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Biomedical subjects

M Yu

Publications and source records attributed to M Yu.

At least 289 records · Page 16Linked to original sources
PubMed1995

Antioxidant activity of allicin, an active principle in garlic.

Garlic has been claimed to be effective against diseases, in the pathophysiology of which oxygen free radicals (OFRs) have been implicated. Effectiveness of garlic could be due to its ability to scavenge OFRs. However, its antioxidant activity is not known. We investigated the ability of allicin (active ingredient of garlic) contained in the commercial preparation Garlicin to scavenge hydroxyl radicals (.OH) using high pressure liquid chromatographic (HPLC) method. .OH was generated by photolysis of H2O2 (1.25-10 mumoles/ml) with ultraviolet light and was trapped with salicylic acid which is hydroxylated to produce .OH adduct products 2,3- and 2,5-dihydroxybenzoic acid (DHBA). H2O2 produced a concentration-dependent .OH as estimated by .OH adduct products 2,3-DHBA and 2,5-DHBA. Allicin equivalent in Garlicin (1.8, 3.6, 7.2, 14.4, 21.6, 28.8 and 36 micrograms) produced concentration-dependent decreases in the formation of 2,3-DHBA and 2,5-DHBA. The inhibition of formation of 2,3-DHBA and 2,5-DHBA with 1.8 micrograms/ml was 32.36% and 43.2% respectively while with 36.0 micrograms/ml the inhibition was approximately 94.0% and 90.0% respectively. The decrease in .OH adduct products was due to scavenging of .OH and not by scavenging of formed .OH adduct products. Allicin prevented the lipid peroxidation of liver homogenate in a concentration-dependent manner. These results suggest that allicin scavenges .OH and Garlicin has antioxidant activity.

Animals↗
PubMed1995

Immunomodulation of pancreatic islet allografts in mice with CTLA4Ig secreting muscle cells.

In an effort to create a model of in vivo production of immunosuppressants, we have transfected C2C12 muscle cells (H-2k) with the cDNA for CTLA4Ig, a fusion protein that prevents the activation of T cells by blocking the costimulatory signal transduced by the T cell receptors CD28 and CTLA4. CTLA4Ig-secreting clones were cotransplanted with islets as composite grafts in the renal subcapsular space of diabetic mice. When the myoblasts were syngeneic to C3H/HeJ hosts (H-2k), there was a significant prolongation of survival of allogeneic C57Bl/6J (H-2b) islets from a mean 11.0 days to 31.7 days. When the graft was completely allogeneic (H-2k myoblasts and islets into H-2b recipients), there was no benefit in survival. A transient blockade of LFA-1 with the mAb M17 was synergistic in this combination: 8 out of 12 C57Bl/6J recipients achieved long-term acceptance. Systemic CTLA4Ig levels were detected up to 60 days after transplantation. In conclusion, we have shown that C2C12 muscle cells can be genetically engineered to secrete functional CTLA4Ig and that they can be used as a gene reservoir for the continuous in vivo production of CTLA4Ig to modulate the survival of islet cell allografts.

Abatacept↗
PubMed1995

Intracellular immunization of human fetal cord blood stem/progenitor cells with a ribozyme against human immunodeficiency virus type 1.

Successful treatment of human immunodeficiency virus infection may ultimately require targeting of hematopoietic stem cells. Here we used retroviral vectors carrying the ribozyme gene to transduce CD34+ cells from human fetal cord blood. Transduction and ribozyme expression had no apparent adverse effect on cell differentiation and/or proliferation. The macrophage-like cells, differentiated from the stem/progenitor cells in vitro, expressed the ribozyme gene and resisted infection by a macrophage tropic human immunodeficiency virus type 1. These results suggest the feasibility of stem cell gene therapy for human immunodeficiency virus-infected patients.

Antigens, CD↗
PubMed1995

Structure and function of a long alternating purine-pyrimidine sequence in the mouse alcohol dehydrogenase Adh-1 gene.

