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Biomedical subjects

A Kim

Publications and source records attributed to A Kim.

At least 91 records · Page 5Linked to original sources

Receptor-mediated maternofetal transfer of immunoglobulins. Inhibition of transport of anti-HIV-1 immunoglobulin by generic immunoglobulins in the in vitro perfused placenta.

OBJECTIVES: The passage of immunoglobulin G (IgG) across the placenta is thought to involve Fc' receptors on the syncytiotrophoblast. To confirm the receptor dependency of this process we have studied the changes in the tissue content and transfer kinetics of immunoglobulins and hyperimmune serum to HIV (HIVIG) during in vitro dual placental perfusion. METHODS: Isolated lobules of term placentae from normal pregnancies were perfused in a model of maternal and fetal circulation. The perfused tissue was compared to fresh tissue samples from the same placenta for the content of IgG, IgG subclasses, IgM, cytokeratin, human placental lactogen and SP1 antigen by immunohistochemistry and by protein elution. RESULTS: The immunoglobulin staining faded by an average of 40% during the 1st hour of perfusion. In contrast, staining for cytokeratin, human placental lactogen and SP1 remained unchanged. During a 4-hour recycling of endogenous immunoglobulins in the maternal circulation, IgG and HIVIG crossed to the fetal side in a steady rate. The transport of HIVIG could be inhibited by preperfusion with an intravenous gammaglobulin preparation (IVIG). DISCUSSION: The transfer of IgG across the placenta occurs in a steady state rate consistent with a receptor-mediated mechanism. Furthermore, inhibition of HIVIG maternofetal transfer by IVIG further establishes the receptor-mediated transfer of immunoglobulins through the placenta.

Female↗

Two cases of atrial flutter with fetal hydrops: successful fetal drug therapy.

We describe two cases of fetal atrial flutter associated with severe fetal hydrops which were unresponsive to digoxin but were successfully treated with flecainide acetate. Two cases of fetal atrial flutter were identified in fetuses with severe fetal hydrops on 3rd trimester ultrasonogram(28 weeks' gestation and 30 weeks' gestation). Following failed digoxin monotherapy, flecainide acetate was added to digoxin. On the 7th day and 13th day after combined therapy, fetal heart rate converted to normal sinus rhythm without recurrence. Our cases showed that the combined therapy of digoxin and flecainide acetate can effectively treat fetal atrial flutter associated with fetal hydrops unresponsive to digoxin monotherapy.

Adult↗

Fate of gamma-irradiated Listeria monocytogenes during refrigerated storage on raw or cooked turkey breast meat.

The radiation resistance and ability of Listeria monocytogenes ATCC 7644, 15313, 43256, and 49594 to multiply on irradiated, air-packed, refrigerated raw or cooked turkey breast meat nuggets (ca. 25 g) and ground turkey breast meat was investigated. Gamma-radiation D values for L. monocytogenes were significantly different on raw and cooked nuggets, 0.56 +/- 0.03 kGy and 0.69 +/- 0.03 kGy, respectively; but they were not significantly different (P < or = 0.05) on raw and cooked ground turkey meat. High populations (approximately 10(9) CFU/g) of L. monocytogenes declined during 14 days of storage at 4 degrees C in both irradiated and nonirradiated samples of raw but not of cooked ground turkey breast meat. A moderate inoculum (approximately 10(3) CFU/g) did not survive a radiation dose of 3 kGy. The population increased in cooked but not in raw samples of irradiated ground turkey meat stored at either 2 or 7 degrees C for 21 days. The D value changed significantly from 0.70 +/- 0.04 to 0.60 +/- 0.02 kGy when the product was cooked to an internal temperature of 80 degrees C before irradiation. Growth on either raw or cooked turkey meat did not alter the radiation resistance of L. monocytogenes. Analyses were performed for pH, aw, moisture, and reducing potential of raw and cooked turkey meat and for pH, amino acid profile, thiamine, and riboflavin contents of aqueous extracts of raw and cooked turkey meats without identifying the factor or factors involved in differences in the survival and multiplication of L. monocytogenes on raw and cooked meat.

