JoVE Visualize What is visualize?
Stop Reading. Start Watching.
Advanced Search
Stop Reading. Start Watching.
Regular Search
Find video protocols related to scientific articles indexed in Pubmed.
The Association of Statin Use and Gonado-Sexual Function in Women: A Retrospective Cohort Analysis.
J Sex Med
PUBLISHED: 11-11-2014
Show Abstract
Hide Abstract
It has been hypothesized that statins reduce sex hormone biosynthesis through hepatic inhibition of cholesterol synthesis, which is a precursor of androstenedione and estradiol. Such a reduction has been associated with menstrual irregularities, menopausal disorders, infertility, and low libido, but studies are conflicting. Few studies have evaluated the clinical effects of statins on gonadal-sexual function in women.
Related JoVE Video
Instrument Development of the UNC Dry Eye Management Scale.
Cornea
PUBLISHED: 09-26-2014
Show Abstract
Hide Abstract
Dry eye disease (DED) is a common ocular disease that can have adverse effects on quality of life. Our aim was to develop a single-item questionnaire that is reliable, patient-driven, and clinic friendly to assess DED symptoms and their effect on quality of life to help support the management of patients with DED.
Related JoVE Video
Requirement for ssbp2 in hematopoietic stem cell maintenance and stress response.
J. Immunol.
PUBLISHED: 09-19-2014
Show Abstract
Hide Abstract
Transcriptional mechanisms governing hematopoietic stem cell (HSC) quiescence, self-renewal, and differentiation are not fully understood. Sequence-specific ssDNA-binding protein 2 (SSBP2) is a candidate acute myelogenous leukemia (AML) suppressor gene located at chromosome 5q14. SSBP2 binds the transcriptional adaptor protein Lim domain-binding protein 1 (LDB1) and enhances LDB1 stability to regulate gene expression. Notably, Ldb1 is essential for HSC specification during early development and maintenance in adults. We previously reported shortened lifespan and greater susceptibility to B cell lymphomas and carcinomas in Ssbp2(-/-) mice. However, whether Ssbp2 plays a regulatory role in normal HSC function and leukemogenesis is unknown. In this study, we provide several lines of evidence to demonstrate a requirement for Ssbp2 in the function and transcriptional program of hematopoietic stem and progenitor cells (HSPCs) in vivo. We found that hematopoietic tissues were hypoplastic in Ssbp2(-/-) mice, and the frequency of lymphoid-primed multipotent progenitor cells in bone marrow was reduced. Other significant features of these mice were delayed recovery from 5-fluorouracil treatment and diminished multilineage reconstitution in lethally irradiated bone marrow recipients. Dramatic reduction of Notch1 transcripts and increased expression of transcripts encoding the transcription factor E2a and its downstream target Cdkn1a also distinguished Ssbp2(-/-) HSPCs from wild-type HSPCs. Finally, a tendency toward coordinated expression of SSBP2 and the AML suppressor NOTCH1 in a subset of the Cancer Genome Atlas AML cases suggested a role for SSBP2 in AML pathogenesis. Collectively, our results uncovered a critical regulatory function for SSBP2 in HSPC gene expression and function.
Related JoVE Video
The BIPM measurements of the Newtonian constant of gravitation, G.
Philos Trans A Math Phys Eng Sci
PUBLISHED: 09-08-2014
Show Abstract
Hide Abstract
This paper is a complement to the two short papers published in 2001 and 2013 in which we presented the results of the two BIPM determinations of the Newtonian constant of gravitation G. While this review contains no new results, it includes more detailed descriptions of certain key parameters that enter into the determination of G. Following a description of the overall method and the two versions of the experiment, we discuss the properties of the torsion strip, including the effects of anelasticity, then the electrostatic torque transducer, the source and test masses, dimensional metrology, angle measurement, the calculation and measurement of the moment of inertia, calculation of the torque, possible magnetic interactions and finally we discuss uncertainties and correlations in the derivation of a value for G.
Related JoVE Video
Dual reporter MESP1(mCherry/w) -NKX2-5(eGFP/w) hESCs enable studying early human cardiac differentiation.
Stem Cells
PUBLISHED: 09-03-2014
Show Abstract
Hide Abstract
Understanding early differentiation events leading to cardiogenesis is crucial for controlling fate of human pluripotent stem cells and developing protocols that yield sufficient cell numbers for use in regenerative medicine and drug screening. Here, we develop a new tool to visualize patterning of early cardiac mesoderm and cardiomyocyte development in vitro by generating a dual MESP1mCherry(/w) -NKX2-5(eGFP/w) reporter line in human embryonic stem cells (hESCs) and using it to examine signals that lead to formation of cardiac progenitors and subsequent differentiation. MESP1 is a pivotal transcription factor for pre-cardiac mesoderm in the embryo, from which the majority of cardiovascular cells arise. Transcription factor NKX2-5 is expressed upon cardiac crescent formation. Induction of cardiac differentiation in this reporter line resulted in transient expression of MESP1-mCherry, followed by continuous expression of NKX2-5-eGFP. MESP1-mCherry cells showed increased expression of mesodermal and EMT (epithelial-mesenchymal-transition) markers confirming their mesodermal identity. Whole-genome microarray profiling and FACS analysis of MESP1-mCherry cells showed enrichment for mesodermal progenitor cell surface markers PDGFR-?, CD13, and ROR2. No enrichment was found for the previously described KDR+PDGFR-?+ progenitors. MESP1-mCherry derivatives contained an enriched percentage of NKX2-5-eGFP and Troponin T expressing cells, indicating preferential cardiac differentiation; this was enhanced by inhibition of the Wnt-pathway. Furthermore, MESP1-mCherry derivatives harboured smooth muscle cells and endothelial cells, demonstrating their cardiac and vascular differentiation potential under appropriate conditions. The MESP1-NKX2-5 hESC reporter line allows us to identify molecular cues crucial for specification and expansion of human cardiac mesoderm and early progenitors and their differentiation to specific cardiovascular derivatives. Stem Cells 2014.
Related JoVE Video
Serial intravascular ultrasound observations from the Tryton first-in-man study.
EuroIntervention
PUBLISHED: 08-21-2014
Show Abstract
Hide Abstract
To report serial intravascular ultrasound (IVUS) findings of bifurcation lesions treated with the dedicated Tryton Side Branch Stent to assess mechanisms of restenosis.
Related JoVE Video
Parascaris univalens-a victim of large-scale misidentification?
Parasitol. Res.
PUBLISHED: 07-31-2014
Show Abstract
Hide Abstract
The equine ascarid parasite Parascaris equorum is well known as a ubiquitous parasite infecting foals. A sibling species, Parascaris univalens, was first described over 130 years ago, but very little attention has been given to its existence and possible implications for anthelmintic resistance, clinical disease, or host age spectrum. P. univalens only possesses one germ line chromosome pair as opposed to two for P. equorum, but the two species are otherwise considered morphologically identical. For the present study, live worms obtained from the University of Kentucky parasitology horse herd were dissected and identified using karyotyping techniques. With no exception, all specimens (n?=?30) were identified to be P. univalens. Further, the karyotyping technique was adapted to ascarid eggs derived from fecal samples and carried out on samples collected from 25 Thoroughbred foals from three farms in Central Kentucky. P. equorum was not identified among these, whereas P. univalens was found in 17 samples, with the remaining being inconclusive. The mitochondrial genome was sequenced, assembled, and annotated from one male worm identified as P. univalens, and comparison with available sequence reads labeled as P. equorum revealed only 0.16 % nucleotide differences. However, it is unlikely that the sequences available in public databases have been unequivocally identified to species level by karyotyping. Taken together, these data suggest that P. univalens is likely the main species now observed in equines and that perhaps the designation Parascaris spp. should be used unless cytological characterization has confirmed the species.
Related JoVE Video
Controlling the physicochemical state of carbon on graphene using focused electron-beam-induced deposition.
ACS Nano
PUBLISHED: 07-07-2014
Show Abstract
Hide Abstract
Focused electron-beam-induced deposition (FEBID) is a promising nanolithography technique using "direct-write" patterning by carbon line and dot deposits on graphene. Understanding interactions between deposited carbon molecules and graphene enables highly localized modification of graphene properties, which is foundational to the FEBID utility as a nanopatterning tool. In this study, we demonstrate a unique possibility to induce dramatically different adsorption states of FEBID-produced carbon deposits on graphene, through density functional theory calculations and complementary Raman experiments. Specifically, an amorphous carbon deposit formed by direct irradiation of high energy primary electrons exhibits unusually strong interactions with graphene via covalent bonding, whereas the FEBID carbon formed due to low-energy secondary electrons is only weakly interacting with graphene via physisorption. These observations not only are of fundamental importance to basic physical chemistry of FEBID carbon-graphene interactions but also enable the use of selective laser-assisted postdeposition ablation to effectively remove the parasitically deposited, physisorbed carbon films for improving FEBID patterning resolution.
Related JoVE Video
p38 MAPK-inhibited dendritic cells induce superior antitumour immune responses and overcome regulatory T-cell-mediated immunosuppression.
Nat Commun
PUBLISHED: 05-27-2014
Show Abstract
Hide Abstract
Dendritic cell (DC)-based cancer immunotherapy is a promising method, but so far has demonstrated limited clinical benefits. Regulatory T cells (Tregs) represent a major obstacle to cancer immunotherapy approaches. Here we show that inhibiting p38 MAPK during DC differentiation enables DCs to activate tumour-specific effector T cells (Teff), inhibiting the conversion of Treg and compromising Treg inhibitory effects on Teff. Inhibition of p38 MAPK in DCs lowers expression of PPAR?, activating p50 and upregulating OX40L expression in DCs. OX40L/OX40 interactions between DCs and Teff and/or Treg are critical for priming effective and therapeutic antitumour responses. Similarly, p38 MAPK inhibition also augments the T-cell stimulatory capacity of human monocyte-derived DCs in the presence of Treg. These findings contribute to ongoing efforts to improve DC-based immunotherapy in human cancers.
Related JoVE Video
Double hit lymphoma: the MD Anderson Cancer Center clinical experience.
Br. J. Haematol.