The mouse alcohol dehydrogenase gene Adh-1 contains an unusually long alternating purine-pyrimidine sequence within its first intron. This alternating sequence differs in length between strains that differ in the extent of Adh-1 expression, and it has been suggested that it plays a role in gene expression. We demonstrate that this alternating sequence can form Z-DNA in vitro. The alternating sequence can act as a positive regulatory element in transient transfection assays in hepatoma cell lines, but not in CV-1 (monkey kidney) cells, suggesting that it can act as a tissue-specific regulatory element. Nuclear run-on experiments showed that the differential expression of Adh-1 from high- and low-activity strains is, however, controlled at the post-transcription level.

Alcohol Dehydrogenase↗
PubMed1995

In vitro and in vivo characterization of a second functional hairpin ribozyme against HIV-1.

We have constructed a hairpin ribozyme targeted to cleave a conserved sequence in the HIV-1 pol gene. The ribozyme was modified to include a structure-stabilizing tetraloop. In vitro studies revealed a cleavage efficiency unprecedented for hairpin ribozymes (Kcat/Km = 75 min-1 microM-1). Stable retroviral vector transduction of this ribozyme gene in T-cell lines resulted in long-term ribozyme expression. As compared to control vector transduced T-cells, the pol ribozyme-transduced cells exhibited significant inhibition of different strains of HIV-1 virus production; this protection was greater when ribozyme expression was driven from an internal pol III promoter (adenovirus VA1) than when driven by a pol II promoter (the MMLV LTR). These results further demonstrate the potential of hairpin ribozymes as anti-HIV gene therapy agents and suggest possibilities for employing combinations of independently targeted hairpin ribozymes.

Base Sequence↗
PubMed1995

Stem cells as vehicles for gene therapy: novel strategy for HIV infection.

Restoration of bone marrow and immune function by means of allogeneic bone marrow transplantation has been attempted in AIDS patients but has not been successful as the donor-derived cells, or their progeny, inevitably became infected. A hairpin ribozyme that specifically cleaves HIV-1 RNA has been developed by F. Wong-Staal et al. and has been demonstrated to confer resistance against HIV-1 infection. Allogeneic transplantation of CD34+ cells or their pluripotent subsets, transduced by vectors bearing this ribozyme gene, can protect the stem cells and their progeny from HIV-1 infection and eventually restores immune function. We have provided evidence that long-term repopulating stem cells can be mobilized into peripheral blood by growth factors. The combination of G-CSF and GM-CSF seems to yield a high frequency of pluripotent stem cells with a CD34+ subset profile that is similar to placental and umbilical cord blood (PUCB). We have then demonstrated a highly efficient transduction of CD34+ cells from PUCB and mobilized leukapheresis products by retroviral vectors bearing the ribozyme gene. Expression of the ribozyme gene, as shown by reverse transcriptase-polymerase chain reaction, was of similar magnitude (70%-90% of cells that grow into colonies). Challenge of the progeny macrophages from such transduced CD34+ cells with monocyte-trophic strains of HIV-1 showed that they were resistant to infection. Thus allogeneic transplantation of CD34+ cells or their pluripotent subsets, transduced with ribozyme gene, can be a promising strategy for the treatment of HIV infection.

Acquired Immunodeficiency Syndrome↗
PubMed1995

Induction of a protective immune response to murine coronavirus with non-internal image anti-idiotypic antibodies.

