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Biomedical subjects

M Kim

Publications and source records attributed to M Kim.

At least 19 recordsLinked to original sources

Acquired resistance to reoviral oncolysis in Ras-transformed fibrosarcoma cells.

Reovirus shows considerable potential as an oncolytic agent for Ras-activated tumors and is currently in clinical trials. Here we ask whether such tumor cell lines can acquire resistance to reoviral oncolysis. We challenged human HT1080 fibrosarcoma cells that carry a Ras mutation by prolonged exposure to reovirus, thereby yielding highly virus-resistant HTR1 cells. These cells are persistently infected with reovirus, exhibit high Ras activity and retain the original Ras gene mutation, showing that resistance to reovirus can be displayed in cells with active Ras. The HTR1 cells also exhibit reduced cellular cathepsin B activity, which normally contributes to viral entry and activation. Persistently infected HTR1 cells were not tumorigenic in vivo, whereas immunologically cured virus-free HTR1 cells were highly tumorigenic. Thus, acquisition of resistance to reovirus may constrain therapeutic strategies. To determine whether reoviral resistance is associated with a general reduction in apoptotic potential, we challenged the HTR1 cells with apoptotic inducers and E1B-defective adenovirus, resulting in significant apoptosis and cell death following both approaches. Therefore, even if resistance to reoviral oncolysis should arise in tumor cells in vivo, other therapeutic strategies may nevertheless remain effective.

Base Sequence↗

The oncolytic effect in vivo of reovirus on tumour cells that have survived reovirus cell killing in vitro.

The use of oncolytic viruses has received considerable attention in recent years and many viruses have proved to be effective against a variety of cancer models and a few are currently being used in clinical trials. However, the possible emergence and outcome of virus-resistant tumour cells has not been addressed. We previously reported the effective use of reovirus against lymphoid malignancies, including the Burkitt's lymphoma cell line Raji. Here we isolated in vitro persistently infected (PI) Raji cells, and cells 'cured' of persistent reovirus infection ('cured' cells). Both PI and cured Raji cells resisted reovirus infection and cell killing in vitro. In vivo, the PI cells were non-tumorigenic in SCID mice, but cured cells regained the parental cells' ability to form tumours. Tumour xenografts from the cured cells, however, were highly susceptible to reovirus oncolysis in vivo. This susceptibility was due to the proteolytic environment within tumours that facilitates reovirus infection and cell killing. Our results show that persistent infection by reovirus impedes tumour development and that although PI cells cleared of reovirus are tumorigenic, they are killed upon rechallenge with reovirus. Both the PI and cured states are therefore not likely to be significant barriers to reovirus oncolytic therapy.

Animals↗

Dietary gamma-aminobutyric acid affects the brain protein synthesis rate in young rats.

The purpose of this study was to determine whether the gamma-aminobutyric acid (GABA) affects the rate of brain protein synthesis in male rats. Two experiments were done on five or three groups of young rats (5 wk) given the diets containing 20% casein administrated 0 mg, 25 mg, 50 mg, 100 mg or 200 mg/100 g body weight GABA dissolved in saline by oral gavage for 1 day (d) (Experiment 1), and given the diets contained 0%, 0.25% or 0.5% GABA added to the 20% casein diet (Experiment 2) for 10 d. The plasma concentration of growth hormone (GH) was the highest in rats administrated 50 mg and 100 mg/100 g body weight GABA. The concentration of serum GABA increased significantly with the supplementation groups. The fractional (Ks) rates of protein synthesis in brain regions, liver and gastrocnemius muscle increased significantly with the 20% casein + 0.25% GABA diet and still more 20% casein + 0.5% GABA compared with the 20% casein diet. In brain regions, liver and gastrocnemius muscle, the RNA activity [g protein synthesized/(g RNA . d)] significantly correlated with the fractional rate of protein synthesis. The RNA concentration (mg RNA/g protein) was not related to the fractional rate of protein synthesis in any organ. Our results suggest that the treatment of GABA to young male rats are likely to increase the concentrations of plasma GH and the rate of protein synthesis in the brain, and that RNA activity is at least partly related to the fractional rate of brain protein synthesis.

Aging↗

Field- and pressure-induced phases in Sr4Ru3O10: a spectroscopic investigation.

We have investigated the magnetic-field- and pressure-induced structural and magnetic phases of the triple-layer ruthenate Sr4Ru3O10. Magnetic-field-induced changes in the phonon spectra reveal dramatic spin-reorientation transitions and strong magnetoelastic coupling in this material. Further, we are able to deduce key magnetoelastic coupling parameters, and evidence that the magnetic moments are localized on the Ru sites. Additionally, pressure-dependent Raman measurements at different temperatures reveal an anomalous negative Gruneisen parameter associated with the B(1g) mode (approximately 380 cm(-1)) at low temperatures (T < 75 K), which can be explained consistently with the field-dependent Raman data.

