Antisense oligonucleotide G3139 is designed for Bcl-2 downregulation and is known to induce toll-like receptor activation. Novel stabilized lipid-polycation-DNA (sLPD) nanoparticles were constructed and evaluated for the delivery of G3139 to human carcinoma KB cells and for bioactivity in vivo. Polyethylenimine (PEI) was incorporated as a DNA condensing agent. The lipid composition used was DOTAP/DDAB/Chol/TPGS/linoleic acid/hexadecenal at molar ratios of 30/30/34/1/5/0.2. The nanoparticles were stabilized by the formation of a reversible covalent bond between the aldehyde group on the cis-11-hexadecenal and amines on the PEI. When sLPDs were used to transfect KB cells, 90.4% Bcl-2 downregulation was observed, compared to no significant down-regulation by free G3139 and 54.6% down regulation by non-stabilized LPD-G3139. The sLPDs were then evaluated for therapeutic efficacy in mice bearing KB subcutaneous tumors and were found to trigger a strong antitumor response, inhibiting tumor growth and prolonging survival with 72% increase in lifespan (ILS). Consistent with previous reports on other G3139 nanoparticles, the increased anti-tumor activities of sLPDs in vivo were found to be associated with increased cytokine induction rather than Bcl-2 down-regulation, suggesting an immunological mechanism.
doi:10.1021/mp100272k
PMCID: PMC3599967
PMID: 21366344
Nanoparticles; antisense oligonucleotide; reversible crosslinking; cancer; immunotherapy; drug delivery
Non-ionic surfactant vesicles, or SPANosomes (SPs), comprised of cationic lipid and sorbitan monooleate (Span 80) were synthesized and evaluated as siRNA vectors. The SPs had a mean diameter of less than 100 nm and exhibited excellent colloidal stability. The SP/siRNA complexes possessed a slightly positive zeta potential of 12 mV and demonstrated a high siRNA incorporation efficiency of greater than 80%. Cryogenic transmission electron microscopy (cryo-TEM) imaging of the SP/siRNA indicated a predominantly core-shell structure. The SP/siRNA complexes were shown to efficiently and specifically silence expression of both green fluorescent protein (GFP) (66% knockdown) and aromatase (77% knockdown) genes in breast cancer cell lines. In addition, the cellular trafficking pathway of the SP/siRNA was investigated by confocal microscopy using molecular beacons as probes for cytosolic delivery. The results showed efficient endosomal escape and cytosolic delivery of the siRNA cargo following internalization of the SP/siRNA complexes. In conclusion, Span 80 is a potent helper lipid and the SPs are promising vehicles for siRNA delivery.
doi:10.1021/mp200426h
PMCID: PMC3273546
PMID: 22149175
Drug delivery; Nanoparticles; Surfactant vesicles; siRNA; Molecular beacon
Objective
To evaluate the association of demographic, behavioral, and socioeconomic factors with all-cause mortality while controlling for health status among a cohort of participants with severe disability related to spinal cord injury (SCI).
Study Design
Prospective cohort study.
Setting
Data were analyzed at a major medical university in the Southeast United States of America.
Methods
Participants included 1361 adults with traumatic SCI of at least one year duration who were recruited through a large specialty hospital in the Southeast United States of America. Three Cox proportional hazards models were generated relating the predictors to all-cause mortality.
Results
Age, disability, smoking, and income were significant in the final model. Both current (hazard ratio [HR]=2.03, 95% confidence interval [CI]=1.46–2.82) and former smokers (HR=1.58, CI=1.16–2.16) were at elevated hazard of mortality, as were those with incomes under $10,000 (HR=2.29, CI=1.53–3.44) and between $10,000–$35,000 (HR=1.47, CI=1.03–2.10).
Conclusions
Even after controlling for health and severity of disability, the coefficients for smoking and income were significant, exceeding that reported previously within the general population. The importance of these factors may be magnified after severe disability, even though life expectancy is already greatly diminished in this population.
doi:10.1038/sc.2012.24
PMCID: PMC3376668
PMID: 22410844
spinal cord injuries; socioeconomic factors; behavior; mortality; life expectancy; health status
Objective
To examine behavioral risk factors in relation to fatigue after spinal cord injury (SCI), specifically cardiovascular related behaviors, prescription medication use, and alcohol and cigarette use.