Neurotropic murine coronaviruses (MHV) provide an excellent animal model to study experimental modulation of the immune response to a viral pathogen with anti-idiotypic antibodies. It is known that among the various types of anti-idiotypic antibodies (anti-Id), those designated beta (beta) or internal image can molecularly mimic the antigen and induce biological activities such as anti-viral protection and neutralization. We have recently shown that polyclonal non-internal image anti-idiotypic antibodies of the gamma-type could induce protective anti-coronavirus immunity. In the present study, a polyclonal anti-Id (Ab2) was induced against a neutralizing murine monoclonal antibody (MAb1), designated 5B170.11. Mice immunized with this affinity-purified rabbit Ab2 alpha, a non-internal image antibody, were partially protected against lethal infection by the JHM strain of MHV. However, other polyclonal and monoclonal non-internal image Ab2 induced against another neutralizing Mab1, designated 4-11G.6, were not able to protect mice against lethal infection with the A59 strain of MHV. These results demonstrate that anti-viral protection by altering the idiotypic network with non-internal image-bearing anti-idiotype reagents can be achieved even with some anti-Id of the alpha-type.

Animals↗
PubMed1995

Octreotide does not prevent bacterial translocation in an infant piglet model of intestinal ischemia-reperfusion.

The process of bacterial translocation (BT) after ischemia/reperfusion (I/R) injury is reported to be mediated by local mucosal factors, the effects of pancreatic enzymes, epithelial disruption, and by dysfunctional intestinal motility. Octreotide (OCT), a somatostatin analog, has been postulated to protect against BT by influencing one or more of these factors. Twenty-two formula-fed piglets (weight, 3.5 +/- 0.5 kg; age, 20 +/- 5 days) were divided into four groups: control (no drug given; no I/R; n = 6), I/R (no drug given; n = 5), I/R plus low-dose OCT (LD OCT, 0.08 microgram/kg; n = 6), and I/R plus high-dose OCT (HD OCT, 8 micrograms/kg; n = 5). All experimental subjects had nonocclusive mesenteric ischemia induced by reversible pericardial tamponade with mesenteric flow decreased to 25 +/- 5% of baseline for 5 hours followed by 15 +/- 5 hours of reperfusion. Mesenteric lymph nodes (MLN), liver, spleen, blood, and peritoneum were harvested for blind microbial analysis. None of the animals in the control group experienced translocation to the tissues tested. All of the animals in the I/R group experienced BT to the MLN. The subjects in the LD OCT and HD OCT groups experienced BT to the MLN 66% and 80% of the time, respectively. Despite the reported clinical evidence that OCT can protect the intestinal mucosa from injury and increase the clearance of bacteria from the gastrointestinal tract, in this study in which variables other than I/R known to promote bacterial translocation were eliminated, OCT failed to modify or prevent the occurrence of translocation to the MLN after I/R injury.

Animals↗
PubMed1995

Nucleocapsid- and virus-like particles assemble in cells infected with recombinant baculoviruses or vaccinia viruses expressing the M and the S segments of Hantaan virus.

The formation of Hantaan (HTN) virus nucleocapsid-like structures (NLS) or virus-like particles (VLP) from expressed gene products was investigated in two eukaryotic systems. Baculovirus expression of the HTN virus small segment (S), which encodes the viral nucleocapsid protein, resulted in assembly of NLS inside infected insect cells. The NLS and authentic ribonucleocapsids, prepared by detergent disruption of HTN virions, had similar sedimentation characteristics and morphologies, and were recognized by HTN virus N-specific antibodies. Co-expression of S and the medium segment (M), which encodes the two viral envelope glycoproteins (G1 and G2), did not efficiently generate VLP in the baculovirus-insect cell system, but VLP were observed in lysates and supernatants of cells infected with a recombinant vaccinia virus co-expressing HTN virus M and S. The VLP sedimented in sucrose to densities consistent with HTN virions, and some of them bore a striking resemblance to Hantaan virions when examined by immunoelectron microscopy.

Animals↗
PubMed1995

Hormonal regulation of nuclear type II estrogen binding sites in the dorsolateral prostate of noble rats.

