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Find video protocols related to scientific articles indexed in Pubmed.
[Characteristics of urinary incontinence in the elderly].
Urologe A
PUBLISHED: 09-18-2014
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Geriatric patients are defined as being over 70 years of age and are vulnerable due to multimedication and multimorbidity. The typical incontinence type in geriatric patients is the overactive bladder syndrome as a result of anatomical alterations and the influence of conditions which typically occur in the elderly, e.g. diabetes mellitus, vaginal atrophy, constipation, neurological affections and dementia. This multimorbidity leads to multimedication but many pharmaceutical compounds aimed at indications of diseases distant from the urinary tract can also influence the continence situation. This has been proven for cardiac medications, such as alpha-blockers and diuretics, neurological drug therapy and analgesics. Diagnostic investigations in geriatric patients are usually non-invasive and include geriatric assessment to quantify incontinence symptoms but invasive diagnostic tools are required if the primary therapy fails or an operative intervention is planned. Pharmacotherapy considers the special requirements of the very old patient with cognitive impairment and vulnerability due to falls or delirium. In the group of anticholinergic drugs, trospium chloride seems to be the favorite substance to treat this group of patients because this hydrophilic compound is considered to be unable to cross the blood-brain barrier and therefore minimizes the risk of side effects in the central nervous system (CNS).
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Secondary damage in left-sided frontal white matter detected by diffusion tensor imaging is correlated with executive dysfunction in patients with acute infarction at the ipsilateral posterior corona radiata.
Eur. J. Med. Res.
PUBLISHED: 08-20-2014
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Executive dysfunction has been observed in patients with left-sided anterior corona radiata infarction. However, whether left-sided posterior corona radiata infarction could cause executive dysfunction is unclear. Also, whether secondary damage in the left frontal white matter following ipsilateral posterior corona radiata infarct is causal or not and contributes to the occurrence and development of executive dysfunction, is still uncertain.
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Identification and characterisation a surface-associated arginine peptidase in Streptococcus suis serotype 2.
Microbiol. Res.
PUBLISHED: 08-18-2014
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Streptococcus suis is an important zoonotic pathogen worldwide and is responsible for disease in swine and humans. In the present study, we identified and characterised a surface-associated peptidase (abpb, amylase-binding protein B) in Streptococcus suis serotype 2 (S. suis 2) that has high hydrolytic activity towards H-Arg-pNa, with maximum activity at pH 7.0. Stimulation of RAW 264.7 macrophages with purified recombinant abpb protein triggered the release of pro-inflammatory cytokines. An abpb-deficient mutant ?Abpb was constructed by homologous recombination to determine the role of abpb in S. suis 2. The mutant ?Abpb showed decreased adherence to Hep-2 cells and attenuated virulence in a mouse model compared to the wild type strains. The results of the infection showed impaired bacterial growth in vivo and poor colonisation of the organs. In a protection assay, the recombinant abpb provided excellent protection against a lethal challenge of S. suis 2. Together, these findings suggest that abpb contributes to the pathogenicity of S. suis 2 and may be another target for S. suis prevention and control.
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Rifampicin improves neuronal apoptosis in LPS-stimulated co?cultured BV2 cells through inhibition of the TLR-4 pathway.
Mol Med Rep
PUBLISHED: 08-11-2014
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Agents inhibiting microglial activation are attracting attention as candidate drugs for neuroprotection in neurodegenerative diseases. Recently, researchers have focused on the immunosuppression induced by rifampicin. Our previous study showed that rifampicin inhibits the production of lipopolysaccharide (LPS)-induced pro-inflammatory mediators and improves neuron survival in inflammation; however, the mechanism through which rifampicin inhibits microglial inflammation and its neuroprotective effects are not completely understood. In this study, we examined the effects of rifampicin on morphological changes induced by LPS in murine microglial BV2 cells. Then we investigated, in BV2 microglia, the effects of rifampicin on two signaling pathway componentss stimulated by LPS, the Toll?like receptor-4 (TLR-4) and the nuclear factor-?B (NF-?B). In addition, we co-cultured BV2 microglia and neurons to observe the indirect neuroprotective effects of rifampicin. Rifampicin inhibited LPS-stimulated expression of the TLR-4 gene. When neurons were co-cultured with LPS-stimulated BV2 microglia, pre-treatment with rifampicin increased neuronal viability and reduced the number of apoptotic cells. Taken together, these findings suggest that rifampicin, with its anti-inflammatory properties, may be a promising agent for the treatment of neurodegenerative diseases.
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Vps15 is required for stress induced and developmentally triggered autophagy and salivary gland protein secretion in Drosophila.
Cell Death Differ.
PUBLISHED: 07-17-2014
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Autophagy is a catabolic process used to deliver cellular material to the lysosome for degradation. The core Vps34/class III phosphatidylinositol 3-kinase (PI3K) complex, consisting of Atg6, Vps15, and Vps34, is highly conserved throughout evolution, critical for recruiting autophagy-related proteins to the preautophagosomal structure and for other vesicular trafficking processes, including vacuolar protein sorting. Atg6 and Vps34 have been well characterized, but the Vps15 kinase remains poorly characterized with most studies focusing on nutrient deprivation-induced autophagy. Here, we investigate the function of Vps15 in different cellular contexts and find that it is necessary for both stress-induced and developmentally programmed autophagy in various tissues in Drosophila melanogaster. Vps15 is required for autophagy that is induced by multiple forms of stress, including nutrient deprivation, hypoxia, and oxidative stress. Furthermore, autophagy that is triggered by physiological stimuli during development in the fat body, intestine, and salivary gland also require the function of Vps15. In addition, we show that Vps15 is necessary for efficient salivary gland protein secretion. These data illustrate the broad importance of Vps15 in multiple forms of autophagy in different animal cells, and also highlight the pleiotropic function of this kinase in multiple vesicle-trafficking pathways.Cell Death and Differentiation advance online publication, 24 October 2014; doi:10.1038/cdd.2014.174.
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G-CSF pretreatment aggravates LPS-associated microcirculatory dysfunction and acute liver injury after partial hepatectomy in rats.
Histochem. Cell Biol.
PUBLISHED: 06-25-2014
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Liver dysfunction is a serious complication in the early phase following major liver resection or liver transplantation and might be aggravated by the translocation of bacteria and lipopolysaccharide (LPS). As a preventive strategy, granulocyte colony-stimulating factor (G-CSF) is prophylactically applied in patients who are subjected to major surgery. However, we previously demonstrated that G-CSF can induce LPS sensitization. In this study, we aimed to evaluate the effects of G-CSF pretreatment on hepatic microcirculatory disturbances and postoperative liver dysfunction after 70 % partial hepatectomy (PH) in rats. PH alone was well tolerated by all animals (100 % survival rate, slight liver damage and inflammation). LPS application after 70 % PH caused moderate inflammation, microcirculatory disturbances and hepatic damage and led to a 24-h survival rate of 30 % after the operations. In the G-CSF-LPS-PH group, all of the rats died within 4 h with severe inflammatory responses and liver damage (i.e., pronounced erythrocyte congestion and neutrophil infiltration). Portal hypertension and microcirculatory disorders (i.e., inhomogeneous perfusion, sinusoidal dilatation and reductions on functional capillary density) were more pronounced in the G-CSF-LPS-PH group. In conclusion, increased circulating LPS levels were associated with an imbalanced inflammatory response and microcirculatory dysfunction that preceded liver damage and subsequent dysfunction following surgery. G-CSF-pretreatment aggravated microcirculatory disturbances and liver damage, which might have been related to G-CSF-induced LPS sensitization.
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[Serum anti-Ku86: a potential biomarker for early detection of hepatocellular carcinoma].
Zhonghua Zhong Liu Za Zhi
PUBLISHED: 05-07-2014
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To investigate the clinical value of serum anti-Ku86 in early detection of hepatocellular carcinoma (HCC).
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Reduced hepatic arterial perfusion impairs the recovery from focal hepatic venous outflow obstruction in liver-resected rats.
Transplantation
PUBLISHED: 04-29-2014
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Extended partial hepatectomy (PH) in patients is leading to portal hyperperfusion but reduced hepatic arterial perfusion (HAP), and is invariably causing focal hepatic venous outflow obstruction (FHVOO). We observed in a rat model that PH in combination with right median hepatic vein ligation (RMHV-L) caused confluent parenchymal necrosis interspersed with viable portal tracts in the obstructed territory and large sinusoidal vascular canals in the border zone. Lack of HAP impaired the spontaneous course of recovery in terms of enlarged parenchymal necrosis, delayed regeneration, and the absence of draining vascular canals. We aimed to investigate whether pharmacological intervention modulates the imbalance between portal venous and hepatic arterial inflow, aggravates the liver damage, and delays the recovery process after FHVOO in liver-resected rats.
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"Teens talk healthy weight": the impact of a motivational digital video disc on parental knowledge of obesity-related diseases in an adolescent clinic.