Design
Cross-sectional.
Setting
A medical university in the Southeastern United States.
Participants
2296 adults at least one year post-SCI from a large specialty hospital in the Southeast responded to a mail-in survey.
Intervention
None
Main Outcomes Measure
The Modified Fatigue Impact Scale-5 item version was used to assess disabling fatigue.
Results
Of participants, 8.3% had disabling fatigue, 45.3% reported fatigue rarely to never impacted their life, and 46.4% reported having some fatigue. Persons who reported having less exercise than others with a similar injury level were 2.49 times as likely to have disabling fatigue as persons who reported more exercise. Those with a fair or poor diet were also more likely to have disabling fatigue. Use of prescription medication to treat pain was associated with disabling fatigue, as was being CAGE positive. Among non-behavioral variables, race and injury severity were significantly associated with disabling fatigue.
Conclusions
We identified several behavioral predictors of disabling fatigue, including cardiovascular risk factors, prescription medication use, and alcohol use. These factors are important as they are able to be modified and could be potential factors for prevention or intervention.
doi:10.1016/j.apmr.2011.09.001
PMCID: PMC3398712
PMID: 22289243
spinal cord injury; fatigue; behavior
The discovery of upregulated glycogen synthase kinase-3 (GSK-3) in various pathological conditions has led to the development of a host of chemically diverse small molecule GSK-3 inhibitors, such as BIP-135. GSK-3 inhibition emerged as an alternative therapeutic target for treating spinal muscular atrophy (SMA) when a number of GSK-3 inhibitors were shown to elevate survival motor neuron (SMN) levels in vitro and to rescue motor neurons when their intrinsic SMN level was diminished by SMN-specific short hairpin RNA (shRNA). Despite their cellular potency, the in vivo efficacy of GSK-3 inhibitors has yet to be evaluated in an animal model of SMA. Herein, we disclose that a potent and reasonably selective GSK-3 inhibitor, namely BIP-135, was tested in a transgenic Δ7 SMA KO mouse model of SMA, and found to prolong the median survival of these animals. In addition, this compound was shown to elevate the SMN protein level in SMA patient-derived fibroblast cells as determined by western blot, and was neuroprotective in a cell-based, SMA-related model of oxidative stress-induced neurodegeneration.
doi:10.1021/cn200085z
PMCID: PMC3279955
PMID: 22348181
GSK-3 inhibitor; BIP-135; median survival; spinal muscular atrophy; survival motor neuron; Δ7 SMA KO mice
The discovery of upregulated glycogen synthase kinase-3
(GSK-3)
in various pathological conditions has led to the development of a
host of chemically diverse small molecule GSK-3 inhibitors, such as
BIP-135. GSK-3 inhibition emerged as an alternative therapeutic target
for treating spinal muscular atrophy (SMA) when a number of GSK-3
inhibitors were shown to elevate survival motor neuron (SMN) levels
in vitro and to rescue motor neurons when their intrinsic SMN level
was diminished by SMN-specific short hairpin RNA (shRNA). Despite
their cellular potency, the in vivo efficacy of GSK-3 inhibitors has
yet to be evaluated in an animal model of SMA. Herein, we disclose
that a potent and reasonably selective GSK-3 inhibitor, namely BIP-135,
was tested in a transgenic Δ7 SMA KO mouse model of SMA and
found to prolong the median survival of these animals. In addition,
this compound was shown to elevate the SMN protein level in SMA patient-derived
fibroblast cells as determined by Western blot, and was neuroprotective
in a cell-based, SMA-related model of oxidative stress-induced neurodegeneration.
doi:10.1021/cn200085z
PMCID: PMC3279955
PMID: 22348181
GSK-3 inhibitor; BIP-135; median survival; spinal muscular atrophy; survival motor neuron; Δ7 SMA KO mice
Purpose
To identify the relationship of race and gender with 3 aspects of life satisfaction and depressive symptoms after spinal cord injury (SCI), evaluating the extent to which socioeconomic factors mediate any observed relationships.