PUBLISHED: 04-16-2014
Show Abstract
Hide Abstract
We report our experience with 129 cases of double hit lymphoma (DHL), defined as B-cell lymphoma with translocations and/or extra signals involving MYC plus BCL2 and/or BCL6. All cases were reviewed for histopathological classification. Median age was 62 years (range, 18-85), 84% of patients had advanced-stage disease, and 87% had an International Prognostic Index score ?2. Fourteen patients (11%) had a history of low-grade follicular lymphoma. MYC translocation was present in 81%, and extra signals of MYC in 25% of patients. IGH-BCL2 translocation was present in 84% and extra signals of BCL2 in 12% of patients. Two-year event-free survival (EFS) rates in all patients and patients who received R-CHOP (rituximab, cyclophosphamide, doxorubicin, vincristine, prednisone), R-EPOCH (rituximab, etoposide, prednisone, vincristine, cyclophosphamide, doxorubicin), and R-HyperCVAD/MA (rituximab, hyperfractionated cyclophosphamide, vincristine, doxorubicin, dexamethasone, alternating with cytarabine plus methotrexate) were 33%, 25%, 67% and 32%, respectively. In patients achieving complete response with initial therapy (n = 71), 2-year EFS rates in patients who did (n = 23) or did not (n = 48) receive frontline stem cell transplantation were 68% and 53%, respectively (P = 0·155). The cumulative incidence of central nervous system involvement was 13% at 3 years. Multivariate analysis identified performance status ?2 and bone marrow involvement as independent adverse prognostic factors for EFS and OS. Further research is needed to identify predictive and/or targetable biological markers and novel therapeutic approaches for DHL patients.
Related JoVE Video
Re-evaluation of the inheritance for root-knot nematode resistance in the Upland cotton germplasm line M-120 RNR revealed two epistatic QTLs conferring resistance.
Theor. Appl. Genet.
PUBLISHED: 03-17-2014
Show Abstract
Hide Abstract
We report a second major QTL for root-knot nematode resistance in the highly resistant Upland cotton line M-120RNR and show epistasis between two resistant QTLs with different mechanisms conferring resistance. In an earlier study, we identified a major QTL on Chromosome 11 associated with resistance to root-knot nematode in the M-120 RNR Upland cotton line (Gossypium hirsutum L.) of the Auburn 623 RNR source. Herein, we re-evaluated the genetics of the resistance to root-knot nematode in the M-120 RNR × Pima S-6 population by linkage mapping using recently published SSR markers. The QTL analysis detected two regions significantly associated with the resistance phenotype. In addition to the QTL previously identified on Chromosome 11 (qMi-C11), a major QTL was identified on Chromosome 14 (qMi-C14). The resistance locus on qMi-C11 originated from the Clevewilt parent, while the qMi-C14 locus originated from the other resistant parent, Mexico Wild Jack Jones. The qMi-C14 locus had logarithms of odds score of 17 and accounted for 45 % of the total phenotype variation in egg production. It was also associated with galling index, but the percent variation explained was only 6 %, suggesting that the qMi-C11 locus had a much stronger effect on root gall suppression than egg production, while the qMi-C14 locus had a stronger effect on egg production than galling. The results also suggest that the transgressive segregation observed in the development of Auburn 623 RNR was due to the pyramiding of at least two main effect QTLs as well as an additive-by-additive epistatic effects between the two resistant loci. The SSRs markers tightly linked to the qMi-C11 and qMi-C14 loci will greatly facilitate the improvement of RKN resistance in cotton via marker-assisted breeding.
Related JoVE Video
Subcutaneous and periorbital emphysema following dental procedure.
Ophthal Plast Reconstr Surg
PUBLISHED: 03-12-2014
Show Abstract
Hide Abstract
Subcutaneous emphysema following a dental procedure is a relatively uncommon phenomenon that may produce a dramatic clinical presentation. The advent of high-powered dental equipment that uses directed and highly pressurized air has increased the risk of this complication, but few cases have been described in the ophthalmic literature. The authors present a case of subcutaneous periorbital emphysema in a young woman following a dental procedure. Aside from a temporary ptosis, there were no other ophthalmic complications. An attempt to decompress the air produced minimal relief. The condition resolved spontaneously within 1 week.
Related JoVE Video
Strategies for rapidly mapping proviral integration sites and assessing cardiogenic potential of nascent human induced pluripotent stem cell clones.
Exp. Cell Res.
PUBLISHED: 03-07-2014
Show Abstract
Hide Abstract
Recent methodological advances have improved the ease and efficiency of generating human induced pluripotent stem cells (hiPSCs), but this now typically results in a greater number of hiPSC clones being derived than can be wholly characterized. It is therefore imperative that methods are developed which facilitate rapid selection of hiPSC clones most suited for the downstream research aims. Here we describe a combination of procedures enabling the simultaneous screening of multiple clones to determine their genomic integrity as well as their cardiac differentiation potential within two weeks of the putative reprogrammed colonies initially appearing. By coupling splinkerette-PCR with Ion Torrent sequencing, we could ascertain the number and map the proviral integration sites in lentiviral-reprogrammed hiPSCs. In parallel, we developed an effective cardiac differentiation protocol that generated functional cardiomyocytes within 10 days without requiring line-specific optimization for any of the six independent human pluripotent stem cell lines tested. Finally, to demonstrate the scalable potential of these procedures, we picked 20 nascent iPSC clones and performed these independent assays concurrently. Before the clones required passaging, we were able to identify clones with a single integrated copy of the reprogramming vector and robust cardiac differentiation potential for further analysis.
Related JoVE Video
Programmed DNA elimination in multicellular organisms.
Curr. Opin. Genet. Dev.
PUBLISHED: 02-19-2014
Show Abstract
Hide Abstract
Genetic information typically remains constant in all cells throughout the life cycle of most organisms. However, there are exceptions where DNA elimination is an integral, developmental program for some organisms, associated with generating distinct germline versus somatic genomes. Programmed DNA elimination occurs in unicellular ciliates and diverse metazoa ranging from nematodes to vertebrates. DNA elimination can occur through chromosome breakage and selective loss of chromosome regions or the elimination of individual chromosomes. Recent studies provide compelling evidence that DNA elimination is a novel form of gene silencing, dosage compensation, and sex determination. Further identification of the eliminated sequences, genome changes, and in depth characterization of this phenomenon in diverse metazoans is needed to shed new light on the functions and mechanisms of this regulated process.
Related JoVE Video
Perceptions of dry eye disease management in current clinical practice.
Eye Contact Lens
PUBLISHED: 02-11-2014
Show Abstract
Hide Abstract
To assess the perceptions of eye care providers regarding the clinical management of dry eye.
Related JoVE Video
SIRPA, VCAM1 and CD34 identify discrete lineages during early human cardiovascular development.
Stem Cell Res
PUBLISHED: 01-30-2014
Show Abstract
Hide Abstract
The study of human cardiogenesis would benefit from a detailed cell lineage fate map akin to that established for the haematopoietic lineages. Here we sought to define cell lineage relationships based on the expression of NKX2-5 and the cell surface markers VCAM1, SIRPA and CD34 during human cardiovascular development. Expression of NKX2-5(GFP) was used to identify cardiac progenitors and cardiomyocytes generated during the differentiation of NKX2-5(GFP/w) human embryonic stem cells (hESCs). Cardiovascular cell lineages sub-fractionated on the basis of SIRPA, VCAM1 and CD34 expression were assayed for differentiation potential and gene expression. The NKX2-5(pos)CD34(pos) population gave rise to endothelial cells that rapidly lost NKX2-5 expression in culture. Conversely, NKX2-5 expression was maintained in myocardial committed cells, which progressed from being NKX2-5(pos)SIRPA(pos) to NKX2-5(pos)SIRPA(pos)VCAM1(pos). Up-regulation of VCAM1 was accompanied by the expression of myofilament markers and reduced clonal capacity, implying a restriction of cell fate potential. Combinatorial expression of NKX2-5, SIRPA, VCAM1 and CD34 can be used to define discrete stages of cardiovascular cell lineage differentiation. These markers identify specific stages of cardiomyocyte and endothelial lineage commitment and, thus provide a scaffold for establishing a fate map of early human cardiogenesis.
Related JoVE Video
Generation of induced pluripotent stem cells from conjunctiva.
Graefes Arch. Clin. Exp. Ophthalmol.
PUBLISHED: 01-14-2014
Show Abstract
Hide Abstract
The objective of this study was to determine whether cells from the conjunctiva could be reprogrammed into induced pluripotent stem (iPS) cells, providing an alternative source of stem cells.
Related JoVE Video
Genome-wide association study for age-related hearing loss (AHL) in the mouse: a meta-analysis.
J. Assoc. Res. Otolaryngol.
PUBLISHED: 01-13-2014
Show Abstract
Hide Abstract
Age-related hearing loss (AHL) is characterized by a symmetric sensorineural hearing loss primarily in high frequencies and individuals have different levels of susceptibility to AHL. Heritability studies have shown that the sources of this variance are both genetic and environmental, with approximately half of the variance attributable to hereditary factors as reported by Huag and Tang (Eur Arch Otorhinolaryngol 267(8):1179-1191, 2010). Only a limited number of large-scale association studies for AHL have been undertaken in humans, to date. An alternate and complementary approach to these human studies is through the use of mouse models. Advantages of mouse models include that the environment can be more carefully controlled, measurements can be replicated in genetically identical animals, and the proportion of the variability explained by genetic variation is increased. Complex traits in mouse strains have been shown to have higher heritability and genetic loci often have stronger effects on the trait compared to humans. Motivated by these advantages, we have performed the first genome-wide association study of its kind in the mouse by combining several data sets in a meta-analysis to identify loci associated with age-related hearing loss. We identified five genome-wide significant loci (<10(-6)). One of these loci confirmed a previously identified locus (ahl8) on distal chromosome 11 and greatly narrowed the candidate region. Specifically, the most significant associated SNP is located 450 kb upstream of Fscn2. These data confirm the utility of this approach and provide new high-resolution mapping information about variation within the mouse genome associated with hearing loss.
Related JoVE Video
From virtual to clinical: The discovery of PGN-1531, a novel antagonist of the prostanoid EP4 receptor.
Bioorg. Med. Chem. Lett.
PUBLISHED: 01-08-2014
Show Abstract
Hide Abstract
In this Letter, we present the results of a hit-finding and lead optimization programme against the EP4 receptor (EP4R). In a short time period, we were able to discover five structurally diverse series of hit compounds using a combination of virtual screening methods. The most favoured hit, compound 6, was demonstrated to be a competitive antagonist of the EP4R. Compound 73 was identified following several rounds of optimization, which centred on improving both the primary EP4R affinity and selectivity against the related EP2R as well as the aqueous solubility. This work culminated in the preparation of PGN-1531, the sodium salt of 73, which showed a marked improvement in solubility (>10 mg/mL). PGN-1531 is a potent and selective antagonist at EP4Rs in vitro and in vivo, with the potential to alleviate the symptoms of migraine that result from cerebral vasodilatation.