J Acad Nutr Diet
PUBLISHED: 04-09-2014
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The purpose of the study was to investigate the impact of a 7-minute educational and motivational weight-management digital video disc (DVD) that uses real patient/parent testimonials and provider-patient interactions, on adolescent and parent knowledge of obesity-related diseases; readiness, motivation, and self-efficacy to lose weight; connectedness to care provider; and likelihood of return to clinic for follow-up care. A randomized controlled trial was conducted among 40 overweight/obese adolescent participants (22.5% male, 77.5% female, mean age=15.43 years) and their parents (n=38) who visited a referral-only adolescent clinic for the first time from October 2009 to March 2010. Adolescents were randomly assigned by a research assistant to standard care alone or standard care plus DVD. Standard care (protocol-driven medical and nutritional assessment and counseling) was provided to all adolescents by a registered dietitian nutritionist and physician or nurse practitioner. Adolescents in the intervention group also viewed the DVD. Adolescents and parents completed assessments pre- and post-clinic visit. Repeated measures analysis of covariance was used to evaluate group differences, while controlling for race/ethnicity and age. Parents who viewed the DVD experienced greater improvements in obesity-related disease knowledge than parents who did not view the DVD. Adolescents in both groups improved on measures of motivation to lose weight and dieting self-efficacy, based on pre and post-test questionnaires. A 7-minute educational and motivational DVD helped improve parent knowledge, but was not more powerful than standard care alone in changing other weight-related outcomes in this adolescent clinic. Because it led to increased parental knowledge, incorporating the DVD into clinical practice could also allow more time for health providers to focus on specific obesity-related treatment/education. Future research might examine whether the DVD has more utility in different settings, such as primary care.
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Investigations into the role of 26S proteasome non-ATPase regulatory subunit 13 in neuroinflammation.
Neuroimmunomodulation
PUBLISHED: 03-08-2014
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To investigate 26S proteasome non-ATPase regulatory subunit 13 (PSMD13) gene silencing as a potential treatment for neuroinflammatory disorders via regulation of microglial activation and production of inflammatory mediators.
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CHD3 facilitates vRNP nuclear export by interacting with NES1 of influenza A virus NS2.
Cell. Mol. Life Sci.
PUBLISHED: 03-05-2014
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NS2 from influenza A virus mediates Crm1-dependent vRNP nuclear export through interaction with Crm1. However, even though the nuclear export signal 1 (NES1) of NS2 does not play a requisite role in NS2-Crm1 interaction, there is no doubt that NES1 is crucial for vRNP nuclear export. While the mechanism of the NES1 is still unclear, it is speculated that certain host partners might mediate the NES1 function through their interaction with NES1. In the present study, chromodomain-helicase-DNA-binding protein 3 (CHD3) was identified as a novel host nuclear protein for locating NS2 and Crm1 on dense chromatin for NS2 and Crm1-dependent vRNP nuclear export. CHD3 was confirmed to interact with NES1 in NS2, and a disruption to this interaction by mutation in NES1 significantly delayed viral vRNPs export and viral propagation. Further, the knockdown of CHD3 would affect the propagation of the wild-type virus but not the mutant with the weakened NS2-CHD3 interaction. Therefore, this study demonstrates that NES1 is required for maximal binding of NS2 to CHD3, and that the NS2-CHD3 interaction on the dense chromatin contributed to the NS2-mediated vRNP nuclear export.
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[Conservative treatment in male urinary incontinence].
Urologe A
PUBLISHED: 03-04-2014
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Prevalence, pathophysiology, diagnostic and therapeutic approaches of urinary incontinence are well studied in women; however, studies on male urinary incontinence focus on incontinence following surgery of the bladder or prostate, predominantly incontinence after radical prostatectomy. Aging men suffer from incontinence, most frequently urge incontinence (overactive bladder, OAB), nearly as often as women do.The domain of conservative therapy of urinary stress incontinence in men is pelvic floor training. It remains unclear whether biofeedback procedures, electrostimulation therapy, or magnetic stimulation therapy can enhance pelvic floor training. There are data suggesting that an off-label therapy with Duloxetin®, a selective serotonin-noradrenaline reuptake inhibitor (SSNRI), improves urinary incontinence following radical prostatectomy. Antimuscarinic agents in combination with bladder training have been proven as safe and effective treatment in men with OAB. Data, however, suggest that men with OAB are far less frequently treated than women.
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Hyperacute ischemic stroke without lesions on diffusion-weighted imaging in a patient treated with rtPA thrombolysis.
Clin Case Rep
PUBLISHED: 01-25-2014
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We report a comatose patient with severe neurological deficits who was without spontaneous language or movement. He had a good response to recombinant tissue plasminogen activator (rtPA) thrombolysis even though there were no detectable lesions on diffusion-weighted imaging (DWI). DWI is very sensitive for diagnosing hyperacute ischemic stroke, and rtPA thrombolysis is the best treatment. However, rtPA thrombolysis in ischemic stroke patients without lesions on DWI has rarely been reported.
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Lower serum uric acid levels in cerebral amyloid angiopathy: a pilot study.
Neurol. Sci.
PUBLISHED: 01-10-2014
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Cerebral amyloid angiopathy (CAA) is a common degenerative disease presenting intracerebral hemorrhage (ICH) in older people. Uric acid (UA) is a natural antioxidant, and may have a beneficial role in neurodegenerative diseases. Nevertheless, the role of UA in CAA remains unknown. In the present study, we compared serum UA levels in CAA-associated ICH patients (n = 82) and age/sex-matched controls (n = 82). Serum UA levels in possible CAA were significantly decreased when compared with healthy controls (232.68 ± 77.70 vs. 309.42 ± 59.83 ?mol/L; p < 0.001). Furthermore, UA levels in patients clinically diagnosed as probable CAA were significantly lower than those in patients diagnosed as possible CAA (193.06 ± 56.98 vs. 232.68 ± 77.70 ?mol/L; p = 0.014). These differences were still significant after adjusting for renal function and dyslipidemia (p < 0.001 and p = 0.002, respectively). However, there were no associations between serum UA levels and the distribution of hemorrhagic lesion, as well as neurological impairment. Our observations indicate that serum UA levels were decreased in CAA patients. UA might play a neuroprotective role in CAA and serve as a potential biomarker for reflecting the severity of A? deposition.
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A quantitative study of gully erosion based on object-oriented analysis techniques: a case study in Beiyanzikou catchment of Qixia, Shandong, China.
ScientificWorldJournal
PUBLISHED: 01-01-2014
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This paper took a subregion in a small watershed gully system at Beiyanzikou catchment of Qixia, China, as a study and, using object-orientated image analysis (OBIA), extracted shoulder line of gullies from high spatial resolution digital orthophoto map (DOM) aerial photographs. Next, it proposed an accuracy assessment method based on the adjacent distance between the boundary classified by remote sensing and points measured by RTK-GPS along the shoulder line of gullies. Finally, the original surface was fitted using linear regression in accordance with the elevation of two extracted edges of experimental gullies, named Gully 1 and Gully 2, and the erosion volume was calculated. The results indicate that OBIA can effectively extract information of gullies; average range difference between points field measured along the edge of gullies and classified boundary is 0.3166 m, with variance of 0.2116 m. The erosion area and volume of two gullies are 2141.6250 m(2), 5074.1790 m(3) and 1316.1250 m(2), 1591.5784 m(3), respectively. The results of the study provide a new method for the quantitative study of small gully erosion.
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Fluorescent protein marker lines in maize: generation and applications.
Int. J. Dev. Biol.
PUBLISHED: 10-30-2013
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Fluorescent proteins (FP) have significantly impacted the way that we study plants in the past two decades. In the post-genomics era, these FP tools are in higher demand by plant scientists for studying the dynamics of protein localization, function, and interactions, and to translate sequence information to biological knowledge that can benefit humans. Although FP tools have been widely used in the model plant Arabidopsis, few FP resources have been developed for maize, one of the most important food crops worldwide, and an ideal species for genetic and developmental biology research. In an effort to provide the maize and cereals research communities with a comprehensive set of FP resources for different purposes of study, we generated more than 100 stable transformed maize FP marker lines, which mark most compartments in maize cells with different FPs. Additionally, we are generating driver and reporter lines, based on the principle of the pOp-LhG4 transactivation system, allowing specific expression or mis-expression of any gene of interest to precisely study protein functions. These marker lines can be used not only for static protein localization studies, but will be useful for studying protein dynamics and interactions using kinetic microscopy methods, such as fluorescence recovery after photobleaching (FRAP), fluorescence correlation spectroscopy (FCS), and fluorescence resonance energy transfer (FRET).
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Endothelial cell stimulation overcomes restriction and promotes productive and latent HIV-1 infection of resting CD4+ T cells.