Methods
Adults with traumatic SCI of at least 1-year duration (N = 1,549) were identified through a Southeastern United States SCI Model System of care, and cross-sectional survey data were collected at a Southeastern United States medical university. Three aspects of life satisfaction (home life satisfaction, vocational satisfaction, global satisfaction) were measured using 20 satisfaction items from the Life Situation Questionnaire-Revised. The Older Adult Health and Mood Questionnaire measured depressive symptoms. MANCOVA assessed mediation of socioeconomic status between race and life satisfaction and depression.
Results
Home life satisfaction and vocational satisfaction were significantly related to race, with White participants scoring higher than Black participants during the first stage of the regression. However, socioeconomic factors mediated the relationships such that race was no longer significant after considering economic factors. Race was significantly associated with global satisfaction after adjusting for socioeconomic factors. Depression was not significantly related to race. Gender was unrelated to all study outcomes. Of the socioeconomic mediators, family income was a significant predictor of each outcome, whereas education was only predictive of vocational satisfaction.
Conclusion
Socioeconomic factors are important mediators of the relationship between race and certain aspects of life satisfaction among persons with SCI. Family income and, to a lesser extent, education should be considered when evaluating race differences in life satisfaction after SCI.
doi:10.1310/sci1803-232
PMCID: PMC3479640
PMID: 23105915
depression; health status disparities; socioeconomic factors; spinal cord injury
Objective
We examined the association of different chronic physical and mental conditions, individually or comorbidly on health-related quality of life (QoL) in Chinese children aged ≤14 years in Hong Kong.
Design
Population-based cross-sectional survey.
Participants
Approximately 7500 Chinese children aged <14 years in Hong Kong.
Interventions
Nil.
Primary and secondary outcome measures
Various health concepts of validated Chinese version of Child Health Questionnaire (CHQ), a health-related QoL questionnaire in children.
Result
There was significant association of physical and mental health conditions, either individually or comorbidly, on the various concepts of CHQ. Children with mental health problems were apparently more affected than those with physical health problems. Chronic renal disease and congenital malformation were the physical health conditions associated with the lowest scores in CHQ concepts in children aged 5–10 years and aged 10–14 years, respectively. Behavioural problem was the mental health condition associated with the lowest score in CHQ concepts in both age groups.
Conclusions
Our study shows important information concerning the prevalence of different health conditions and its association, either individually or comorbidly on the QoL in a representative sample of Chinese children in HK.
doi:10.1136/bmjopen-2012-001183
PMCID: PMC3549227
PMID: 23293240
PUBLIC HEALTH
Objective
To evaluate the association of health status, secondary health conditions, hospitalizations, and risk of mortality and life expectancy (LE) after spinal cord injury (SCI).
Design
Prospective cohort study.
Setting
Preliminary data were collected from a specialty hospital in the Southeastern United States, with mortality follow-up and data analysis conducted at a medical university.
Participants
A total of 1361 adults with traumatic SCI, all at least 1 year post-injury at the time of assessment, were enrolled in the study. There were 325 deaths. After elimination of those with missing data on key variables, there were 267 deaths and 12,032 person-years.
Interventions
None
Main Outcome Measures
Mortality status was determined by routine follow-up using the National Death Index through December 31, 2008. A logistic regression model was developed to estimate the probability of dying in any given year using person years.
Results
A history of chronic pressure ulcers, amputations, a depressive disorder, symptoms of infections, and being hospitalized within the past year were all predictive of mortality. LE estimates were generated using the example of a male with non-cervical, non-ambulatory SCI. Using 3 age examples (20, 40, 60), the greatest estimated lost LE was associated with chronic pressure ulcers (50.3%), followed by amputations (35.4%), 1 or more recent hospitalizations (18.5%), and the diagnosis of probable major depression (18%). Symptoms of infections was associated with a 6.7% reduction in LE for a 1 standard deviation increase in infectious symptoms.
