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Biomedical subjects

J D Lee

Publications and source records attributed to J D Lee.

At least 163 records · Page 9Linked to original sources

The effects of Vero cell co-culture on human zygotes resulting from in vitro fertilization and oocytes following subzonal insemination.

A variety of co-culture systems have been devised to enhance the human embryo development in vitro. Vero cells were selected because they can be highly controlled and are easy to handle. To evaluate the embryotrophic effects of Vero cell monolayers, when they were co-cultured with human in vitro fertilized zygotes or subzonal inseminated oocytes. Total 1695 two-pronuclear embryos resulting from in vitro fertilization were cultured with Vero cells or medium alone for 24 hours. Similarly, sixty-six two-pronuclear embryos resulted from subzonal insertion of sperm (SUZI) with co-culture starting immediately following SUZI were compared with fifty-two two-pronuclear embryos resulted from SUZI, without co-culture. The numbers of blastomere and morphology of embryos were compared between the co-culture group and control group using student's t-test. Cell numbers in each embryo were greater in the IVF/co-culture group than in the control group (4.01 +/- 1.32 vs. 3.86 +/- 1.45, p < 0.05). The rates of poor quality embryo with major fragmentation were lower in the co-culture group than in the control group(11.5% vs. 19.9%, p < 0.001, for IVF embryos; 9% vs. 27%, p < 0.005, for SUZI oocytes). Co-culture SUZI oocytes on Vero cells prior to fertilization did not positively influence embryo cleavage, but improved embryo quality. We conclude that Vero cells can enhance human embryo development; however, the period for one-day or two-day co-culture is too short to provide a maximal support. Short term co-culture did not increase implantation rates. Immediate co-culture following SUZI might somewhat rescue the microinseminated oocytes. However, a longer duration of co-culture is necessary to exert the maximal effects on embryo development and implantation.

Adult↗

Idiopathic hypereosinophilic syndrome with eosinophilic myositis, peripheral neuropathy and central nervous system involvement.

Idiopathic hypereosinophilic syndrome (HES) is a rare disorder marked by a sustained overproduction of eosinophils and a predilection for damage to multiple organ systems. Its neurologic involvement ranges from the central to the peripheral nervous system, and can be associated with eosinophilic myositis. We report a 68-year-old woman who had eosinophilia, eosinophilic dermatitis and eosinophilic pneumonia. She also suffered from numbness and weakness of the lower limbs. Because of long-lasting (> 6 mo) eosinophilia (> 1.5 x 10(9)/L) in the peripheral blood and the fact that no other underlying causes of eosinophilia and neurologic involvement could be identified, a diagnosis of idiopathic hypereosinophilic syndrome was made. The muscle biopsy showed infiltration of inflammatory cells, including a few eosinophils (Liu's stain). Magnetic resonance images, motor evoked potentials, somatosensory evoked potentials and nerve conduction velocities also showed abnormalities in the central and peripheral nervous systems. The pathogenesis and treatments of HES are discussed in this report.

Central Nervous System Diseases↗

Bioflocculant produced by Aspergillus sp. JS-42.

A bioflocculant from a fungus, Aspergillus sp. JS-42, was purified by precipitations with acetone and cetylpyridinium chloride. The flocculating activity was not significantly affected by pH from 3 to 8, but was stimulated by the addition of CaCl2, and was effective only when the reaction mixture contained an adequate amount of flocculant. The flocculant could efficiently flocculate all tested solids suspended in aqueous solution, including various microorganisms, organic acids, and inorganic materials.

Aspergillus↗

Primary structure of BMK1: a new mammalian map kinase.

Mitogen-activated protein (MAP) kinases comprise a family of conserved, eukaryotic enzymes that mediate responses to a wide variety of extracellular stimuli. We have identified a new human MAP kinase gene here termed BMK1. BMK1 encodes a protein of 816 amino acid residues and has at least three different forms of mRNA. BMK1 messages are abundant in heart, placenta and kidney but not detectable in liver. Although BMK1 has the dual phosphorylation site of MAP kinases characterized by the TEY sequence found in ERK1 and ERK2, it has a distinct C-terminal and loop-12 structure when compared to other mammalian MAP kinases. This suggests BMK1 may regulate signaling events distinct from those controlled by the ERK group of enzymes.

