tbuckley/cabot-search huggingface dataset:
95. Effective Genetic Engineering of Human Embryonic Stem Cells Using the Sleeping Beauty Transposon System
Authors: Andrew Wilber, Jonathan L. Linehan, Xinghui Tian, R. Scott McIvor, Dan S. Kaufman
Journal: Molecular Therapy (2006)

Human embryonic stem (ES) cells hold great promise for the study of human development and the generation of new therapeutic approaches to tissue regeneration and their genetic modification will play a key role in this development. While viral vectors have been readily used for gene transfer into human ES cells, only limited success has been achieved using non-viral vectors for this purpose. The Sleeping Beauty (SB) transposon system mediates gene insertion into chromosomes, providing a means of durable gene expression in target cells and tissues. To determine whether the SB transposon system can mediate stable gene expression in human ES cells, undifferentiated H9 human ES cells were nucleofected with an SB transposon encoding the firefly luciferase (luc) gene. The vector was designed to be transcriptionally regulated by promoters containing or depleted of CpG dinucleotides, and was delivered with or without a DNA or RNA source of transposase. Transfected human ES cells were maintained in culture, and luc expression was monitored by bioluminescence imaging. Only human ES cells cotransfected with transposon and transposase encoding sequences exhibited luc expression after 2 weeks. Importantly, we found that after 6 weeks, incorporation of a CpG-free promoter increased the average luc activity by 6- to 10-fold compared to a promoter containing CpG sequences. After 5 months, stable expression of luc was observed in human ES cells co-transfected with a transposon containing the CpG-free promoter along with transposase-encoding DNA or RNA. Human ES cells engineered with the CpG-free promoter demonstrated the ability to differentiate into hematopoietic cells in vitro and into teratomas in vivo and maintained luc expression with minimal incidence of epigenetic silencing. We conclude that the SB transposon system provides an effective approach for genetic manipulation of human ES cells and for durable expression using appropriately engineered transposon cargo. Proof-of-principal studies are currently in progress to test gain-of-function and loss-of-function outcomes using the SB system in human ES cells whereby any candidate gene can be evaluated for its role in the differentiation of human ES cells into committed lineages.
Targeting Gastrin-Releasing Peptide Receptors on Small Cell Lung Cancer Cells with a Bispecific Molecule that Activates Polyclonal T Lymphocytes
Authors: Jiehua Zhou, Jian Chen, Ruikun Zhong, Michael Mokotoff, Leonard D. Shultz, Edward D. Ball
Journal: Clinical Cancer Research (2006)

Gastrin-releasing peptide (GRP) is a growth factor for small cell lung cancer (SCLC). GRP belongs to the bombesin peptide family and has significant homology to bombesin. We constructed a bispecific molecule, OKT3xAntag2, by conjugating a monoclonal antibody OKT3 (anti-CD3) with a bombesin/GRP antagonist (Antag2) and evaluated cytotoxicity against SCLC cells.We tested binding of the bispecific molecule to SCLC cell lines and T cells by flow cytometry, antibody-dependent cellular cytotoxicity (ADCC) of SCLC cells in vitro and in a murine SCLC xenograft model. We studied SCLC apoptosis and necrosis during ADCC and the activity and cleavage of caspase-3, caspase-9, and poly(ADP-ribose) polymerase (PARP).The bispecific molecule functions as a cross-linker between T cells and SCLC cells, induces T cell activation, and mediates ADCC of SCLC cells; 40% to 80% growth inhibition of SCLC cells mediated by the bispecific molecule at low effector to target cell ratios was achieved. Activation of T cells by the bispecific molecule resulted in significant increases in IFNgamma production and apoptosis and necrosis of SCLC cells associated with cleavage of PARP and caspase-3. Targeted immunotherapy with the bispecific molecule-armed human T cells significantly reduced SCLC tumor burdens in a mouse model.The bispecific molecule OKT3xAntag2 mediates growth inhibition and apoptosis of SCLC cells by activated T cells through activation and cleavage of caspase-3 and PARP in vitro and in vivo. Clinical trials of this bispecific molecule through adoptive transfer of ex vivo activated T cells in GRP receptor-positive tumors, such as SCLC, are warranted.
Psychological aspects of pain.
Authors: R G Large
Journal: Annals of the Rheumatic Diseases (1996)