J. Virol.
PUBLISHED: 07-03-2013
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Highly active antiretroviral therapy (HAART) is able to suppress human immunodeficiency virus type 1 (HIV-1) to undetectable levels in the majority of patients, but eradication has not been achieved because latent viral reservoirs persist, particularly in resting CD4(+) T lymphocytes. It is generally understood that HIV-1 does not efficiently infect resting CD4(+) T cells, and latent infection in those cells may arise when infected CD4(+) T lymphoblasts return to resting state. In this study, we found that stimulation by endothelial cells can render resting CD4(+) T cells permissible for direct HIV infection, including both productive and latent infection. These stimulated T cells remain largely phenotypically unactivated and show a lower death rate than activated T cells, which promotes the survival of infected cells. The stimulation by endothelial cells does not involve interleukin 7 (IL-7), IL-15, CCL19, or CCL21. Endothelial cells line the lymphatic vessels in the lymphoid tissues and have frequent interactions with T cells in vivo. Our study proposes a new mechanism for infection of resting CD4(+) T cells in vivo and a new mechanism for latent infection in resting CD4(+) T cells.
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Clopidogrel with aspirin in acute minor stroke or transient ischemic attack.
N. Engl. J. Med.
PUBLISHED: 06-26-2013
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Stroke is common during the first few weeks after a transient ischemic attack (TIA) or minor ischemic stroke. Combination therapy with clopidogrel and aspirin may provide greater protection against subsequent stroke than aspirin alone.
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Fluoroquinolone-resistant Haemophilus parasuis isolates exhibit more putative virulence factors than their susceptible counterparts.
J. Clin. Microbiol.
PUBLISHED: 06-19-2013
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The prevalence of 23 putative virulence factors among fluoroquinolone-susceptible and -resistant Haemophilus parasuis isolates was analyzed. Putative hemolysin precursor, fimbrial assembly chaperone, and type I site-specific restriction modification system R subunit genes were more prevalent among fluoroquinolone-resistant H. parasuis isolates than among fluoroquinolone-susceptible H. parasuis isolates. Fluoroquinolone resistance may be associated with an increase in the presence of some virulence factors.
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Characterization of Streptococcus suis serotype 2 isolates from China.
Vet. Microbiol.
PUBLISHED: 06-04-2013
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Streptococcus suis (S. suis) is a major pathogen in the pig industry and an important zoonotic agent that causes severe invasive diseases in humans. Previous studies based on multilocus sequence typing (MLST) and the associations between sequence types and genotypes or virulence suggested that North American S. suis serotype 2 isolates are composed of multiple populations. This study investigated the population structure of S. suis serotype 2 isolates in China. We constructed a phylogenetic tree for S. suis serotype 2 isolates based on 16S rRNA gene typing and MLST, studied associations between clades and sources, analyzed the genotype distributions of virulence markers [muramidase-released protein (MRP), extracellular protein factor (EF), and suilysin (SLY)] in different clades, computed the selection pressures for these virulence marker genes, and verified the associations between clades and virulence. There were two primary clades (populations) in the phylogenetic tree of S. suis serotype 2. The two populations were associated with different tissue tropisms. The genotypic distributions and selection pressures of MRP, EF, and SLY were different between the two populations, which suggested that they had different evolutionary paths. The two populations also displayed differences in virulence in experimentally infected mice. The results provide insights into the population structure of S. suis serotype 2 isolates in China and suggest that S. suis serotype 2 clade 1 is an overlooked population that deserves further evaluation.
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Genetic analysis of the FBXO42 gene in Chinese Han patients with Parkinsons disease.
BMC Neurol
PUBLISHED: 05-03-2013
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Parkinsons disease (PD), the second most common neurodegenerative disease, is characterized by loss of dopaminergic neurons in the substantia nigra. The clinical manifestations of PD encompass a variety of motor and non-motor symptoms. Mutations in the F-box protein 7 gene (FBXO7) have been identified to cause Parkinsonian-pyramidal syndrome, an autosomal recessive form of Parkinsonism. The F-box protein 42 gene (FBXO42), a paralog of the FBXO7 gene, is involved in the ubiquitin-proteasome system that may play a role in the pathogenesis of PD.
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Genetic analysis of the leucine-rich repeat and lg domain containing Nogo receptor-interacting protein 1 gene in essential tremor.
J. Mol. Neurosci.
PUBLISHED: 03-24-2013
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Variants in the leucine-rich repeat and lg domain containing nogo receptor-interacting protein 1 gene (LINGO1) have been identified to be associated with the increased risk of essential tremor (ET), especially among Caucasians. To explore whether the LINGO1 gene plays a role in ET susceptibility, we performed a systematic genetic analysis of the coding region in the LINGO1 gene. Four nucleotide variants have been genotyped, including three known variants (rs2271398, rs2271397, and rs3743481), and a novel G ? C transition (ss491228439). Extended analysis showed no significant difference in genotypic and allelic distributions between 151 patients and 301 control subjects for these four variants (all P > 0.05). However, further sex-stratified analysis revealed that the C allele of rs2271397 and ss491228439 contributed the risk of ET in female (P = 0.017, OR = 2.139, 95 % CI 1.135 ~ 4.030 for rs2271397 and P = 0.038, OR = 1.812, 95 % CI 1.027 ~ 3.194 for ss491228439). Haplotype analysis indicated that A465-C474-C714 haplotype was significantly associated with increased risk of ET in female (P = 0.041, OR = 1.800, 95 % CI 1.020 ~ 3.178). Our results indicate that the LINGO1 variants are associated with ET in Chinese Han female patients.
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The fibrin-derived peptide b?15-42 attenuates liver damage in a rat model of liver ischemia/reperfusion injury.
Shock
PUBLISHED: 03-19-2013
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The inflammatory response after liver ischemia/reperfusion (I/R) contributes to increased risk of liver failure after liver surgery. Strategies aimed to preventing inflammation could be beneficial in reducing liver I/R injury. Recent studies have demonstrated that peptide B?15-42 is able to decrease the injury of I/R in heart and kidney by inhibition of leukocyte migration and preserving endothelial barrier function. Prompted by these results, we hypothesized that B?15-42 could also possess anti-inflammatory abilities to protect from or reduce hepatic I/R injury. Therefore, in this study, we aimed to evaluate the effects of B?15-42 in a model of liver I/R injury in rats. Rats were treated with B?15-42 at initiation of reperfusion and 2 h thereafter. Rats were killed at 0.5, 6, 24, and 48 h after reperfusion. Hepatic mRNA levels of fibrinogen-? (Fg?), Fg?, Fg? were significantly increased after I/R. Treatment with Fg-derived B?15-42 ameliorated liver I/R injury, as indicated by lower serum aminotransferase levels and fewer I/R-associated histopathologic changes. B?15-42 treatment decreased leukocyte infiltration and expression of hepatic inflammatory cytokines. Moreover, B?15-42 significantly reduced high-mobility group box 1 release and altered mitogen-activated protein kinase activation. In conclusion, B?15-42 treatment protected against liver warm I/R injury. The mechanism of protective action of B?15-42 seemed to involve its ability to reduce hepatic inflammatory response through preventing high-mobility group box 1 release and altering mitogen-activated protein kinase activation.
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Liver transplantation and inflammation: is lipopolysaccharide binding protein the link?
Cytokine
PUBLISHED: 03-05-2013
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Lipopolysaccharide (LPS) binding protein (LBP) is an acute phase protein, which upregulated in response to surgical interventions. LBP plays an important role in modulating LPS-induced inflammatory response. We investigated the expression of LBP and the translocation of LPS in rat models of hepatic ischemia reperfusion injury and liver transplantation (LTx). We also elucidated the effect of LBP on the inflammatory response.
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[Imaging diagnostics of the male pelvic floor].
Urologe A
PUBLISHED: 02-28-2013
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The understanding of the female pelvic floor during the last 20 years was very much influenced by the new techniques of sonographic and magnetic resonance imaging (MRI). Functional imaging of the male pelvic floor is, however, still in its infancy. In analogy to ultrasound examinations of the female pelvic floor, perineal ultrasound can be also be applied to men. The mobility of the proximal urethra, scarring of the bladder neck or implanted suburethral meshes can be easily visualized. Studies on healthy men provide information about different muscular structures during micturition. Morphology and function of the external sphincter can be visualized with transrectal or intraurethral ultrasound and also with a perineal approach. Using functional MRI the complex interactions of bladder, urethra, external sphincter and pelvic floor muscles can be evaluated. Functional MRI is so far not generally available but enables a better understanding of the function of the male pelvic floor. Imaging of the male pelvic floor makes a substantial contribution for improving surgical procedures for male incontinence in the future.
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EIF4G1 Ala502Val and Arg1205His variants in Chinese patients with Parkinson disease.
Neurosci. Lett.