Animals↗

Dose from syringe procedures during technetium-99m radiopharmaceutical preparation.

UNLABELLED: We examined the dose from 99mTc contained in syringes and shielded vials to assess in detail the dose burdens in a central pharmacy. METHODS: Absorbed-dose rates at the end of the plunger of a shielded syringe, when 99mTc is contained either in the syringe or in a shielded vial from which the activity would be drawn, were measured with CaF2 dosimeters. The dose rates also were calculated with a Monte Carlo model. RESULTS: When activity was contained in either 3-ml or 10-ml disposable syringes shielded by lead glass, the absorbed-dose rates were 1.35-1.62 mGy hr-1 GBq-1 (5-6 mrad hr-1 mCi-1). When the activity was contained in either a shielded elution or product kit vial, the absorbed-dose rates at the end of the syringe plunger were about 0.40 mGy hr-1 GBq-1 (1.4-1.5 mrad hr-1 mCi-1). These results were reproduced with reasonable accuracy by Monte Carlo simulations. CONCLUSION: The dose burden per unit of activity handled from 99mTc in procedures using syringes is likely to be two to five times larger than the dose burden from calibrating generator eluate. The Monte Carlo simulations suggest that lead K x-rays may be responsible for a significant fraction of the total dose to the fingers and hand of the pharmacist when lead-glass syringe shields are used.

Equipment Design↗

Thy/Liv-SCID-Hu mice implanted with human intestine: an in vivo model for investigation of mucosal transmission of HIV.

Mucosal transmission is a major route by which individuals become infected with HIV. Investigation into the mechanism by which mucosal transmission of HIV occurs would be greatly facilitated by the development of a small animal model infectible with HIV by the mucosal route. We have previously described a SCID-hu mouse model, in which human thymic and liver tissues are implanted under both kidney capsules (thy/liv-SCID-hu mice), which are populated in the periphery with high numbers of human T cells and that develop disseminated HIV-1 infection after intraperitoneal injection. To expand further the usefulness of the thy/liv-SCID-hu mouse as a model for studying mucosal transmission of HIV, thy/liv-SCID-hu mice were subcutaneously implanted with human intestinal tissue in a manner that maintained the lumen. Four months later, the histological appearance of the implanted intestine resembled that of normal human bowel tissue and the lamina propria was populated with human T cells. Six weeks after introduction of HIV into the lumen of the intestinal implant, the mice developed disseminated HIV infection. Scattered HIV-infected cells were detected in the lamina propria of the implant, indicating that HIV infection in these mice was mediated by transmission of the virus across the mucosa of the human intestinal implant. Thus, our modified thy/liv-SCID-hu mice transplanted with human bowel tissue should provide a novel model for investigating mucosal transmission of HIV.

Animals↗

Mechanisms of reduced striatal NMDA excitotoxicity in type I nitric oxide synthase knock-out mice.

We investigated the role of neuronal (type I) nitric oxide synthase (nNOS) in NMDA-mediated excitotoxicity in wild-type (SV129 and C57BL/6J) and type I NOS knock-out (nNOS-/-) mice and examined its relationship to apoptosis. Excitotoxic lesions were produced by intrastriatal stereotactic NMDA microinjections (10-20 nmol). Lesion size was dose- and time-dependent, completely blocked by MK-801 pretreatment, and smaller in nNOS knock-out mice compared with wild-type littermates (nNOS+/+, 11.7 +/- 1.7 mm3; n = 8; nNOS-/-, 6. 4 +/- 1.8 mm3; n = 7). The density and distribution of striatal NMDA binding sites, determined by NMDA receptor autoradiography, did not differ between strains. Pharmacological inhibition of nNOS by 7-nitroindazole (50 mg/kg, i.p.) decreased NMDA lesion size by 32% in wild-type mice (n = 7). Neurochemical and immunohistochemical measurements of brain nitrotyrosine, a product of peroxynitrite formation, were increased markedly in wild-type but not in the nNOS-/- mice. Moreover, elevations in 2,3- and 2,5-dihydroxybenzoic acid levels were significantly reduced in the mutant striatum, as a measure of hydroxyl radical production. The importance of apoptosis to NMDA receptor-mediated toxicity was evaluated by DNA laddering and by quantitative histochemistry [terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate-biotin nick end-labeling (TUNEL) staining]. DNA laddering was first detected within lesioned tissue after 12-24 hr. TUNEL-positive cells were first observed at 12 hr, increased in number at 48 hr and 7 d, and were located predominantly in proximity to the lesion border. The density was significantly lower in nNOS-/- mice. Hence, oligonucleosomal DNA breakdown suggesting apoptosis develops as a late consequence of NMDA microinjection and is reduced in nNOS mutants. The mechanism of protection in nNOS-/- mice may relate to decreased oxygen free radical production and related NO reaction products and, in part, involves mechanisms of neuronal death associated with the delayed appearance of apoptosis.