Large'An unpleasant sensory and emotional experi- ence associated with actual or potential tissue damage, or described in terms of such damage'.This is the definition of pain accepted by the International Association for the Study of Pain (IASP) and it is a modification of Merskey's original definition.' 2 The striking character- etic dystrophy, now called 'complex regional pain syndrome'-the avoidance of movement and stimulation may be central to the pathophysiology of the disorder.Much of the
A PLEA FOR EAREY VAGINAL HYSTERECTOMY FOR CANCER OF THE UTERUS.Read in the Section of Obstetrics and Diseases of Women, at the Forty-first Annual Meeting of the American Medical Association, held in Nashville, Tenn., May, 1890.
Authors: FRANKLIN H. MARTIN
Journal: JAMA (1891)

It is, at present, a generally conceded fact that cancer can be permanently eradicated from the human body if the parts of primary development and subsequent invasion can, early in the progress of disease, be absolutely separated and removed from the surrounding normal tissue. This implies that the operation for cancer which will go the farthest beyond the diseased limits, is the one most liable to accomplish absolute removal, consequently the one by which the most satisfactory curative results must be expected. With this fact well established, we can have no hesitancy in making the assertion that the operation which will obtain the best ultimate results in cancer of any portion of the uterus is the one which will accomplish its total removal, together with as much of the tissue which lies in close proximity to it as can be removed without unduly increasing immediate mortality. Hence my first proposition
Self administered tampons can be used to diagnose sexually transmitted diseases
Authors: F. J Bowden, B. A Paterson, S. M Garland, S. Tabrizi, C. K Fairley
Journal: BMJ (1997)

Editor–Lars Ostergaard and colleagues highlight the importance of self administered techniques for diagnosing and screening for chlamydial infection in women.1 They used the combined results from three samples (first void urine, midstream urine, and a sample obtained with a 5 ml vaginal pipette) but were concerned about whether women in the general community would comply with this method.

We have used a tampon method administered by patients to detect several genital infections, including infection with Chlamydia trachomatis , …
Antietileptics: age-old search for effective therapy continues
Authors: Terra Ziporyn
Journal: JAMA (1985)

Blood of the tortoise. Testicles of the hippopotamus. Feces of the land crocodile. Root of strychnos gathered at the time of the waning of the moon, put into a piece of linen, and hung around the neck. Scrapings from the selenite stone found by night at the waxing of the moon. Filings of iron sharpened on whetstone from Naxos. What all these substances have in common is that at one time they were all used as drugs to treat epilepsy (Owsei Temkin,<i>The Falling Sickness</i>, Johns Hopkins University Press, Baltimore, 1979). Today carbamazepine, clonazepam, chlorazepate dipotassium, and valproic acid are the latest additions to the modern pharmacologic armamentarium against epilepsy. Together with drugs such as phenytoin, phenobarbital, and primidone, they have greatly relieved seizure frequency for about 85% of the estimated 2.5 million patients with epilepsy in the United States. "But for the other 15%, the rules don't work," says
Personal Digital Educators
Authors: James J. Cimino, Suzanne Bakken
Journal: New England Journal of Medicine (2005)

"What's his infection sensitive to?" In 1995, getting the answer to this question required a house officer to find an unoccupied computer at the nursing station, sign on, and traverse the necessary screens to get the culture result. Armed with the answer — say, "levofloxacin" — she would ask the follow-up question: "What's the renal dosing?" The next step would typically entail a search for a tattered copy of a drug reference manual.In 2000, the house officer still needed to turn to the computer for the first answer, but for the second, she might have skipped the scavenger hunt . . .
Enhancement of Xenograft Tumor Radiosensitivity by the Histone Deacetylase Inhibitor MS-275 and Correlation with Histone Hyperacetylation
Authors: Kevin Camphausen, Tamalee Scott, Mary Sproull, Philip J. Tofilon
Journal: Clinical Cancer Research (2004)