Related JoVE Video
Autoimmunity in the pathogenesis and treatment of keratoconjunctivitis sicca.
Curr Allergy Asthma Rep
PUBLISHED: 01-08-2014
Show Abstract
Hide Abstract
Dry eye is a chronic corneal disease that impacts the quality of life of many older adults. Keratoconjunctivitis sicca (KCS), a form of aqueous-deficient dry eye, is frequently associated with Sjögren's syndrome and mechanisms of autoimmunity. For KCS and other forms of dry eye, current treatments are limited, with many medications providing only symptomatic relief rather than targeting the pathophysiology of disease. Here, we review proposed mechanisms in the pathogenesis of autoimmune-based KCS: genetic susceptibility and disruptions in antigen recognition, immune response, and immune regulation. By understanding the mechanisms of immune dysfunction through basic science and translational research, potential drug targets can be identified. Finally, we discuss current dry eye therapies as well as promising new treatment options and drug therapy targets.
Related JoVE Video
Pyoverdine synthesis by the Mn(II)-oxidizing bacterium Pseudomonas putida GB-1.
Front Microbiol
PUBLISHED: 01-01-2014
Show Abstract
Hide Abstract
When iron-starved, the Mn(II)-oxidizing bacteria Pseudomonas putida strains GB-1 and MnB1 produce pyoverdines (PVDGB-1 and PVDMnB1), siderophores that both influence iron uptake and inhibit manganese(II) oxidation by these strains. To explore the properties and genetics of a PVD that can affect manganese oxidation, LC-MS/MS, and various siderotyping techniques were used to identify the peptides of PVDGB-1 and PVDMnB1 as being (for both PVDs): chromophore-Asp-Lys-OHAsp-Ser-Gly-aThr-Lys-cOHOrn, resembling a structure previously reported for P. putida CFML 90-51, which does not oxidize Mn. All three strains also produced an azotobactin and a sulfonated PVD, each with the peptide sequence above, but with unknown regulatory or metabolic effects. Bioinformatic analysis of the sequenced genome of P. putida GB-1 suggested that a particular non-ribosomal peptide synthetase (NRPS), coded by the operon PputGB1_4083-4086, could produce the peptide backbone of PVDGB-1. To verify this prediction, plasmid integration disruption of PputGB1_4083 was performed and the resulting mutant failed to produce detectable PVD. In silico analysis of the modules in PputGB1_4083-4086 predicted a peptide sequence of Asp-Lys-Asp-Ser-Ala-Thr-Lsy-Orn, which closely matches the peptide determined by MS/MS. To extend these studies to other organisms, various Mn(II)-oxidizing and non-oxidizing isolates of P. putida, P. fluorescens, P. marincola, P. fluorescens-syringae group, P. mendocina-resinovorans group, and P. stutzerii group were screened for PVD synthesis. The PVD producers (12 out of 16 tested strains) were siderotyped and placed into four sets of differing PVD structures, some corresponding to previously characterized PVDs and some to novel PVDs. These results combined with previous studies suggested that the presence of OHAsp or the flexibility of the pyoverdine polypeptide may enable efficient binding of Mn(III).
Related JoVE Video
Meta-analysis identifies gene-by-environment interactions as demonstrated in a study of 4,965 mice.
PLoS Genet.
PUBLISHED: 01-01-2014
Show Abstract
Hide Abstract
Identifying environmentally-specific genetic effects is a key challenge in understanding the structure of complex traits. Model organisms play a crucial role in the identification of such gene-by-environment interactions, as a result of the unique ability to observe genetically similar individuals across multiple distinct environments. Many model organism studies examine the same traits but under varying environmental conditions. For example, knock-out or diet-controlled studies are often used to examine cholesterol in mice. These studies, when examined in aggregate, provide an opportunity to identify genomic loci exhibiting environmentally-dependent effects. However, the straightforward application of traditional methodologies to aggregate separate studies suffers from several problems. First, environmental conditions are often variable and do not fit the standard univariate model for interactions. Additionally, applying a multivariate model results in increased degrees of freedom and low statistical power. In this paper, we jointly analyze multiple studies with varying environmental conditions using a meta-analytic approach based on a random effects model to identify loci involved in gene-by-environment interactions. Our approach is motivated by the observation that methods for discovering gene-by-environment interactions are closely related to random effects models for meta-analysis. We show that interactions can be interpreted as heterogeneity and can be detected without utilizing the traditional uni- or multi-variate approaches for discovery of gene-by-environment interactions. We apply our new method to combine 17 mouse studies containing in aggregate 4,965 distinct animals. We identify 26 significant loci involved in High-density lipoprotein (HDL) cholesterol, many of which are consistent with previous findings. Several of these loci show significant evidence of involvement in gene-by-environment interactions. An additional advantage of our meta-analysis approach is that our combined study has significantly higher power and improved resolution compared to any single study thus explaining the large number of loci discovered in the combined study.
Related JoVE Video
Bilateral Concordance of the Fundus Hyperautofluorescent Ring in Typical Retinitis Pigmentosa Patients.
Ophthalmic Genet.
PUBLISHED: 10-12-2013
Show Abstract
Hide Abstract
Abstract Background: It has long been assumed that in retinitis pigmentosa, disease presentation and progression are symmetrical. This study investigated whether hyperautofluorescent ring size, one known marker of disease progression, is symmetrical in typical RP patients. Materials and Methods: A total of 88 patients with typical retinitis pigmentosa were enrolled in the study. Each presented with a hyperautofluorescent ring when imaged at baseline with fundus autofluorescence (AF). Vertical and horizontal diameters were analyzed according to mode of inheritance and age group. Seven of 88 patients had data missing in one eye and were excluded from further analysis. Results: There was no significant relationship between hyperautofluorescent ring diameter and inheritance mode. There was a tendency toward smaller ring size with age and 3.7% of subjects displayed marked asymmetry in ring size between right and left eyes, although their electroretinogram results did not differ. Overall, when patients were considered as a group, there was a high correlation between right and left eyes horizontal and vertical diameters (r?=?0.99, p?
Related JoVE Video
Transcription in Pronuclei and One- to Four-Cell Embryos Drives Early Development in a Nematode.
Curr. Biol.
PUBLISHED: 10-04-2013
Show Abstract
Hide Abstract
A long-standing view of development is that transcription is silenced in the oocyte until early divisions in the embryo. The point at which major transcription is reactivated varies between organisms, but is usually after the two-cell stage. However, this model may not be universal.
Related JoVE Video
Complete Genome Sequence of Staphylococcus aureus Tager 104, a Sequence Type 49 Ancestor.
Genome Announc
PUBLISHED: 09-14-2013
Show Abstract
Hide Abstract
We report here the complete genome sequence of Staphylococcus aureus Tager 104, originally isolated from a cutaneous abscess in 1947 by Morris Tager. Sequence typing of the strain revealed its membership in sequence type 49 (ST49), a previously unknown multilocus sequence type (MLST) in clinical samples.
Related JoVE Video
Ceftriaxone increases glutamate uptake and reduces striatal tyrosine hydroxylase loss in 6-OHDA Parkinsons model.
Mol. Neurobiol.
PUBLISHED: 09-06-2013
Show Abstract
Hide Abstract
Excess glutamatergic neurotransmission may contribute to excitotoxic loss of nigrostriatal neurons in Parkinsons disease (PD). Here, we determined if increasing glutamate uptake could reduce the extent of tyrosine hydroxylase (TH) loss in PD progression. The beta-lactam antibiotic, ceftriaxone, increases the expression of glutamate transporter 1 (GLT-1), a glutamate transporter that plays a major role in glutamate clearance in central nervous system and may attenuate adverse behavioral or neurobiological function in other neurodegenerative disease models. In association with >80 % TH loss, we observed a significant decrease in glutamate uptake in the established 6-hydroxydopamine (6-OHDA) PD model. Ceftriaxone (200 mg/kg, i.p.) increased striatal glutamate uptake with >5 consecutive days of injection in nonlesioned rats and lasted out to 14 days postinjection, a time beyond that required for 6-OHDA to produce >70 % TH loss (?9 days). When ceftriaxone was given at the time of 6-OHDA, TH loss was ?57 % compared to ?85 % in temporally matched vehicle-injected controls and amphetamine-induced rotation was reduced about 2-fold. This attenuation of TH loss was associated with increased glutamate uptake, increased GLT-1 expression, and reduced Serine 19 TH phosphorylation, a calcium-dependent target specific for nigrostriatal neurons. These results reveal that glutamate uptake can be targeted in a PD model, decrease the rate of TH loss in a calcium-dependent manner, and attenuate locomotor behavior associated with 6-OHDA lesion. Given that detection of reliable PD markers will eventually be employed in susceptible populations, our results give credence to the possibility that increasing glutamate uptake may prolong the time period before locomotor impairment occurs.
Related JoVE Video
Therapeutic margins in a novel preclinical model of retinitis pigmentosa.
J. Neurosci.
PUBLISHED: 08-16-2013
Show Abstract
Hide Abstract
The third-most common cause of autosomal recessive retinitis pigmentosa (RP) is due to defective cGMP phosphodiesterase-6 (PDE6). Previous work using viral gene therapy on PDE6-mutant mouse models demonstrated photoreceptors can be rescued if administered before degeneration. However, whether visual function can be rescued after degeneration onset has not been addressed. This is a clinically important question, as newly diagnosed patients exhibit considerable loss of rods and cones in their peripheral retinas. We have generated and characterized a tamoxifen inducible Cre-loxP rescue allele, Pde6b(Stop), which allows us to temporally correct PDE6-deficiency. Whereas untreated mutants exhibit degeneration, activation of Cre-loxP recombination in early embryogenesis produced stable long-term rescue. Reversal at later time-points showed partial long-term or short-lived rescue. Our results suggest stable restoration of retinal function by gene therapy can be achieved if a sufficient number of rods are treated. Because patients are generally diagnosed after extensive loss of rods, the success of clinical trials may depend on identifying patients as early as possible to maximize the number of treatable rods.
Related JoVE Video
Racial and ethnic differences in the physiology and clinical symptoms of menopause.
Semin. Reprod. Med.