PUBLISHED: 02-18-2013
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Growing evidences show that genetic abnormalities play an important role in the etiopathogenesis of Parkinson disease (PD). At least 18 genetic loci and 13 disease-related genes for parkinsonism have been identified. Recently, the p.Ala502Val and p.Arg1205His variants in the eukaryotic translation initiation factor 4-gamma 1 gene (EIF4G1) were found to be associated with PD. To evaluate whether the EIF4G1 p.Ala502Val and p.Arg1205His variants are related to PD in Chinese Han population, we conducted genetic examination of these two variants in 425 PD patients from Mainland China and none was found in our patients. We did identify a known non-pathogenic polymorphism c.3660C>T (p.Ala1220Ala, rs143852330) in a 73-year-old male patient. Our results, consistent with other recent reports, suggest that the EIF4G1 p.Ala502Val and p.Arg1205His variants are a rare cause of PD, at least in Chinese population.
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Genetic analysis of the FBXO48 gene in Chinese Han patients with Parkinson disease.
Neurosci. Lett.
PUBLISHED: 02-13-2013
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The F-box only protein 48 gene (FBXO48) is located in 2p13.3, the disease gene locus of Parkinson disease type 3 (PARK3), and it is one of the paralogs of the F-box only protein 7 gene (FBXO7), which is a causative gene of the Parkinson disease type 15 (PARK15; also known as Parkinsonian-pyramidal disease, PPD). To determine whether genetic mutation in the coding region of the FBXO48 gene plays a role in the etiology of PD, we screened DNA samples from 350 Chinese Han patients with PD. No mutation in the coding region of the FBXO48 gene was identified in our PD cohort, suggesting that mutations in the coding region of the FBXO48 gene play little or no role in the development of PD.
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The multiple roles for Sox2 in stem cell maintenance and tumorigenesis.
Cell. Signal.
PUBLISHED: 01-26-2013
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The Sry-containing protein Sox2 initially was known to regulate the self-renewal of the mouse and human embryonic stem cells (ESCs). It is also important for the maintenance of stem cells in multiple adult tissues including the brain and trachea, and it is one of the key transcription factors for establishing induced pluripotent stem cells. Recently, overexpression and gene amplification of Sox2 have been associated with the development of squamous cell carcinoma in multiple tissues such as the lung and esophagus. These different roles for Sox2 involve a complicated regulatory networks consisting of microRNAs, kinases and signaling molecules. While the levels of Sox2 are modulated transcriptionally and translationally, post-translational modification is also important for the various functions of Sox2. In clinics, high levels of Sox2 are correlated with poor prognosis and increased proliferation of cancer stem cells. Therefore targeting Sox2 can be potentially explored for a new therapeutic avenue to treat cancers. This review will focus on the different roles for Sox2 in stem cell maintenance and its oncogenic roles in the context of signal transcription and microRNA regulation. We will also review the main upstream and downstream targets of Sox2, which can be potentially used as therapeutic measures to treat cancer with abnormal levels of Sox2.
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Molecular characterisation of resistance to fluoroquinolones in Haemophilus parasuis isolated from China.
Int. J. Antimicrob. Agents
PUBLISHED: 01-25-2013
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There are many studies on fluoroquinolone (FQ) resistance in the literature, but little is known regarding the molecular characterisation of FQ-resistant Haemophilus parasuis. In this study, a total of 138 H. parasuis isolates were examined, among which 83 (60.1%) were resistant to enrofloxacin (EFX) and 8 (5.8%) were resistant to levofloxacin (LFX) as determined by Etest. Ten point mutations in the quinolone resistance-determining regions of gyrA, gyrB, parC and parE were detected by PCR and DNA sequencing. Interestingly, 100% of the resistant isolates contained mutations at 87D of gyrA, but other mutations occurred less frequently. Furthermore, it was found that there was synergy between 73S?R/I in parC and other point mutations with respect to FQ resistance. To examine the effect of different point mutations on FQ resistance, the minimum inhibitory concentrations of EFX and LFX were determined for strains generated by site-directed mutagenesis, and three point mutations (gyrA 87D?N, parC 73S?R and parE 551T?A) were shown to be involved in FQ resistance.
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Chronic lithium treatment protects against liver ischemia/reperfusion injury in rats.
Liver Transpl.
PUBLISHED: 01-18-2013
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Lithium has long been widely used in the treatment of bipolar mood disorders. Recent studies have demonstrated that lithium is able to decrease ischemia/reperfusion (I/R) injury in the brain, kidneys, and heart. Because lithium may act on a number of stress and survival pathways, it is of great interest to explore this compound also in the setting of liver I/R injury. In this study, we aimed to evaluate the effects of lithium in a model of liver I/R injury in rats. Chronic treatment with lithium (2 mmol/kg for 3 days before ischemia) decreased I/R injury, whereas acute treatment with a single dose of lithium (2 mmol/kg 1 hour before ischemia) did not confer any protection in a partial hepatic I/R model. Furthermore, rats subjected to chronic lithium treatment had a significantly better survival rate (60%) than saline-treated rats (27%) in a total hepatic I/R survival model. Chronic lithium treatment protected against liver I/R injury, as indicated by lower serum aminotransferase levels, fewer I/R-associated histopathological changes, lower hepatic inflammatory cytokine levels, less neutrophil infiltration, and lower hepatic high-mobility group box expression and serum levels. The mechanism of action of lithium appears to involve its ability to inhibit glycogen synthase kinase 3? activation, modulate mitogen-activated protein kinase activation, inhibit hepatic apoptosis, and induce autophagy. On the basis of these data, we conclude that lithium treatment may be a simple and applicable preconditioning intervention for protecting against liver I/R injury.
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Granulocyte colony stimulating factor induces lipopolysaccharide (LPS) sensitization via upregulation of LPS binding protein in rat.
PLoS ONE
PUBLISHED: 01-12-2013
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Liver is the main organ for lipopolysaccharide (LPS) clearance. Sensitization to LPS is associated with the upregulation of LPS-binding protein (LBP) in animal models. Therefore, we hypothesized that LBP could induce LPS sensitization through enhancing hepatic uptake of LPS. In this study, we examined the role of LBP in pathogenesis of LPS induced systemic inflammatory response syndrome (SIRS). LBP expression was upregulated after granulocyte colony stimulating (G-CSF) pretreatment. The effect of LBP was further confirmed by blockade of LBP using LBP blocking peptide--LBPK95A. After G-CSF pretreatment, upregulation of LBP was observed in bone marrow cells and liver. The G-CSF induced LBP upregulation caused LPS hypersensitization in rats as indicated by higher mortality and severer liver damage. Of note, LBP blockade increased the survival rate and attenuated the liver injury. The LBP induced LPS hypersensitization was associated with increased hepatic uptake of LPS and augmented hepatic expression of LPS receptors, such as toll-like receptor (TLR)-4. Furthermore, LBP mediated early neutrophil infiltration, which led to increased monocyte recruitment in liver after LPS administration. In conclusion, G-CSF induced LBP expression could serve as a new model for investigation of LPS sensitization. We demonstrated the crucial role of LBP upregulation in pathogenesis of LPS induced SIRS.
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4-Hydroxybenzyl modification of the highly teratogenic retinoid, 4-[(1E)-2-(5,5,8,8-tetramethyl-5,6,7,8-tetrahydro-2-naphthalenyl)-1-propen-1-yl]benzoic acid (TTNPB), yields a compound that induces apoptosis in breast cancer cells and shows reduced terato
Chem. Res. Toxicol.
PUBLISHED: 10-19-2011
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Retinoids are a class of compounds with structural similarity to vitamin A. These compounds inhibit the proliferation of many cancer cell lines but have had limited medical application as they are often toxic at therapeutic levels. Efforts to synthesize retinoids with a greater therapeutic index have met with limited success. 4-[(1E)-2-(5,5,8,8-tetramethyl-5,6,7,8-tetrahydro-2-naphthalenyl)-1-propen-1-yl]benzoic acid (TTNPB) is one of the most biologically active all-trans-retinoic acid (atRA) analogues and is highly teratogenic. In this study, we show that modification of the TTNPB carboxyl group with an N-(4-hydroxyphenyl)amido (4HPTTNPB) or a 4-hydroxybenzyl (4HBTTNPB) group changes the activity of the compound in cell culture and in vivo. Unlike TTNPB, both compounds induce apoptosis in cancer cells and bind poorly to the retinoic acid receptors (RARs). Like the similarly modified all-trans-retinoic acid (atRA) analogues N-(4-hydroxyphenyl)retinamide (4-HPR/fenretinide) and 4-hydroxybenzylretinone (4-HBR), 4HBTTNPB is a potent activator of components of the ER stress pathway. The amide-linked analogue, 4HPTTNPB, is less toxic to developing embryos than the parent TTNPB, and most significantly, the 4-hydroxybenzyl-modified compound (4HBTTNPB) that cannot be hydrolyzed in vivo to the parent TTNPB compound is nearly devoid of teratogenic liability.
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Large-scale identification of bacteria-host crosstalk by affinity chromatography: capturing the interactions of Streptococcus suis proteins with host cells.