Histone deacetylase (HDAC) inhibitors are undergoing clinical evaluation in cancer therapy. Because HDAC modulation has been shown to enhance the radiosensitivity of tumor cells in vitro, we investigated the effects of the HDAC inhibitor MS-275 on the radioresponse of DU145 prostate carcinoma xenografts.As an indicator of HDAC inhibition in vivo, the histone acetylation status in tumor lysates was determined after two, four, and six injections of MS-275 delivered at 12-hour intervals, as well as 24 and 48 hours after the last injection. Tumor growth delay studies were then performed using this DU-145 xenograft model with radiation administered to leg tumors after the fourth dose of MS-275, which corresponded to the time of maximum histone hyperacetylation.An increase in histone hyperacetylation was detected in each tumor after two injections of MS-275 with a maximum hyperacetylation occurring after four to six injections. In tumor growth delay studies, the combination of MS-275 and radiation resulted in a greater than additive inhibition of tumor growth as compared with the individual modalities. As alternative sources for an indicator of drug radiosensitizing activity, histone hyperacetylation was determined in a series of normal tissues, including lymphocytes. Each of the normal tissues also had a maximal histone hyperacetylation after four to six injections of MS-275.These studies show that MS-275 enhances the radiosensitivity of DU145 xenografts and suggest that histone hyperacetylation status can serve as a useful marker for drug radiosensitizing activity.
Quantitative Buffy Coat Analysis-Reply
Authors: Stephen Wardlaw
Journal: JAMA (1983)

<h3>In Reply.—</h3> The apparent complexity of quantitative buffy coat analysis that Fischer and Addison refer to in their letter was certainly true in our prototype instrumentation and methodology. A system is currently being manufactured that can be easily operated by personnel who have no laboratory training. The instrument incorporates a microprocessor that uses the described algorithms instantly to display results after the measurements are made. Results are available in less than 12 minutes, with about one minute of actual<i>hands-on</i>time. We polled five senior hematology technologists, and they stated that the same tests, when performed by their most skilled technologists using the methods described by Fischer and Addison, would take at least 25 minutes,<i>the majority of that being hands-on time</i>. In our article, the data were taken in a manner that would show the<i>worst case</i>performance of the system. Data were gathered over a period of weeks,
How Little Kidney Tissue Is Enough?
Authors: Leon G. Fine
Journal: New England Journal of Medicine (1991)

One of the intriguing mysteries of biologic design is why there is such redundancy in the function of the kidneys. If a single kidney is removed (as it is from a living renal-transplant donor), the remaining kidney promptly grows larger and increases its glomerular and tubular functions to approach the levels previously achieved by two kidneys. When nephrons are gradually destroyed by glomerular or tubulointerstitial diseases, as little as 10 percent of the original nephron population can maintain acceptable fluid and electrolyte balance, at a level that is certainly sufficient to sustain life. It is only when renal function falls . . .
Bacterial lipopolysaccharides prime human neutrophils for enhanced production of leukotriene B4.
Authors: M E Doerfler, R L Danner, J H Shelhamer, J E Parrillo
Journal: Journal of Clinical Investigation (1989)

Neutrophils can be "primed" for an enhanced respiratory burst by lipopolysaccharide (LPS) in concentrations measurable in patients with septic shock. Leukotriene B4 (LTB4) is the primary eicosanoid product of neutrophils and is felt to be a mediator of host defense and inflammation. We investigated the in vitro effects of LPS on neutrophil production of LTB4 and the omega-oxidation metabolites of LTB4. Incubation of neutrophils with LPS in concentrations ranging from 0.01 to 100 ng/ml did not result in production of LTB4 or metabolites in the absence of a second stimulus. Priming neutrophils with LPS and then stimulating with opsonized zymosan, phorbol-myristate-acetate or a low concentration of the calcium ionophore A23187 resulted in enhanced production of LTB4. LPS priming of neutrophils occurred in a concentration dependent manner. LPS did not result in LTB4 production in response to the chemoattractant peptide FMLP. LPS priming of neutrophils had no effect on cytosolic calcium concentrations of resting or zymosan-stimulated cells. These results suggest that LPS might effect host defense and tissue injury by potentiating the effect of other stimulants on neutrophil production of LTB4. This LPS induced enhancement may represent an important pathogenetic pathway in patients with gram negative sepsis.
Late-onset hydroa vacciniforme: two case reports
Authors: S-N. Wong, S.H. Tan, S.W. Khoo
Journal: British Journal of Dermatology (2001)

Hydroa vacciniforme (HV) is a rare blistering photodermatosis that heals with vacciniform scarring, with onset usually in childhood and spontaneous resolution by early adulthood. We report two cases of HV seen at the National Skin Centre, Singapore, both with a late onset at 20 years of age, during compulsory military service, and one with a less typical non-vesicular presentation, that initially caused some diagnostic difficulties.
Insulin regulates apolipoprotein B turnover and phosphorylation in rat hepatocytes.
Authors: T K Jackson, A I Salhanick, J Elovson, M L Deichman, J M Amatruda
Journal: Journal of Clinical Investigation (1990)