PUBLISHED: 08-09-2013
Show Abstract
Hide Abstract
More than 4 million menopausal women are from ethnic minority groups. Over the past 25 years, recognition of the importance of social, emotional, and physical changes of midlife to womens long-term health and well-being has emerged. Multiple factors influence how a woman perceives menopausal changes and what she addresses as associated symptoms. Factors such as educational level to socioeconomic status, health-related factors, stress, and marital status influence these choices. Increasingly, researchers are reporting on the impact of race and ethnicity on menopausal symptoms. Understanding similarities and differences among womens perceptions, attitudes, and expectations surrounding menopause improves delivery of culturally appropriate care and promotes lifestyles that may decrease symptoms and increase quality of life. Historically, the majority of the research in this area has been conducted in Western countries with clinical samples of women predominantly from European backgrounds. Thus, this population has shaped the emerging clinical picture of the midlife menopausal transition. Recently, studies of non-European women, both in the United States and internationally, indicate significant variations in their experiences during this transition, but these cultural differences have not broadened the understanding of the meaning of this universal experience. To date, there are still large knowledge gaps in race, ethnic, and cultural differences in menopausal health. The content of this review summarizes the current body of knowledge on racial differences in the menopause experience.
Related JoVE Video
hsa-miR29b, a critical downstream target of non-canonical Wnt signaling, plays an anti-proliferative role in non-small cell lung cancer cells via targeting MDM2 expression.
Biol Open
PUBLISHED: 07-15-2013
Show Abstract
Hide Abstract
In non-small cell lung cancer cell lines, activation of ?-catenin independent signaling, via Wnt7a/Frizzled9 signaling, leads to reversal of cellular transformation, reduced anchorage-independent growth and induction of epithelial differentiation. miRNA expression profiling on a human lung adenocarcinoma cell line (A549) identified hsa-miR29b as an important downstream target of Wnt7a/Frizzled9 signaling. We show herein that hsa-miR29b expression is lost in non-small cell lung cancer (NSCLC) cell lines and stimulation of ?-catenin independent signaling, via Wnt7a expression, in NSCLC cell lines results in increased expression of hsa-miR29b. Surprisingly, we also identify specific regulation of hsa-miR29b by Wnt7a but not by Wnt3, a ligand for ?-catenin-dependent signaling. Interestingly, knockdown of hsa-miR29b was enough to abrogate the tumor suppressive effects of Wnt7a/Frizzled9 signaling in NSCLC cells, suggesting that hsa-miR29b is an important mediator of ?-catenin independent signaling. Finally, we show for the first time that hsa-miR29b plays an important role as a tumor suppressor in lung cancer by targeting murine double mutant 2 (MDM2), revealing novel nodes for Wnt7a/Frizzled9-mediated regulation of NSCLC cell proliferation.
Related JoVE Video
Passive gravitational sedimentation of peripheral blood increases the sensitivity of microscopic detection of malaria.
Asian Pac J Trop Med
PUBLISHED: 06-15-2013
Show Abstract
Hide Abstract
To determine if passive gravitational sedimentation of blood samples, followed by buffy coat thin smear preparation could increase the sensitivity of malaria diagnosis when compared to conventional thin smear preparation without the additional cost of centrifuges or molecular diagnostics.
Related JoVE Video
Connective tissue growth factor regulates adipocyte differentiation of mesenchymal stromal cells and facilitates leukemia bone marrow engraftment.
Blood
PUBLISHED: 06-05-2013
Show Abstract
Hide Abstract
Mesenchymal stromal cells (MSCs) are a major component of the leukemia bone marrow (BM) microenvironment. Connective tissue growth factor (CTGF) is highly expressed in MSCs, but its role in the BM stroma is unknown. Therefore, we knocked down (KD) CTGF expression in human BM-derived MSCs by CTGF short hairpin RNA. CTGF KD MSCs exhibited fivefold lower proliferation compared with control MSCs and had markedly fewer S-phase cells. CTGF KD MSCs differentiated into adipocytes at a sixfold higher rate than controls in vitro and in vivo. To study the effect of CTGF on engraftment of leukemia cells into BM, an in vivo model of humanized extramedullary BM (EXM-BM) was developed in NOD/SCID/IL-2rg(null) mice. Transplanted Nalm-6 or Molm-13 human leukemia cells engrafted at a threefold higher rate in adipocyte-rich CTGF KD MSC-derived EXM-BM than in control EXM-BM. Leptin was found to be highly expressed in CTGF KD EXM-BM and in BM samples of patients with acute myeloid and acute lymphoblastic leukemia, whereas it was not expressed in normal controls. Given the established role of the leptin receptor in leukemia cells, the data suggest an important role of CTGF in MSC differentiation into adipocytes and of leptin in homing and progression of leukemia.
Related JoVE Video
WNT3A Promotes Hematopoietic or Mesenchymal Differentiation from hESCs Depending on the Time of Exposure.
Stem Cell Reports
PUBLISHED: 06-04-2013
Show Abstract
Hide Abstract
We investigated the role of canonical WNT signaling in mesoderm and hematopoietic development from human embryonic stem cells (hESCs) using a recombinant human protein-based differentiation medium (APEL). In contrast to prior studies using less defined culture conditions, we found that WNT3A alone was a poor inducer of mesoderm. However, WNT3A synergized with BMP4 to accelerate mesoderm formation, increase embryoid body size, and increase the number of hematopoietic blast colonies. Interestingly, inclusion of WNT3A or a GSK3 inhibitor in methylcellulose colony-forming assays at 4 days of differentiation abrogated blast colony formation but supported the generation of mesospheres that expressed genes associated with mesenchymal lineages. Mesospheres differentiated into cells with characteristics of bone, fat, and smooth muscle. These studies identify distinct effects for WNT3A, supporting the formation of hematopoietic or mesenchymal lineages from human embryonic stem cells, depending upon differentiation stage at the time of exposure.
Related JoVE Video
Generation of induced pluripotent stem cells from human foetal fibroblasts using the Sleeping Beauty transposon gene delivery system.
Differentiation
PUBLISHED: 05-13-2013
Show Abstract
Hide Abstract
Transposon gene delivery systems offer an alternative, non-viral-based approach to generate induced pluripotent stem cells (iPSCs). Here we used the Sleeping Beauty (SB) transposon to generate four human iPSC lines from foetal fibroblasts. In contrast to other gene delivery systems, the SB transposon does not exhibit an integration bias towards particular genetic elements, thereby reducing the risk of insertional mutagenesis. Furthermore, unlike the alternative transposon piggyBac, SB has no SB-like elements within the human genome, minimising the possibility of mobilising endogenous transposon elements. All iPSC lines exhibited the expected characteristics of pluripotent human cells, including the ability to differentiate to derivatives of all three germ layers in vitro. Re-expression of the SB transposase in the iPSCs after reprogramming resulted in the mobilisation of some of the transposons. These results indicate that the SB transposon system is a useful addition to methods for generating human iPSCs, both for basic and applied biomedical research, and in the context of future therapeutic application.
Related JoVE Video
Isogenic human pluripotent stem cell pairs reveal the role of a KCNH2 mutation in long-QT syndrome.
EMBO J.
PUBLISHED: 05-03-2013
Show Abstract
Hide Abstract
Patient-specific induced pluripotent stem cells (iPSCs) will assist research on genetic cardiac maladies if the disease phenotype is recapitulated in vitro. However, genetic background variations may confound disease traits, especially for disorders with incomplete penetrance, such as long-QT syndromes (LQTS). To study the LQT2-associated c.A2987T (N996I) KCNH2 mutation under genetically defined conditions, we derived iPSCs from a patient carrying this mutation and corrected it. Furthermore, we introduced the same point mutation in human embryonic stem cells (hESCs), generating two genetically distinct isogenic pairs of LQTS and control lines. Correction of the mutation normalized the current (IKr) conducted by the HERG channel and the action potential (AP) duration in iPSC-derived cardiomyocytes (CMs). Introduction of the same mutation reduced IKr and prolonged the AP duration in hESC-derived CMs. Further characterization of N996I-HERG pathogenesis revealed a trafficking defect. Our results demonstrated that the c.A2987T KCNH2 mutation is the primary cause of the LQTS phenotype. Precise genetic modification of pluripotent stem cells provided a physiologically and functionally relevant human cellular context to reveal the pathogenic mechanism underlying this specific disease phenotype.
Related JoVE Video
Maritime adaptations and dietary variation in prehistoric Western Alaska: stable isotope analysis of permafrost-preserved human hair.
Am. J. Phys. Anthropol.
PUBLISHED: 03-28-2013
Show Abstract
Hide Abstract
The reconstruction of diet and subsistence strategies is integral in understanding early human colonizations and cultural adaptations, especially in the Arctic-one of the last areas of North America to be permanently inhabited. However, evidence for early subsistence practices in Western Alaska varies, particularly with regards to the emergence, importance, and intensity of sea mammal hunting. Here, we present stable carbon and nitrogen isotope data from permafrost-preserved human hair from two new prehistoric sites in Western Alaska, providing a direct measure of diet. The isotope evidence indicates a heavy reliance on sea mammal protein among the earlier Norton-period group (1,750 ± 40 cal BP), confirming that the complex hunting technologies required to intensively exploit these animals were most likely already in place in this region by at least the beginning of 1st millennium AD. In contrast, analysis of the more recent Thule-period hair samples (650 ± 40 cal BP; 570 ± 30 cal BP) reveals a more mixed diet, including terrestrial animal protein. Sequential isotope analysis of two longer human hair locks indicates seasonal differences in diet in a single Norton-period individual but demonstrates little dietary variation in a Thule-period individual. These analyses provide direct evidence for dietary differences among Alaskas early Eskimo groups and confirm the antiquity of specialized sea mammal hunting and procurement technologies. The results of this study have implications for our understanding of human adaptation to maritime and high-latitude environments, and the geographical and temporal complexity in early Arctic subsistence.
Related JoVE Video
Ocular disease, knowledge and technology applications in patients with diabetes.
Am. J. Med. Sci.