Journal Article↗

Cancer gene therapy using a novel secretable trimeric TRAIL.

Tumor necrosis factor (TNF)-related apoptosis-inducing ligand (TRAIL), a member of the TNF family, is a type II transmembrane cytokine molecule. Soluble TRAIL has been shown to induce apoptosis in a wide variety of cancer cells in vitro and to suppress tumor growth specifically without damaging normal cells and tissues in vivo. In our previous report, we have demonstrated that an artificial gene encoding the polypeptide composed of the three functional elements (a secretion signal, a trimerization domain and an apoptosis-inducing moiety of TRAIL gene sequence) expresses and secretes highly apoptotic trimeric TRAIL into the culture supernatant. Here, as an approach to TRAIL-based cancer gene therapy, we developed an adenoviral vector delivering the gene that encodes our secretable trimeric TRAIL (stTRAIL). This adenovirus (Ad-stTRAIL) potently induced apoptosis in vitro in cancer cell lines such as HeLa, MDA-MB-231, A549, HCT116 and U-87MG. In an animal xenograft tumor model bearing a human glioma cell line U-87MG, intratumoral delivery of Ad-stTRAIL dramatically suppressed tumor growth without showing detectable adverse side effects. Histological analysis revealed that Ad-stTRAIL suppresses tumor growth by inducing apoptotic cell death. Contrary to the known rapid clearance of systemically delivered TRAIL protein from the blood circulation, stTRAIL expressed by Ad-stTRAIL in tumor tissues persisted for more than 4 days. In a comparison of tumor suppressor activity between Ad-stTRAIL and Ad-flTRAIL (delivering the full-length TRAIL gene) after mixing infected cells with uninfected cells and implanting these mixed cells in nude mice, Ad-stTRAIL showed higher tumor suppressor activity than that of Ad-flTRAIL. Our data reveal that a gene therapy using Ad-stTRAIL has a promising potential to treat human cancers including gliomas.

Adenoviridae↗

Development of an impregnated reagent and automation of solid-phase analytical derivatization for carbonyls: proof of principle.

This study undertakes reduction of scale and automation of a solid-phase analytical derivatization of carbonyls with 2,4-dinitrophenylhyrazine on a styrene-divinylbenzene resin (XAD-2). Three processes are tested. In the batch process, an aqueous phase consisting of 50 microL of sample and 150 microL of reagent solution is contacted with 6 mg XAD-2 by shaking. An impregnated reagent consisting of 2,4-dinitrophenylhydrazine hydrochloride (DNPH) deposited on XAD-2 enables two additional processes. In-vial derivatization with an impregnated reagent requires shaking 50 microL of sample with 6 mg of the impregnated reagent and reduced the reaction time from 10 to 5 min. The third process involves packing impregnated reagent a mini-column and flowing 50 microL of sample through under positive pressure supplied by a Harvard Pump. This reduces sample preparation time to 1 min. Studies are conducted with worst-case model analytes: butanone, 2-pentanone, and malonyldialdehyde. The carbonyl of the two ketones is hindered, and, thus, these two compounds react very slowly with DNPH in aqueous solution. Malonyldialdehyde is highly water soluble, and it does not react in aqueous phase but also would not sorb onto XAD-2 for reaction. Nevertheless, derivatization/extraction of all model compounds any of the three procedures result in reproducible and high yields.

Automation, Laboratory↗

Prevalence of workplace exacerbation of asthma symptoms in an urban working population of asthmatics.

OBJECTIVES: We used an interviewer-administered questionnaire to investigate workplace exacerbation of asthma symptoms (WEAS) among low-income, minority, working asthmatics admitted Bellevue Hospital Center in New York City from 2001 to 2002. We hypothesized that a high prevalence of WEAS would be found in this population among all jobs held and a subset of individual occupational classifications. MEASUREMENTS AND MAIN RESULTS: Of 301 subjects, 51% reported WEAS in their current or most recent job; 71% reported WEAS in any job. Prevalences (95% confidence intervals) of WEAS in common job classifications were 61% (49-73%) in janitorial jobs, 50% (33-67%) in garment and textile manufacturing jobs, and 38% (23-55%) in construction jobs. CONCLUSION: WEAS is prevalent in this urban minority population.

Adult↗

Natural history of patients with recurrent chronic hepatitis C virus and occult hepatitis B co-infection after liver transplantation.