Conclusion
Several secondary health conditions represent risk factors for mortality and diminish LE after SCI. The presence of 1 or more of these factors should be taken as an indicator of the need for intervention.
doi:10.1016/j.apmr.2011.05.024
PMCID: PMC3385509
PMID: 22032212
spinal cord injury; mortality; risk; health; economics; life expectancy
Background
N-lactosyl-dioleoylphosphatidylethanolamine (Lac-DOPE) was synthesized and evaluated as a liver-specific targeting ligand via asialoglycoprotein receptors for liposomal delivery of doxorubicin.
Methods
Lactosylated liposomes encapsulating calcein (Lac-L-calcein) or doxorubicin (Lac-L-DOX) composed of egg phosphatidylcholine, cholesterol, monomethoxy polyethylene glycol 2000-distearoyl phosphatidylethanolamine, and Lac-DOPE at 50:35:5:10 (mol/mol) were prepared by polycarbonate membrane extrusion and evaluated in human hepatocellular carcinoma HepG2 cells. Cellular uptake of Lac-L-calcein was monitored by confocal microscopy and by flow cytometry. The cytotoxicity of Lac-L-DOX was evaluated by MTT assay. The pharmacokinetic properties of Lac-L-DOX were studied in normal mice, and its biodistribution and antitumor activity were studied in nude mice with HepG2 xenografts.
Results
The size of Lac-L-DOX was less than 100 nm and the liposomes demonstrated excellent colloidal stability. In vitro uptake of Lac-L-calcein by HepG2 cells was four times greater than that of non-targeted L-calcein. In the presence of 20 mM lactose, the uptake of Lac-L-calcein was inhibited, suggesting that asialoglycoprotein receptors mediated the observed cellular uptake. Lac-L-DOX exhibited enhanced in vivo cytotoxicity compared with the nontargeted liposomal doxorubicin (L-DOX), and its pharmacokinetic parameters indicate that Lac-L-DOX has a long blood circulation time (t1/2 8.73 hours). Tissue distribution and therapeutic efficacy studies in nude mice bearing HepG2 xenografts show that Lac-L-DOX had significantly stronger tumor inhibitory activity compared with L-DOX and free doxorubicin, along with a higher accumulation of drug within the tumor site and greater cellular uptake by tumor cells.
Conclusion
These data suggest that lactosylated liposomes are promising drug delivery vehicles for hepatocellular carcinoma.
doi:10.2147/IJN.S33965
PMCID: PMC3476751
PMID: 23093902
liposomes; hepatocellular carcinoma; asialoglycoprotein receptor; drug targeting
doi:10.1310/sci1802-113
PMCID: PMC3469270
PMID: 23074361
Lipoplexes and polyplexes represent the two major nanocarrier systems for nucleic acid delivery. Previous studies examining their uptake and intracellular unpacking rely on organic fluorophores fraught with low signal intensity and photobleaching. In this work quantum dots-mediated Förster resonance energy transfer (QD-FRET) was first used to study and compare the cellular uptake and the intracellular fate of oligodeoxynucelotide (ODN)-based lipoplexes and polyplexes. QD605-Amine and Cy5-labeled ODN (Cy5-GTI2040) were chosen as the FRET pair. By adjusting the lipids/ODN ratio of lipoplexes and the nitrogen/phosphate (N/P) ratio of polyplexes, lipoplexes and polyplexes with comparable physical properties were produced. The biological activities of dual-labeled lipoplexes and polyplexes remained unaltered compared to their unlabeled counterparts as evidenced by their comparable antisense activities against protein R2 in KB cells. Flow cytometry and confocal microscopy revealed similar pattern of uptake for these two types of nanoparticles, although polyplexes had a higher dissociation rate than lipoplexes in KB cells. We demonstrate that QD-FRET is a sensitive tool to study the uptake and intracellular unpacking of lipoplexes and polyplexes, which may help optimize their formulations for various theranostics applications.
doi:10.1021/mp100466m
PMCID: PMC3185110
PMID: 21740056
quantum dots; FRET; lipoplexes; polyplexes; multifunctional nanoparticles; oligodeoxynucleotide (ODN); KB cells
Objective
To identify the stability of socio-environmental, behavioral, and health predictors of mortality over an eight year time frame.