PUBLISHED: 03-28-2013
Show Abstract
Hide Abstract
An estimated 25.8 million children and adults in the United States, approximately 8.3% of the population, have diabetes. Diabetes prevalence varies by race and ethnicity. African Americans have the highest prevalence (12.6%), followed closely by Hispanics (11.8%), Asian Americans (8.4%) and whites (7.1%). The purpose of this article was to discuss the ocular complications of diabetes, the cultural and racial differences in diabetes knowledge and the role of telemedicine as a means to reach the undeserved who are at risk of complications. Information on the pathophysiology of ocular disease in patients with diabetes and the role of telemedicine in diabetes care was derived from a literature review. National Institutes of Health online resources were queried to present data on the racial and cultural understandings of diabetes and diabetes-related complications. The microvascular ocular complications of diabetes are discussed for retinopathy, cataracts, glaucoma and ocular surface disease. Racial and cultural differences in knowledge of recommended self-care practices are presented. These differences, in part, may explain health disparities and the increased risk of diabetes and its complications in rural minority communities. Finally, advances in telemedicine technology are discussed that show improvements in metabolic control and cardiovascular risk in adults with type 2 diabetes. Improving provider and patient understanding of diabetes complications may improve management and self-care practices that are important for diabetes control. Telemedicine may improve access to diabetes specialists and may improve self-management education and diabetes control particularly in rural and underserved communities.
Related JoVE Video
Nonstereotyped lymphoma B cell receptors recognize vimentin as a shared autoantigen.
J. Immunol.
PUBLISHED: 03-27-2013
Show Abstract
Hide Abstract
Ag activation of the BCR may play a role in the pathogenesis of human follicular lymphoma (FL) and other B cell malignancies. However, the nature of the Ag(s) recognized by tumor BCRs has not been well studied. In this study, we used unbiased approaches to demonstrate that 42 (19.35%) of 217 tested FL Igs recognized vimentin as a shared autoantigen. The epitope was localized to the N-terminal region of vimentin for all vimentin-reactive tumor Igs. We confirmed specific binding to vimentin by using recombinant vimentin and by performing competitive inhibition studies. Furthermore, using indirect immunofluorescence staining, we showed that the vimentin-reactive tumor Igs colocalized with an anti-vimentin mAb in HEp-2 cells. The reactivity to N-terminal vimentin of IgG FL Igs was significantly higher than that of IgM FL Igs (30.4 versus 10%; p = 0.0022). However, vimentin-reactive FL Igs did not share CDR3 motifs and were not homologous. Vimentin was expressed in the T cell-rich regions of FL, suggesting that vimentin is available for binding with tumor BCRs within the tumor microenvironment. Vimentin was also frequently recognized by mantle cell lymphoma and multiple myeloma Igs. Our results demonstrate that vimentin is a shared autoantigen recognized by nonstereotyped FL BCRs and by the Igs of mantle cell lymphoma and multiple myeloma and suggest that vimentin may play a role in the pathogenesis of multiple B cell malignancies. These findings may lead to a better understanding of the biology and natural history of FL and other B cell malignancies.
Related JoVE Video
Primary Hodgkin lymphoma of the central nervous system: Two case reports and review of the literature.
Neuropathology
PUBLISHED: 03-05-2013
Show Abstract
Hide Abstract
CNS involvement by systemic Hodgkin lymphoma (HL) is quite rare, but the disease limited to the CNS is an exceptionally rare entity. The incidence of CNS-HL has been estimated at 0.2-0.5% of cases, but a more recent study has modified that figure to less than 0.02%. Like the conventional form, the diagnosis of primary CNS-HL rests upon distinct morphological and immunohistochemical characteristics, including diagnostic Reed-Sternberg cells, in addition to staging studies demonstrating a lack of disease elsewhere. The paucity of cases in the literature precludes reliable clinical and demographic data, as well as a consensus on treatment and prognosis. We present two cases of primary cerebellar HL, one with 10-year follow-up, and a relevant review of the literature.
Related JoVE Video
Hidden in plain sight: discovery of sheath-forming, iron-oxidizing Zetaproteobacteria at Loihi Seamount, Hawaii, USA.
FEMS Microbiol. Ecol.
PUBLISHED: 02-28-2013
Show Abstract
Hide Abstract
Lithotrophic iron-oxidizing bacteria (FeOB) form microbial mats at focused flow or diffuse flow vents in deep-sea hydrothermal systems where Fe(II) is a dominant electron donor. These mats composed of biogenically formed Fe(III)-oxyhydroxides include twisted stalks and tubular sheaths, with sheaths typically composing a minor component of bulk mats. The micron diameter Fe(III)-oxyhydroxide-containing tubular sheaths bear a strong resemblance to sheaths formed by the freshwater FeOB, Leptothrix ochracea. We discovered that veil-like surface layers present in iron-mats at the Loihi Seamount were dominated by sheaths (40-60% of total morphotypes present) compared with deeper (> 1 cm) mat samples (0-16% sheath). By light microscopy, these sheaths appeared nearly identical to those of L. ochracea. Clone libraries of the SSU rRNA gene from this top layer were dominated by Zetaproteobacteria, and lacked phylotypes related to L. ochracea. In mats with similar morphologies, terminal-restriction fragment length polymorphism (T-RFLP) data along with quantitative PCR (Q-PCR) analyses using a Zetaproteobacteria-specific primer confirmed the presence and abundance of Zetaproteobacteria. A Zetaproteobacteria fluorescence in situ hybridization (FISH) probe hybridized to ensheathed cells (4% of total cells), while a L. ochracea-specific probe and a Betaproteobacteria probe did not. Together, these results constitute the discovery of a novel group of marine sheath-forming FeOB bearing a striking morphological similarity to L. ochracea, but belonging to an entirely different class of Proteobacteria.
Related JoVE Video
Genome-wide association mapping of blood cell traits in mice.
Mamm. Genome
PUBLISHED: 01-11-2013
Show Abstract
Hide Abstract
Genetic variations in blood cell parameters can impact clinical traits. We report here the mapping of blood cell traits in a panel of 100 inbred strains of mice of the Hybrid Mouse Diversity Panel (HMDP) using genome-wide association (GWA). We replicated a locus previously identified in using linkage analysis in several genetic crosses for mean corpuscular volume (MCV) and a number of other red blood cell traits on distal chromosome 7. Our peak for SNP association to MCV occurred in a linkage disequilibrium (LD) block spanning from 109.38 to 111.75 Mb that includes Hbb-b1, the likely causal gene. Altogether, we identified five loci controlling red blood cell traits (on chromosomes 1, 7, 11, 12, and 16), and four of these correspond to loci for red blood cell traits reported in a recent human GWA study. For white blood cells, including granulocytes, monocytes, and lymphocytes, a total of six significant loci were identified on chromosomes 1, 6, 8, 11, 12, and 15. An average of ten candidate genes were found at each locus and those were prioritized by examining functional variants in the HMDP such as missense and expression variants. These results provide intermediate phenotypes and candidate loci for genetic studies of atherosclerosis and cancer as well as inflammatory and immune disorders in mice.
Related JoVE Video
Persistent antigen at vaccination sites induces tumor-specific CD8? T cell sequestration, dysfunction and deletion.
Nat. Med.
PUBLISHED: 01-02-2013
Show Abstract
Hide Abstract
To understand why cancer vaccine-induced T cells often do not eradicate tumors, we studied immune responses in mice vaccinated with gp100 melanoma peptide in incomplete Freunds adjuvant (peptide/IFA), which is commonly used in clinical cancer vaccine trials. Peptide/IFA vaccination primed tumor-specific CD8(+) T cells, which accumulated not in tumors but rather at the persisting, antigen-rich vaccination site. Once there, primed T cells became dysfunctional and underwent antigen-driven, interferon-? (IFN-?)- and Fas ligand (FasL)-mediated apoptosis, resulting in hyporesponsiveness to subsequent vaccination. Provision of CD40-specific antibody, Toll-like receptor 7 (TLR7) agonist and interleukin-2 (IL-2) reduced T cell apoptosis but did not prevent vaccination-site sequestration. A nonpersisting vaccine formulation shifted T cell localization toward tumors, inducing superior antitumor activity while reducing systemic T cell dysfunction and promoting memory formation. These data show that persisting vaccine depots can induce specific T cell sequestration, dysfunction and deletion at vaccination sites; short-lived formulations may overcome these limitations and result in greater therapeutic efficacy of peptide-based cancer vaccines.
Related JoVE Video
Protective cardiovascular and renal actions of vitamin D and estrogen.
Front Biosci (Schol Ed)
PUBLISHED: 01-02-2013
Show Abstract
Hide Abstract
Both basic science and clinical studies support the concept that vitamin D deficiency is involved in the pathogenesis of cardiovascular and renal diseases through its association with diabetes, obesity, and hypertension. Understanding the underlying mechanisms may provide a rationale for advocating adequate intake of vitamin D and calcium in all populations, thereby preventing many chronic diseases. This review explores the effect of vitamin D deficiency in the development of cardiovascular and renal diseases, and the role of vitamin D supplementation on cardiovascular outcomes. In addition, it highlights the importance of vitamin D intake for the prevention of adverse long-term health consequences, and in ways to facilitate the management of cardiovascular disease. This is particularly true for African American and postmenopausal women, who are at added risk for cardiovascular disease. We suggest that the negative cardiovascular effects of low vitamin D in postmenopausal women could be improved by a combined treatment of vitamin D and sex steroids acting through endothelium-dependent and/or -independent mechanisms, resulting in the generation of nitric oxide and calcitonin gene-related peptide (CGRP).
Related JoVE Video
Infection prevention promotion program based on the PRECEDE model: improving hand hygiene behaviors among healthcare personnel.
Infect Control Hosp Epidemiol
PUBLISHED: 12-19-2011
Show Abstract
Hide Abstract
Healthcare-associated infections (HAIs) result in significant morbidity and mortality. Hand hygiene remains a cornerstone intervention for preventing HAIs. Unfortunately, adherence to hand hygiene guidelines among healthcare personnel is poor.
Related JoVE Video
shRNA knockdown of guanylate cyclase 2e or cyclic nucleotide gated channel alpha 1 increases photoreceptor survival in a cGMP phosphodiesterase mouse model of retinitis pigmentosa.