We previously demonstrated that simultaneous treatment of Noble (NBL) rats with estradiol (E2) and testosterone (T) for 16 weeks induces a proliferative response selectively in the dorsolateral prostates (DLP) of all treated animals [1, 2]. The unique sensitivity of rat DLP to the conjoint androgen-estrogen-induced mitogenic action may be attributable to the presence of a moderate affinity, high capacity, nuclear estrogen binder (type II sites) found exclusively in this prostatic lobe [2, 3]. Little is known about whether prostatic type II site levels are under hormonal regulation. The aim of this study is to determine whether testicular steroids play a role in regulating the basal and/or induced levels of type II site expression in rat DLP. In the first experiment, rats were castrated and immediately treated with 5 alpha-dihydrotestosterone (DHT) and/or E2 for 6 weeks to determine whether these steroids, separately or jointly, could sustain DLP type II site levels in castrates. Treatments of castrated rats with DHT and DHT+E2 were found to be effective in maintaining DLP type II site levels and gland wet weights at values close to those found in intact untreated controls, while treatments with E2 failed to maintain these parameters at levels observed in intact animals. In the second experiment, intact rats were treated with an androgen (T or DHT) or E2, alone or in combination, for 16 weeks to ascertain which hormonal regimen could induce a higher level of type II site expression in the DLP. Treatments of rats with an androgen (T or DHT) or E2 alone did not change DLP type II site levels even though T treatment caused a slight increase in gland weight, while E2 treatment induced prostatic atrophy. Contrary to single hormone treatments, combined T + E2 and DHT+E2 treatments were effective in inducing a doubling of type II sites and increases in wet weight of the DLPs. These data indicate that testicular androgen is the primary factor responsible for maintaining a basal level of type II site expression in rat DLP, while conjoint androgenic-estrogenic action is needed for the induction of a higher level of type II site expression in the tissue.

Animals↗
PubMed1995

Identification and tissue distribution of a novel variant of 11 beta-hydroxysteroid dehydrogenase 1 transcript.

A novel variant of 11 beta-hydroxysteroid dehydrogenase 1 (11 beta-HSD1) mRNA was identified from the ovine liver by reverse transcription-polymerase chain reaction (RT/PCR), and was named 11 beta-HSD1C mRNA. Sequence analysis of the RT-PCR product revealed that 11 beta-HSD1C mRNA was the product of an alternative exon-splicing within the 11 beta-HSD1 gene in which exon 5 was spliced out. Although it caused a deletion of 48 amino acids in the deduced 11 beta-HSD1 protein, this alternative splicing did not result in a shift within the predicted open reading frame of 11 beta-HSD1 cDNA. Thus, 11 beta-HSD1C mRNA was predicted to code for a protein of 244 amino acids. Using RT-PCR, we also examined the expression of 11 beta-HSD1C mRNA in ovine fetal organs and in maternal myometrium, endometrium, chorion, amnion and placenta. The 11 beta-HSD1C mRNA was expressed ubiquitously, similar to 11 beta-HSD1A mRNA, but at a lower abundance. Furthermore, since levels of 11 beta-HSD1C mRNA were directly related to those of 11 beta-HSD1A mRNA, there is no tissue-specificity for this shorter transcript and the only factor regulating its production appears to be 11 beta-HSD1A mRNA itself. To determine whether 11 beta-HSD1C mRNA encoded a functional enzyme, we inserted the cDNA into the expression vector pRc/CMV, and transfected the construct into Chinese hamster ovary cells. The transfected cells expressed a mRNA of expected size but contained no detectable 11 beta-HSD activity. When combined with cellular extracts of 11 beta-HSD1A cDNA transfected cells, they also did not alter either the dehydrogenase or reductase activity. The functional significance of the 11 beta-HSD1 transcript lacking exon 5 (11 beta-HSD1C mRNA) remains to be determined.

11-beta-Hydroxysteroid Dehydrogenases↗
PubMed1995

Isolation and characterization of cytotoxic and antibacterial tetrasaccharide glycosides from Ipomoea stans.