Our laboratory has previously shown that insulin inhibits the secretion of newly-synthesized and immunoreactive apo B from rat hepatocytes. We have also shown that apo B is secreted as a phosphoprotein and that phosphorylation is increased in hypoinsulinemic nonketotic diabetes. The present studies were conducted to determine whether the ability of insulin to inhibit apo B secretion is related to alterations in apo B turnover and whether insulin itself affects apo B phosphorylation. Pulse-chase studies with [35S]methionine in primary cultures of hepatocytes from normal rats in the absence and presence of insulin show that the secretion of apo B100 and apo B48 are inhibited by insulin and that this inhibition may be due in part to enhanced intracellular degradation. In addition, there is a second intracellular apo B48 pool which is not insulin regulated or degraded. In experiments in which hepatocytes were incubated with [32P]orthophosphate, insulin decreased 32P incorporation into apo B100 (42%) with only small effects on apo B48 (11%). The small insulin effect on apo B48 may relate to an insulin-insensitive apo B48 intracellular pool. These studies show that insulin can affect the intracellular turnover, secretion, degradation, and phosphorylation of apo B and emphasize the differential regulation of apo B100 and apo B48 with regard to these parameters in rat liver.
Genetic polymorphisms and childhood acute lymphoblastic leukemia: GWAS of the ESCALE study (SFCE)
Authors: L Orsi, J Rudant, A Bonaventure, S Goujon-Bellec, E Corda, T-J Evans, A Petit, Y Bertrand, B Nelken, A Robert, G Michel, N Sirvent, P Chastagner, S Ducassou, X Rialland, D Hémon, E Milne, R J Scott, A Baruchel, J Clavel
Journal: Leukemia (2012)
Smoking and Carcinoma of the Lung
Authors: R. Doll, A. B. Hill
Journal: BMJ (1950)

DNA repair is crucial for genome stability in the developing cortex, as somatic de novo mutations cause neurological disorders. However, how DNA repair contributes to neuronal development is largely unknown. To address this issue, we studied the spatiotemporal roles of DNA polymerase β (Polβ), a key enzyme in DNA base excision repair pathway, in the developing cortex using distinct forebrain-specific conditional knock-out mice, Emx1-Cre/Polβfl/fl and Nex-Cre/Polβfl/fl mice. Polβ expression was absent in both neural progenitors and postmitotic neurons in Emx1-Cre/Polβfl/fl mice, whereas only postmitotic neurons lacked Polβ expression in Nex-Cre/Polβfl/fl mice. We found that DNA double-strand breaks (DSBs) were frequently detected during replication in cortical progenitors of Emx1-Cre/Polβfl/fl mice. Increased DSBs remained in postmitotic cells, which resulted in p53-mediated neuronal apoptosis. This neuronal apoptosis caused thinning of the cortical plate, although laminar structure was normal. In addition, accumulated DSBs also affected growth of corticofugal axons but not commissural axons. These phenotypes were not observed in Nex-Cre/Polβfl/fl mice. Moreover, cultured Polβ-deficient neural progenitors exhibited higher sensitivity to the base-damaging agent methylmethanesulfonate, resulting in enhanced DSB formation. Similar damage was found by vitamin C treatment, which induces TET1-mediated DNA demethylation via 5-hydroxymethylcytosine. Together, genome stability mediated by Polβ-dependent base excision repair is crucial for the competence of neural progenitors, thereby contributing to neuronal differentiation in cortical development.  SIGNIFICANCE STATEMENT DNA repair is crucial for development of the nervous system. However, how DNA polymerase β (Polβ)-dependent DNA base excision repair pathway contributes to the process is still unknown. We found that loss of Polβ in cortical progenitors rather than postmitotic neurons led to catastrophic DNA double-strand breaks (DSBs) during replication and p53-mediated neuronal apoptosis, which resulted in thinning of the cortical plate. The DSBs also affected corticofugal axon growth in surviving neurons. Moreover, induction of base damage and DNA demethylation intermediates in the genome increased DSBs in cultured Polβ-deficient neural progenitors. Thus, genome stability by Polβ-dependent base excision repair in neural progenitors is required for the viability and differentiation of daughter neurons in the developing nervous system.
Determinants of lung bacterial clearance in normal mice.
Authors: S J Jay, W G Johanson, A K Pierce, J S Reisch
Journal: Journal of Clinical Investigation (1976)