J. Cell. Mol. Med.
PUBLISHED: 12-13-2011
Show Abstract
Hide Abstract
In vertebrate rods, dark and light conditions produce changes in guanosine 3,5-cyclic monophosphate (cGMP) and calcium (Ca(2+) ) levels, which are regulated by the opposing function of several proteins. During the recovery of a bright flash, guanylate cyclase (GUCY) helps raise cGMP to levels that open cGMP-gated calcium sodium channels (CNG) to increase Na(+) and Ca(2+) influx in the outer segment. In contrast, light activates cGMP phosphodiesterase 6 (PDE6) causing rapid hydrolysis of cGMP, CNG closure, and reduced Na(+) and Ca(2+) levels. In?Pde6b?mouse models of retinitis pigmentosa (RP), photoreceptor death is preceded by abnormally high cGMP and Ca(2+) levels, likely because of continued synthesis of cGMP by guanylate cyclases and unregulated influx of Ca(2+) to toxic levels through CNG channels. To reverse the effects of?Pde6b?loss of function, we employed an shRNA knockdown approach to reduce the expression of?Gucy2e?or?Cnga1?in?Pde6b(H620Q) photoreceptors prior to degeneration.?Gucy2e- or?Cnga1-shRNA lentiviral-mediated knockdown GUCY2E and CNGA1 expression increase visual function and photoreceptor survival in?Pde6b(H620Q) mice. We demonstrated that effective knockdown of?GUCY2E?and?CNGA1?expression to counteract loss of PDE6 function may develop into a valuable approach for treating some patients with RP.
Related JoVE Video
Effect of nicotine on body composition in mice.
J. Endocrinol.
PUBLISHED: 12-02-2011
Show Abstract
Hide Abstract
Nicotine induces weight loss in both humans and rodents consuming a regular diet; however, the effect of nicotine on body weight and fat composition in rodents consuming a high-fat diet (HFD) has not been well studied. Thus, this study examined the effect of nicotine vs saline on body weight and fat composition in mice fed with either an HFD (62% of kcal from fat) or a standard normal chow diet (NCD) for 7 weeks. Nicotine dose dependently reduced body weight gain in mice that consumed both diets, but this effect was significantly greater in mice on the HFD. Caloric intake was decreased in nicotine-treated mice. Estimates of energy intake suggested that decreased caloric intake accounted for all the reduced weight gain in mice on an NCD and 66% of the reduced weight gain on an HFD. Computed tomography analysis for fat distribution demonstrated that nicotine was effective in reducing abdominal fat in mice that consumed the HFD, with nicotine treatment leading to lower visceral fat. The effect of nicotine on weight loss in mice on an HFD was completely blocked by mecamylamine, a nonselective nicotinic acetylcholine receptor (nAChR) antagonist, but only partially blocked by the ?4?2 nAChR partial agonist/antagonist, varenicline. We conclude that nicotine is effective in preventing HFD-induced weight gain and abdominal fat accumulation.
Related JoVE Video
Systems genetics of susceptibility to obesity-induced diabetes in mice.
Physiol. Genomics
PUBLISHED: 10-18-2011
Show Abstract
Hide Abstract
Inbred strains of mice are strikingly different in susceptibility to obesity-driven diabetes. For instance, deficiency in leptin receptor (db/db) leads to hyperphagia and obesity in both C57BL/6 and DBA/2 mice, but only on the DBA/2 background do the mice develop beta-cell loss leading to severe diabetes, while C57BL/6 mice are relatively resistant. To further investigate the genetic factors predisposing to diabetes, we have studied leptin receptor-deficient offspring of an F2 cross between C57BL/6J (db/+) males and DBA/2J females. The results show that the genetics of diabetes susceptibility are enormously complex and a number of quantitative trait loci (QTL) contributing to diabetes-related traits were identified, notably on chromosomes 4, 6, 7, 9, 10, 11, 12, and 19. The Chr. 4 locus is likely due to a disruption of the Zfp69 gene in C57BL/6J mice. To identify candidate genes and to model coexpression networks, we performed global expression array analysis in livers of the F2 mice. Expression QTL (eQTL) were identified and used to prioritize candidate genes at clinical trait QTL. In several cases, clusters of eQTLs colocalized with clinical trait QTLs, suggesting a common genetic basis. We constructed coexpression networks for both 5 and 12 wk old mice and identified several modules significantly associated with clinical traits. One module in 12 wk old mice was associated with several measures of hepatic fat content as well as with other lipid- and diabetes-related traits. These results add to the understanding of the complex genetic interactions contributing to obesity-induced diabetes.
Related JoVE Video
Structural basis for nematode eIF4E binding an m(2,2,7)G-Cap and its implications for translation initiation.
Nucleic Acids Res.
PUBLISHED: 09-29-2011
Show Abstract
Hide Abstract
Metazoan spliced leader (SL) trans-splicing generates mRNAs with an m(2,2,7)G-cap and a common downstream SL RNA sequence. The mechanism for eIF4E binding an m²²?G-cap is unknown. Here, we describe the first structure of an eIF4E with an m(2,2,7)G-cap and compare it to the cognate m?G-eIF4E complex. These structures and Nuclear Magnetic Resonance (NMR) data indicate that the nematode Ascaris suum eIF4E binds the two different caps in a similar manner except for the loss of a single hydrogen bond on binding the m(2,2,7)G-cap. Nematode and mammalian eIF4E both have a low affinity for m(2,2,7)G-cap compared with the m?G-cap. Nematode eIF4E binding to the m?G-cap, m(2,2,7)G-cap and the m(2,2,7)G-SL 22-nt RNA leads to distinct eIF4E conformational changes. Additional interactions occur between Ascaris eIF4E and the SL on binding the m(2,2,7)G-SL. We propose interactions between Ascaris eIF4E and the SL impact eIF4G and contribute to translation initiation, whereas these interactions do not occur when only the m(2,2,7)G-cap is present. These data have implications for the contribution of 5-UTRs in mRNA translation and the function of different eIF4E isoforms.
Related JoVE Video
Prevalence of occult nostril asymmetry in the oversized nasal tip: a quantitative photographic analysis.
Arch Facial Plast Surg
PUBLISHED: 09-21-2011
Show Abstract
Hide Abstract
To objectively determine the prevalence of occult nasal base asymmetry in adults with wide nasal tips using a standard photographic editing program.
Related JoVE Video
Lentivirus-mediated expression of cDNA and shRNA slows degeneration in retinitis pigmentosa.
Exp. Biol. Med. (Maywood)
PUBLISHED: 09-01-2011
Show Abstract
Hide Abstract
Mutations in Pde6b lead to high levels of signaling molecules cyclic guanosine monophosphate (cGMP) and Ca(2+), which ultimately result in photoreceptor cell death in certain forms of retinitis pigmentosa (RP). The level of cGMP, which is controlled by opposing activities of guanylate cyclase (GUCY) and photoreceptor phosphodiesterase-6 (PDE6), regulates the opening of cyclic nucleotide-gated ion channels [CNG] and thereby controls Ca(2+) influx into the outer segments. Using a lentiviral gene therapy approach, we have previously shown that degeneration can be temporarily slowed either by introducing wild-type PDE6? or knocking down expression of GUCY2E and CNGA1 in photoreceptors of Pde6b(H620Q), a mouse model for RP. Rescue was transient with either approach. Therefore, we tested a novel combination therapy using bipartite lentiviral vectors designed to both introduce wild-type PDE6? expression and knockdown GUCY2E or CNGA1. Immunoblot analysis shows simultaneous increases in PDE6? and decreases in GUCY2E or CNGA1 in retinas transduced by the vectors, indicating successful transduction. In Pde6b(H620Q) mutants, we observe rescue of photoreceptor function and an increase in photoreceptor rows as compared with untreated controls. However, no evidence of prolonged rescue beyond the limit of the previously tested single therapy was observed.
Related JoVE Video
Effect of the ILE86TER mutation in the ? subunit of cGMP phosphodiesterase (PDE6) on rod photoreceptor signaling.
Cell. Signal.
PUBLISHED: 08-18-2011
Show Abstract
Hide Abstract
The light-dependent decrease in cyclic guanosine monophosphate (cGMP) in the rod outer segment is produced by a phosphodiesterase (PDE6), consisting of catalytic ? and ? subunits and two inhibitory ? subunits. The molecular mechanism of PDE6? regulation of the catalytic subunits is uncertain. To study this mechanism in vivo, we introduced a modified Pde6g gene for PDE6? into a line of Pde6g(tm1)/Pde6g(tm1) mice that do not express PDE6?. The resulting ILE86TER mice have a PDE6? that lacks the two final carboxyl-terminal Ile(86) and Ile(87) residues, a mutation previously shown in vitro to reduce inhibition by PDE6?. ILE86TER rods showed a decreased sensitivity and rate of activation, probably the result of a decreased level of expression of PDE6 in ILE86TER rods. More importantly, they showed a decreased rate of decay of the photoresponse, consistent with decreased inhibition of PDE6 ? and ? by PDE6?. Furthermore, ILE86TER rods had a higher rate of spontaneous activation of PDE6 than WT rods. Circulating current in ILE86TER rods that also lacked both guanylyl cyclase activating proteins (GCAPs) could be increased several fold by perfusion with 100?M of the PDE6 inhibitor 3-isobutyl-1-methylxanthine (IBMX), consistent with a higher rate of dark PDE6 activity in the mutant photoreceptors. In contrast, IBMX had little effect on the circulating current of WT rods, unlike previous results from amphibians. Our results show for the first time that the Ile(86) and Ile(87) residues are necessary for normal inhibition of PDE6 catalytic activity in vivo, and that increased basal activity of PDE can be partially compensated by GCAP-dependent regulation of guanylyl cyclase.
Related JoVE Video
Characterization of recombinant human IL-15 deamidation and its practical elimination through substitution of asparagine 77.
Pharm. Res.
PUBLISHED: 07-28-2011
Show Abstract
Hide Abstract
The use of recombinant human interleukin (rhIL)-15 as a potential therapeutic immune modulator and anticancer agent requires pure, stable preparations. However, purified rhIL-15 preparations readily accumulated heterogeneities. We sought to improve rhIL-15 stability through process, formulation, and targeted amino acid changes.
Related JoVE Video
Influence of infection of cotton by rotylenchulus reniformis and meloidogyne incognita on the production of enzymes involved in systemic acquired resistance.