J. Proteome Res.
PUBLISHED: 10-18-2011
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Protein-protein interactions between bacteria and their hosts are responsible for all types of infection processes. The investigation of the bacteria-host crosstalk can provide a comprehensive understanding of the pathogenesis of bacterial disease. Despite scattered efforts in this field, a systematic identification of interactions between host and bacterial proteins remains unavailable. Here, we develop ACSP (affinity chromatography-based surface proteomics), which combines affinity chromatography and shotgun proteomics (LC-MS/MS), to investigate the interactions on a large-scale. Using ACSP, the potential surface interacting proteins (SIPs) of Streptococcus suis serotype 2 (SS2) were captured by the chromatographic resin, which was immobilized with the native surface molecules of Hep-2 cells. And then 40 potential SIPs were identified from the preys by LC-MS/MS, including 3 SIPs that have been previously reported in the literature. We selected 8 important SIPs and confirmed their ability to adhere to Hep-2 cells. Additionally, 3 newly identified SIPs, or their polyclonal antibodies, were found to significantly inhibit the adherence of SS2 to Hep-2 cells, indicating their essential role in the interaction between SS2 and Hep-2 cells. Using this example, we show that ACSP represents a new valuable tool for investigating the bacteria-host interactions.
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Effects of the C-terminal truncation in NS1 protein of the 2009 pandemic H1N1 influenza virus on host gene expression.
PLoS ONE
PUBLISHED: 06-28-2011
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The 2009 pandemic H1N1 influenza virus encodes an NS1 protein with 11 amino acids (aa) truncation at the C-terminus. The C-terminal tail of influenza virus NS1 protein constitutes a nucleolar localization signal (NoLS) and is the binding domain of the cellular pre-mRNA processing protein, poly(A)-binding protein II (PABII). Here, our studies showed that the C-terminal-truncated NS1 of the 2009 pandemic virus was inefficient at blocking host gene expression, extension of the truncated NS1 to its full length increased the inhibition of host gene expression. Mechanistically, this increased inhibition of host gene expression by the full-length NS1 was not associated with nucleolar localization, but was due to the restoration of NS1s binding capacity to PABII. Furthermore, in vitro and in vivo characterization of two recombinant viruses encoding either the C-terminal 11-aa truncated or full-length NS1 of the 2009 pandemic virus showed that the C-terminal 11-aa truncation in NS1 did not significantly alter virus replication, but increased virus pathogenicity in mice.
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Early release of macrophage migration inhibitory factor after liver ischemia and reperfusion injury in rats.
Cytokine
PUBLISHED: 06-22-2011
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Macrophage migration inhibitory factor (MIF) is an important mediator of ischemia/reperfusion (I/R) injury in heart, brain and intestine. We previously demonstrated that MIF was released during warm/cold ischemia in vitro. However, the role of MIF in liver I/R injury remains unclear. We aimed to test the hypothesis that MIF acts as an early proinflammatory cytokine and could mediate the inflammatory injury in liver I/R. Rats (n=6 per group) were subjected to 90 min warm ischemia followed by 0.5h, 6h and 24h reperfusion, respectively to liver transplantation (LTx) after 6h of cold ischemia followed by 24h of reperfusion. The expression of MIF, its receptor (cluster of differentiation 74 (CD74)) and the downstream inflammatory cytokines (tumor necrosis factor-alpha (TNF-?) and interleukin-1 beta (IL-1?)) were analyzed. Peritoneal macrophages were cultured for 6h alone or in the presence of effluent from cold-preserved livers or effluent depleted of MIF. Warm I/R increased hepatic MIF-mRNA and protein expression. MIF-protein was released into peripheral circulation in vivo with a maximum at 0.5h after reperfusion. Induction of MIF-expression was associated with the expression of proinflammatory cytokines and its receptor in both models. MIF released by isolated cold preserved livers, induced TNF-? and IL-1? production by cultured peritoneal macrophages. Intrahepatic upregulation of MIF, release into systemic circulation and the associated upregulation of the proinflammatory mediators suggest a role of MIF in mediating the inflammatory response to I/R injury. Blocking experiments will help to elucidate its role as potential molecular target for preventing hepatic I/R injury.
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Comparative genomic analysis of Streptococcus suis reveals significant genomic diversity among different serotypes.
BMC Genomics
PUBLISHED: 06-20-2011
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Streptococcus suis (S. suis) is a major swine pathogen and an emerging zoonotic agent. Serotypes 1, 2, 3, 7, 9, 14 and 1/2 are the most prevalent serotypes of this pathogen. However, almost all studies were carried out on serotype 2 strains. Therefore, characterization of genomic features of other serotypes will be required to better understand their virulence potential and phylogenetic relationships among different serotypes.
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[Simultaneous determination of six flavonoids in Apocynum venetum by HPLC].
Zhongguo Zhong Yao Za Zhi
PUBLISHED: 06-11-2011
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To develop a high-performance liquid chromatography (HPLC) method for the simultaneous determination of rutin, hyperoside, isoquercetin, astragulin, quercetin, and kaempferol in Apocynum venetum and its extracts.
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ROP GTPases act with the receptor-like protein PAN1 to polarize asymmetric cell division in maize.
Plant Cell
PUBLISHED: 06-07-2011
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Plant Rho family GTPases (ROPs) have been investigated primarily for their functions in polarized cell growth. We previously showed that the maize (Zea mays) Leu-rich repeat receptor-like protein PANGLOSS1 (PAN1) promotes the polarization of asymmetric subsidiary mother cell (SMC) divisions during stomatal development. Here, we show that maize Type I ROPs 2 and 9 function together with PAN1 in this process. Partial loss of ROP2/9 function causes a weak SMC division polarity phenotype and strongly enhances this phenotype in pan1 mutants. Like PAN1, ROPs accumulate in an asymmetric manner in SMCs. Overexpression of yellow fluorescent protein-ROP2 is associated with its delocalization in SMCs and with aberrantly oriented SMC divisions. Polarized localization of ROPs depends on PAN1, but PAN1 localization is insensitive to depletion and depolarization of ROP. Membrane-associated Type I ROPs display increased nonionic detergent solubility in pan1 mutants, suggesting a role for PAN1 in membrane partitioning of ROPs. Finally, endogenous PAN1 and ROP proteins are physically associated with each other in maize tissue extracts, as demonstrated by reciprocal coimmunoprecipitation experiments. This study demonstrates that ROPs play a key role in polarization of plant cell division and cell growth and reveals a role for a receptor-like protein in spatial localization of ROPs.
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Preliminary experience of a PDCA-cycle and quality management based training curriculum for rat liver transplantation.
J. Surg. Res.
PUBLISHED: 05-29-2011
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As repeatedly operating rat liver transplantation (LTx) until animals survive is inefficient in respect to time and use of living animals, we developed a new training concept. METHODS AND CONCEPTS: Training was divided into four phases: pretraining-phase, basic-microsurgical-training phase, advanced-microsurgical-training phases, and expert-microsurgical-training phase. Two "productivity-phases" were introduced right after the basic- and advanced-microsurgical-training phases, respectively, to allow the trainee to accumulate experience and to be scientifically productive before proceeding to a more complex procedure. PDCA cycles and quality criteria were employed to control the learning-process and the surgical quality. Predefined quality criteria included survival rate, intraoperative, postoperative, and histologic parameters.
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Comparative genomics study of multi-drug-resistance mechanisms in the antibiotic-resistant Streptococcus suis R61 strain.
PLoS ONE
PUBLISHED: 05-23-2011
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Streptococcus suis infections are a serious problem for both humans and pigs worldwide. The emergence and increasing prevalence of antibiotic-resistant S. suis strains pose significant clinical and societal challenges.
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Complete genome sequence of Streptococcus suis serotype 3 strain ST3.
J. Bacteriol.
PUBLISHED: 05-13-2011
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Streptococcus suis is a zoonotic pathogen causing economic loss in the swine industry and is also a threat to human health. To date, the mechanism of pathogenesis is not fully understood. Here, we report the complete genome sequence of S. suis strain ST3 of serotype 3, which provides opportunities to reveal genetic basis of infection of S. suis non-serotype 2 strains.
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Determinants of childrens use of and time spent in fast-food and full-service restaurants.
J Nutr Educ Behav
PUBLISHED: 05-10-2011
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Identify parental and childrens determinants of childrens use of and time spent in fast-food (FF) and full-service (FS) restaurants.
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Early detection of secondary damage in ipsilateral thalamus after acute infarction at unilateral corona radiata by diffusion tensor imaging and magnetic resonance spectroscopy.
BMC Neurol
PUBLISHED: 05-05-2011
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Traditional magnetic resonance (MR) imaging can identify abnormal changes in ipsilateral thalamus in patients with unilateral middle cerebral artery (MCA) infarcts. However, it is difficult to demonstrate these early changes quantitatively. Diffusion tensor imaging (DTI) and proton magnetic resonance spectroscopy (MRS) are potentially sensitive and quantitative methods of detection in examining changes of tissue microstructure and metabolism. In this study, We used both DTI and MRS to examine possible secondary damage of thalamus in patients with corona radiata infarction.