The determinants of the lung clearance of Streptococcus pneumoniae, Klebsiella pneumoniae, Escherichia coli, and Staphylococcus aureus were studied in normal mice after exposure to an aerosol of viable bacteria and 99mTc-labeled dead bacteria. The fraction of bacteria in lungs that remained viable 4 h after exposure were: S. pneumoniae, 7.3%; K. pneumoniae, 121%; E. coli, 88.5%; S. aureus, 27.6%. The rate of physical removal of bacterial particles (Kmc) was determined from the change in lung 99mTc counts with time: Kmc ranged between 7 and 12%/h and and was similar in all species. The rate of mucociliary clearance and of intrapulmonary bacterial killing (Kk + Kmc) was calculated from the change in bacterial counts with time in animals that had received tetracycline to inhibit bacterial multiplication. Kk, the rate of intrapulmonary killing, was obtained by subtraction of Kmc from (Kk + Kmc). The calculated values for Kk were: S. pneumoniae, - 87%/h; K. pneumoniae, - 17%/h; E. coli, - 18%/h; S. aureus, - 22%/h. The rate of intrapulmonary bacterial multiplication (Kg) was estimated from the relationship of bacterial counts in tetracycline and nontetracycline-treated animals, assuming that tetracycline altered only Kg. Kg, expressed as the doubling time, was: S. pneumoniae, 310 min; K. pneumoniae, 217 min; E.coli, 212 min; S. aureus, infinity (no multiplication). The data indicate that the marked differences in the clearance of these species from the normal mouse lung result from the interaction of differing rates of in vivo bacterial multiplication and killing.
Medical Devices: Significant vs Nonsignificant Risk
Authors: Harriet Segal
Journal: JAMA (1995)

<h3>To the Editor.</h3> —After reading the recently published article regarding medical devices,<sup>1</sup>I feel compelled to respond. As an administrative assistant to an institutional review board (IRB) and voting member of that committee, I find it inconceivable that an effective IRB could be as easily manipulated and misled as the one described in the article by Drs Sherertz and Streed. Some manufacturers may feel they can intimidate an IRB or its institution with the implied threat that funding will be awarded elsewhere to a more cooperative facility. But, an IRB makes its own determinations, after serious deliberations, on all matters, and most certainly does not rely on a manufacturer's (biased) recommendations. I feel the authors should have stated that "Unfortunately,<i>some</i>IRBs are sometimes confused...." When an IRB is established in an institution, it is imperative that there is support staff with expertise to offer assistance and guidance on
In vitro hepatitis B virus infection of human bone marrow cells.
Authors: J B Zeldis, H Mugishima, H N Steinberg, E Nir, R P Gale
Journal: Journal of Clinical Investigation (1986)

Infection of humans with hepatitis B virus (HBV) frequently results in suppression of hematopoiesis; in some cases this may lead to severe bone marrow failure. The mechanism whereby HBV infection affects hematopoiesis is unknown. In vitro exposure of human bone marrow to HBV results in a dose-dependent inhibition of erythroid (erythroid burst forming units, BFU-E; erythroid colony-forming units CFU-E), myeloid (colony-forming units-granulocyte macrophage CFU-GM), and lymphoid (CFU-[T-lymphocytic]-TL) hematopoietic stem cells. Inactivation or immunoabsorption of HBV from sera resulted in loss of HBV-induced inhibition of hematopoietic stem cells. De novo gamma interferon was not detectable in the supernatants of cultures of bone marrow cells with HBV. Antibodies to gamma interferon did not affect the suppression of hematopoietic stem cells by HBV. Hepatitis B surface antigen (HBsAg) was detected by immune electron microscopy in nuclei of greater than 70% of immature hematopoietic cells including myeloblasts, normoblasts, and lymphoblasts; granulocytes had mostly cytoplasmic HBsAg. Hepatitis B virus core antigen (HBcAg) was also detected in about 5% of HBV infected bone marrow cells by immunoperoxidase staining. These data indicate that HBV can infect hematopoietic cells and their progenitors, thus suggesting a wider range of tropism for HBV than previously reported. These results may provide a basis to study HBV infection in vitro, and the effects of HBV on hematopoiesis.