Study Design
Cohort study.
Setting
Data were analyzed at a large medical university in the Southeast United States of America (USA).
Methods
Adults with residual impairment from a spinal cord injury (SCI) who were at least one year post-injury at assessment were recruited through a large specialty hospital in the Southeast USA. 1209 participants were included in the final analysis. A piecewise exponential model with 2 equal time intervals (eight years total) was used to assess the stability of the hazard and the predictors over time.
Results
The hazard did significantly change over time, where the hazard in the first time interval was significantly lower than the second. There were no interactions between the socio-environmental, behavior, or health factors and time, although there was a significant interaction between age at injury (a demographic variable) and time.
Conclusion
These results suggest there is stability in the association between the predictors and mortality, even over an eight year time period. Results reinforce the use of historic variables for prediction of mortality in persons with SCI.
doi:10.1038/sc.2011.158
PMCID: PMC3321101
PMID: 22231541
spinal cord injury; mortality; proportional hazards models
PMCID: PMC3594959
PMID: 23483860
Objective. We examined the hypothesis that foetal exposure to maternal passive smoking is associated with childhood asthma, allergic rhinitis, and eczema. Methods. The study was a population-based cross-sectional survey of Hong Kong Chinese children aged ≤14 years carried out in 2005 to 2006. Results. Foetal exposure to maternal passive smoking was significantly associated with wheeze ever (OR 2.05; 95% CI 1.58–2.67), current wheeze (OR 2.06; 95% CI 1.48–2.86), allergic rhinitis ever (OR 1.22; 95% CI 1.09–1.37), and eczema ever (OR 1.61; 95% CI 1.38–1.87). Foetal exposure to maternal active smoking was significantly associated with asthma ever (OR 2.10; 95% CI 1.14–3.84), wheeze ever (OR 2.46; 95% CI 1.27–4.78), and current wheeze (OR 2.74; 95% CI 1.24–6.01) but not with allergic rhinitis ever (OR 1.01; 95% CI 0.70–1.46) or eczema ever (OR 1.38; 95% CI 0.87–2.18). The dose response relationship between wheeze ever and current wheeze with increasing exposure, from no exposure to maternal passive smoking and then to maternal active smoking, further supports causality. Conclusion. There is significant association between foetal exposure to maternal passive smoking and maternal active smoking with childhood asthma and related atopic illnesses. Further studies are warranted to explore the potential causal relationship.
doi:10.1100/2012/542983
PMCID: PMC3425811
PMID: 22927783
Objective
To investigate the relationship between prescription medication use for pain and spasticity and ambulation distances, while controlling for pain severity, injury severity, age, gender, and race among participants with spinal cord injury (SCI).
Design
Secondary analysis of survey data.
Setting
A specialty hospital in the southeast USA.
Participants
407 adults with traumatic SCI identified through inpatient and outpatient hospital databases.
Interventions
Not applicable.
Main Outcomes Measures
A questionnaire measured prescription medication use, ambulation distance, and other demographic data. A composite score of four 10-point scales from the Brief Pain Inventory was used to measure pain severity. Multinomial logistic regression was used to calculate the odds ratio of ambulation distance using ≥1000 feet as the reference group.
Results
Persons with SCI who were heavy prescription medication users (defined as weekly or daily use for pain or spasticity) were more likely to be limited to distances of less than 150 feet (Odds ratio [OR]=2.82, confidence interval [CI]: 1.57, 5.04) and between 150 feet–999 feet (OR=2.52, CI: 1.45, 4.39).