Animals↗

Severe combined immunodeficiency mice engrafted with human T cells, B cells, and myeloid cells after transplantation with human fetal bone marrow or liver cells and implanted with human fetal thymus: a model for studying human gene therapy.

To develop an in vivo model wherein human hematopoiesis occurs, we transplanted severe combined immunodeficiency (SCID) mice with either human fetal bone marrow (HFBM) or human fetal liver (HFL). After transplantation of SCID mice with cultured HFBM (BM-SCID-hu mice) or HFL cells (Liv-SCID-hu mice), significant engraftment of the mouse bone marrow (BM) and population of the peripheral blood with human leukocytes was detected. Human colony-forming unit-granulocyte macrophage and burst forming unit-erythroid were detected in the BM of the BM-SCID-hu and Liv-SCID-hu mice up to 8 months after transplantation. When the HFBM or HFL cells were transduced with a retroviral vector before transplantation, integrated retroviral sequences were detected in human precursor cells present in the SCID mouse BM and in leukocytes circulating in the peripheral blood (PB) up to 7 months after transplantation. The PB of the BM-SCID-hu mice also became populated with human T cells after implantation with human thymic tissue, which provided a human microenvironment wherein human pre-T cells from the BM could mature. When the HFBM was retrovirally transduced before transplantation, integrated retrovirus was detected in sorted CD4+CD8+ double positive and CD4+ single positive cells from the thymic implant and CD4+ cells from the PB. Taken together, these data indicated that the BM of our BM-SCID-hu and Liv-SCID-hu mice became engrafted with retrovirally transduced human hematopoietic precursors that undergo the normal human hematopoietic program and populate the mouse PB with human cells containing integrated retroviral sequences. In addition to being a model for studying in vivo human hematopoiesis, these mice should also prove to be a useful model for investigating in vivo gene therapy using human stem/precursor cells.

Animals↗

A new model of spinal cord edema.

Edema of the spinal cord has not been well understood. Brain edema produced by Marmarou's infusion method is essentially similar to vasogenic edema. This infusion method for producing edema was applied to a cat spinal cord. After laminectomy, a 30-gauge needle was inserted into the intumescentia cervicalis. A total amount of 10 microliters of 2% Evans' blue or autoserum were infused using an infusion pump at a rate of 5 microliters/hr. Macroscopally, Evans' blue was observed in the vicinity of infused site at the same level of the needle insertion and was seen spreading mainly longitudinally in the lateral column for a certain distance. The extracellular space was markedly distended in the in fused white mater and filled with electron-dense materials which were thought to be proteins in the electron microscopic study. The fine structural features were similar to the findings which were seen in Marmarou's infusion type of brain edema. Using this model, it seems to be feasible to produce reproducible spinal cord edema at any location in order to investigate not only the morphological aspect but also physiological aspect of the edema.

Animals↗

Effect of ovariectomy on the local residual ridge remodeling.