Three new tetrasaccharide glycosides, differing from one reported previously in the type of short-chain fatty acids ester-linked to the tetrasaccharide core, have been isolated and identified from an oligosaccharide fraction of Ipomea stans. Preliminary screening tests showed that the fraction containing these compounds had pronounced cytotoxicity towards three human tumor cell lines as well as specific antibiotic activity against two bacterial strains.

Anti-Bacterial Agents↗
PubMed1995

Platelet membrane glycoproteins and microvesicles in blood from postoperative salvage: a study in cardiac bypass patients.

BACKGROUND: Transfusion of blood collected by intraoperative and postoperative salvage systems has been linked to the development of thrombocytopenia and disseminated intravascular coagulation. Although functional defects have been reported in platelets from unwashed salvaged blood, platelet membrane glycoprotein (GP) composition, a potentially important determinant of function and survival, has not been studied. STUDY DESIGN AND METHODS: Platelets from 22 patients whose blood was salvaged at the completion of surgery were analyzed and compared to platelets obtained from the venous blood from the same patient. Platelet membranes were stained with fluorescein isothiocyanate-conjugated CD41a monoclonal antibody (anti-GPIIb/IIIa) to identify platelets, a phycoerythrin-conjugated monoclonal antibody, CD62 (anti-P-selectin) to identify activated platelets, and CD42b (anti-GPIb) or anti-GPIb/IX to assess GPIb. Samples were analyzed with a flow cytometer using software. RESULTS: Platelets obtained from salvaged blood demonstrated lower GPIb expression (CD42b and GPIb/IX monoclonal antibody binding), higher P-selectin expression, and greater numbers of platelet-derived microvesicles. CONCLUSION: The clinical significance of transfusing blood containing activated platelets and microvesicles merits investigation.

Blood Platelets↗
PubMed1995

Evaluation of the clinical usefulness of thermodilution volumetric catheters.

OBJECTIVE: To determine if treatment modalities (fluid, inotropes, and blood) would be altered based on preload measurements of right ventricular end-diastolic volume index measured by fast response thermodilution catheter, as compared with pulmonary artery occlusion pressure (PAOP). DESIGN: A prospective clinical trial. SETTING: An 11-bed surgical intensive care unit (ICU) at The Queen's Medical Center, an affiliate of the University of Hawaii Surgical Residency program. PATIENTS: Surgical ICU patients who required pulmonary artery catheters, except those patients with arrhythmias or history of tricuspid valve disease. INTERVENTIONS: During the first 48 hrs after catheter insertion, hemodynamic data were obtained at least every 4 hrs. Treatment of low preload was initiated only if clinical indications were present. These indications included a mean arterial pressure of < 70 mm Hg, heart rate of > 120 beats/min, urine output of < 40 mL/hr, stroke volume of < 40 mL/m2 with oxygen delivery of < 450 mL/min/m2, and lactic acidosis. Volume infusion was considered if PAOP was < 18 mm Hg and right ventricular end-diastolic volume index was < 140 mL/m2. Treatment was given tohigh preload, defined as a PAOP of > 18 mm Hg to prevent pulmonary edema. When PAOP and right ventricular end-diastolic volume index gave conflicting information, other clinical parameters were assessed to determine treatment. MEASUREMENTS AND MAIN RESULTS: Twenty-seven patients requiring 70 catheters were evaluated for the study. Thirteen patients with 46 pairs of data points completed the study. Fourteen patients were excluded from analysis due to irregular heart rate, poor quality of cardiac output at the time of volume infusion, or lack of major volume manipulation. PAOP and right ventricular end-diastolic volume index measurements agreed in 42 of 46 instances (PAOP of < 18 mm Hg, right ventricular end-diastolic volume index of < 140 mL/m2), leading to fluid treatment. In one instance, PAOP was > 18 mm Hg, right ventricular end-diastolic volume index was < 140 mL/m2, and the patient had normal blood pressure and good urine output. PAOP was used in this instance as a guide to diurese the patient, which led to improvement of heart rate and stroke volume index. Three measurements in two patients with high intra-abdominal pressure indicated a PAOP of > 18 mm Hg with right ventricular end-diastolic volume index of < 140 mL/m2. A rigid abdomen accompanied hypotension, tachycardia and low urine output. Thus, a fluid bolus was administered, resulting in improved blood pressure, stroke volume, and heart rate. PAOP were obtained at end-expiration. Positive end-expiratory pressure (PEEP) was removed for < 1 sec, if patients were on PEEP > or = 10 cm H2O, to avoid the effects of high intrapleural pressure on PAOP readings. Cardiac output was measured at end-expiration, and stroke volume index and right ventricular end diastolic volume index were derived. CONCLUSIONS: In this small sample of surgical patients with sepsis, adult respiratory distress syndrome, and hemorrhagic shock (n = 13), the additional information derived from right ventricular end-diastolic volume index did not change treatment in 43 of 46 instances. However, patients with increased intra-abdominal pressures may show misleadingly high PAOP despite low preload. These patients clearly benefitted from the additional information derived from ventricular volume measurements. Additionally, clinicians who are reluctant to take off-PEEP PAOP may also find this catheter useful.