Amino Acid Sequence↗

Reduction of dopamine beta-hydroxylase gene expression by calcium in bovine chromaffin cells.

We determined the effect of Ca++ influx on dopamine beta-hydroxylase (DBH) gene expression in primary cultured bovine chromaffin cells. The Ca++ ionophore A23187 (100nM) specifically reduced DBH mRNA to 15 +/- 10% of the untreated control while tyrosine hydroxylase was induced (142 +/- 15%) and phenylethanolamine N-methyltransferase was unchanged (107 +/- 11%). This effect on DBH was also observed with ionomycin, reversed by EGTA and unaffected by cycloheximide. Depolarization by potassium also resulted in DBH reduction which was reversed by the voltage dependent Ca++ channel blockers nifedipine and verapamil. DBH mRNA level reduced by A23187 could be re-induced by forskolin and DBH induced by forskolin could be reduced by the ionophore, suggesting that the cAMP and Ca++ pathways might act independently in regulating DBH gene expression.

Animals↗

Intrapleural chemotherapy for patients with incompletely resected malignant mesothelioma: the UCLA experience.

From 1986 to 1993, 15 patients with malignant pleural mesothelioma were treated by pleurectomy/decortication and intrapleural cisplatin (100 mg/m2) and cytosine arabinoside (1,200 mg). All patients were without known extrathoracic disease and had a mean age of 63 +/- 7.9 years (range 51-78); 13 were male. Histologic subtype of disease were epithelial (47%), sarcomatoid (27%), and mixed-biphasic (27%). The major morbidity and mortality rates were 13% and 0%, respectively. The mean length of hospital stay was 6.5 +/- 2.1 days. Postoperatively, adjuvant chemotherapy and radiation therapy were given to 46% and 73% of the patients respectively. Median survival from date of treatment was 11.5 months. Those patients with an epithelial histologic subtype experienced significantly improved survival compared to those of sarcomatoid subtype (P = 0.024). Whether adjuvant chemotherapy or radiation therapy were given had no significant effect on survival. These data suggest that although this treatment regimen can be administered with very limited morbidity and no mortality, the role of this approach in the treatment of malignant pleural mesothelioma appears limited and cannot currently be recommended.

Aged↗

Intravenous dopexamine in the treatment of acute congestive heart failure: results of a multicenter, double-blind, placebo-controlled withdrawal study.

Acute hemodynamic effects of intravenous infusion of dopexamine were evaluated by a placebo-controlled withdrawal study in patients with acute congestive heart failure. Twenty patients were enrolled at 10 centers in Japan. All patients had a pulmonary capillary or diastolic pressure of 15 mmHg or greater and a cardiac index of 2.5 l/min/m2 or less. Phase I: Intravenous dopexamine was introduced in a single-blind, uncontrolled fashion at the rate of 0.5 micrograms/kg/min and was titrated up to achieve a 30% or more increase in the cardiac index. Two patients withdrew from the study due to sinus tachycardia and ventricular ectopy or exacerbation of heart failure. Phase II: The remaining 18 responders who were free of limiting side effects were randomized in double-blind fashion to continue dopexamine or to switch to placebo for an additional 60 minutes. At the end of phase II, the hemodynamic improvement obtained in phase I of the study disappeared completely after substitution of placebo but was maintained in dopexamine-treated patients. Our findings suggest that dopexamine, when given in appropriate doses to selected patients, shows balanced vasodilator action suitable for the treatment of acute congestive heart failure.

Acute Disease↗

Serum levels of insulin-like growth factor I and insulin-like growth factor-binding protein-1 and -3 in women with regular menstrual cycles.

