estrous cycle when using female mice in sepsis research.Abbreviations: CLP, cecal ligation and punctureSepsis is a complex, multifactorial disease that remains a common cause of morbidity and mortality after events such as surgery, trauma, and burns.^2,27 Sepsis is difficult to investigate with a single animal model, but many researchers consider cecal ligation and puncture (CLP) in rodents the ‘gold standard’ in creating polymicrobial peritonitis.^5,18 However, several innate and environmental factors cause variability in CLP studies.One variable is the use of analgesia in rodents undergoing CLP. The use of perioperative analgesia for studies using major surgery is the standard of veterinary care. The 2011 Guide also reminds us that the appropriate use of analgesics in research animals is an “ethical and moral imperative.”²⁰ However, sepsis research focuses on the immune response during the course of the disease, and some analgesics, including nonsteroidal antiinflammatory drugs and opioids such as morphine and fentanyl, are known to have immunomodulatory effects.^31,37Buprenorphine is a safe and effective opioid analgesic¹⁶ for rodents and is considered to be minimally immunosuppressive.³¹ Our laboratory has shown that a specific dosing regimen of buprenorphine did not significantly affect survival or immune parameters in female ICR and C57BL/6 mice undergoing CLP,^7,19 whereas male C57BL/6 mice showed a dose-responsive decrease in survival. There were no obvious changes in immune parameters to explain the differences between male and female mice. However, there are known sex-associated differences in responses to opioids. Several reports show that opioids have greater potency, efficacy, and overall analgesia in male rodents than in female, although results looking specifically at buprenorphine are mixed.^8,35In addition to differences in response to opioids, gender may also play a role in the response to sepsis. In humans, several studies have shown that women are less susceptible to sepsis than men.^24,38 In contrast, a different group showed that women with sepsis have a worse survival rate than do men.²⁶ Similarly, contradictory literature in mice has shown either a higher or lower survival in female mice as compared with male in sepsis studies.^23,41 One possible factor for these contradictory results in both rodent and human studies is that female subjects were not grouped by phase of the estrous or menstrual cycle. In both mice and humans, fluctuations of estrogen and progesterone occur regularly through their respective cycles. Other hormonal factors such as prolactin, follicle stimulating hormone, and luteinizing hormone vary as well.¹⁴ Studies show that estrogen, prolactin, and other hormones can affect the immune system with and without sepsis,^1,22,42 leading to the concern that the changing hormonal status of female subjects may affect the responses to opioids and to sepsis. Although several sepsis and trauma studies in rodents look at specific stages of estrous^34,41 or the effects of exogenous estrogen,^9,10,33,39 few target CLP specifically, and none look at this issue in conjunction with buprenorphine.The purpose of our study was to investigate the effects of buprenorphine and the estrous cycle on a CLP model of sepsis. For these investigations, we chose to use female BALB/c mice to assess survival and immune responses. In addition, we performed survival experiments in male mice for comparison. Coupled with our previous studies, these findings will give a comprehensive profile of the major mouse strains used in sepsis research.     

Complete mitochondrial genome sequence of Pyropia yezoensis (Bangiales,Rhodophyta) from Korea

Species of the genera Porphyra and Pyropia are the major seaweed cultivars in Korea, Japan, and China. Genomic data from Porphyra/Pyropia species is now being used for molecular breeding and bioengineering. We determined the complete mitochondrial genome (mtDNA) sequence of Pyropia yezoensis (Bangiales, Rhodophyta) from Korea. Pyropia yezoensis mtDNA was 35,596 bp in size and exhibited high sequence similarity to Py. yezoensis mtDNA reported from China (NC_017837; 41,688 bp). However, the Korean Py. yezoensis differed from the Chinese strain in the presence/absence of introns and intronic open reading frames of the ribosomal RNA large subunit gene (rnl) and cytochrome c oxidase subunit 1 gene (cox1). These differences result in large variations between the mtDNA of the two strains of Py. yezoensis. Moreover, the Korean strain had a single copy of the trnfM–trnQ region, as compared to a duplicate copy of Py. yezoensis (NC_017837). Pyropia mtDNA exhibited unique genetic features at the intra- and interspecies levels. Therefore, mtDNA genomics can provide novel knowledge for red algal molecular systematics and help to differentiate between cultivars of Pyropia species with new molecular markers.     

Gestational Diabetes Is Characterized by Reduced Mitochondrial Protein Expression and Altered Calcium Signaling Proteins in Skeletal Muscle

The rising prevalence of gestational diabetes mellitus (GDM) affects up to 18% of pregnant women with immediate and long-term metabolic consequences for both mother and infant. Abnormal glucose uptake and lipid oxidation are hallmark features of GDM prompting us to use an exploratory proteomics approach to investigate the cellular mechanisms underlying differences in skeletal muscle metabolism between obese pregnant women with GDM (OGDM) and obese pregnant women with normal glucose tolerance (ONGT). Functional validation was performed in a second cohort of obese OGDM and ONGT pregnant women. Quantitative proteomic analysis in rectus abdominus skeletal muscle tissue collected at delivery revealed reduced protein content of mitochondrial complex I (C-I) subunits (NDUFS3, NDUFV2) and altered content of proteins involved in calcium homeostasis/signaling (calcineurin A, α1-syntrophin, annexin A4) in OGDM (n = 6) vs. ONGT (n = 6). Follow-up analyses showed reduced enzymatic activity of mitochondrial complexes C-I, C-III, and C-IV (−60–75%) in the OGDM (n = 8) compared with ONGT (n = 10) subjects, though no differences were observed for mitochondrial complex protein content. Upstream regulators of mitochondrial biogenesis and oxidative phosphorylation were not different between groups. However, AMPK phosphorylation was dramatically reduced by 75% in the OGDM women. These data suggest that GDM is associated with reduced skeletal muscle oxidative phosphorylation and disordered calcium homeostasis. These relationships deserve further attention as they may represent novel risk factors for development of GDM and may have implications on the effectiveness of physical activity interventions on both treatment strategies for GDM and for prevention of type 2 diabetes postpartum.     

Binding Mode Analyses and Pharmacophore Model Development for Stilbene Derivatives as a Novel and Competitive Class of α-Glucosidase Inhibitors

Stilbene urea derivatives as a novel and competitive class of non-glycosidic α-glucosidase inhibitors are effective for the treatment of type II diabetes and obesity. The main purposes of our molecular modeling study are to explore the most suitable binding poses of stilbene derivatives with analyzing the binding affinity differences and finally to develop a pharmacophore model which would represents critical features responsible for α-glucosidase inhibitory activity. Three-dimensional structure of S. cerevisiae α-glucosidase was built by homology modeling method and the structure was used for the molecular docking study to find out the initial binding mode of compound 12, which is the most highly active one. The initial structure was subjected to molecular dynamics (MD) simulations for protein structure adjustment at compound 12-bound state. Based on the adjusted conformation, the more reasonable binding modes of the stilbene urea derivatives were obtained from molecular docking and MD simulations. The binding mode of the derivatives was validated by correlation analysis between experimental K_i value and interaction energy. Our results revealed that the binding modes of the potent inhibitors were engaged with important hydrogen bond, hydrophobic, and π-interactions. With the validated compound 12-bound structure obtained from combining approach of docking and MD simulation, a proper four featured pharmacophore model was generated. It was also validated by comparison of fit values with the K_i values. Thus, these results will be helpful for understanding the relationship between binding mode and bioactivity and for designing better inhibitors from stilbene derivatives.