Blood Pressure↗
PubMed1995

Frequency of mortality and myocardial infarction during maximizing oxygen delivery: a prospective, randomized trial.

OBJECTIVES: To determine the frequency of myocardial infarction and mortality during treatment that increased oxygen delivery (DO2) to > or = 600 mL/min/m2. To define the characteristics of patients achieving a high DO2 without inotropes in order to guide future studies. DESIGN: A prospective, randomized, controlled trial. SETTING: Two surgical intensive care units at The Queen's Medical Center in the University of Hawaii Surgical Residency Program. PATIENTS: Eighty-nine surgical patients (> or = 18 yrs of age), who were admitted to a surgical intensive care unit and who required pulmonary artery catheter monitoring, were selected for the study. Diagnoses included sepsis, septic shock, adult respiratory distress syndrome, or hypovolemic shock. Patients facing imminent death were excluded from the study. INTERVENTIONS: The treatment group received fluid boluses, blood products, and inotropes, as needed, to achieve a DO2 of > or = 600 mL/min/m2 in the first 24 hrs. Using the same interventions, we treated the control group to reach a DO2 of 450 to 550 mL/min/m2. MEASUREMENTS AND MAIN RESULTS: Hemodynamic measurements were obtained every 4 hrs until the pulmonary artery catheter was removed. DO2 and oxygen consumption were calculated by standard formulas. Serial creatine kinase myocardial fraction and electrocardiograms were documented for the first 48 hrs after study entry and for any new onset of arrhythmia or increasing hemodynamic instability. The patients who generated a high DO2 (> or = 600 mL/min/m2) with only preload treatment were reflective of patients with better cardiac reserve and low mortality rates. These patients, from both treatment and control groups, were excluded in the final analysis. The treatment group who received inotropes to achieve the high DO2 had a 14% mortality rate. Those patients who failed to achieve the high DO2 had a 67% mortality rate, and the control group who achieved a normal DO2 had a 62% mortality rate (p = .005). The frequency of myocardial infarction after study entry was 5.6% (five of 89 patients). This rate was not higher among the groups who received inotropes. Logistic regression analysis showed that age of > or = 50 yrs could be used to classify patients as not self-generating, with an 83% chance of being correct. CONCLUSIONS: The group that required catecholamines to achieve a DO2 of > or = 600 mL/min/m2 had a lower mortality rate, with no increase in the frequency of myocardial infarction. Future prospective, controlled trials examining select groups of patients (age > or = 50 yrs) may demonstrate a difference between control and treatment groups by eliminating the majority of patients who generate the high DO2 with only preload augmentation.

Aged↗