Conclusion
Heavy prescription medication use for pain and spasticity was inversely related to a person’s ability to achieve community ambulation distances of ≥1000 feet.
doi:10.1016/j.apmr.2011.03.006
PMCID: PMC3149790
PMID: 21807143
spinal cord injury; ambulation; walking; medication adherence; pain; muscle spasticity
Liposome nanoparticles (LNs) with a targeting ligand were used in a semi-continuous flow electroporation (SFE) device to enhance in vitro delivery of exogenous oligonucleotides (ODN). Nanoparticles comprising transferrin-targeted lipoplex encapsulating ODN G3139 were mixed with K562 cells (a chronic myeloid leukemia cell line) and incubated for half an hour to accomplish nanoparticle binding. The mixture was then flowed through a SFE channel where electric pulses were given. Better ODN delivery efficiency was achieved with an increase of ~24% to the case in combination of non-targeted LNs and SFE, and ~60% to the case using targeted LNs alone, respectively. The MTS assay results confirmed cell viability greater than 75%.
doi:10.1016/j.bios.2010.06.025
PMCID: PMC3369826
PMID: 20630739
Electroporation; Lipoplex; Microfluidics; Gene Delivery; Antisense ODN
Purpose:
To identify the relationship of race and gender with 3 aspects of life satisfaction and depressive symptoms after spinal cord injury (SCI), evaluating the extent to which socioeconomic factors mediate any observed relationships.
Methods:
Adults with traumatic SCI of at least 1-year duration (N = 1,549) were identified through a Southeastern United States SCI Model System of care, and cross-sectional survey data were collected at a Southeastern United States medical university. Three aspects of life satisfaction (home life satisfaction, vocational satisfaction, global satisfaction) were measured using 20 satisfaction items from the Life Situation Questionnaire-Revised. The Older Adult Health and Mood Questionnaire measured depressive symptoms. MANCOVA assessed mediation of socioeconomic status between race and life satisfaction and depression.
Results:
Home life satisfaction and vocational satisfaction were significantly related to race, with White participants scoring higher than Black participants during the first stage of the regression. However, socioeconomic factors mediated the relationships such that race was no longer significant after considering economic factors. Race was significantly associated with global satisfaction after adjusting for socioeconomic factors. Depression was not significantly related to race. Gender was unrelated to all study outcomes. Of the socioeconomic mediators, family income was a significant predictor of each outcome, whereas education was only predictive of vocational satisfaction.
Conclusion:
Socioeconomic factors are important mediators of the relationship between race and certain aspects of life satisfaction among persons with SCI. Family income and, to a lesser extent, education should be considered when evaluating race differences in life satisfaction after SCI.
doi:10.1310/sci1803-232
PMCID: PMC3479640
PMID: 23105915
depression; health status disparities; socioeconomic factors; spinal cord injury
Tang, Tracy | Tang, Jean Y. | Li, Dongwei | Reich, Mike | Callahan, Christopher A. | Fu, Ling | Yauch, Robert L. | Wong, Frank | Kotkow, Karen | Chang, Kris | Shpall, Elana | Wu, Angela | Rubin, Lee L. | Marsters, James C | Epstein, Ervin H. | Caro, Ivor | de Sauvage, Frederic J.
Inappropriate activation of the Hedgehog (Hh) signaling pathway in skin is critical for the development of basal cell carcinomas (BCCs). We have investigated the anti-BCC efficacy of topically-applied CUR61414, an inhibitor of the Hedgehog signal transduction molecule Smoothened. In mice, topical CUR61414 significantly inhibited skin Hh signaling, blocked the induction of hair follicle anagen, and shrank existing BCCs. However, we observed no clinical activity of this formulation in human superficial or nodular BCCs in a phase I clinical study. Our data highlight some of the challenges of translating preclinical experience into successful human results for a topical anti-cancer agent.
doi:10.1158/1078-0432.CCR-10-3370
PMCID: PMC3113453
PMID: 21558397
Cummings, A. C. | Lee, S. L. | McCauley, J. L. | Jiang, L. | Crunk, A. | McFarland, L. L. | Gallins, P. J. | Fuzzell, D. | Knebusch, C. | Jackson, C. E. | Scott, W. K. | Pericak-Vance, M. A. | Haines, J. L.