OBJECTIVE: To investigate the cyclic changes of serum insulin-like growth factor (IGF-I), IGF-binding protein-1 (IGFBP-1) and IGFBP-3 levels during ovulatory menstrual cycle. DESIGN: A prospective study following a preset protocol. SETTING: A tertiary-care academic medical center. PARTICIPANTS: Thirty young female adults with regular menstrual cycles were recruited (18 with normal luteal phase and 12 with inadequate luteal function confirmed by serum P levels). MAIN OUTCOME MEASURES: Serum concentrations of IGF-I, IGFBP-1, and IGFBP-3 from women with regular menstrual cycles were assayed. Circulating E2 and P levels also were determined to certify the ovulatory cycles. RESULTS: In women with normal luteal function, there was a peak of serum IGFBP-1 levels before ovulation concomitant with the preovulatory E2 peak. The nadir of serum IGFBP-1 levels was in the midluteal phase. Circulating IGFBP-1 elevated rapidly during late luteal phase and reached a peak on the 1st day of menstruation then declined slightly until a preovulatory IGFBP-1 peak occurred. In women with inadequate luteal function (midluteal serum levels of P < 10 ng/mL [conversion factor to SI unit, 3.180]), the preovulatory increase in serum IGFBP-1 was not significant and the circulating IGFBP-1 levels fluctuated throughout the menstrual cycle except for a unique peak of serum IGFBP-1 on the 1st day of menstruation. By contrast, there were no cyclic changes of serum IGF-I and IGFBP-3 levels in women with regular menstrual cycles, including both normal nad inadequate luteal functions. CONCLUSIONS: The preovulatory increase in serum IGFBP-1 levels may be of follicular origin and associated with the subsequent luteal function in females with ovulatory cycles. However, the involvement of IGFBP-1 in the process of follicular maturation and luteogenesis, as well as the regulation of luteal function, needs to be explored further.

Adult↗

Interleukin-4 inhibits the expression of tumour necrosis factors alpha and beta, interleukins-1 beta and -6 and interferon-gamma.

T cell-derived IL-4 inhibits the expression of several inflammatory cytokine genes. In this study, IL-4 downregulated TNF-alpha and -beta, IL-1 beta IL-6 and IFN-gamma steady-state mRNA and protein production by blood leukocytes. In a detailed kinetic analysis, cytokine mRNA levels were significantly decreased in the presence of IL-4, suggesting an early and prolonged downregulatory effect. These findings support the general thesis that IL-4 is an important regulator of the inflammatory immune response.

Cytokines↗

Detection of malignant melanoma by Tc-99m HMPAO.

A number of radiopharmaceutical agents have been evaluated for the detection of malignant melanoma. Because anamine is necessary for melanin synthesis, melanotic tumors may possess numerous amine receptors. Tc-99m HMPAO consists of macrocyclic amine radicals. The purpose of this study was to evaluate Tc-99m HMPAO as an effective agent for localizing malignant melanomas on the radioisotope imaging. Six patients, with malignant melanomas were studied. Tc-99m HMPAO SPECT images were obtained in all patients. SPECT was used for the localization of the primary focus, and whole body scans were used for the evaluation of unexpected metastatic lesions. The results were compared with MRI and CT. Tc-99m HMPAO SPECT revealed a high correlation with CT and MRI for the primary focus. Abnormally increased activity at the unexpected metastatic lymph nodes in two patients were also noticed. This was confirmed by CT and MRI for those sites. Tc-99m HMPAO appears to be a useful agent for the localization of both the primary focus and unexpected metastatic lesion in patients with malignant melanoma.

Aged↗

Evaluation of embolization effect of hepatocellular carcinoma by hepatic arterial flow study with Tc-99m MAA SPECT.

The degree of hepatocellular carcinoma vascularity in eight patients was assessed with Tc-99m MAA radionuclide angiography before and after treatment. For chemoembolization therapy, lipiodol and doxorubicin hydrochloride were administered intraarterially along with Gelfoam particles. The ratio of radioactivity in the hepatocellular carcinoma compared to that in the surrounding uninvolved tissue before therapy was 6.5-19.0 with a mean of 12.0. After therapy, the ratio dropped to 0.5-1.3. Because survival improved significantly after the therapeutic regimen used in these patients, radionuclide Tc-99m MAA angiography appears to be an accurate method for demonstrating successful embolization of the vascular supply in hepatocellular carcinoma.

Adult↗

Molecular cloning of the cDNA and gene for an elastinolytic aspartic proteinase from Aspergillus fumigatus and evidence of its secretion by the fungus during invasion of the host lung.