It is uncertain whether occult hepatitis B virus co-infection will hasten progressive liver disease in chronic hepatitis C patients after liver transplantation. This study evaluated fibrosis progression and severe fibrosis in 118 consecutive hepatitis B surface antigen-negative patients with virological and histological evidence of recurrent chronic hepatitis C infection co-infected with occult hepatitis B virus after liver transplantation. HBV DNA was detected from serum at the time of recurrent chronic hepatitis C infection by polymerase chain reaction. Each subject underwent a repeat liver biopsy 5 years post-liver transplantation. Occult hepatitis B virus co-infection was present in 41 of the 118 (34.7%) patients. At 5 years post-liver transplantation, 13 of the 41 occult hepatitis B virus co-infected patients compared with 16 of the 77 patients without occult hepatitis B virus co-infection developed fibrosis progression (31.7% vs. 20.8%, respectively, p = 0.39). Eight of 41 the occult hepatitis B virus co-infected patients compared with 13 of the 77 patients without occult hepatitis B virus co-infection had severe fibrosis (19.5% vs. 16.9%, respectively, p = 0.97). In conclusion, occult hepatitis B virus co-infection in patients with recurrent chronic hepatitis C infection was not associated with accelerated fibrosis progression or severe fibrosis after liver transplantation.

Adult↗

Saccular aneurysms on straight and curved vessels are subject to different hemodynamics: implications of intravascular stenting.

Our aim was to examine hemodynamic implications of intravascular stenting in the canine venous pouch (sidewall or straight-vessel) and rabbit elastase (curved-vessel) aneurysm models. Flow dynamics in stented (Wallstent) and nonstented versions were studied by using computational fluid dynamics simulations and in vitro flow visualization, with a focus on stent placement effects on aneurysmal flow stagnancy and flow impingement. Results show that sidewall and curved aneurysm models have fundamentally different hemodynamics (shear-driven versus inertia-driven) and thus stent placement outcomes.

Animals↗

APOE epsilon2/epsilon4 polymorphism and cerebral microbleeds on gradient-echo MRI.

The association of APOE genotypes with cerebral microbleeds (CMBs) was examined on the basis of the location of CMBs in 414 patients who were admitted primarily because of stroke. With respect to possession of the epsilon2 or epsilon4 allele, the adjusted odds ratio was 1.94 (1.05 to 3.58) for lobar CMBs but 1.21 (0.69 to 2.11) for nonlobar CMBs. This suggests that the pathogenesis of CMBs may differ depending on their location.

Age Factors↗

JNK pathway mediates apoptotic cell death induced by tumor suppressor LKB1 in Drosophila.

Although recent progresses have unveiled the diverse in vivo functions of LKB1, detailed molecular mechanisms governing these processes still remain enigmatic. Here, we showed that Drosophila LKB1 negatively regulates organ growth by caspase-dependent apoptosis, without affecting cell size and cell cycle progression. Through genetic screening for LKB1 modifiers, we discovered the JNK pathway as a novel component of LKB1 signaling; the JNK pathway was activated by LKB1 and mediated the LKB1-dependent apoptosis. Consistently, LKB1-null mutant was defective in embryonic apoptosis and displayed a drastic hyperplasia in the central nervous system; these phenotypes were fully rescued by ectopic JNK activation as well as wild-type LKB1 expression. Furthermore, inhibition of LKB1 resulted in epithelial morphogenesis failure, which was associated with a decrease in JNK activity. Collectively, our studies unprecedentedly elucidate JNK as the downstream mediator of the LKB1-dependent apoptosis, and provide a new paradigm for understanding the diverse LKB1 functions in vivo.

AMP-Activated Protein Kinase Kinases↗

Viral evolution of hepatitis C in injection drug users.

Injection drug users represent the largest cohort of patients with established hepatitis C virus (HCV) infection as well as the group that is at highest risk for new infections. Most published studies have focused on the clinical consequences of established HCV infection and have not examined the consequences of new infection. The aim of the current study was to measure the virological consequences of HCV in patients with ongoing injection drug use that might pose a risk for new and/or for superinfection with additional strains of HCV. We examined the following groups: (a) those with resolved HCV infection with ongoing injection drug use, (b) those with chronic infection who continued to inject and (c) those with chronic infection who no longer injected. Our study demonstrated a spectrum of responses. The majority of patients appeared to be 'protected' from new infection. None of six patients with resolved infection had detectable HCV RNA by quantitative or qualitative PCR when followed for 1 year. Similarly, despite ongoing injection drug use, no patient with persistent infection had a 'switch' in HCV genotype indicative of possible superinfection. Virological analysis of HCV quasispecies to detect possible infection with new variants of HCV in patients with apparently 'stable' infection, indicated divergence of virus over time, divergence that was unrelated to injection drug behaviour. Thus, patients with ongoing or prior HCV infection appear to develop immunity that protects against further infection with HCV despite repeated exposure.

Adult↗

Covalent and noncovalent chemical modifications of arginine residues decrease dopamine transporter activity.