J. Nematol.
PUBLISHED: 07-26-2011
Show Abstract
Hide Abstract
Systemic acquired resistance (SAR), which results in enhanced defense mechanisms in plants, can be elicited by virulent and avirulent strains of pathogens including nematodes. Recent studies of nematode reproduction strongly suggest that Meloidogyne incognita and Rotylenchulus reniformis induce SAR in cotton, but biochemical evidence of SAR was lacking. Our objective was to determine whether infection of cotton by M. incognita and R. reniformis increases the levels of P-peroxidase, G-peroxidase, and catalase enzymes which are involved in induced resistance. A series of greenhouse trials was conducted; each trial included six replications of four treatments applied to one of three cotton genotypes in a randomized complete block design. The four treatments were cotton plants inoculated with i) R. reniformis, ii) M. incognita, iii) BTH (Actigard), and iv) a nontreated control. Experiments were conducted on cotton genotypes DP 0935 B2RF (susceptible to both nematodes), LONREN-1 (resistant to R. reniformis), and M-120 RNR (resistant to M. incognita), and the level of P-peroxidase, G-peroxidase, and catalase activity was measured before and 2, 4, 6, 10, and 14 d after treatment application. In all cotton genotypes, activities of all three enzymes were higher (P ? 0.05) in leaves of plants infected with M. incognita and R. reniformis than in the leaves of control plants, except that M. incognita did not increase catalase activity on LONREN-1. Increased enzyme activity was usually apparent 6 d after treatment. This study documents that infection of cotton by M. incognita or R. reniformis increases the activity of the enzymes involved in systemic acquired resistance; thereby providing biochemical evidence to substantiate previous reports of nematode-induced SAR in cotton.
Related JoVE Video
Induction of Systemic Acquired Resistance by Rotylenchulus reniformis and Meloidogyne incognita in Cotton Following Separate and Concomitant Inoculations.
J. Nematol.
PUBLISHED: 07-25-2011
Show Abstract
Hide Abstract
Systemic acquired resistance (SAR) can be elicited by virulent and avirulent pathogenic strains and SAR against plant-parasitic nematodes has been documented. Our objective was to determine whether co-infection of cotton by Meloidogyne incognita and Rotylenchulus reniformis affects the population level of either nematode compared to infection by each species individually. Split-root trials were conducted in which plants were inoculated with i) R. reniformis only, ii) M. incognita only, iii) both R. reniformis and M. incognita, or iv) no nematodes. Half of the root system was inoculated with R. reniformis or M. incognita on day 0 and the other half with M. incognita or R. reniformis on day 0 or day 14 depending on the experiment. Experiments were conducted on cotton cultivar DP 0935 B2RF (susceptible to both nematodes), LONREN-1 (germplasm line resistant to R. reniformis), and M-120 RNR (germplasm line resistant to M. incognita), and tests were terminated 8 wk after the last inoculation. Both soil (vermiform) and roots (egg) extracted from each half of the root system to determine the total nematode population levels, and root galling was rated on a 0 to 10 scale. Mixed models analysis and comparison of least squares means indicated no differences in root galling (except on LONREN-1) or population levels when the two nematode species were introduced on the same day. When M. incognita was introduced 14 d after R. reniformis, reduction in galling (36% on DP 0935 and 33% on LONREN-1) and M. incognita population levels (35% on DP 0935 and 45% on LONREN-1) were significant (P ? 0.05). When R. reniformis was inoculated 14 d after M. incognita, reduction in R. reniformis population levels (18% on DP 0935 and 26% on M-120) were significant. This study documents for the first time that infection of cotton by a nematode can elicit SAR to another nematode species.
Related JoVE Video
NKX2-5(eGFP/w) hESCs for isolation of human cardiac progenitors and cardiomyocytes.
Nat. Methods
PUBLISHED: 07-14-2011
Show Abstract
Hide Abstract
NKX2-5 is expressed in the heart throughout life. We targeted eGFP sequences to the NKX2-5 locus of human embryonic stem cells (hESCs); NKX2-5(eGFP/w) hESCs facilitate quantification of cardiac differentiation, purification of hESC-derived committed cardiac progenitor cells (hESC-CPCs) and cardiomyocytes (hESC-CMs) and the standardization of differentiation protocols. We used NKX2-5 eGFP(+) cells to identify VCAM1 and SIRPA as cell-surface markers expressed in cardiac lineages.
Related JoVE Video
Allelic variation on murine chromosome 11 modifies host inflammatory responses and resistance to Bacillus anthracis.
PLoS Pathog.
PUBLISHED: 06-24-2011
Show Abstract
Hide Abstract
Anthrax is a potentially fatal disease resulting from infection with Bacillus anthracis. The outcome of infection is influenced by pathogen-encoded virulence factors such as lethal toxin (LT), as well as by genetic variation within the host. To identify host genes controlling susceptibility to anthrax, a library of congenic mice consisting of strains with homozygous chromosomal segments from the LT-responsive CAST/Ei strain introgressed on a LT-resistant C57BL/6 (B6) background was screened for response to LT. Three congenic strains containing CAST/Ei regions of chromosome 11 were identified that displayed a rapid inflammatory response to LT similar to, but more severe than that driven by a LT-responsive allele of the inflammasome constituent NRLP1B. Importantly, increased response to LT in congenic mice correlated with greater resistance to infection by the Sterne strain of B. anthracis. The genomic region controlling the inflammatory response to LT was mapped to 66.36-74.67 Mb on chromosome 11, a region that encodes the LT-responsive CAST/Ei allele of Nlrp1b. However, known downstream effects of NLRP1B activation, including macrophage pyroptosis, cytokine release, and leukocyte infiltration could not fully explain the response to LT or the resistance to B. anthracis Sterne in congenic mice. Further, the exacerbated response in congenic mice is inherited in a recessive manner while the Nlrp1b-mediated response to LT is dominant. Finally, congenic mice displayed increased responsiveness in a model of sepsis compared with B6 mice. In total, these data suggest that allelic variation of one or more chromosome 11 genes in addition to Nlrp1b controls the severity of host response to multiple inflammatory stimuli and contributes to resistance to B. anthracis Sterne. Expression quantitative trait locus analysis revealed 25 genes within this region as high priority candidates for contributing to the host response to LT.
Related JoVE Video
Deep small RNA sequencing from the nematode Ascaris reveals conservation, functional diversification, and novel developmental profiles.
Genome Res.
PUBLISHED: 06-17-2011
Show Abstract
Hide Abstract
Eukaryotic cells express several classes of small RNAs that regulate gene expression and ensure genome maintenance. Endogenous siRNAs (endo-siRNAs) and Piwi-interacting RNAs (piRNAs) mainly control gene and transposon expression in the germline, while microRNAs (miRNAs) generally function in post-transcriptional gene silencing in both somatic and germline cells. To provide an evolutionary and developmental perspective on small RNA pathways in nematodes, we identified and characterized known and novel small RNA classes through gametogenesis and embryo development in the parasitic nematode Ascaris suum and compared them with known small RNAs of Caenorhabditis elegans. piRNAs, Piwi-clade Argonautes, and other proteins associated with the piRNA pathway have been lost in Ascaris. miRNAs are synthesized immediately after fertilization in utero, before pronuclear fusion, and before the first cleavage of the zygote. This is the earliest expression of small RNAs ever described at a developmental stage long thought to be transcriptionally quiescent. A comparison of the two classes of Ascaris endo-siRNAs, 22G-RNAs and 26G-RNAs, to those in C. elegans, suggests great diversification and plasticity in the use of small RNA pathways during spermatogenesis in different nematodes. Our data reveal conserved characteristics of nematode small RNAs as well as features unique to Ascaris that illustrate significant flexibility in the use of small RNAs pathways, some of which are likely an adaptation to Ascaris life cycle and parasitism. The transcriptome assembly has been submitted to NCBI Transcriptome Shotgun Assembly Sequence Database(http://www.ncbi.nlm.nih.gov/genbank/TSA.html) under accession numbers JI163767–JI182837 and JI210738–JI257410.
Related JoVE Video
Biodiversity and emerging biogeography of the neutrophilic iron-oxidizing Zetaproteobacteria.
Appl. Environ. Microbiol.
PUBLISHED: 06-10-2011
Show Abstract
Hide Abstract
Members of the neutrophilic iron-oxidizing candidate class Zetaproteobacteria have predominantly been found at sites of microbially mediated iron oxidation in marine environments around the Pacific Ocean. Eighty-four full-length (>1,400-bp) and 48 partial-length Zetaproteobacteria small-subunit (SSU) rRNA gene sequences from five novel clone libraries, one novel Zetaproteobacteria isolate, and the GenBank database were analyzed to assess the biodiversity of this burgeoning class of the Proteobacteria and to investigate its biogeography between three major sampling regions in the Pacific Ocean: Loihi Seamount, the Southern Mariana Trough, and the Tonga Arc. Sequences were grouped into operational taxonomic units (OTUs) on the basis of a 97% minimum similarity. Of the 28 OTUs detected, 13 were found to be endemic to one of the three main sampling regions and 2 were ubiquitous throughout the Pacific Ocean. Additionally, two deeply rooted OTUs that potentially dominate communities of iron oxidizers originating in the deep subsurface were identified. Spatial autocorrelation analysis and analysis of molecular variance (AMOVA) showed that geographic distance played a significant role in the distribution of Zetaproteobacteria biodiversity, whereas environmental parameters, such as temperature, pH, or total Fe concentration, did not have a significant effect. These results, detected using the coarse resolution of the SSU rRNA gene, indicate that the Zetaproteobacteria have a strong biogeographic signal.
Related JoVE Video
A systems genetic analysis of high density lipoprotein metabolism and network preservation across mouse models.
Biochim. Biophys. Acta
PUBLISHED: 05-05-2011
Show Abstract
Hide Abstract
We report a systems genetic analysis of high density lipoprotein (HDL) levels in an F2 intercross between inbred strains CAST/EiJ and C57BL/6J. We previously showed that there are dramatic differences in HDL metabolism in a cross between these strains, and we now report co-expression network analysis of HDL that integrates global expression data from liver and adipose with relevant metabolic traits. Using data from a total of 293 F2 intercross mice, we constructed weighted gene co-expression networks and identified modules (subnetworks) associated with HDL and clinical traits. These were examined for genes implicated in HDL levels based on large human genome-wide associations studies (GWAS) and examined with respect to conservation between tissue and sexes in a total of 9 data sets. We identify genes that are consistently ranked high by association with HDL across the 9 data sets. We focus in particular on two genes, Wfdc2 and Hdac3, that are located in close proximity to HDL QTL peaks where causal testing indicates that they may affect HDL. Our results provide a rich resource for studies of complex metabolic interactions involving HDL. This article is part of a Special Issue entitled Advances in High Density Lipoprotein Formation and Metabolism: A Tribute to John F. Oram (1945-2010).
Related JoVE Video
Modulation of neurotransmitter release in orexin/hypocretin-2 receptor knockout mice: a microdialysis study.