STATEMENT OF PROBLEM: Osteoporosis and edentulism are two disease processes that affect a large group of elderly people in the United States (24 and 25 million, respectively). These two diseases are independent of each other; however, they have several pathologic symptoms in common, such as reduction in bone mass. PURPOSE: The purpose of this study was to determine whether estrogen deficiency or its replacement therapy have any effect on the phenomenon of residual ridge remodeling. MATERIAL AND METHODS: Three animal groups were formed that consisted of six female Sprague-Dawley rats each. The two groups had ovariectomy and received either a vehicle solution or a daily dose (1.5 micrograms/day) of 17 beta-estradiol delivered through osmotic pumps. The control group underwent sham surgery and received a vehicle solution. Animals were pair fed throughout the experiment. Unilateral molar extraction was performed in the maxilla, which produced a suitable site for examination of histologic characteristics and molecular biologic analyses. At the 4-week postextraction period the bone remodeling activity was noted at the surface of the residual ridge in the control group. RESULTS: The ovariectomized group showed increased bone resorption activity, whereas the surface of the residual ridge alveolar bone of the ovariectomized and estrogen-treated group was covered by a layer of hyaline tissue. Poly(A)+ ribonucleic acid samples were isolated from the remodeling residual ridge tissues. Expression of alpha 2(I), alpha 1(II), alpha 1(IX), and alpha 2(X) collagens were examined by ribonucleic acid transfer dot blots. Compared with the control group, ovariectomized animals showed a reduction in bone formation with decreased expressions of type I and II collagens. In contrast, the estrogen-treatment group showed decreased formation of type I collagen with a much increased expression of type II collagen. Further examination of type II collagen formation on the ovariectomized and estrogen-treated group by means of in situ hybridization revealed the notable labeling by the type IIA collagen probe, which was associated with the surface tissue of the residual ridge alveolar bone. CONCLUSION: These findings suggest that estrogen deficiency and its replacement therapy seem to affect the activity of residual ridge bone remodeling at the molecular level.

Alveolar Process↗

Three-vessel view of the fetal upper mediastinum: an easy means of detecting abnormalities of the ventricular outflow tracts and great arteries during obstetric screening.

The three-vessel view is a transverse view of the fetal upper mediastinum is as simple to obtain as the four-chamber view. It demonstrates the main pulmonary artery, ascending aorta and superior vena cava in cross- or oblique sections. The purposes of this study were to describe the normal anatomy of the three-vessel view and to analyze what anatomical changes would occur in this view when there are lesions of the ventricular outflow tracts and/or great arteries. Sonograms of 29 fetuses with lesions involving the ventricular outflow tracts and/or great arteries were reviewed. Three-vessel views were evaluated in terms of vessel size, number, arrangement and alignment. Twenty-eight of 29 fetuses showed an abnormal three-vessel view that included abnormal vessel size (n = 12), abnormal alignment (n = 8), abnormal arrangement (n = 7) and abnormal vessel number (n = 3). The vessel size was abnormal in obstructive lesions of the right (n = 4) or the left (n = 8) side of the heart. An abnormal alignment was seen in tetralogy of Fallot (n = 6) and double-outlet right ventricle (n = 2) that showed anterior displacement of the aorta. An abnormal arrangement was seen in complete (n = 4) and corrected (n = 1) transposition, double-outlet right ventricle (n = 1) and pulmonary atresia with ventricular septal defect (n = 1). Only two vessels were seen in truncus arteriosus (n = 1). Four vessels were seen in persistent left superior vena cava (n = 2). A fetus with pulmonary atresia and intact ventricular septum showed a normal three-vessel view. In conclusion, most of the lesions involving the ventricular outflow tracts and/or great arteries showed an abnormal three-vessel view.

Aortic Valve Stenosis↗

Urban-rural differences in prehospital care of major trauma.