Hydrolysis of structural proteins in the lung by extracellular proteinases secreted by Aspergillus fumigatus is thought to play a significant role in invasive aspergillosis. This fungus was found previously to secrete an elastinolytic serine proteinase and a metalloproteinase. We report that A. fumigatus also secretes an aspartic proteinase (aspergillopepsin F) that can catalyze hydrolysis of the major structural proteins of basement membrane, elastin, collagen, and laminin. The pH optimum for the enzymatic activity was 5.0 with elastin-Congo red as the substrate, and the activity was not significantly inhibited by pepstatin A, diazoacetyl norleucine methylester, and 1,2-epoxy-3-(p-nitrophenoxy) propane. The cDNA and gene encoding this aspartic proteinase were cloned and sequenced. The open reading frame, interrupted by three introns, would encode a protein of 393 amino acids composed of a putative 21-amino-acid signal peptide and a 49-amino-acid propeptide preceding the 323-amino-acid mature protein. The amino acid sequence of A. fumigatus aspartic proteinase has 70, 66, and 67% homology to the sequences of those from Aspergillus oryzae, Aspergillus awamori, and Aspergillus saitoi, respectively. The active-site motif (DTG) and the catalytic aspartic residues characteristic of aspartic proteinases are found in the presently described enzyme, indicating that it belongs to a family of aspartic proteinases. Polyclonal antibodies were produced in rabbits against both the mature and precursor forms of the aspartic proteinase expressed in Escherichia coli. Immunoblotting with both antibodies detected a 39-kDa mature enzyme in the culture supernatant of A. fumigatus. The aspartic proteinase activity was inhibited by the antibodies, suggesting that the aspartic proteinase in the culture supernatant corresponds to the product of the cloned gene. Immunogold electron microscopy showed that the aspartic proteinase was secreted by A. fumigatus invading neutropenic mouse lung and its secretion was directed toward the germ tubes of penetrating hyphae.

Amino Acid Sequence↗

Tracheopathia osteoplastica simulating asthmatic symptoms. Diagnosis by bronchoscopy and computerized tomography.

Tracheopathia osteoplastica (TO) is a relatively rare benign disease of the trachea and major bronchi, characterized by cartilaginous and bony submucosal nodules covered by intact mucosa, which may cause narrowing and rigidity of the upper airways. The diagnosis of TO is rarely considered because of a lack of awareness of this entity, rather than the reported rare occurrence. We intend to report herein a case initially misinterpreted as bronchial asthma but later disclosed through computerized tomography (CT) and bronchoscopic biopsies as TO.

Asthma↗

Localization of intracellular monoclonal antibody specific for mycobacteria in experimentally induced pulmonary tuberculous lesion.

To determine the intracellular localization of intravenously injected infection-specific monoclonal antibodies (mAbs) in the infected cells, immunohistochemical staining was carried out in an animal model having pulmonary tuberculous lesions induced by inoculation of heat-killed Mycobacterium tuberculosis H37Rv. One milligram of intact mouse mAb against mycobacteria (group I, n = 10) and F(ab')2 (group II, n = 6) was intravenously injected to the rabbits of each group. Immunohistochemical staining using an antimouse Ab was performed at days 1, 3, 5, 7 and 8 in group I and at days 1, 2 and 3 in group II by the streptavidin-biotin method. For the control study, 1 ml of nonspecific polyclonal human IgG (group III, n = 10) and 100 micrograms of normal rabbit IgG F(ab')2 (group IV, n = 6) was injected to rabbits and guinea pigs having tuberculous inflammation, respectively. Both groups (group I and II) showed a positive antigen (Ag)-Ab reaction within the cytoplasm of monocytes. A weak but positive reaction was observed intracellularly in group III; however, no positive reaction was seen in group IV. Our results suggest that an intracellular Ag-Ab reaction plays an important role in the localization of infection by immunoscintigraphy using specific mAb fragments.

Animals↗

Two mechanisms for IgG uptake in cultured human trophoblast: evidence for a novel high affinity Fc receptor.