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Complex alternative splicing of the smarca2 gene suggests the importance of smarca2-B variants.
J Cancer
PUBLISHED: 04-11-2011
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BRM is an ATPase component of the SWI/SNF complex that regulates chromatin remodeling and cell proliferation and is considered a tumor suppressor. In this study we characterized transcripts from the Smarca2 gene that encodes the BRM protein. We found that the human Smarca2 gene (hSmarca2), like its mouse counterpart (mSmarca2), also initiated a short transcript from intron 27 of the long transcript. We name the long and short transcripts as Smarca2-a and Smarca2-b, respectively. Like its human counterpart, mSmarca2-a also underwent alternative splicing at the 54-bp exon 29. The hSmarca2-b had two alternative initiation sites and underwent alternative splicing at three different 3 sites of exon 1 and at exons 2, 3 and/or 5. We identified nine hSmarca2-b mRNA variants that might produce five different proteins. mSmarca2-b also underwent alternative splicing at exon 3 and/or exon 5, besides alternatively retaining part of intron 1 in exon 1. Smarca2-b was expressed more abundantly than Smarca2-a in many cell lines and was more sensitive to serum starvation. Moreover, cyclin D1 also regulated the expression of both Smarca2-a and Smarca2-b in a complex manner. These data suggest that the functions of the Smarca2 gene may be very complex, not just simply inhibiting cell proliferation, and in certain situations may be elicited mainly by expressing the much less known Smarca2-b, not the better studied Smarca2-a and its products BRM proteins.
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E3 ligase Rad18 promotes monoubiquitination rather than ubiquitin chain formation by E2 enzyme Rad6.
Proc. Natl. Acad. Sci. U.S.A.
PUBLISHED: 03-21-2011
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In ubiquitin conjugation, different combinations of E2 and E3 enzymes catalyse either monoubiquitination or ubiquitin chain formation. The E2/E3 complex Rad6/Rad18 exclusively monoubiquitinates the proliferating cell nuclear antigen (PCNA) to signal for "error prone" DNA damage tolerance, whereas a different set of conjugation enzymes is required for ubiquitin chain formation on PCNA. Here we show that human E2 enzyme Rad6b is intrinsically capable of catalyzing ubiquitin chain formation. This activity is prevented during PCNA ubiquitination by the interaction of Rad6 with E3 enzyme Rad18. Using NMR and X-ray crystallography we show that the R6BD of Rad18 inhibits this activity by competing with ubiquitin for a noncovalent "backside" binding site on Rad6. Our findings provide mechanistic insights into how E3 enzymes can regulate the ubiquitin conjugation process.
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Prolonged cold ischemia does not trigger lethal rejection or accelerate the acute rejection in two allogeneic rat liver transplantation models.
J. Surg. Res.
PUBLISHED: 03-16-2011
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Prolonged cold ischemia (CI) might induce lethal liver graft rejection and/or accelerate the course of lethal rejection in BN-Lew and ACI-Lew liver transplantation (LT) models, which have been successfully challenged by liver-size reduction.
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Complete genome sequence of Streptococcus suis serotype 14 strain JS14.
J. Bacteriol.
PUBLISHED: 03-11-2011
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Streptococcus suis is an important zoonotic agent leading to a variety of diseases in swine and can be transmitted to human beings upon close contact. Here, we report the complete genome sequence of S. suis serotype 14 strain JS14 which was isolated from a diseased pig in Jiangsu Province, China.
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HMGB1 in ischemic and non-ischemic liver after selective warm ischemia/reperfusion in rat.
Histochem. Cell Biol.
PUBLISHED: 03-06-2011
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High mobility group box 1 (HMGB1) acts as an early mediator in inflammation and organ injury. Ischemia reperfusion (I/R) injury induces HMGB1 translocation and expression in ischemic areas. However, it is unknown whether selective warm liver I/R injury also induces the expression of HMGB1 in non-ischemic lobes. The present study aimed to test the hypothesis that selective liver I/R injury also causes HMGB1 translocation and up-regulates its expression in non-ischemic liver areas. In the present study, selective I/R injury was induced by clamping the median and left lateral liver lobes for 90 min followed by 0.5, 6 and 24 h reperfusion. We used male inbred Lewis rats; six animals for each point in time and six animals for the normal control group. Selective hepatic I/R injury induced morphological changes not only in ischemic lobes but also in non-ischemic lobes. HMGB1 translocation and expression was increased in a time-dependent manner in the ischemic lobes, and increased in with delayed onset in the non-ischemic lobes. Serum HMGB1 levels were increased after reperfusion. Furthermore, liver I/R injury up-regulated the expression of HMGB1 receptors (Toll-like receptor 4 and receptor for advanced glycation end products and pro-inflammatory cytokines (Tumor necrosis factor-alpha and interleukin-6) in both ischemic lobes, however, the up-regulation of these cytokines was more prominent in the ischemic lobes. In conclusion, selective warm I/R induces a substantial "sympathetic/bystander" effect on the non-ischemic lobes in terms of HMGB1 translocation and local cytokine production.
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Synthesis, biological evaluation and molecular modeling of novel triazole-containing berberine derivatives as acetylcholinesterase and ?-amyloid aggregation inhibitors.
Bioorg. Med. Chem.
PUBLISHED: 02-14-2011
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A series of novel triazole-containing berberine derivatives were synthesized via the azide-alkyne cycloaddition reaction. Their biological activity as inhibitors of both acetylcholinesterase (AChE) and butyrylcholinesterase (BuChE) were evaluated. Among them, compound 16d, which featured a diisopropylamino substitution at the 4-position of triazole ring, was found to be a potent inhibitor of AChE, with IC(50) value of 0.044 ?M. Compound 18d, which beares a butyl at the 4-position of the triazole ring, showed the highest potency of ?-amyloid aggregation inhibition (77.9% at 20 ?M). Molecular modeling studies indicated that the triazole moiety of berberine derivatives displayed a face-to-face ?-? stacking interaction in a sandwich form with the Trp84 (4.09 Å) and Phe330 (4.33 Å) in catalytic sites of AChE.
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Emergence of novel reassortant H3N2 influenza viruses among ducks in China.
Arch. Virol.
PUBLISHED: 01-24-2011
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During 2006-2009 influenza virus surveillance, three H3N2 viruses were isolated from ducks in Central China. Sequence and phylogenetic analyses revealed that most segments of these three isolates had high identity with H3N2 swine isolates in South China. However, for M, the three viruses, along with H1N1 swine isolates of North America, formed a cluster; for PB2, two of these isolates fell into the cluster of the H5N1 duck isolates, indicating a reassortment among H3N2, H1N1 swine viruses and H5N1 avian virus. The emergence of H3N2 virus with incorporation of an H5N1 virus gene raises new concerns about the generation of novel viruses that could affect humans.
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HMGB1 translocation and expression is caused by warm ischemia reperfusion injury, but not by partial hepatectomy in rats.
Exp. Mol. Pathol.
PUBLISHED: 01-13-2011
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Mechanical injury or ischemia/reperfusion (I/R) injury induces high mobility of group box 1 (HMGB1) translocation and release. However, the surgical procedure itself can initiate pathophysiologic processes causing damage to the respective organ. A liver resection, as an example, leads to portal hyperperfusion injury of the remnant liver. Therefore, we aimed to elucidate the impact of different hepatic surgical injury models on cellular localization and expression of HMGB1. Focal warm I/R injury was induced by clamping the vascular blood supply to the median and left lateral liver lobes for 90 min followed by 0.5 h, 6 h and 24 h reperfusion, as reported previously. Liver injury by PH was induced by subjecting rats to 30%, 70% or 90% partial hepatectomy (PH) followed by a 24 h observation period. Additional 12 rats were subjected to 90% PH and sacrificed at 1 h and 6 h to investigate the expression and release pattern of HMGB1. Elevation of serum liver enzymes indicating hepatic injury peaked at 6 h and recovered thereafter in models, warm I/R injury and PH. Liver injury was confirmed by liver histology. HMGB1 was translocated from the nucleus to the cytoplasm in livers subjected to warm I/R; but not in livers subjected to PH. Both protein and mRNA expression of HMGB1 were significantly up-regulated in livers subjected to warm I/R. In contrast, neither 30% PH, 70% PH nor 90% PH caused an elevation of hepatic HMGB1 mRNA and protein expression. High serum levels of HMGB1 (30 ng/ml) were measured at 0.5 h reperfusion period after warm I/R, much lower levels thereafter (<5 ng/ml). Similar low serum levels were measured at all time points after 90% PH. Subsequently expression levels of TNF-a should be changed to tumor necrosis factor-alpha (TNF-?) reached a peak (26-fold elevation) at 6 h and decreased down to 5-fold at 24 h after warm I/R. TNF-? expression levels after PH never exceeded a 5-fold elevation. In conclusion, HMGB1 translocation and expression depends on the type of liver injury as it is induced by ischemia, but not by liver resection/hyperperfusion. These results suggest that HMGB1 may be used as molecular marker to visualize ischemic damage. Mechanic injury in hepatic surgery is associated with focal warm ischemia, and thereby HMGB1 translocation reflects surgical quality in experimental PH. Expression of hepatic TNF-? follows the kinetic pattern of HMGB1, pointing to a muss less pronounced inflammatory response after successful PH compared to warm I/R injury.