Rotating disk electrode voltammetry was used to measure dopamine (DA) transport in rat striatum and in human embryonic kidney cells expressing the rat dopamine transporter (DAT). The goals of this study were to determine 1) if arginine (Arg) selective agents could alter DA transport, and 2) if DA analogs and DAT inhibitors could attenuate the effects of these agents on the DAT. Phenylglyoxal (PG), Hill coefficient 2.5, and other Arg selective agents decreased DA transport velocities. DA, Hill coefficient 1.0, and its analogs 3-hydroxyphenethylamine and 4-hydroxyphenethylamine attenuated the effects of PG on the DAT while phenethylamine did not. The tropane-based DAT inhibitors cocaine, WIN 35065-2, and WIN 35428 also attenuated the effects of PG. Benztropine, GBR 12935, and GBR 12909 did not. Thus, Arg residues are important for DAT activity and the results suggest that DA and cocaine both interact with Arg residues. Structure-activity studies suggest that DA interacts with Arg through its catechol hydroxyl groups and cocaine through the ester linkage attached to carbon 2 of the tropane ring. The results that 1). DA and cocaine may interact with the same functionally important Arg residue at the DAT, and 2). some members of the tropane and 1,4-dialkylpiperazine classes of DAT inhibitors may interact differently with DAT-derived Arg residue(s) furthers the notion that DAT activity sparing antagonists of cocaine can be designed.

Animals↗

Enhanced transfollicular delivery of adriamycin with a liposome and iontophoresis.

To find a better way to deliver drugs into hair follicles, we tried two approaches: single topical application using various liposomes; and iontophoresis combined with topical application of ionic liposome. After delivery of adriamycin (ADR) to wax-depilated rat skin, the transport of the drug was examined under fluorescence microscopy. Most liposomal ADR showed more effective transdermal and transfollicular penetration than free ADR. Among tested liposomes, the non-ionic GDL liposome (GDL/CH/POE-10 = glycerol dilaulate/cholesterol/polyoxyethylene-10) was the most selective to hair follicles against skin, while the cationic liposome (GDL/CH/POE-10/DOTAP, dioleoyl trimethylammonium propane) containing monocationic DOTAP was less selective; however, it was better at improving the delivery amount and penetration of ADR into the follicles and skin. The DMPC/DMPG (7/3) formulation of anionic PC liposome (DMPC/DMPG = dimyristoyl-phosphocholine/-phospoglycerol) showed results similar to the cationic liposome. The DMPC/DMPG (3/7) formulation yielded poor results, however, probably because of its increased viscosity and anionic property. Although ADR delivery was enhanced by liposomal formulations, topical applications had some limitations in delivery capacity and speed. To accelerate delivery, iontophoresis was combined with the cationic liposome at positive 0.2-0.4 mA/cm(2) for 20-30 min. The resulting delivery of ADR through follicular routes was excellent. This combination method diffused ADR 3.0-fold more efficiently, rapidly and deeply than single topical application of cationic liposomal ADR. This system also achieved a 3.5-fold higher diffusive follicular delivery than a free ADR/iontophoresis combination. Furthermore, it was demonstrated that the tetracationic lipid DOSPER and hydrophile spermine could serve as a cationic additive instead of the monocationic DOTAP in the liposome. These results suggest that the combinative system of the topically applied cationic liposome followed by iontophoresis has a significant synergistic effect on the transfollicular delivery of ADR.

Administration, Cutaneous↗

Suppressive effect of functional drinking yogurt containing specific egg yolk immunoglobulin on Helicobacter pylori in humans.

Helicobacter pylori is a human pathogen that infects over 50% of the population worldwide. It is the most important etiologic agent of gastroduodenal ulcers and malignancies. Helicobacter pylori urease enzyme is considered the main factor for the organism's colonization in the gastroduodenal mucosa. Hens immunized with the purified urease produce a highly specific anti-H. pylori urease immunoglobulin (IgY-urease) in their egg yolks. Immunoglobulin Y-urease was stable at 60 to 65 degrees C for 30 min and at pH 4.0 for 7 h. Its activity was lost at 80 degrees C for 20 min and at pH 2 for 4 h. Specially designed functional drinking yogurt containing Lactobacillus acidophilus and Bifidobacterium spp. with 1% egg yolk IgY-urease was produced commercially. Immunoglobulin Y-urease activity showed stability in the product up to 7 d, and then decreased to 85% after 3 wk of storage. A clinical study was conducted to determine the effectiveness of IgY-urease yogurt to suppress infection in humans. Forty-two volunteers who tested positive for H. pylori using a 13C-urea breath test were recruited. A total of 450 mL of IgY-urease (test group) or IgY-urease-free yogurt (control group) was consumed in 150-mL portions 3 times daily for 4 wk. Volunteers were tested after 2 and 4 wk; urea breath test values significantly decreased in the test group compared with the control group. The results indicate that suppression of H. pylori infection in humans could be achieved by consumption of drinking yogurt fortified with IgY-urease.

Adult↗