J. Neurosci. Res.
PUBLISHED: 04-29-2011
Show Abstract
Hide Abstract
Orexinergic neurons are discretely localized within the lateral hypothalamus and have widespread projections to the whole brain. Here, the role of orexin/hypocretin-2 receptors (OX2) in modulating extracellular concentrations of neurotransmitters was evaluated in the hypothalamus and the prefrontal cortex (PFC) of OX2 knockout (KO) mice by using a microdialysis technique. In the hypothalamus, basal concentrations of norephinephrine (NE), acetylcholine (ACh), and histamine (Hist) were significantly higher in KO mice, whereas KCl perfusion (147 mM) resulted in significantly lesser increases in NE, ACh, and Hist release in KO compared with wild-type (WT) mice. No differences in basal concentrations or evoked release of serotonin (5-HT) or dopamine (DA) were found in the hypothalamus between genotypes. In the PFC, no differences in the basal concentrations of the studied neurotransmitters were found between genotypes. After KCl perfusion, significantly higher increases in NE, 5-HT, and DA release were found in KO compared with WT mice. No differences in the evoked release of ACh and Hist in the PFC were found between genotypes. The present results demonstrate that genetic deletion of OX2 receptors differentially modulates extracellular concentrations of distinct neurotransmitters in the somatodendritic region vs. a nerve terminal region of the orexinergic neurons. In the hypothalamus, an inhibitory role of the OX2 receptors in modulating basal concentrations of NE, ACh, and Hist was revealed, which probably accounts for the reduced responsiveness to KCl as well. In the PFC, the evoked release of the monoamines NE, 5-HT, and DA seems to be controlled negatively by OX2 receptors.
Related JoVE Video
A targeted NKX2.1 human embryonic stem cell reporter line enables identification of human basal forebrain derivatives.
Stem Cells
PUBLISHED: 03-23-2011
Show Abstract
Hide Abstract
We have used homologous recombination in human embryonic stem cells (hESCs) to insert sequences encoding green fluorescent protein (GFP) into the NKX2.1 locus, a gene required for normal development of the basal forebrain. Generation of NKX2.1-GFP(+) cells was dependent on the concentration, timing, and duration of retinoic acid treatment during differentiation. NKX2.1-GFP(+) progenitors expressed genes characteristic of the basal forebrain, including SHH, DLX1, LHX6, and OLIG2. Time course analysis revealed that NKX2.1-GFP(+) cells could upregulate FOXG1 expression, implying the existence of a novel pathway for the generation of telencephalic neural derivatives. Further maturation of NKX2.1-GFP(+) cells gave rise to ?-aminobutyric acid-, tyrosine hydroxylase-, and somatostatin-expressing neurons as well as to platelet-derived growth factor receptor ?-positive oligodendrocyte precursors. These studies highlight the diversity of cell types that can be generated from human NKX2.1(+) progenitors and demonstrate the utility of NKX2.1(GFP/w) hESCs for investigating human forebrain development and neuronal differentiation.
Related JoVE Video
Characterization of the Salmonella enterica serovar Typhimurium ydcI gene, which encodes a conserved DNA binding protein required for full acid stress resistance.
J. Bacteriol.
PUBLISHED: 03-11-2011
Show Abstract
Hide Abstract
Salmonella enterica serovar Typhimurium possesses a stimulon of genes that are differentially regulated in response to conditions of low fluid shear force that increase bacterial virulence and alter other phenotypes. In this study, we show that a previously uncharacterized member of this stimulon, ydcI or STM1625, encodes a highly conserved DNA binding protein with related homologs present in a range of gram-negative bacterial genera. Gene expression analysis shows that ydcI is expressed in different bacterial genera and is involved in its autoregulation in S. Typhimurium. We demonstrate that purified YdcI protein specifically binds a DNA probe consisting of its own promoter sequence. We constructed an S. Typhimurium ?ydcI mutant strain and show that this strain is more sensitive to both organic and inorganic acid stress than is an isogenic WT strain, and this defect is complemented in trans. Moreover, our data indicate that ydcI is part of the rpoS regulon related to stress resistance. The S. Typhimurium ?ydcI mutant was able to invade cultured cells to the same degree as the WT strain, but a strain in which ydcI expression is induced invaded cells at a level 2.8 times higher than that of the WT. In addition, induction of ydcI expression in S. Typhimurium resulted in the formation of a biofilm in stationary-phase cultures. These data indicate the ydcI gene encodes a conserved DNA binding protein involved with aspects of prokaryotic biology related to stress resistance and possibly virulence.
Related JoVE Video
Pluripotent stem cell models of cardiac disease and their implication for drug discovery and development.
Trends Mol Med
PUBLISHED: 02-10-2011
Show Abstract
Hide Abstract
Recent advances in pluripotent stem cell biology now make it possible to generate human cardiomyocytes in vitro from both healthy individuals and from patients with cardiac abnormalities. This offers unprecedented opportunities to study cardiac disease development in a dish and establish novel platforms for drug discovery, either to prevent disease progression or to reverse it. In this review paper, we discuss some of the genetic diseases that affect the heart and illustrate how these new paradigms could assist our understanding of cardiac pathogenesis and aid in drug discovery. In particular, we highlight the limitations of other commonly used model systems in predicting the consequences of drug exposure on the human heart.
Related JoVE Video
Amyloid beta dimers/trimers potently induce cofilin-actin rods that are inhibited by maintaining cofilin-phosphorylation.
Mol Neurodegener
PUBLISHED: 01-24-2011
Show Abstract
Hide Abstract
Previously we reported 1 ?M synthetic human amyloid beta1-42 oligomers induced cofilin dephosphorylation (activation) and formation of cofilin-actin rods within rat hippocampal neurons primarily localized to the dentate gyrus.
Related JoVE Video
In vivo effects of antiviral protein kinase C modulators on zebrafish development and survival.
ISRN Toxicol
PUBLISHED: 01-01-2011
Show Abstract
Hide Abstract
Clinical interventions using protein kinase C (PKC) modulators have been proposed for eradication of HIV-1-infected cellular reservoirs which persist in patients despite prolonged antiretroviral therapy. The effects of some of these agents have not been assessed in a developing vertebrate model. This study examines the developmental and toxicological effects of these compounds on zebrafish embryos and larvae. Treatment of zebrafish through the first week of development with various PKC pathway modulators did not elicit gross physical defects or elevated incidences of death at lower doses. Higher concentrations resulted in rapid death for both later-stage embryos and larvae. Each compound had a threshold dose for lethality. The defined nonlethal doses may be useful toward assessing the effects of modulating PKC activity on zebrafish development. They may further provide some guidance for the potential dosing of PKC modulators in clinical trials toward the goal of HIV-1 reservoir eradication.
Related JoVE Video
Continuity and change in the eastern Aleutian archaeological sequence.
Hum. Biol.
PUBLISHED: 12-07-2010
Show Abstract
Hide Abstract
The eastern Aleutian prehistoric archaeological sequence is key for understanding population movements, cultural exchanges, and adaptations to environmental changes over a wide area of the north Pacific and Bering Sea during the Holocene. An important question is, Can the settlement history of the eastern Aleutians be understood as a single continuous tradition lasting some 9,000 years, or were there major population and cultural influxes along with periods of widespread population abandonment? We review the available archaeological evidence with reference to recent mtDNA and nucleic DNA studies of prehistoric and contemporary Arctic and Subarctic populations and conclude that the evidence points to an overall cultural continuity with notable incursions and excursions of people and cultural elements into and out of the eastern Aleutians.
Related JoVE Video
In situ growth of silver nanoparticles in porous membranes for surface-enhanced raman scattering.
ACS Appl Mater Interfaces
PUBLISHED: 11-02-2010
Show Abstract
Hide Abstract
We demonstrate the in situ growth of silver nanoparticles in porous alumina membranes (PAMs) for use as a surface-enhanced Raman scattering (SERS) detection substrate. This fabrication method is simple, cost-effective, and fast, while providing control over the size of silver nanoparticles through the entire length of the cylindrical nanopores with uniform particle density inside the pores unachievable by the traditional infiltration technique. The in situ growth of silver nanoparticles was conducted from electroless-deposited nanoscale seeds on the interior of the PAM and resulted in the formation of numerous hot spots, which facilitated significantly higher SERS enhancement for these substrates compared with previously reported porous substrates.
Related JoVE Video
PGC-1?, a potential therapeutic target for early intervention in Parkinsons disease.
Sci Transl Med
PUBLISHED: 10-08-2010
Show Abstract
Hide Abstract
Parkinsons disease affects 5 million people worldwide, but the molecular mechanisms underlying its pathogenesis are still unclear. Here, we report a genome-wide meta-analysis of gene sets (groups of genes that encode the same biological pathway or process) in 410 samples from patients with symptomatic Parkinsons and subclinical disease and healthy controls. We analyzed 6.8 million raw data points from nine genome-wide expression studies, and 185 laser-captured human dopaminergic neuron and substantia nigra transcriptomes, followed by two-stage replication on three platforms. We found 10 gene sets with previously unknown associations with Parkinsons disease. These gene sets pinpoint defects in mitochondrial electron transport, glucose utilization, and glucose sensing and reveal that they occur early in disease pathogenesis. Genes controlling cellular bioenergetics that are expressed in response to peroxisome proliferator-activated receptor ? coactivator-1? (PGC-1?) are underexpressed in Parkinsons disease patients. Activation of PGC-1? results in increased expression of nuclear-encoded subunits of the mitochondrial respiratory chain and blocks the dopaminergic neuron loss induced by mutant ?-synuclein or the pesticide rotenone in cellular disease models. Our systems biology analysis of Parkinsons disease identifies PGC-1? as a potential therapeutic target for early intervention.
Related JoVE Video
Gadolinium released from MR contrast agents is deposited in brain tumors: in situ demonstration using scanning electron microscopy with energy dispersive X-ray spectroscopy.
Acta Radiol
PUBLISHED: 09-26-2010
Show Abstract
Hide Abstract
gadolinium (Gd)-containing MRI contrast agents (GdCA) are widely used in studies of brain tumors, and a number of reports suggest that under certain conditions, such as renal failure, Gd may be released from GdCA into patients tissues. Whether this may happen in abnormal tissues in the absence of renal failure has not been studied.
Related JoVE Video
Effect of long-term storage in TRIzol on microarray-based gene expression profiling.
Cancer Epidemiol. Biomarkers Prev.
PUBLISHED: 08-30-2010
Show Abstract
Hide Abstract
Although TRIzol is widely used for preservation and isolation of RNA, there is suspicion that prolonged sample storage in TRIzol may affect array-based gene expression profiling (GEP) through premature termination during reverse transcription.
Related JoVE Video

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.

How does it work?

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.