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Symmetry and asymmetry of the RING-RING dimer of Rad18.
J. Mol. Biol.
PUBLISHED: 01-13-2011
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The human ubiquitin-conjugating enzyme Rad6 (E2), with ubiquitin ligase enzyme Rad18 (RING E3), monoubiquitinates proliferating cell nuclear antigen at stalled replication forks in DNA translesion synthesis. Here, we determine the structure of the homodimeric Rad18 RING domains by X-ray crystallography and classify it to RING-RING dimers that dimerize through helices adjacent to the RING domains and through the canonical RING domains. Using NMR spectroscopy and site-directed mutagenesis, we demonstrate that the Rad6b binding site, for the Rad18 RING domain, strongly resembles that of other E2/E3 RING/U-box complexes. We show that the homodimeric Rad18 RING domain can recruit two Rad6b E2 enzymes, whereas the full-length Rad18 homodimer binds only to a single Rad6b molecule. Such asymmetry is a common feature of RING-RING heterodimers and has been observed for the CHIP U-box homodimer. We propose that asymmetry may be a common feature of dimeric RING E3 ligases.
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Remote focusing for programmable multi-layer differential multiphoton microscopy.
Biomed Opt Express
PUBLISHED: 11-08-2010
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We present the application of remote focusing to multiphoton laser scanning microscopy and utilize this technology to demonstrate simultaneous, programmable multi-layer imaging. Remote focusing is used to independently control the axial location of multiple focal planes that can be simultaneously imaged with single element detection. This facilitates volumetric multiphoton imaging in scattering specimens and can be practically scaled to a large number of focal planes. Further, it is demonstrated that the remote focusing control can be synchronized with the lateral scan directions, enabling imaging in orthogonal scan planes.
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Effect on virulence and pathogenicity of H5N1 influenza A virus through truncations of NS1 eIF4GI binding domain.
J. Infect. Dis.
PUBLISHED: 09-22-2010
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To study the effect of NS1 eIF4GI binding domain on virulence and pathogenicity of H5N1 influenza A virus, 5 recombinant H5N1 viruses encoding eIF4GI binding domain-truncated NS1 proteins and parental NS1 (NS1?wt) were generated by an 8?plasmid-based reverse genetics system. The results indicated that the recombinants with the addition of 5?amino acid and the deletion position of 85-89 in NS1?wt were attenuated in replication in vitro and in vivo, compared with the recombinant wild?type virus rNS1?wt, whereas the deletion position 85-94 or the entire eIF4GI binding domain in NS1?wt displayed a significantly attenuated phenotype in chicken and mice. We also showed that the eIF4GI binding domain-truncated mutants were impaired in their ability to inhibit interferon production in vitro, and they did not replicate as efficiently as the parental recombinant strain in embryonated hen eggs, in Madin ?Darby Canine Kidney cells, or in vivo in chickens and in a mouse model. Therefore, these attenuated NS1?truncated viruses may have a great potential as live attenuated vaccine candidates against H5N1 influenza A virus.
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Mothers and meals. The effects of mothers meal planning and shopping motivations on childrens participation in family meals.
Appetite
PUBLISHED: 09-01-2010
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Participation in family meals has been associated with benefits for health and social development of children. The objective of the study was to identify the impact of mothers work of caring through planning regularly scheduled meals, shopping and cooking, on childrens participation in family meals. Parents of children aged 9-11 or 13-15 years from 300 Houston families were surveyed about parents work, meal planning for and scheduling of meals, motivations for food purchases, importance of family meals, and childrens frequency of eating dinner with their families. The children were interviewed about the importance of eating family meals. Hypotheses were tested using path analysis to calculate indirect and total effects of variables on the outcome variable of frequency of children eating dinner with their family. Mothers belief in the importance of family meals increased likelihood of children eating dinner with families by increasing likelihood that mothers planned dinner and that dinners were regularly scheduled. Mothers perception of time pressures on meal preparation had a negative, indirect effect on the frequency of childrens participation in family dinners by reducing mothers meal planning.
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NMR characterization of foldedness for the production of E3 RING domains.
J. Struct. Biol.
PUBLISHED: 07-23-2010
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We summarize the use of NMR spectroscopy in the production and the screening of stability and foldedness of protein domains, and apply it to the RING domains of E3 ubiquitin-ligases. RING domains are involved in specific interactions with E2 ubiquitin-conjugating enzymes and thus play an essential role in the ubiquitination pathway. Protein production of the Zn(2+) containing and cysteine rich RING domains for molecular studies frequently turns out to be problematic. We compared the expression and solubility of 14 E3 RING/U-box domains fused to the N-terminal tags of His(6), His(6)-GB1, His(6)-Trx and His(6)-GST at small scale and analyzed, by NMR spectroscopy, their correct folding after purification. The addition of GST, Trx or GB1 to the N-terminal His(6) tag significantly improved both the expression and solubility of target proteins as compared to His(6) tag alone. More importantly most of the immobilized metal affinity chromatography (IMAC) purified proteins were largely unfolded as judged by analysis of the (1)H-(15)N HSQC spectra. We demonstrate that imidazole causes a concentration dependent decrease in stability of RING proteins ascribed to metal depletion and resulting in unfolding or precipitation. In contrast, using glutathione affinity chromatography, the His(6)-GST fused RING and U-box domains were purified as correctly folded proteins with high yields. Our data clearly demonstrate that IMAC should be avoided and that GST-fusion affinity chromatography is generally applicable for expression and purification of Zn(2+) containing proteins.
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Release of danger signals during ischemic storage of the liver: a potential marker of organ damage?
Mediators Inflamm.
PUBLISHED: 07-07-2010
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Liver grafts suffer from unavoidable injury due to ischemia and manipulation before implantation. Danger signals such as high-mobility group box -1(HMGB1) and macrophage migration inhibitory factor (MIF) play a pivotal role in the immune response. We characterized the kinetics of their release into the effluent during cold/warm ischemia and additional manipulation-induced mechanical damage. Furthermore, we evaluated the relationship between HMGB1/MIF release and ischemic/mechanical damage. Liver enzymes and protein in the effluent increased with increasing ischemia time. HMGB1/MIF- release correlated with the extent of hepatocellular injury. With increasing ischemia time and damage, HMGB1 was translocated from the nucleus to the cytoplasma as indicated by weak nuclear and strong cytoplasmic staining. Enhancement of liver injury by mechanical damage was indicated by an earlier HMGB1 translocation into the cytoplasm and earlier release of danger signals into the effluent. Our results suggest that determination of HMGB1 and MIF reflects the extent of ischemic injury. Furthermore, HMGB1 and MIF are more sensitive than liver enzymes to detect the additional mechanical damage inflicted on the organ graft during surgical manipulation.
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Berberine derivatives, with substituted amino groups linked at the 9-position, as inhibitors of acetylcholinesterase/butyrylcholinesterase.
Bioorg. Med. Chem. Lett.
PUBLISHED: 06-04-2010
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Berberine derivatives with substituted amino groups linked at the 9-position using different carbon spacers were designed, synthesized, and biologically evaluated as inhibitors of acetylcholinesterase. Compound 10b, with a cyclohexylamino group linked to berberine by a three carbon spacer, gave the most potent inhibitor activity with an IC(50) of 0.020 ?M for AChE. Kinetic studies revealed mixed inhibition of AChE, and molecular modeling simulations of the AChE-inhibitor complex confirmed that compounds bound to both the catalytic active site and the peripheral anionic site.
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Proteomics analysis of differential expression of chicken brain tissue proteins in response to the neurovirulent H5N1 avian influenza virus infection.
J. Proteome Res.
PUBLISHED: 05-05-2010
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A certain H5N1 avian influenza virus has gained the ability to cause the classic central nervous system dysfunction in poultry and migratory birds. This study presents the proteomics analysis on the change of proteins to H5N1 avian influenza virus with neurovirulence infection in chicken brain tissue. By using 2-DE, coupled with MALDI-TOF MS/MS, we identified a set of differentially expressed cellular proteins, including 18 up-regulated proteins and 13 down-regulated proteins. The most significant changes were found in cytoskeleton proteins, proteins associated with the ubiquitin-proteasome pathway, and neural signal transduction proteins. Some identified proteins such as CRMP and SEP5 were found to participate in the pathogenesis progress of Parkinsons and Huntingtons diseases, which also developed encephalitis accompanied with CNS dysfunction. The obtained data can provide insight into the virus-chicken brain tissue interaction and reveal the potential mechanism of the neuropathogenesis when the host was infected by the neurovirulent avian influenza virus.