Summary
Parkinson disease (PD) is a common complex neurodegenerative disorder with an underlying genetic etiology that has been difficult to dissect. Although some PD risk genes have been discovered, most of the underlying genetic etiology remains unknown. To further elucidate the genetic component, we have undertaken a genome-wide linkage screen in an isolated founder population of Amish living in the Midwestern United States. We performed tests for linkage and for association using a marker set of nearly 6,000 single nucleotide polymorphisms. Parametric multipoint linkage analysis generated a LOD score of 2.44 on chromosome 6 in the SYNE1 gene, approximately 8 Mbp from the PARK2 gene. In a different region on chromosome 6 (~67 Mbp from PARK2) an association was found for rs4302647 (p< 4.0×10−6), which is not within 300 kb of any gene. While the association exceeds Bonferroni correction, it may yet represent a false positive due to the small sample size and the low minor allele frequency. The minor allele frequency in affecteds is 0.07 compared to 0.01 in unaffecteds. Taken together, these results support involvement of loci on chromosome 6 in the genetic etiology of PD.
doi:10.1111/j.1469-1809.2011.00643.x
PMCID: PMC3077806
PMID: 21488853
Leeds, J. A. | LaMarche, M. J. | Brewer, J. T. | Bushell, S. M. | Deng, G. | Dewhurst, J. M. | Dzink-Fox, J. | Gangl, E. | Jain, A. | Lee, L. | Lilly, M. | Manni, K. | Mullin, S. | Neckermann, G. | Osborne, C. | Palestrant, D. | Patane, M. A. | Raimondi, A. | Ranjitkar, S. | Rann, E. M. | Sachdeva, M. | Shao, J. | Tiamfook, S. | Whitehead, L. | Yu, D.
Recently, we identified aminothiazole derivatives of GE2270 A. These novel semisynthetic congeners, like GE2270 A, target the essential bacterial protein elongation factor Tu (EF-Tu). Medicinal chemistry optimization of lead molecules led to the identification of preclinical development candidates 1 and 2. These cycloalklycarboxylic acid derivatives show activity against difficult to treat Gram-positive pathogens and demonstrate increased aqueous solubility compared to GE2270 A. We describe here the in vitro and in vivo activities of compounds 1 and 2 compared to marketed antibiotics. Compounds 1 and 2 were potent against clinical isolates of methicillin-resistant Staphylococcus aureus and vancomycin-resistant enterococci (MIC90 ≤ 0.25 μg/ml) but weaker against the streptococci (MIC90 ≥ 4 μg/ml). Like GE2270 A, the derivatives inhibited bacterial protein synthesis and selected for spontaneous loss of susceptibility via mutations in the tuf gene, encoding EF-Tu. The mutants were not cross-resistant to other antibiotic classes. In a mouse systemic infection model, compounds 1 and 2 protected mice from lethal S. aureus infections with 50% effective doses (ED50) of 5.2 and 4.3 mg/kg, respectively. Similarly, compounds 1 and 2 protected mice from lethal systemic E. faecalis infections with ED50 of 0.56 and 0.23 mg/kg, respectively. In summary, compounds 1 and 2 are active in vitro and in vivo activity against difficult-to-treat Gram-positive bacterial infections and represent a promising new class of antibacterials for use in human therapy.
doi:10.1128/AAC.00582-11
PMCID: PMC3195004
PMID: 21825297
Ichida, Justin K. | Blanchard, Joel | Lam, Kelvin | Son, Esther Y. | Chung, Julia E. | Egli, Dieter | Loh, Kyle M. | Carter, Ava C. | Di Giorgio, Francesco P. | Koszka, Kathryn | Huangfu, Danwei | Akutsu, Hidenori | Liu, David R. | Rubin, Lee L. | Eggan, Kevin
Summary
The combined activity of three transcription factors can reprogram adult cells into induced pluripotent stem (iPS) cells. However, the transgenic methods used to deliver reprogramming factors have raised concerns regarding the future utility of the resulting stem cells. These uncertainties could be overcome if each transgenic factor were replaced with a small molecule that either directly activated its expression from the somatic genome or in some way compensated for its activity. To this end, we have used high-content chemical screening to identify small molecules that can replace Sox2 in reprogramming. We show that one of these molecules functions in reprogramming by inhibiting Tgf-β signaling in a stable and trapped intermediate cell type that forms during the process. We find that this inhibition promotes the completion of reprogramming through induction of the transcription factor Nanog.