The mechanism of IgG transport by the placental trophoblast was examined by studying IgG uptake by purified trophoblast maintained in culture. This model retains the ability to bind and endocytose human IgG from human serum. Comparison of the relative IgG uptake by the trophoblast among the four subclasses of both human and mouse IgG indicates that the trophoblast IgG receptor has different affinities from those described for the three known human Fc gamma receptors, FcR gamma I, FcR gamma II, and FcR gamma III. These results suggest the presence of a novel trophoblast Fc gamma receptor. Although Fc gamma RIII has been reported to be present on trophoblasts, immunocytochemical studies failed to detect binding to the cell surface of antibody-specific for Fc gamma RIII, 3G8 MAb. In addition, blocking studies with MAb 3G8 did not interfere with IgG uptake. Scatchard analysis of human IgG uptake revealed a biphasic curve consistent with two distinct mechanisms for the transport of IgG by the trophoblast. The first is a higher affinity system (Ka = 1.7 x 10(7) M-1, 1.7 x 10(4) binding sites/cell) which exhibits IgG subclass and species specificity, and the second is a low affinity system (Ka = 6.9 x 10(3) M-1, 7.5 x 10(7) binding sites/cell).

Animals↗

Effects of labor on serum levels of insulin and insulin-like growth factor-binding proteins at the time of delivery.

BACKGROUND: The purposes of this study were to explore whether serum levels of insulin, insulin-like growth factor-I (IGF-I), insulin-like growth factor-binding protein-1 (IGFBP-1) and IGFBP-3 in both maternal and fetal compartments were affected by the stress of labor, and to investigate the relationship between the fetal birthweight and serum levels of insulin, IGF-I and IGFBPs. METHODS: Blood samples were collected at the time of delivery from 147 parturients with vaginal delivery and 128 cases of Cesarean section (112 cases without labor and 16 cases with arrest of cervical dilatation during the active phase of labor). Serum concentrations of insulin, IGF-I, IGFBP-1 and IGFBP-3 were determined by radioimmunoassays (insulin, IGFBP-1 and IGFBP-3) and immunoradiometric assay (IGF-I). RESULTS: Maternal circulating IGFBP-1 levels in parturients with normal spontaneous delivery (NSD) and in subjects receiving Cesarean section (CS) due to arrest of cervical dilatation during active phase of labor were higher than those undergoing scheduled CS without labor. By contrast, insulin levels in both maternal and umbilical cord serum were higher in parturients with CS without labor than those with NSD. No difference in maternal serum IGFBP-3 levels was observed between NSD and CS at the time of delivery. As for all measurements (insulin, IGF-I, IGFBP-1 and IGFBP-3), serum levels in pregnant women (from both NSD and CS) were strikingly higher than those in the fetus. Serum levels of IGFBP-1 in umbilical cords from both groups of NSD (p < 0.02) and scheduled CS (p < 0.01) were inversely correlated with birthweight (BW). By contrast, serum concentration of insulin and IGF-I in umbilical cords from NSD (p < 0.005 and p < 0.01; respectively) and scheduled CS (p < 0.01 and p < 0.05; respectively) were positively related to BW. CONCLUSIONS: From the present results, we conclude that insulin appears to be a regulator for circulating IGFBP-1 during pregnancy. The fetal growth may not be well reflected by maternal serum IGFBP-1 levels, nor by IGFBP-3. By contrast, cord serum IGFBP-1 from CS group without labor may preeminently reflect fetal weight. In additional, serum concentration of insulin and IGF-I in umbilical cord may also be good indicators to reflect the result of neonatal birthweight.

Adult↗

The susceptibility of five stains mice to infections with Angiostrongylus cantonensis.

The susceptibility to the infection of Angiostrongylus cantonensis was studied in five inbred stains of mice including four H-2 haplotypes. Three weeks after infection, C57 BL/10J (H-2b) and C57BL/6J (H-2b) stains of mice showed the lowest, CBA (H-2k) and A/J (H-2a) stains showed intermediate, and the BALB/cJ (H-2d) mice showed the greatest susceptibility to infection. BALB/cJ mice also displayed the most body weight loss, the highest number of worms and showed the least number of days of survival. The s susceptibility of C57BL/6J (H-2b) and C57BL/10J (H-2b) strains of mice sharing same H-2 haplotype was the same despite their different genetic backgrounds. These data suggested that the H-2 gene might influence the susceptibility of mice to A. cantonensis.

Angiostrongylus cantonensis↗