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Response of swine spleen to Streptococcus suis infection revealed by transcription analysis.
BMC Genomics
PUBLISHED: 05-04-2010
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Streptococcus suis serotype 2 (SS2), a major swine pathogen and an emerging zoonotic agent, has greatly challenged global public health. Systematical information about host immune response to the infection is important for understanding the molecular mechanism of diseases.
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Using multiple household food inventories to measure food availability in the home over 30 days: a pilot study.
Nutr J
PUBLISHED: 04-15-2010
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The consumption of foods, especially by children, may be determined by the types of foods that are available in the home. Because most studies use a single point of data collection to determine the types of foods in the home, which can miss the change in availability when resources are not available, the primary objective of this study was to determine the extent to which the weekly availability of household food items changed over one month by 1) developing the methodology for the direct observation of the presence and amount of food items in the home; 2) conducting five in-home household food inventories over a thirty-day period in a small convenience sample; and 3) determining the frequency that food items were present in the participating households.
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Evaluation of the protective efficacy of a newly identified immunogenic protein, HP0272, of Streptococcus suis.
FEMS Microbiol. Lett.
PUBLISHED: 04-08-2010
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Streptococcus suis serotype 2 (SS2) infection is a major cause of sudden death in pigs and is of concern for humans as it has strong zoonotic capabilities. Developing novel effective vaccines would be beneficial to control SS2 infection. HP0272 is a novel immunogenic surface protein; its protective efficacy remains to be evaluated. The present mouse model found that the purified recombinant HP0272 could elicit a significant humoral antibody response, and to confer complete protection against a lethal dose of SS2 infection. In addition, real-time PCR confirmed that in vivo-induced antigen existed in most SS2 field pathogenic strains, and in half of all reference strains of different serotypes of S. suis. The results indicate that HP0272 has the potential as an effective component of a new vaccine.
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Viral reservoir is suppressed but not eliminated by CD8 vaccine specific lymphocytes.
Vaccine
PUBLISHED: 03-02-2010
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It has long been postulated that while CD8 lymphocytes are capable of suppressing human immunodeficiency virus (HIV)-1 replication it is unlikely that the viral reservoirs once formed can be cleared. Our previous studies demonstrate that co-immunizing cynomologous macaques with a simian/human immunodeficiency virus (SHIV) DNA-based vaccines induces a strong cellular immune response that is able to suppress viral replication. We further demonstrated that interleukin (IL)-12 could significantly enhance the vaccine specific CD8 lymphocyte response. In this manuscript cynomologous macaques were vaccinated with a SHIV DNA-based vaccine co-delivered with IL-12. The macaques were then challenged with SHIV89.6p. Two years post-immunization and viral challenge we transiently depleted CD8(+) T cells. Plasma viral load increased, demonstrating the central role of CD8(+) T cells in viral suppression yet an inability to clear the viral reservoirs. Furthermore, in the data presented here, we found a higher number of IFN-gamma producing vaccine specific cells did not enhance suppression of viral replication.
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Effect of needle length when immunizing obese adolescents with hepatitis B vaccine.
Pediatrics
PUBLISHED: 02-08-2010
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Several studies have noted that obese adolescents and adults achieve lower titers of antibody in response to vaccines such as hepatitis B virus (HBV) vaccine. The objective of this study was to determine whether use of a longer (1.5-in) rather than a standard (1-in) needle to penetrate the thicker deltoid fat pad among obese youth would result in higher antibody titers after immunization against HBV.
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IgA1 protease contributes to the virulence of Streptococcus suis.
Vet. Microbiol.
PUBLISHED: 01-08-2010
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Streptococcus suis (S. suis) is a major swine pathogen and emerging zoonotic agent. However, the current understanding of the S. suis pathogenesis of infection remains limited. In the present study, the contribution to the pathogenesis of S. suis was evaluated on IgA1 protease (or iga gene), which has been regarded as a virulence factor of gram-negative pathogenic bacteria and of certain gram-positive pathogenic bacteria. In contrast to the wild type (WT) strain of S. suis serotype 2, the isogenic iga mutant (?iga) constructed by allelic replacement showed significantly decreased lethality to pigs. The present study suggests that IgA1 protease might contribute to S. suis pathogenesis.
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Synthesis, biological evaluation, and molecular modeling of berberine derivatives as potent acetylcholinesterase inhibitors.
Bioorg. Med. Chem.
PUBLISHED: 10-22-2009
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By targeting the dual active sites of acetylcholinesterase (AChE), a new series of berberine derivatives was designed, synthesized, and evaluated as AChE inhibitors. Most of the derivatives inhibited AChE in the sub-micromolar range. Compound 8c, berberine linked with phenol by a 4-carbon spacer, showed the most potent inhibition of AChE. A kinetic study of AChE and BuChE indicated that a mix-competitive binding mode existed for these berberine derivatives. Molecular modeling studies confirmed that these hybrids target both the catalytic active site (CAS) and the peripheral anionic site (PAS) of AChE. This is the first report where AChE inhibitory activity has been associated with berberine as a lead molecule.
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[Prokaryotic expression of recombinant Streptococcus suis IgG binding protein and its binding activity with IgG].
Sheng Wu Gong Cheng Xue Bao
PUBLISHED: 07-30-2009
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Streptococcus suis (S. suis) IgG-binding protein (SPG) was present in all S. suis strains examined. It showed binding activities with IgG from various host species. Little was known about the biological role of this protein, but it was commonly believed that it acted as virulence factor. In this study, the genes encoding SPG were amplified respectively from the total DNA of the S. suis serotype 1/2, 1, 2 and 9 with PCR and expressed in Escherichia coli BL21 by plasmid pET28a as vector. The recombinant proteins were first purified with affinity chromatography (Ni-NTA), and further purified by sephadexG-200 gel chromatography. The recombinant SPG proteins were identified to have binding activities with IgG of different host species, and for human and porcine IgG they showed better binding activities. But the SPG from different serotypes of S. suis showed no great differences in their binding activities with IgG from the same host species.
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Ectopic expression of multiple microbial rhodopsins restores ON and OFF light responses in retinas with photoreceptor degeneration.
J. Neurosci.
PUBLISHED: 07-24-2009
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By expressing channel rhodopsin-2 (ChR2) in inner retinal neurons, previous studies have demonstrated restoration of ON responses in the retina after the death of rod and cone photoreceptors. In this study, we report that the expression of halorhodopsin (HaloR), a light-driven chloride pump, can effectively restore OFF responses in inner retinal neurons of mice with retinal degeneration. We show that HaloR-expressing retinal ganglion cells respond to light with rapid hyperpolarization and suppression of spike activity. After termination of the light stimulus, their membrane potential exhibits a rapid rebound overshoot with robust sustained or transient spike firing. Furthermore, we show that coexpression of ChR2/HaloR in retinal ganglion cells can produce ON, OFF, and even ON-OFF responses, depending on the wavelength of the light stimulus. Our results suggest that the expression of multiple microbial rhodopsins such as ChR2 and HaloR is a possible strategy to restore both ON and OFF light responses in the retina after the death of rod and cone photoreceptors.
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A comprehensive framework of E2-RING E3 interactions of the human ubiquitin-proteasome system.
Mol. Syst. Biol.
PUBLISHED: 07-07-2009
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Covalent attachment of ubiquitin to substrates is crucial to protein degradation, transcription regulation and cell signalling. Highly specific interactions between ubiquitin-conjugating enzymes (E2) and ubiquitin protein E3 ligases fulfil essential roles in this process. We performed a global yeast-two hybrid screen to study the specificity of interactions between catalytic domains of the 35 human E2s with 250 RING-type E3s. Our analysis showed over 300 high-quality interactions, uncovering a large fraction of new E2-E3 pairs. Both within the E2 and the E3 cohorts, several members were identified that are more versatile in their interaction behaviour than others. We also found that the physical interactions of our screen compare well with reported functional E2-E3 pairs in in vitro ubiquitination experiments. For validation we confirmed the interaction of several versatile E2s with E3s in in vitro protein interaction assays and we used mutagenesis to alter the E3 interactions of the E2 specific for K63 linkages, UBE2N(Ubc13), towards the K48-specific UBE2D2(UbcH5B). Our data provide a detailed, genome-wide overview of binary E2-E3 interactions of the human ubiquitination system.
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What is Visualize?

JoVE Visualize is a tool created to match the last 5 years of PubMed publications to methods in JoVE's video library.

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We use abstracts found on PubMed and match them to JoVE videos to create a list of 10 to 30 related methods videos.

Video X seems to be unrelated to Abstract Y...

In developing our video relationships, we compare around 5 million PubMed articles to our library of over 4,500 methods videos. In some cases the language used in the PubMed abstracts makes matching that content to a JoVE video difficult. In other cases, there happens not to be any content in our video library that is relevant to the topic of a given abstract. In these cases, our algorithms are trying their best to display videos with relevant content, which can sometimes result in matched videos with only a slight relation.