doi:10.1016/j.stem.2009.09.012
PMCID: PMC3335195
PMID: 19818703
Oakes, SR | Vaillant, F | Lim, E | Lee, L | Breslin, K | Feleppa, F | Deb, S | Ritchie, ME | Takano, E | Ward, T | Fox, SB | Generali, D | Smyth, GK | Strasser, A | Huang, DCS | Visvader, JE | Lindeman, GJ
doi:10.1186/1897-4287-10-S2-A25
PMCID: PMC3326698
Goldblatt, D. | Plikaytis, B. D. | Akkoyunlu, M. | Antonello, J. | Ashton, L. | Blake, M. | Burton, R. | Care, R. | Durant, N. | Feavers, I. | Fernsten, P. | Fievet, F. | Giardina, P. | Jansen, K. | Katz, L. | Kierstead, L. | Lee, L. | Lin, J. | Maisonneuve, J. | Nahm, M. H. | Raab, J. | Romero-Steiner, S. | Rose, C. | Schmidt, D. | Stapleton, J. | Carlone, G. M.
Lot 89SF has been the reference standard serum pool used in pneumococcal enzyme-linked immunosorbent assays (ELISAs) since 1990. In 2005, it was estimated that there remained between 2 and 5 years' supply of lot 89SF. Since lot 89SF was the reference standard used in the evaluation of the seven-valent pneumococcal conjugate vaccine Prevnar (PCV7), the link to clinical efficacy would be severed if stocks became completely depleted. Furthermore, demonstration of immune responses comparable to those elicited by PCV7 is a licensure approach used for new pneumococcal conjugate vaccines, so a replacement reference standard was required. A total of 278 volunteers were immunized with the 23-valent unconjugated polysaccharide vaccine Pneumovax II, and a unit of blood was obtained twice within 120 days following immunization. Plasma was prepared, pooled, and confirmed to be free from hepatitis B virus (HBV), hepatitis C virus (HCV), and HIV. The pooled serum was poured at 6 ml per vial into 15,333 vials and lyophilized. Immunological bridging of 007sp to 89SF was used to establish equivalent reference values for 13 pneumococcal capsular serotypes (1, 3, 4, 5, 6A, 6B, 7F, 9V, 14, 18C, 19A, 19F and 23F) by five independent laboratories. Antibody concentrations in 007sp were established relative to the lot 89SF reference preparation using the WHO reference ELISA. Subsequently, 12 existing WHO calibration sera had concentrations reassigned for 13 pneumococcal serotypes using new serum 007sp as the reference, and these were compared to concentrations relative to the original reference serum. Agreement was excellent for the 12 WHO calibration sera. The 007sp preparation has replaced 89SF as the pneumococcal reference standard. Sufficient quantity of this new preparation is available such that, with judicious use, it should be available for at least 25 years.
doi:10.1128/CVI.05252-11
PMCID: PMC3187044
PMID: 21852547
Objective
To identify the relationship between spasticity and life satisfaction as measured by 3 multi-item factor scales and a rating of overall quality of life among participants with spinal cord injury (SCI).
Study Design
Cross-sectional survey; secondary analysis of existing data by linear regression analysis between spasticity and quality of life.
Setting
Large specialty hospital in the Southeastern United States.
Methods
Participants included 1,549 adults with traumatic spinal cord injuries, at least 18 years of age and a minimum of 1 year post-injury at survey. Outcome measures included: (1) Home Life Satisfaction, (2) Global Satisfaction, (3) Vocational Satisfaction, (4) Overall Quality of Life and (5) three subscales from the Patient Reported Impact of Spasticity Measure.
Results
Three aspects of spasticity (Daily Activities, Positive Impact, and spasticity at its worst) all were negatively correlated with Home Life Satisfaction, Global Satisfaction, and Overall Quality of Life. Only the Daily Activities scale and the spasticity at its worst rating had a significant negative correlation with Vocational Satisfaction.
Conclusion
Spasticity is negatively associated with quality of life after SCI. These negative outcomes need to be considered in an individual’s rehabilitation and treatment methods.
doi:10.1038/sc.2011.49
PMCID: PMC3166438
PMID: 21606929
spinal cord injury; spasticity; life satisfaction; quality of life