OBJECTIVES: To compare differences in response times, scene times, and transport times by advanced life-support-trained paramedics to trauma incidents in urban and rural locations. METHODS: This report was a prospective cohort study of professional emergency medical services conducted in a five-county area in the state of Washington. Ninety-eight percent of trauma transports are provided by professional paramedics trained in advanced life support. Subjects were included in this study if they qualified as a major trauma victim and were transported or found dead at the scene by one of the region's advanced life support transport agencies between August 1, 1991, and January 31, 1992. The severity of injury was rated using the Prehospital Index. Incident locations were defined as "rural" if they occurred in a US Census division (a geographic area) in which more than 50% of the residents resided in a rural location. RESULTS: During the 6-month data collection period, advanced life support agencies responded to a total of 459 major trauma victims in the region. A geographic locations was determined for 452 of these subjects. Of these, 42% of subjects were injured in urban areas and the remainder in rural areas. The severity of injuries, as determined both by the triage classification (p = 0.17) and the distribution of Prehospital Index scores (p = 0.92), was similar for urban and rural major trauma patients. Twenty-six (5.7%) subjects died at the scene. About one quarter of both groups had a severe injury, as indicated by Prehospital Index score of more than 3. The mean response time for urban locations was 7.0 minutes (median = 6 minutes) compared with 13.6 minutes (median = 12 minutes) for rural locations (p < 0.0001). The mean scene time in rural areas was slightly longer than in urban areas (21.7 vs. 18.7 minutes, p = 0.015). Mean transport times from the scene to the hospital were also significantly longer for rural incidents (17.2 minutes vs. 8.2 minutes, p < 0.0001). Rural victims were over seven times more likely to die before arrival (relative risk = 7.4, 95% confidence interval 2.4-22.8) if the emergency medical services' response time was more than 30 minutes. CONCLUSIONS: Response and transport times among professional, advanced life-support-trained paramedics responding to major trauma incidents are longer in rural areas, compared with urban areas.

Algorithms↗

Fidelity and mutational spectrum of Pfu DNA polymerase on a human mitochondrial DNA sequence.

The study of rare genetic changes in human tissues requires specialized techniques. Point mutations at fractions at or below 10(-6) must be observed to discover even the most prominent features of the point mutational spectrum. PCR permits the increase in number of mutant copies but does so at the expense of creating many additional mutations or "PCR noise". Thus, each DNA sequence studied must be characterized with regard to the DNA polymerase and conditions used to avoid interpreting a PCR-generated mutation as one arising in human tissue. The thermostable DNA polymerase derived from Pyrococcus furiosus designated Pfu has the highest fidelity of any DNA thermostable polymerase studied to date, and this property recommends it for analyses of tissue mutational spectra. Here, we apply constant denaturant capillary electrophoresis (CDCE) to separate and isolate the products of DNA amplification. This new strategy permitted direct enumeration and identification of point mutations created by Pfu DNA polymerase in a 96-bp low melting domain of a human mitochondrial sequence despite the very low mutant fractions generated in the PCR process. This sequence, containing part of the tRNA glycine and NADH dehydrogenase subunit 3 genes, is the target of our studies of mitochondrial mutagenesis in human cells and tissues. Incorrectly synthesized sequences were separated from the wild type as mutant/wild-type heteroduplexes by sequential enrichment on CDCE. An artificially constructed mutant was used as an internal standard to permit calculation of the mutant fraction. Our study found that the average error rate (mutations per base pair duplication) of Pfu was 6.5 x 10(-7), and five of its more frequent mutations (hot spots) consisted of three transversions (GC-->TA, AT-->TA, and AT-->CG), one transition (AT-->GC), and one 1-bp deletion (in an AAAAAA sequence). To achieve an even higher sensitivity, the amount of Pfu-induced mutants must be reduced.

DNA Mutational Analysis↗

Saquinavir-mediated inhibition of human immunodeficiency virus (HIV) infection in SCID mice implanted with human fetal thymus and liver tissue: an in vivo model for evaluating the effect of drug therapy on HIV infection in lymphoid tissues.

Treatment with protease inhibitors alone or in combination with inhibitors of reverse transcriptase potently suppresses levels of human immunodeficiency virus (HIV) RNA in plasma and thereby may significantly delay the progression of HIV-mediated disease. To investigate the effect of treatment with the protease inhibitor saquinavir on HIV replication in the lymphoid tissues, we used a SCID-hu mouse model that we developed, in which human thymic and liver tissues (hu-thy/liv) were implanted under both kidney capsules in SCID mice (thy/liv-SCID-hu mice). These mice are populated in the periphery with large numbers of human T cells and develop disseminated HIV infection after intraimplant injection. thy/liv-SCID-hu mice with established HIV infection that were treated for 1 month with saquinavir had a significantly lower viral load present in the implanted hu-thy/liv and mouse spleen than did the untreated HIV-infected thy/liv-SCID-hu mice. To examine the capacity of acute treatment with saquinavir to prevent HIV infection, some thy/liv-SCID-hu mice were inoculated with HIV and then immediately started on saquinavir. Although treated mice had markedly lower viral loads in the thy/liv implants and spleens, HIV infection was not completely prevented. Thus, the effect of antiviral therapy on HIV infection in the major site of HIV replication, the lymphoid tissues, can be readily evaluated in our thy/liv-SCID-hu mice. These mice should prove to be a useful model for determining the in vivo effectiveness of different therapeutic interventions on acute and chronic HIV infection.

Administration, Oral↗

Cellular antioxidant properties of human natural killer enhancing factor B.

The protein, NKEF (natural killer enhancing factor), has been identified as a member of an antioxidant family of proteins capable of protecting against protein oxidation in cell-free assay systems. The mechanism of action for this family of proteins appears to involve scavenging or suppressing formation of protein thiyl radicals. In the present study we investigated the antioxidant protective properties of the NKEF-B protein overexpressed in an endothelial cell line (ECV304). Nkef-B-transfected cells displayed significantly lower levels of reactive oxygen species (ROS) compared with control or vector-transfected cells. Tert-Butylhydroperoxide-induced ROS was 15% lower in nkef-B-transfected cells and cytotoxicity was slightly, though not significantly, lower. NKEF-B had no effect on ROS induced by menadione or xanthine plus xanthine oxidase. NKEF-B overexpression resulted in slightly (approximately 10%) lower levels of cellular glutathione (GSH) and had no effect on rate or extent of GSH depletion following either diethylmaleate (DEM) or buthionine sulfoximine (BSO) treatment. Lipid peroxidation, assessed as thiobarbituric acid-reactive substances, was 40% lower in nkef-B-transfected cells compared with vector-only-transfected cells. DEM-induced lipid peroxidation was suppressed by NKEF-B at DEM concentrations of 20 microM to 1 mM. At 10 mM DEM, lipid peroxidation was unaffected by NKEF-B. NKEF-B expression also protected cells against menadione-induced inhibition of [3H]-thymidine uptake. The NKEF-B protein appears most effective in suppressing basal low-level oxidative injury such as that produced during normal metabolism. These results indicate that overexpression of the NKEF-B protein promotes resistance to oxidative stress in this endothelial cell line.

Antioxidants↗

Perforated appendicitis within an inguinal hernia: case report and review of the literature.

The finding of the vermiform appendix within an inguinal hernia sac is not uncommon. However, it is rare to find a perforated appendix within an inguinal hernia. An unusual case of an incarcerated and perforated appendix within an inguinal hernia complicated by an intra-abdominal abscess is reported herein. Perforated appendix as a cause of abscess was revealed during abdominal exploration. Clinicians are encouraged to be aware of this unusual entity, which is rarely recognized before exploration.

Abdominal Abscess↗

About the origin of retroviruses and the co-evolution of the gypsy retrovirus with the Drosophila flamenco host gene.

The gypsy element of Drosophila melanogaster is the first retrovirus identified so far in invertebrates. According to phylogenetic data, gypsy belongs to the same group as the Ty3 class of LTR-retrotransposons, which suggests that retroviruses evolved from this kind of retroelements before the radiation of vertebrates. There are other invertebrate retroelements that are also likely to be endogenous retroviruses because they share with gypsy some structural and functional retroviral-like characteristics. Gypsy is controlled by a Drosophila gene called flamenco, the restrictive alleles of which maintain the retrovirus in a repressed state. In permissive strains, functional gypsy elements transpose at high frequency and produce infective particles. Defective gypsy proviruses located in pericentromeric heterochromatin of all strains seem to be very old components of the genome of Drosophila melanogaster, which indicates that gypsy invaded this species, or an ancestor, a long time ago. At that time, Drosophila melanogaster presumably contained permissive alleles of the flamenco gene. One can imagine that the species survived to the increase of genetic load caused by the retroviral invasion because restrictive alleles of flamenco were selected. The characterization of a retrovirus in Drosophila, one of the most advanced model organisms for molecular genetics, provides us with an exceptional clue to study how a species can resist a retroviral invasion.

Animals↗